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Hepatocellular carcinoma (HCC) is generally accompanied by high mortality and low cure rate. CCAAT enhancer-binding proteins (CEBPs) are transcriptional regulators that play a key role in maintaining liver function. Altered expression of C/EBPα and C/EBPβ occurs in many tumours including HCC. saRNAs are small double-stranded RNAs that enhance target gene expression at the transcriptional level. In this report, we activate CEPBA with saRNAs and suppress CEBPB with siRNAs in cells that represent three different degrees of HCC. We performed functional assays to investigate the effects of enhancing C/EBPα and its downstream targets, p21 and albumin across these lines. We also used Mass-spectrometry (MS) subsequent to a ChIP pull-down assay to characterise the components of the protein complex involved in regulating saRNA function. Putative saRNA interacting protein candidates that were identified by MS were knocked-down with siRNAs to investigate its impact on saRNA activity. We confirmed CEBPA-saRNA decreased proliferation and migration in the differentiated lines (HepG3/Hep3B). The undifferentiated line (PLCPRF5) showed saRNA-induced increase in CEBPA but with no loss in proliferation. This effect was reversed when CEBPB was suppressed with CEBPB-siRNA. When interrogating saRNA mode of action; three saRNA interacting proteins, CTR9, HnRNPA2/B1 and DDX5 were identified by MS. Targeted knock-down of these two proteins (by siRNA) abrogated saRNA activity. This study provides insight into how different HCC lines are affected by CEBPA-saRNAs and that endogenous abundance of CEBPB and saRNA accessory proteins may dictate efficacy of CEBPA-saRNA when used in a therapeutic context.

Recent studies have reported that small double-strand RNAs (dsRNAs) can activate endogenous genes via an RNA-based promoter targeting mechanism termed RNA activation (RNAa). In the present study, we showed that dsVDUP1-834, a novel small activating RNA (saRNA) targeting promoter of vitamin D3 up-regulated protein 1 (VDUP1) gene, up-regulated expression of VDUP1 at both mRNA and protein levels in A549 lung cancer cells. We also demonstrated that dsVDUP1-834 inhibited cell proliferation in A549 lung cancer cells. Further studies showed that dsVDUP1-834 induced cell-cycle arrest by increasing p27 and p53 and decreasing cyclin A and cyclin B1. In addition, knockdown of VDUP1 abrogated dsVDUP1-834-induced up-regulation of VDUP1 gene expression and related effects. The activation of VDUP1 by dsVDUP1-834 was accompanied by an increase in dimethylation of histone 3 at lysine 4 (H3K4me2) and acetylation of histone 3 (H3ac) and a decrease in dimethylation of histone 3 at lysine 9 (H3K9me2) at the target site of VDUP1 promoter. Moreover, the enrichment of Ago2 was detected at the dsVDUP1-834 target site, and Ago2 knockdown significantly suppressed dsVDUP1-834-mediated inhibition of cell proliferation and modulation of cell-cycle regulators. Taken together, the results presented in this report demonstrate that dsVDUP1-834 induces VDUP1 gene expression by epigenetic changes, resulting in cell growth inhibition and cell-cycle arrest. Our results suggest that targeted induction of VDUP1 by dsVDUP1-834 might be a promising therapeutic strategy for the treatment of lung cancer.

The aim of this study was to identify and evaluate the cases of scabies caused by Sarcoptes scabiei and endoparasites in wild and captive vicuñas (Vicugna vicugna) from the Cusco Region, Peru. Scrapings of skin lesions were made and faecal samples were taken during the capture of vicuñas during the chaccus. The frequency of S. scabiei was 1.9% (54/2826), being 6.1% (48/777) in the wild and 0.2% (6/2049) in captivity. Likewise, F. hepatica (2.0%), Strongylus type eggs (42.1%), Nematodirus sp (6.8%), N. spathiger (26.5%), Trichuris sp (4.0%), Eimeria spp (85.0%), and Moniezia spp (2.7%) were found in a sample of 147 animals, being most cases monoparasitism.sarna, El objetivo del presente estudio fue identificar y evaluar los casos de sarna causada por Sarcoptes scabiei y de endoparásitos en vicuñas silvestres y en cautiverio (Vicugna vicugna) de la Región Cusco, Perú. Se realizaron raspados de lesiones en piel y se tomaron muestras fecales durante la captura de vicuñas durante los chaccus. La frecuencia de S. scabiei fue de 1.9% (54/2826), siendo de 6.1% (48/777) en silvestría y de 0.2% (6/2049) en cautiverio. Asimismo, se identificaron a F. hepatica (2.0%), huevos tipo Strongylus (42.1%), Nematodirus sp (6.8%), N. spathiger (26.5%), Trichuris sp (4.0%), Eimeria spp (85.0%), y Moniezia spp (2.7%) en una muestra de 147 animales, siendo la mayor parte casos de monoparasitismo.

En el presente informe técnico se describen los principales resultados obtenidos en estudios de eficacia de formulaciones inyectables de ivermectina (IVM) y doramectina (DRM) contra Psoroptes ovis en ovinos infestados experimentalmente, su relación con la farmacocinética, y las recomendaciones prácticas que se sugieren para el uso de estos productos a partir de los resultados encontrados. Los estudios en los que se basa este informe fueron realizados por el Grupo de Salud Animal de la EEA INTA Bariloche y el Laboratorio de Farmacología del Centro de Investigación Veterinaria de Tandil (CIVETAN). Estación Experimental Agropecuaria Bariloche Fil: Larroza, Marcela Patricia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Área Producción Animal. Grupo Sanidad Animal; Argentina Fil: Soler, Paula. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Área Producción Animal. Grupo Sanidad Animal; Argentina Fil: Robles, Carlos Alejandro. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Area de Produccion Animal. Grupo de Sanidad Animal; Argentina Fil: Martinez, Agustin. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Área Producción Animal. Grupo Sanidad Animal; Argentina Fil: Cabrera, Francisco Raúl. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Area de Produccion Animal. Grupo de Sanidad Animal; Argentina Fil: Canton, Candela. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Laboratorio de Farmacología; Argentina Fil: Ballent, Mariana. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Laboratorio de Farmacología; Argentina Fil: Lanusse, Carlos Edmundo. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Laboratorio de Farmacología; Argentina Fil: Lifschitz, Adrián Luis. Universidad Nacional del Centro de la Provincia de Buenos Aires. Facultad de Ciencias Veterinarias. Laboratorio de Farmacología; Argentina

ObjectiveThe study aimed to investigate the role of Long non-coding RNA (LncRNA) H19 in the pathogenesis of Diarrhea Irritable Bowel Syndrome (IBS-D), and further to the regulatory effect of LncRNA H19 on AQP1, 3 in the intestinal mucosa of IBS-D patients, so as to seek a new way to elucidate the mechanism of IBS in clinic.MethodsThe levels of LncRNA H19, AQP1, and AQP3 were detected in colonic tissues of IBS-D patients, compared with that in healthy controls. Through RNA gene interference and activation methods, small activating RNA (saRNA) and small interfering (siRNA) were transfered into Caco-2 cells in vitro experiment, and sub-group for two control group, siH19 empty vector group, siH19 interference group, overexpression H19 vector group, and overexpression H19 empty vector group. Quantitative real-time reverse transcription-polymerase chain reaction (qRT-PCR) and Western blot were applied to evaluate the expression levels of LncRNA H19 and the amount of AQP1 and AQP3 protein expression, respectively.ResultsCompared with healthy volunteers, the levels of LncRNA H19, AQP1, and AQP3 in the colonic mucosa of IBS-D patients were significantly decreased (P < 0.05). The results in vitro transfection experiment revealed that the level of LncRNA H19 in the siH19 interference group was significantly declined (P < 0.05), while there was a remarkable increase in the overexpression H19 vector group (P < 0.05), compared with the corresponding control groups. The expression of AQP1 and AQP3 in Caco-2 cells was of positive correlation with the level of LncRNA H19.ConclusionThat the down-regulation of LncRNA H19 resulted in the expression changes of AQP1 and AQP3 may play an important role in the occurrence and development of IBS-D.

Objetivo. Determinar la prevalencia de la escabiosis y la tungiasis en Colombia. Métodos. Estudio observacional descriptivo a partir de los datos del Sistema Integrado de Información de la Protección Social (SISPRO) del periodo comprendido entre el año 2009 al 2019. Las variables analizadas fueron departamento de residencia de las personas, tipo de afiliación al Sistema General de Seguridad Social en Salud (SGSSS), edad y sexo. Resultados. La prevalencia de escabiosis encontrada fue de 5,3 por 100 000 habitantes y la de tungiasis de 0,09 por 100 000 habitantes. Las prevalencias más altas se encuentran en Vaupés, Amazonas, Guainía y Cauca. En los últimos seis años el régimen subsidiado reporta las mayores prevalencias para las dos enfermedades. En general, la prevalencia de escabiosis es similar entre hombres y mujeres y más alta en adultos mayores, mientras que en tungiasis es mayor en las mujeres y en personas entre los 5 a 14 años y por encima de los 65 años. Conclusiones. La distribución de la escabiosis y la tungiasis en los diferentes departamentos, así como los resultados en las demás variables analizadas, apoya los estudios que describen la asociación entre estas enfermedades con la pobreza y el mayor riesgo de contraerlas en la población de niños, adultos mayores y mujeres. Aunque existen limitaciones con la utilización de las bases de datos de SISPRO y sea necesario mejorar la calidad de los registros desde la fuente de los datos, esta información permite obtener estimaciones importantes sobre la magnitud de las enfermedades. Magíster en Epidemiología Maestría

Background Sarcoptic mange is a globally distributed parasitic disease caused by the burrowing mite Sarcoptes scabiei. This mite has a certain degree of host specificity, although interspecific transmission can occur among phylogenetically related species or through prey–predator mediated exposure. In 2018, a wild boar (Sus scrofa) with lesions compatible with sarcoptic mange was hunted in Ports de Tortosa i Beseit Natural Park (PTB, north-eastern Spain), where an active epizootic outbreak of sarcoptic mange is affecting Iberian ibexes (Capra pyrenaica) since 2014. Methods A complete necropsy, skin scrapings and skin digestions with hydroxide potassium were performed to confirm the diagnosis. Routine histopathological analysis, toluidine blue staining and immunohistochemistry were used to characterize the lesions and the inflammatory infiltrate. Finally, 10 specific S. scabiei microsatellites were molecularly genotyped through polymerase chain reactions in mites obtained from the affected wild boar. For phylogenetic comparison, mites obtained from sympatric Iberian ibexes and allopatric wild boars and Iberian ibexes from southern Spain were analysed. Results Sarcoptes scabiei was visually and molecularly identified in the infested wild boar from PTB, causing skin lesions with dermal inflammatory infiltrate rich in T and B cells, which indicate an adaptive immune response. Three S. scabiei genetic clusters were identified: one included mites from southern Iberian ibexes, another included mites from southern wild boars, and a third one distinctively grouped the wild boar from PTB with the sympatric ibexes. Conclusions To the authors’ knowledge, this is the first reported case of sarcoptic mange in wild boar in Spain and the first documented case of S. scabiei cross-transmission from a wild ruminant host to a wild boar. The wild boar presented an ordinary scabies type reaction, which is typical of the self-limiting infestations reported in other cases of interspecific transmission. Graphical abstract

This review will explore the four major pillars required for design and development of an saRNA vaccine: antigen design, vector design, non-viral delivery systems, and manufacturing (both saRNA and lipid nanoparticles (LNP)). In will report on the major innovations, preclinical and clinical data reported in the last five years and will discuss future prospects.

Self-amplifying RNA (saRNA) is a cutting-edge platform for both nucleic acid vaccines and therapeutics. saRNA is self-adjuvanting, as it activates types I and III interferon (IFN), which enhances the immunogenicity of RNA vaccines but can also lead to inhibition of translation. In this study, we screened a library of saRNA constructs with cis-encoded innate inhibiting proteins (IIPs) and determined the effect on protein expression and immunogenicity. We observed that the PIV-5 V and Middle East respiratory syndrome coronavirus (MERS-CoV) ORF4a proteins enhance protein expression 100- to 500-fold in vitro in IFN-competent HeLa and MRC5 cells. We found that the MERS-CoV ORF4a protein partially abates dose nonlinearity in vivo, and that ruxolitinib, a potent Janus kinase (JAK)/signal transducer and activator of transcription (STAT) inhibitor, but not the IIPs, enhances protein expression of saRNA in vivo. Both the PIV-5 V and MERS-CoV ORF4a proteins were found to enhance the percentage of resident cells in human skin explants expressing saRNA and completely rescued dose nonlinearity of saRNA. Finally, we observed that the MERS-CoV ORF4a increased the rabies virus (RABV)-specific immunoglobulin G (IgG) titer and neutralization half-maximal inhibitory concentration (IC50) by ∼10-fold in rabbits, but not in mice or rats. These experiments provide a proof of concept that IIPs can be directly encoded into saRNA vectors and effectively abate the nonlinear dose dependency and enhance immunogenicity., Graphical Abstract, Self-amplifying RNA is a cutting-edge vaccine platform but is known to activate interferons; this activation can augment immunogenicity but also inhibit translation of the antigen. Blakney et al. found that a cis-encoded innate inhibiting protein, MERS-CoV ORF4a, enhances both protein expression and immunogenicity of an saRNA rabies vaccine.

Highlights • Alphavirus self-amplifiying RNA (saRNA) induces more potent immune responses than conventional mRNA. • saRNA delivery can be enhanced by electroporation or conjugation with cationic lipid or polymers. • saRNA vaccines induce protective responses against human pathogens in preclinical models. • saRNA replication mediates innate immune signals that contribute to the strength of immune responses. • saRNA vaccines could be generated in a quick way to face emergent pathogens like SARS-CoV-2., DNA or mRNA vaccines have potential advantages over conventional vaccines since they are easier to manufacture and have higher safety profiles. In particular, self-amplifying RNA (saRNA) derived from alphavirus expression vectors has shown to be very efficient to induce humoral and cellular responses against many antigens in preclinical models, being superior to non-replicating mRNA and DNA. This is mainly due to the fact that saRNA can provide very high expression levels and simultaneously induces strong innate responses, potentiating immunity. saRNA can be administered as viral particles or DNA, but direct delivery as RNA represents a safer and more simple approach. Although saRNA can be delivered as naked RNA, in vivo transfection can be enhanced by electroporation or by complexing it with cationic lipids or polymers. Alphavirus saRNA could have broad application to vaccinate against human pathogens, including emerging ones like SARS-CoV-2, for which clinical trials have been recently initiated.

Este artículo se encuentra disponible en la siguiente URL: https://asescu.com/portfolio-items/boletin-de-cunicultura-no196-sector-cunicola-al-dia/?portfolioCats=20 La sarna es una enfermedad de la piel altamente contagiosa causada por una o varias especies de ácaros, que puede producir inmunosupresión y reacciones inflamatorias. En el conejo, Sarcoptes scabiei y Psoroptes equi cuniculi son los ácaros que producen esta patología en mayor medida. El diagnóstico y tratamiento de esta enfermedad son relativamente sencillos una vez se evidencia la sintomatología en los animales; pero si no se instauran tratamientos y medidas de control en las granjas afectadas, la infestación por sarna puede provocar graves pérdidas económicas.

Sheep psoroptic mange is a form of highly itchy and contagious dermatitis caused by the mite Psoroptes ovis. Here, we reporteda case of sheep psoroptic mange outbreak at a property in São Vicente do Sul, Rio Grande do Sul (RS), Brazil. Poor nutritional status, restless behavior, pruritus, wool fall, yellowish crusted skin lesions with edge exudate, erythema surrounding the lesions and presence of wool trapped between the teeth were observed in the herd. Complete blood count test showed marked eosinophilia, and parasitological examination of a skin scrape revealed P. ovis. Histopathological examination of a skin biopsy sample revealed eosinophilic pustular dermatitis. The treatment consisted of two administrations of 1% ivermectin at 1 mL/33 kg of body weight subcutaneously over an interval of 10 days. To the best of our knowledge, this is the first report of the occurrence of psoroptic mange at a sheep farm of RS with description of the clinical signs and laboratory and histopathological findings. RESUMO: A sarna psoróptica ovina é uma dermatite altamente pruriginosa e contagiosa causada pelo ácaro Psoroptes ovis. Este trabalho registra o caso da doença em uma propriedade no município de São Vicente do Sul, Rio Grande do Sul (RS), Brasil. No rebanho foram observados mau estado de nutrição, comportamento inquieto, prurido, queda de lã, lesões crostosas amareladas na pele com exsudato nas bordas, eritema perilesional e presença de lã presa entre os dentes. O hemograma mostrou marcada eosinofilia e um raspado cutâneo demonstrou a presença de P. ovis no exame parasitológico. No exame histopatológico, oriundo de uma biópsia de pele, observou-se dermatite pustular eosinofílica. O tratamento consistiu de duas administrações de ivermectina a 1% na dose de 1ml/33kg de peso corporal por via subcutânea em um intervalo de 10 dias. Este é primeiro estudo relatando a ocorrência, sinais clínicos e achados laboratoriais/histopatológicos da sarna psoróptica em uma propriedade de criação de ovinos no RS.

Resumen: La escabiosis afecta a más de 200 millones de personas en el mundo, y ocasiona un importante impacto socioeconómico. El mecanismo de contagio es por contacto directo prolongado. El contagio por fómites es infrecuente, aunque puede ser importante en la sarna noruega. La terapia con permetrina tópica al 5% es recomendada como tratamiento de primera línea. Puede indicarse durante el embarazo y la lactancia, y parece ser segura en niños 15 kg. Diverse systematic reviews and meta-analysis have concluded that oral ivermectin is as effective and safe as topical permethrin. Mass drug administration of oral ivermectin is an excellent option for the management of scabies in communities with high prevalence, or for scabies outbreaks in institutions.

BACKGROUND: Oligonucleotide drug development has revolutionised the drug discovery field allowing the notoriously "undruggable" genome to potentially become "druggable". Within this field, 'small' or 'short' activating RNAs (saRNA) are a more recently discovered category of short double stranded RNA with clinical potential. SaRNAs promote endogenous transcription from target loci, a phenomenon widely observed in mammals known as RNA activation (RNAa). The ability to target a particular gene is dependent on the sequence of the saRNA. Hence, the potential clinical application of saRNA is to increase target gene expression in a sequence specific manner. SaRNA based oligonucleotide therapeutics present great promise in expanding the "druggable" genome with particular areas of interest including transcription factor activation and haploinsufficency. Review and Conclusion: In this mini-review, we describe the pre-clinical development of the first saRNA drug to enter the clinic. This saRNA, referred to as MTL-CEBPA, induces transcription of the transcription factor CCAAT/enhancer-binding protein alpha (CEBPα), a tumour suppressor and critical regulator of hepatocyte function. MTL-CEBPA is presently in Phase I clinical trials for hepatocellular carcinoma (HCC). The clinical development of MTL-CEBPA will demonstrate "proof of concept", showing that saRNAs can provide the basis for drugs which enhance targeted gene expression and consequently improve disease outcome in patients.

Background RNA is increasingly recognized as a powerful molecule that can be used to control gene expression. Sophisticated, well-engineered RNA-based regulators are being developed as oligotherapeutics. Methods In particular, small activating RNAs (saRNAs) are promising therapeutic options for targeting human diseases. Numerous saRNAs targeting multiple cancers have been developed in preclinical models. One saRNA targeting C/EBPα is currently undergoing clinical trials in liver cancer. Results and conclusion In this review, we describe the current working model of the intracellular mechanism of saRNA, discuss the recent progress of saRNA therapeutics in preclinical and clinical trials, and current advances in targeted delivery using aptamers in detail.

A infecção pelo VIH predispõe à ocorrência de múltiplas dermatoses que se manifestam frequentemente de forma atípica. Reporta-se o caso de um homem com 57 anos e imunossupressão grave no contexto de infecção pelo VIH que desenvolveu dermatose pruriginosa disseminada aquando de internamento hospitalar prolongado. Tendo por base as características do doente, as queixas e a distribuição corporal das lesões, e apesar de não existirem lesões típicas nem contexto epidemiológico sugestivo, foi admitido o diagnóstico de escabiose, com realização de biópsias cutâneas e dois ciclos de tratamento com benzoato de benzilo, apenas com melhoria transitória. Na repetição de biópsia, viria a confirmar-se hiperinfestação por Sarcoptes scabiei pelo que foi realizado ciclo de tratamento com ivermectina com resolução completa das lesões. A propósito deste caso clínico, os autores revêm aspectos clínicos e o tratamento da sarna crostosa. info:eu-repo/semantics/publishedVersion

The aim of this study was to evaluate self-replicating RNA lipid nanoparticles (saRNA LNPs) to neutralize SARS-CoV-2 variants delta (B.1.617 lineage) and alpha (B.1.1.7 lineage). Before immunization of mice with saRNA LNPs, we saw high expression of S-protein at both mRNA and protein levels after transfection of HEK293T/17 cells with saRNA LNPs. After oral immunization of BALB/c mice with 0.1 - 10 µg saRNA LNPs , a high quantity of SARS-CoV-2 specific IgG and IgA antibodies were seen with a dose-dependent pattern. Importantly, the ratio of IgG2a/IgG1 in serum of vaccinated mice showed Th1/Th2 skewing response. We also found that the secreted antibodies could neutralize SARS-CoV-2 variants delta (B.1.617 lineage) and alpha (B.1.1.7 lineage). Re-stimulated splenocytes of vaccinated mice showed high secretion of IFN-γ, IL-6, and TNF- α . The authors think that although the preclinical study confirmed the efficacy of saRNA LNPs against SARS-CoV-2, the actual efficacy and safety of the oral vaccine must be evaluated in clinical trials.

The present study aims to determine the epidemiological, clinical and laboratorial profile of the dermatopathies that affect dogs and cats living in a semi-arid area in the Northeast of Brazil. Seventy-eight dogs and cats consulted at the Veterinary Hospital with dermatological complaints were included in this study. Skin lesions were characterized with respect to morphology, appearance, and distribution, an epidemiological questionnaire was applied, and samples were collected for complementary examination. The diagnosis was confirmed by parasitological and microbiological tests. There was a predominance of the canine species (93.6%), of young animals (46.3%), and of animals of undefined breeds (61.5%). It was observed that 29.5% of the affections were of fungal aetiology, 14.1% were bacterial, 3.9% were scabies. In 5.1% of the cases there were associations between different pathogens, and in 47.4% the laboratory examination was negative for the pathogens investigated. The most frequent clinical manifestations included alopecia (74.4%), pruritus (61.6%) and erythema (43.6%), distributed mainly in the dorsal-ventral region (36.1%) or even disseminated (43.1%). With respect to One Health, 51.3% (40/78) of the owners reported that they did not know what "zoonoses" were. Dermatopathies have been shown to be important disorders, especially the mycotic and bacterial diseases. Atypical cases of infections motivated by pathogens little described in the literature as etiologic agents of dermatopathies in dogs and cats were observed. El presente estudio tuvo como objetivo determinar el perfil epidemiológico, clínico y de laboratorio de las dermatopatías que afectan a perros y gatos domiciliados en una zona semiárida del noreste de Brasil. En este estudio se incluyeron setenta y ocho perros y gatos tratados en el Hospital Veterinario con quejas dermatológicas. Las lesiones cutáneas se caracterizaron en términos de morfología, apariencia y distribución, se aplicó un cuestionario epidemiológico y se recolectaron muestras para su posterior examen. Diagnóstico confirmado en pruebas parasitológicas y microbiológicas. Hubo predominio de la especie canina (93,6%), de animales jóvenes (46,3%) y de raza no definida (61,5%). Se observó que el 29,5% de las afecciones fueron de etiología fúngica, 14,1% bacterianas, 3,9% sarna. En el 5,1% de los casos hubo asociaciones entre diferentes patógenos y en el 47,4% la prueba de laboratorio fue negativa para los patógenos estudiados. Las manifestaciones clínicas más frecuentes incluyeron alopecia (74,4%), prurito (61,6%) y eritema (43,6%), distribuidos principalmente en la región dorsoventral (36,1%) o incluso diseminados (43,1%). En cuanto a Unica Salud, el 51,3% (40/78) de los tutores informaron que no sabían qué eran las “zoonosis”. Se demostró que las dermatopatías son afecciones importantes, especialmente enfermedades micóticas y bacterianas. Se observaron casos atípicos de infecciones causadas por patógenos poco descritos en la literatura como agentes etiológicos de dermatopatías en perros y gatos. O presente trabalho objetivou determinar o perfil epidemiológico, clínico e laboratorial das dermatopatias que acometem os cães e gatos domiciliados em área semiárida do Nordeste do Brasil. Setenta e oito cães e gatos atendidos no Hospital Veterinário com queixas dermatológicas foram incluídos neste estudo. As lesões de pele foram caracterizadas quanto à morfologia, aspecto e distribuição, foi aplicado um questionário epidemiológico, e colhidas amostras para exame complementar. O diagnóstico confirmado em testes parasitológicos e microbiológicos. Houve predominância da espécie canina (93,6%), de animais jovens (46,3%) e sem raça definida (61,5%). Observou-se que 29,5% das afecções eram de etiologia fúngica, 14,1% bacteriana, 3,9% de sarnas. Em 5,1% dos casos havia associações entre diferentes patógenos e em 47,4% o exame laboratorial foi negativo para os patógenos pesquisados. As manifestações clínicas mais frequentes incluíram alopecia (74,4%), prurido (61,6%) e eritema (43,6%), distribuídas principalmente na região dorso-ventral (36,1%) ou ainda disseminada (43,1%). Sobre Saúde Única, 51,3% (40/78) dos tutores informaram que não sabiam o que eram “zoonoses”. Dermatopatias demonstraram-se como importantes afecções, sobretudo as doenças micóticas e bacterianas. Foram observados casos atípicos de infecções motivadas por patógenos pouco descritos na literatura como agentes etiológicos de dermatopatias em cães e gatos.

Small active RNA (saRNA)-induced gene activation (RNAa) is a novel strategy to treat cancer. Our previous work proved that the p21-saRNA-322 successfully hindered colorectal cancer growth by activating p21 gene. However, the barrier for successful saRNA therapy is lack of efficient drug delivery. In the present study, a rectal delivery system entitled p21-saRNA-322 encapsulated tumor-selective lipopolyplex (TSLPP-p21-saRNA-322) which consist of PEI/p21-saRNA-322 polyplex core and hyaluronan (HA) modulated lipid shell was developed to treat colorectal cancer. Our results showed that this system maintained at the rectum for more than 6 h and preferentially accumulated at tumor site. CD44 knock down experiment instructed that the superb cellular uptake of TSLPP-p21-saRNA-322 attributed to HA-CD44 recognition. An orthotopic model of bio-luminescence human colorectal cancer in mice was developed using microsurgery and TSLPP-p21-saRNA-322 demonstrated a superior antitumor efficacy in vitro and in vivo . Our results provide preclinical proof-of-concept for a novel method to treat colorectal cancer by rectal administration of TSLPP formulated p21-saRNA-322.

The present study evaluated the efficacy of a single oral dose of doramectin in the control of Psoroptes ovis and Leporacarus gibbus in naturally infested rabbits. Sixteen adult rabbits were selected and distributed in two experimental groups. The treated group received 200 μg/Kg of oral doramectin and the control group received the same volume of saline solution. The diagnosis of the mites was made with a stereoscopic microscope. Hairs from the dorsal part of the neck, lumbar right, lumbar left, ventral side of the tail and ventral abdomen were evaluated for L. gibbus, and ear wax evaluated for P. ovis. The evaluation of the efficiency and the clinical assessment of the lesions was made in days 0, +3, +7, +14, +21, +28, and +35 after treatment. An efficacy of 75% and 87,5% was observed for L. gibbus in days +3 and +7 after treatment, an efficacy of 100% was observed in days +14, + 21, +28 e +35. An efficacy of 100% for the control of P. ovis was observed following day +7. The clinical lesion score of the control group remained unaltered, except for one animal which conditions worsened during experimentation. In the treated group animals, regression of the lesions was observed following day +3, and on day +21 no signal of infestation by P. ovis was present. None of the animals from the treated group presented secondary collateral effects caused by the doramectin, which proved itself as an optimal alternative for mite control in naturally infested rabbits. RESUMO: O objetivo do trabalho foi avaliar a eficácia da doramectina administrada por via oral no controle de Psoroptes ovis e Leporacarus gibbus em coelhos naturalmente infestados. Foram selecionados 16 coelhos adultos, distribuídos em dois grupos experimentais, compondo oito animais por grupo. O grupo tratado foi medicado com 200μg/kg de doramectina por via oral, enquanto que no grupo controle foi administrado o mesmo volume de solução salina. O diagnóstico dos ácaros foi realizado com auxílio de microscópio estereoscópico. Foram coletados pelos das regiões do pescoço dorsal, lombar direita, lombar esquerda, cauda ventral e abdômen ventral para avaliação de L. gibbus e para P. ovis foi coletado cerúmen das orelhas com auxílio de zaragatoas. A avaliação da eficácia e a avaliação clínica das lesões, mensuradas em escores (grau 0 a 4), foi realizada nos dias 0, +3, +7, +14, +21, +28 e +35, após o tratamento. Foi observada eficácia de 75% e 87,5% no controle de L. gibbus nos dias +3 e +7 após o tratamento, sendo observada eficácia de 100% nos dias +14, + 21, +28 e +35. Foi observada eficácia de 100% no controle de P. ovis a partir do dia +7, permanecendo até o final do período observacional. O escore das lesões clínicas no grupo controle permaneceu de forma inalterada, exceto em um animal que piorou ao longo dos dias experimentais, enquanto nos animais do grupo tratado regrediu a partir do dia +3 e já no dia +21 após o tratamento, os animais apresentavam-se sem sinais da infestação por P. ovis. Nenhum animal do grupo tratado apresentou quaisquer efeitos colaterais secundários causados pela doramectina, que se mostrou uma ótima alternativa para o controle dos ácaros em coelhos naturalmente parasitados.

Transfection of small non-coding RNAs (sncRNAs) molecules has become a routine technique widely used for silencing gene expression by triggering post-transcriptional and transcriptional RNA interference (RNAi) pathways. Moreover, in the past decade, small activating (saRNA) sequences targeting promoter regions were also reported, thereby a RNA-based gene activation (RNAa) mechanism has been proposed. In this regard, Turner and colleagues recently discovered an endogenous microRNA (miRNA) which binds its promoter in order to upregulate its own expression. Interestingly, several miRNA-induced RNA activation (miRNAa) phenomena have since then been identified. My objective here is to introduce the reader into the emergent miRNAa research field, as well as bring together important discoveries about this unexplored transcriptional activation pathway. Fil: Vaschetto, Luis Maria Benjamin. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Diversidad y Ecología Animal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto de Diversidad y Ecología Animal; Argentina

Sarcoptes scabiei es un ectoparásito obligado de más de 17 familias de mamíferos. E s el agente causante de la sarna , patología caracterizada por erupciones cutáneas, un prurito intenso y la presencia de lesiones en la epidermis, causadas por la infección parásita, entre las que se encuentran el surco y la pápula o eminencia acarina. Pese a que los casos de sarna se han reducido en el último siglo y que la idea de que se trata de una enfermedad más propia del siglo pasado, su prevalencia es de 300 millones de afectados en todo el mundo. Por lo tanto, no solo se trata de una enfermedad vigente, sino que también se encuentr a desatendida, consecuencia directa de la persistente creencia de su erradicación. Se requerirá, por lo tanto, volver a prestarle la importancia que realmente precisa y continuar investigando y desarrollando metodologías de diagnóstico, tratamiento y preve nción rápidos y precisos que eviten brotes y epidemias y que contribuyan a la disminución de la prevalencia mundial registrada. En esta revisión ofrecemos una perspectiva global, no solo los antecedentes, orígenes y etiología de la enfermedad, sino que tam bién ofreceremos una recopilación de los métodos de diagnóstico que actualmente se emplean, así como los tratamientos y las medidas de control más vigentes. Por otra parte, también se discutirá la incertidumbre que engloba el origen de Sarcoptes scabiei , así como el debate existente sobre el posible origen zoonótico, importante a tener en cuenta en el desarrollo de estrategias preventivas y de control . Universidad de Sevilla. Grado en Farmacia

Self-amplifying RNA (saRNA) is a promising biotherapeutic tool that has been used as a vaccine against both infectious diseases and cancer. saRNA has been shown to induce protein expression for up to 60 days and elicit immune responses with lower dosing than messenger RNA (mRNA). Because saRNA is a large (~9500 nt), negatively charged molecule, it requires a delivery vehicle for efficient cellular uptake and degradation protection. Lipid nanoparticles (LNPs) have been widely used for RNA formulations, where the prevailing paradigm is to encapsulate RNA within the particle, including the first FDA-approved small-interfering siRNA therapy. Here, we compared LNP formulations with cationic and ionizable lipids with saRNA either on the interior or exterior of the particle. We show that LNPs formulated with cationic lipids protect saRNA from RNAse degradation, even when it is adsorbed to the surface. Furthermore, cationic LNPs deliver saRNA equivalently to particles formulated with saRNA encapsulated in an ionizable lipid particle, both in vitro and in vivo. Finally, we show that cationic and ionizable LNP formulations induce equivalent antibodies against HIV-1 Env gp140 as a model antigen. These studies establish formulating saRNA on the surface of cationic LNPs as an alternative to the paradigm of encapsulating RNA.

La sarna del manzano ocasionada por Venturia inaequalis es la enfermedad de mayor importancia en Uruguay y en la mayoría de las regiones productoras de manzana con clima húmedo. Su control se basa en el uso de fungicidas preventivos anticipándose a los periodos de infección, realizándose aplicaciones curativas cuando no fue posible prever la infección. Las ascosporas se producen en pseudotecios en las hojas caídas al suelo en la temporada anterior, madurando escalonadamente desde el inicio de la brotación de los manzanos hasta mediados de diciembre, liberándose cada vez que las hojas se mojan. No se conocen trabajos que indiquen que tiempo debe transcurrir entre eventos de liberación para que la acumulación de ascosporas maduras sea significativa. Es sabido que la luz influye favoreciendo la liberación, sin embargo, existe información contradictoria en referencia a la existencia o ausencia de liberación nocturna. Debido a esta falta de información se realizan aplicaciones cuando ocurren eventos consecutivos de lluvias y cuando ocurren eventos nocturnos, las que podrían evitarse en caso de constatarse la ausencia de liberación en esas situaciones. Los objetivos de este trabajo fueron conocer el comportamiento de descargas de ascoporas de V. inaequalis en días sucesivos y en horas nocturnas mediante liberaciones forzadas y determinar el comportamiento de la liberación natural de ascosporas en el campo. La liberación de ascosporas presentó un pico máximo en la primera lluvia forzada y fue disminuyendo en los días siguientes. Se verificó que existe liberación en horas de la noche. En relación a las liberaciones naturales, en ambos años estudiados el inicio de la descarga coincidió con la brotación de los manzanos, mientras que el final ocurrió a inicios de noviembre, un mes antes de lo indicado por el sistema de alarma en Uruguay. Estos resultados permiten especular con la posibilidad de adelantar el final de las aplicaciones de fungicidas en cada temporada disponiendo de datos reales de la concentración de ascosporas en el aire.

Recent studies have reported that chemically synthesized small activating RNA (saRNA) targeting the promoter regions of a gene can activate its expression in different cell lines. This technique can be a powerful therapeutic method for diseases caused by complete inactivation or reduced expression of specific genes. E-cadherin is a typical tumor suppressor gene. Loss of E-cadherin mediates the transition from benign lesions to invasive, metastatic cancer. In this study, several 21-nt small double-stranded RNAs (dsRNAs) targeting the promoter regions of human E-cadherin were designed and synthesized and the features of their function were investigated to study the regulatory role of dsRNA on E-cadherin expression. A new saRNA (dsEcad‑661) that can enhance E-cadherin expression by targeting non-coding regulatory regions in gene promoters was identified. Using dsRNA with modified base quantity and cholesterol-conjugated dsRNA, we found the antisense strand may be the guide strand of saRNA in the upregulation of E-cadherin. These findings provide several important pieces of evidence that may improve understanding of the function of saRNA and may promote its development for clinical application.

Background Epidermal growth factor receptor-tyrosine kinase inhibitors (TKI-EGFRs) present a new prospect for the treatment of lung cancer. However, in clinical application, the majority of patients become TKI resistant within a year. More and more studies have shown that a loss of phosphatase and tensin homolog (PTEN) expression is associated with TKI resistance. An alternative method of upregulating PTEN expression may reverse TKI resistance. Methods We designed five candidate small activating ribonucleic acids (saRNAs) to target PTEN, and transfected them into H-157 cells to screen out functional saRNA. We used reverse transcriptase-polymerase chain reaction and Western blot to evaluate the effect of saRNA to PTEN expression. We then analyzed the growth and apoptosis of cells transfected with saRNA under the treatment of TKI to investigate whether saRNAs can reverse TKI resistance by upregulating PTEN expression. Results The functional saRNA we designed could upregulate PTEN expression. The H-157 cells transfected with saRNA grew slower in the presence of TKI drugs than the cells that were not transfected with saRNA. The apoptosis rate was also obviously higher. Conclusions Our study proves that loss of PTEN expression is an important mechanism of TKI resistance. It is possible to control TKI resistance by upregulating PTEN expression using RNA activation technology.

// Changlin Li 1 , Wencong Jiang 1, 2 , Qingting Hu 1, 3 , Long-cheng Li 4 , Liang Dong 1 , Ruibao Chen 1 , Yinghong Zhang 1 , Yuzhe Tang 1 , J. Brantley Thrasher 1 , Chang-Bai Liu 3 , Benyi Li 1, 2, 3 1 Department of Urology, University of Kansas Medical Center, Kansas City, KS 66160, USA 2 Department of Urology, The Affiliated Hospital, Guangdong Medical University, Zhanjiang 524001, China 3 Institute of Cell Therapy, China Three Gorges University, Yichang 443002, China 4 Laboratory of Molecular Medicine, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences, Beijing 100073, China Correspondence to: Benyi Li, e-mail: bli@kumc.edu Keywords: DPYSL3, CRMP4, metastasis, RNAa, saRNA Received: January 25, 2016 Accepted: February 23, 2016 Published: March 23, 2016 ABSTRACT To explore a novel strategy in suppressing tumor metastasis, we took the advantage of a recent RNA activation (RNAa) theory and used small double-strand RNA molecules, termed as small activating RNAs (saRNA) that are complimentary to target gene promoter, to enhance transcription of metastasis suppressor gene. The target gene in this study is Dihydro-pyrimidinase-like 3 ( DPYSL3 , protein name CRMP4), which was identified as a metastatic suppressor in prostate cancers. There are two transcriptional variants of DPYSL3 gene in human genome, of which the variant 2 is the dominant transcript (DPYSL3v2, CRMP4a) but is also significantly down-regulated in primary prostate cancers. A total of 8 saRNAs for DPYSL3v1 and 14 saRNAs for DPYSL3v2 were tested in multiple prostate cancer cell lines. While none of the saRNAs significantly altered DPYSL3v1 expression, 4 saRNAs showed a strong enhancing effect on DPYSL3v2 expression, resulting in reduced cell mobility in vitro . To achieve a prostate cancer-specific delivery for in vivo testing, we conjugated the most potent sa V2-9 RNA molecule with the prostate-specific membrane antigen (PSMA)-targeting aptamer A10-3.2. The conjugates successful increased DPYSL3v2 gene expression in PSMA-positive but not PSMA-negative prostate cancer cells. In nude mice bearing orthotopic xenograft of prostate cancer, a 10-day consecutive treatment with the sa V2-9 conjugates significantly suppress distal metastasis compared to the control saRNAs. Analysis of xenograft tissues revealed that DPYSL3v2 expression was largely increased in sa V2-9 conjugate-treated group compared to the control group. In conclusion, DPYSL3v2 promoter-targeted saRNA molecules might be used as an adjunctive therapy to suppress prostate cancer metastasis.