Complete genome amplification of Equine influenza virus subtype 2

This work reports a method for rapid amplification of the complete genome of equine influenza virus subtype 2 (H3N8). A ThermoScriptTM reverse transcriptase instead of the avian myeloblastosis virus reverse transcriptase or Moloney murine leukemia virus reverse transcriptase was used. This enzyme has demonstrated higher thermal stability and is described as suitable to make long cDNA with a complex secondary structure. The product obtained by this method can be cloned, used in later sequencing reactions or nested-PCR with the purpose of achieving a rapid diagnosis and characterization of the equine influenza virus type A. This detection assay might be a valuable tool for diagnosis and screening of field samples as well as for conducting molecular studies.En este trabajo comunicamos un método rápido que permite la amplificación del genoma completo del subtipo 2 (H3N8) del virus de la influenza equina. Se utilizó la enzima transcriptasa reversa ThermoScriptTM en lugar de la transcriptasa reversa del virus de la mieloblastosis aviar o la transcriptasa reversa del virus de la leucemia murina de Moloney. Esta enzima ha demostrado tener una alta estabilidad térmica y la capacidad de hacer largas copias de ADN con una estructura secundaria compleja. El producto obtenido por esta técnica puede ser clonado y utilizado posteriormente en reacciones de secuenciación o de PCR anidada con la finalidad de lograr un diagnóstico rápido y la caracterización del virus de la influenza equina tipo A. Este ensayo de detección puede llegar a ser una valiosa herramienta para el diagnóstico y el análisis de muestras de campo, así como para la realización de estudios moleculares

MANAJEMEN PEMBINAAN PERPUSTAKAAN KELURAHAN DI KOTA BANDUNG

Dinas Perpustakaan dan Kearsipan (Dispusip) Kota Bandung merupakan suatu bentuk dari perpustakaan umum yang memiliki fungsi membina perpustakaan. Pemberdayaan perpustakaan kelurahan di Kota Bandung saat ini belum terealisasi sesuai dengan jumlah kelurahan yang terdapat di kota Bandung sebanyak 151 kelurahan. Perpustakaan kelurahan dapat menjalankan fungsinya dengan baik sesuai dengan fungsi perpustakaan dengan penerapan fungsi manajemen perpustakaan. Masalah yang menjadi kajian pada penelitian ini bagaimana implementasi manajemen pembinaan perpustakaan kelurahan di Kota Bandung oleh Dispusip Kota Bandung. Tujuan penelitian ini yaitu untuk mendeskripsikan mengenai perencanaan, pengorganisasian, penggerakan dan pengawasan pembinaan perpustakaan kelurahan oleh Dispusip Kota Bandung. Metode yang digunakan pada penelitian ini adalah metode kualitatif dengan pendekatan studi kasus. Partisipan dalam penelitian ini terdiri atas tiga informan yang bertindak sebagai pustakawan dan key informan sebagai Kepala Bidang Pengembangan Perpustakaan dan Kearsipan menggunakan teknik purposive sampling. Instrumen penelitian yang digunakan adalah wawancara, observasi dan studi dokumentasi. Teknik analisis data yang digunakan adalah data reduction, data display, dan conclusion drawing (verification). Berdasarkan hasil penelitian yaitu sebuah konstuksi model manajemen pembinaan perpustakaan kelurahan yang menunjukkan bahwa kegiatan pembinaan perpustakaan kelurahan oleh Dispusip Kota Bandung relevan dengan fungsi manajemen. Fungsi perencanaan, pengorganisasian, penggerakan kegiatan manajemen pembinaan perpustakaan kelurahan di Kota Bandung terealisasi cukup baik. Namun pada fungsi pengawasan belum berjalan secara optimal, hal ini dikarenakan oleh keterbatasan sumber daya manusia yang dimiliki oleh Dispusip Kota Bandung. Program pembinaan ini diharapkan dapat mewujudkan penyelenggaraan perpustakaan kelurahan dan dapat memberikan kontribusi terhadap perbaikan/ penyempurnaan manajemen pembinaan perpustakaan oleh Dispusip Kota Bandung. ;--- Dinas Perpustakaan dan Kearsipan (Dispusip) Kota Bandung is a form of public library, which function is to develop urban village libraries. Currently, the empowerment of urban village libraries in 151 village in Bandung City is not realized yet. Urban village libraries can perform its functions properly in accordance with the implementation of library management functions. The problem of this study is how Dispusip Kota Bandung manages the implementation of urban village libraries coaching in Bandung City. The purpose of this study is to describe the planning, organizing, actuating, and controlling of urban village libraries coaching by Dispusip Kota Bandung. The research method used in this study is qualitative method with a case study approach. The subjects of this study consist of three informants who act as a librarian and a key informant as a Head of Library and Archive Development by using purposive sampling technique. The research instruments used in this study are interviews, observations, and documentation studies. This study uses data analysis technique of data reduction, data display, and conclusion drawing (verification). Based on the study of a model construction of urban village libraries coaching, the result is that the coaching activities of urban village libraries done by Dispusip Kota Bandung is relevant to the management functions. The functions of planning, organizing, and actuating of village libraries coaching activities in Bandung City are realized quite well. However, the function of controlling has not run optimally. This is due to the limitations of human resources owned by Dispusip Kota Bandung. This coaching program is expected to be able to realize the enforcement of urban village libraries and to give contribution to the improvement of library coaching management done by Dispusip Kota Bandung

High Mutational Heterogeneity, and New Mutations in the Human Coagulation Factor V Gene. Future Perspectives for Factor V Deficiency Using Recombinant and Advanced Therapies

Enfermedad de Owren; Análisis de mutaciones; ParahemofiliaMalaltia d'Owren; Anàlisi de mutacions; ParahemofíliaOwren’s disease; Mutation analysis; ParahemophiliaFactor V is an essential clotting factor that plays a key role in the blood coagulation cascade on account of its procoagulant and anticoagulant activity. Eighty percent of circulating factor V is produced in the liver and the remaining 20% originates in the α-granules of platelets. In humans, the factor V gene is about 80 kb in size; it is located on chromosome 1q24.2, and its cDNA is 6914 bp in length. Furthermore, nearly 190 mutations have been reported in the gene. Factor V deficiency is an autosomal recessive coagulation disorder associated with mutations in the factor V gene. This hereditary coagulation disorder is clinically characterized by a heterogeneous spectrum of hemorrhagic manifestations ranging from mucosal or soft-tissue bleeds to potentially fatal hemorrhages. Current treatment of this condition consists in the administration of fresh frozen plasma and platelet concentrates. This article describes the cases of two patients with severe factor V deficiency, and of their parents. A high level of mutational heterogeneity of factor V gene was identified, nonsense mutations, frameshift mutations, missense changes, synonymous sequence variants and intronic changes. These findings prompted the identification of a new mutation in the human factor V gene, designated as Jaén-1, which is capable of altering the procoagulant function of factor V. In addition, an update is provided on the prospects for the treatment of factor V deficiency on the basis of yet-to-be-developed recombinant products or advanced gene and cell therapies that could potentially correct this hereditary disorder.This study was supported by the Andalusian Association of Hemophilia (ASANHEMO FV 2016–20 grant) and Octapharma S.A. (OCPH-2019-20 grant)

A Digital Pattern Methodology supporting Railway Industries in Portfolio Management

The object of this paper is the development of a decision support system involved in the bidding for invitations to tender in the railway field. The proposed methodology is based on the characterization of the whole train and its components, through several attributes according to a digital pattern approach. In particular some key components were chosen such as the traction motor, the bogie and the auxiliary equipment converter. The system measures the extent to which the products offered by the company fit the one required by the customer, comparing the homologous attributes. Such analysis is called ‘adopt/adapt/innovate’ (AAI). In this way it is possible to identify products already designed that fully or partly fit what required, obtaining huge benefits in terms of effectiveness and efficiency

Neuronal guidance molecule netrin-1 attenuates inflammatory cell trafficking during acute experimental colitis

Background: Inflammatory bowel diseases, encompassing Crohn’s disease and ulcerative colitis, are characterised by persistent leucocyte tissue infiltration leading to perpetuation of an inappropriate inflammatory cascade. The neuronal guidance molecule netrin-1 has recently been implicated in the orchestration of leucocyte trafficking during acute inflammation. We therefore hypothesised that netrin-1 could modulate leucocyte infiltration and disease activity in a model of inflammatory bowel disease. Design: DSS-colitis was performed in mice with partial genetic netrin-1 deficiency (Ntn-1+/- mice) or wild-type mice treated with exogenous netrin-1 via osmotic pump to examine the role of endogenous and therapeutically administered netrin-1. These studies were supported by in vitro models of transepithelial migration and intestinal epithelial barrier function. Results: Consistent with our hypothesis, we observed induction of netrin-1 during intestinal inflammation in vitro or in mice exposed to experimental colitis. Moreover, mice with partial netrin-1 deficiency demonstrated an exacerbated course of DSS-colitis compared to littermate controls, with enhanced weight loss and colonic shortening. Conversely, mice treated with exogenous mouse netrin-1 experienced attenuated disease severity. Importantly, permeability studies and quantitative assessment of apoptosis reveal that netrin-1 signalling events do not alter mucosal permeability or intestinal epithelial cell apoptosis. In vivo studies of leucocyte transmigration demonstrate suppression of neutrophil trafficking as a key function mediated by endogenous or exogenously administered netrin-1. Finally, genetic studies implicate the A2B adenosine receptor in netrin-1-mediated protection during DSS-colitis. Conclusions: The present study identifies a previously unrecognised role for netrin-1 in attenuating experimental colitis through limitation of neutrophil trafficking

Expression of taste receptors in Solitary Chemosensory Cells of rodent airways

<p>Abstract</p> <p>Background</p> <p>Chemical irritation of airway mucosa elicits a variety of reflex responses such as coughing, apnea, and laryngeal closure. Inhaled irritants can activate either chemosensitive free nerve endings, laryngeal taste buds or solitary chemosensory cells (SCCs). The SCC population lies in the nasal respiratory epithelium, vomeronasal organ, and larynx, as well as deeper in the airway. The objective of this study is to map the distribution of SCCs within the airways and to determine the elements of the chemosensory transduction cascade expressed in these SCCs.</p> <p>Methods</p> <p>We utilized a combination of immunohistochemistry and molecular techniques (rtPCR and in situ hybridization) on rats and transgenic mice where the Tas1R3 or TRPM5 promoter drives expression of green fluorescent protein (GFP).</p> <p>Results</p> <p>Epithelial SCCs specialized for chemoreception are distributed throughout much of the respiratory tree of rodents. These cells express elements of the taste transduction cascade, including Tas1R and Tas2R receptor molecules, α-gustducin, PLCβ2 and TrpM5. The Tas2R bitter taste receptors are present throughout the entire respiratory tract. In contrast, the Tas1R sweet/umami taste receptors are expressed by numerous SCCs in the nasal cavity, but decrease in prevalence in the trachea, and are absent in the lower airways.</p> <p>Conclusions</p> <p>Elements of the taste transduction cascade including taste receptors are expressed by SCCs distributed throughout the airways. In the nasal cavity, SCCs, expressing Tas1R and Tas2R taste receptors, mediate detection of irritants and foreign substances which trigger trigeminally-mediated protective airway reflexes. Lower in the respiratory tract, similar chemosensory cells are not related to the trigeminal nerve but may still trigger local epithelial responses to irritants. In total, SCCs should be considered chemoreceptor cells that help in preventing damage to the respiratory tract caused by inhaled irritants and pathogens.</p

Ketogal Safety Profile in Human Primary Colonic Epithelial Cells and in Mice

In our previous studies, a ketorolac–galactose conjugate (ketogal) showed prolonged anti-inflammatory and analgesic activity, causing less gastric ulcerogenic effect and renal toxicity than its parent drug ketorolac. In order to demonstrate the safer profile of ketogal compared to ketorolac, histopathological changes in the small intestine and liver using three staining techniques before and after repeated oral administration in mice with ketorolac or an equimolecular dose of its galactosylated prodrug ketogal were assessed. Cytotoxicity and oxidative stress parameters were evaluated and compared in ketorolac-and ketogal-treated Human Primary Colonic Epithelial cells at different concentrations and incubation times. Evidence of mitochondrial oxidative stress was found after ketorolac treatment; this was attributable to altered mitochondrial membrane depolarization and oxidative stress parameters. No mitochondrial damage was observed after ketogal treatment. In ketorolac-treated mice, severe subepithelial vacuolation and erosion with inflammatory infiltrates and edematous area in the intestinal tissues were noted, as well as alterations in sinusoidal spaces and hepatocytes with foamy cytoplasm. In contrast, treatment with ketogal provided a significant improvement in the morphology of both organs. The prodrug clearly demonstrated a safer profile than its parent drug both in vitro and ex vivo, confirming that ketogal is a strategic alternative to ketorolac

MOQUILLO EN ZORRO COLORADO: La ocurrencia de moquillo en mamíferos silvestres refuerza la importancia de la vacunación periódica en cánidos domésticos a fin de conservar la salud de la fauna autóctona.

El moquillo canino es una enfermedad infectocontagiosa que afecta principalmente a perros, pero también a mamíferos silvestres. En 2022, se diagnosticó moquillo en un zorro colorado en Junín de los Andes, Neuquén. El aumento del número de perros con signos compatibles con esta enfermedad durante el 2021 nos pone en alerta por la posible transmisión del virus entre estas especies. La forma de prevenirla es la vacunación completa y anual, siendo una responsabilidad de todo ciudadano que posea perros bajo su cuidado, no solamente por el bien de los animales de compañía sino para el cuidado de la fauna silvestre en general