Aortic stenting in the growing sheep causes aortic endothelial dysfunction but not hypertension: Clinical implications for coarctation repair

Stent implantation is the treatment of choice for adolescents and adults with aortic coarctation (CoAo). Despite excellent short-term results, 20%-40% of the patients develop arterial hypertension later in life, which was attributed to inappropriate response of the aortic baroreceptors to increased stiffness of the ascending aorta (ASAO), either congenital or induced by CoAo repair. In particular, it has been hypothesized that stent itself may cause or sustain hypertension. Therefore, we aimed to study the hemodynamic and structural impact following stent implantation in the normal aorta of a growing animal

Paraneoplastic epidermolysis bullosa acquisita associated with thyroid carcinoma

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Angiotensin II Facilitates Breast Cancer Cell Migration and Metastasis

Breast cancer metastasis is a leading cause of death by malignancy in women worldwide. Efforts are being made to further characterize the rate-limiting steps of cancer metastasis, i.e. extravasation of circulating tumor cells and colonization of secondary organs. In this study, we investigated whether angiotensin II, a major vasoactive peptide both produced locally and released in the bloodstream, may trigger activating signals that contribute to cancer cell extravasation and metastasis. We used an experimental in vivo model of cancer metastasis in which bioluminescent breast tumor cells (D3H2LN) were injected intra-cardiacally into nude mice in order to recapitulate the late and essential steps of metastatic dissemination. Real-time intravital imaging studies revealed that angiotensin II accelerates the formation of metastatic foci at secondary sites. Pre-treatment of cancer cells with the peptide increases the number of mice with metastases, as well as the number and size of metastases per mouse. In vitro, angiotensin II contributes to each sequential step of cancer metastasis by promoting cancer cell adhesion to endothelial cells, trans-endothelial migration and tumor cell migration across extracellular matrix. At the molecular level, a total of 102 genes differentially expressed following angiotensin II pre-treatment were identified by comparative DNA microarray. Angiotensin II regulates two groups of connected genes related to its precursor angiotensinogen. Among those, up-regulated MMP2/MMP9 and ICAM1 stand at the crossroad of a network of genes involved in cell adhesion, migration and invasion. Our data suggest that targeting angiotensin II production or action may represent a valuable therapeutic option to prevent metastatic progression of invasive breast tumors

Studio della farmacocinetica del Propofol, progettazione e validazione di un sistema di target controlled infusion nel gatto

Total intravenous anesthesia (TIVA) is an actual topic and a still developing research area. Technological progress makes equipment for infusion increasingly sophisticated, reliable and safe. Latest infusion systems have been developed incorporating microprocessors and algorithms based on the drug’s pharmacokinetic and pharmacodynamic parameters for the population involved. The aim of this research project was to develop and validate a system for target-controlled infusion (TCI) for administering propofol to domestic cats. Knowledge on behavior of propofol, administered as a continuous infusion in cats are, at present, still few and sometimes contradictory. It is well known that cats differ substantially from other animals in the ability to form glucuronide conjugates of some xenobiotics, particularly low molecular weight phenolic derivatives. A system for propofol infusion, allowing to predict as accurately as possible, the actual behavior of the drug and the concentrations at any time-point of anesthesia, may help reducing perianesthetic risk and improve quality of anesthesia. As no publications could be found reporting three-compartment pharmacokinetics for propofol in cats, the first part of the this work was dedicated to the development of a pharmacokinetic model, on 7 cats, premedicated with medetomidine, ketamine and methadone, anesthetized with propofol and maintained with isoflurane anesthesia, during gonadectomy. A two-compartment and a three-compartment pharmacokinetic linear model were obtained, the latter being considered the most appropriate to describe plasma concentration-time profile, based on appearance of observed and predicted concentrations and determined by least square linear regression and applying Akaike's information criterion. The tri-compartmental pharmacokinetic data obtained in the first part of the project were used for implementation of a TCI system prototype (TCI-1), based on the free software, Computer Control Infusion Pump (CCIP) Ver 2.0E, available online (http: / / www.cuhk.edu.hk / med / ans / softwares.htm). We analyzed the performance of the TCI-1 system during the course of anesthesia for gonadectomy in 9 premedicated cats, and target concentrations for induction, maintenance and extubation were reported. The mean concentration, as measured by HPLC, in venous blood, two minutes after intubation was 4.92 ± 2.43 μg ml-1 and for maintenance, 4.74 ± 2.70 μg ml-1. The indices for system prediction error, MDPE%, MDAPE%, divergence (% / hour) and wobble, calculated as described by Varvel et al. (1992), reported as median [10 / 90 percentile], were 63.50 [31.24 / 151.93], 63.50 [32.28 / 151.93], -22.77 [- 331 / 26.57], 20.32 [9.8 / 43.10], respectively. Despite it has demonstrated to be manageable in its clinical use, the system has significantly underestimated the actual concentrations, as expected for a single bolus derived pharmacokinetic model. The next step was to optimize the TCI-1 system, modifying the pharmacokinetic data set through computer simulations. The new system (TCI-2.0) was assessed for predictive performance, in 6 cats, calculating MDPE%, MDAPE%, divergence (%/hours) and wobble, and it was compared to the results obtained with the TCI-1 system. Mean concentration, measured by HPLC, in venous blood of cats, two minutes after intubation was 3.23 ± 0.87 μg kg-1. Mean maintenance target concentration, calculated as the mean of targets set introduced in the system, during anesthesia, was 6.44 ± 1.27 μg ml-1, while average concentrations measured by HPLC during the same time period , was 5.56 ± 2.46 μg ml-1. Values of MDPE%, MDAPE%, divergence and wobble, reported as median [10 / 90 percentile], were -1.85 [-35.19 / 31.08], 29.67 [16.85 / 43.93], -39.08 [-50.02 / 8.23], 18.93 [10.49 / 25.01], respectively. Prediction curves, obtained by the system TCI-2.0 were similar to actually measured plasma concentrations. The resulting values for MDPE and MDAPE fall within the range described in human and veterinary literature for the validation of TCI systems in the clinical setting (MDPE <± 20% and MDAPE <30-40%). Difference in accuracy and bias observed between the two systems, TCI-1 and TCI-2.0 were statistically significant. Timing and dose of induction, maintenance, and extubation were also compared. Differences between the two systems, for doses and timing of administration and extubation, were not found to be statistically significant. Obtained results are, in the author’s opinion, of significant scientific relevance, as they provide: • A three-compartment pharmacokinetic model for propofol in cats, never described previously in the literature. • A TCI system for propofol administration in cats, validated for prediction performance and easy to use in a clinical setting. • reference values for plasma concentrations of propofol required for intubation, maintenance of anesthesia and extubation, in cats premedicated with medetomidine, ketamine and methadone, anesthetized with target infusion of propofol.L’anestesia totalmente endovenosa (TIVA) è ancora un campo in via di sviluppo. Il progresso tecnologico rende le apparecchiature per infusione sempre più sofisticate, affidabili e sicure. I sistemi infusionali più recenti sono sviluppati incorporando in un microprocessore i parametri farmacocinetici e farmacodinamici dell’anestetico impiegato, per la popolazione in oggetto. Il fine di questo progetto di ricerca è stato quello di sviluppare e validare un sistema di infusione controllata a target (TCI) per il propofol nel gatto domestico. In questa specie, le conoscenze relative al comportamento del propofol nell’ambito dell’infusione continua sono ancora poche e spesso contraddittorie. È noto che il gatto differisca sostanzialmente dagli altri animali nell’abilità di formare coniugati glucuronidi di alcuni xenobiotici ed in particolare dei derivati fenolici a basso peso molecolare. Un metodo di infusione del propofol in TIVA, che permetta di prevedere in modo più accurato possibile l’effettivo comportamento del farmaco e la concentrazione in ogni istante, può permettere di ridurre i rischi perianestetici e di migliorare la qualità dell’anestesia. Non esistendo nella letteratura una farmacocinetica tricompartimentale del propofol nel gatto, la prima parte del lavoro è stata dedicata allo studio del modello farmacocinetico su 7 gatti sottoposti a gonadectomia, premedicati con medetomidina, ketamina e metadone, anestetizzati con propofol e mantenuti in anestesia con isofluorano. È stato possibile ottenere un modello farmacocinetico bi-compartimentale ed uno tri-compartimentale. Il modello più appropriato per rappresentare la curva di concentrazione plasmatica nel tempo è stato quello tri-compartimentale, sulla base dell’aspetto delle concentrazioni osservate e predette, della riduzione della somma dei quadrati residui e dell’applicazione del test di verifica delle informazioni di Akaike. I dati di farmacocinetica tri-compartimentale estrapolati nella prima parte del progetto sono stati utilizzati per programmare il sistema di TCI prototipo (TCI-1), basato sul software gratuito Computer Control Infusion Pump (CCIP) Ver 2.0E, disponibile in rete (http://www.cuhk.edu.hk/med/ans/softwares.htm). È stata analizzata la performance del sistema TCI-1 durante l’anestesia per interventi chirurgici di gonadectomia, su 9 gatti premedicati, e sono state studiate le concentrazioni target di induzione, mantenimento e risveglio. La concentrazione plasmatica, misurata con HPLC, nel sangue venoso, due minuti dopo l’intubazione è stata in media di 4,92 ± 2,43 μg ml-1, quella di mantenimento 4,74 ±2,70 μg ml-1. Gli indici di errore del sistema, MDPE%, MDAPE%, divergenza (%/ora) e wobble, calcolati secondo quanto descritto da Varvel et al. (1992), riportati come mediana [10 / 90 percentile], sono stati pari a 63,50 [31,24 / 151,93], 63,50 [32,28 / 151,93], -22,77 [-331 / 26,57], 20,32 [9,8 / 43,10] , rispettivamente. Il sistema, nonostante si sia rivelato maneggevole nell’utilizzo clinico, ha sensibilmente sottostimato le concentrazioni effettive, come previsto per una farmcocinetica derivata da bolo singolo. La fase successiva è stata quella di ottimizzazione del sistema TCI-1, modificando il set farmacocinetico attraverso simulazioni al computer. Il nuovo sistema (TCI-2.0) è stato sottoposto a valutazione delle performances predittive, su 6 gatti, calcolando MDPE%, MDAPE%, divergenza (%/ora) e wobble ed è stato messo a confronto, per i risultati ottenuti, con al sistema TCI-1. La concentrazione plasmatica, misurata con HPLC, nel sangue venoso dei gatti, due minuti dopo l’intubazione è stata in media di 3,23 ± 0,87 μg kg-1. La concentrazione plasmatica target media di mantenimento, calcolata come la media dei target impostati nel sistema durante l’anestesia, è stata pari a 6,44 ±1,27 μg ml-1, la media delle concentrazioni misurate con HPLC, nello stesso periodo temporale, è risultata pari a 5,56 ±2,46 μg ml-1. I valori di MDPE%, MDAPE%, divergenza e wobble, riportati come mediana [10 / 90 percentile], sono stati pari a -1,85 [-35,19 / 31,08], 29,67 [16,85 / 43,93], -39,08 [-50,02 / 8,23], 18,93 [10,49 / 25,01], rispettivamente. Le curve di predizione ottenute dal sistema TCI-2.0 sono state vicine a quelle relative ai valori di concentrazione plasmatica realmente misurati. I valori ottenuti per MDPE e MDAPE rientrano nei range descritti in medicina umana e veterinaria, per la validazione di sistemi TCI in ambito clinico (MDPE < ±20% e MDAPE < 30-40%). La differenza di accuratezza e deviazione osservata tra i due sistemi TCI-1 e TCI-2.0 è stata significativa. Sono stati confrontati anche i tempi e le dosi di induzione, mantenimento ed estubazione. Non sono risultate esserci, tra i due sistemi differenze significative per dosi e tempi di somministrazione ed estubazione. I risultati ottenuti sono, secondo il parere degli autori, di significativa rilevanza scientifica, poiché forniscono: • Un modello farmacocinetico tri-compartimentale del propofol nel gatto, non descritto prima in letteratura. • Un sistema TCI con propofol nel gatto, validato per performances di predizione , facilmente fruibile in ambito clinico. • Dei valori di riferimento per le concentrazioni plasmatiche di propofol, necessarie per l’intubazione, il mantenimento e l’estubazione in gatti premedicati con medetomidina, ketamina e metadone, anestetizzati con infusione di propofol a target

Breed and adaptive response modulate bovine peripheral blood cells' transcriptome

Background: Adaptive response includes a variety of physiological modifications to face changes in external or internal conditions and adapt to a new situation. The acute phase proteins (APPs) are reactants synthesized against environmental stimuli like stress, infection, inflammation. Methods: To delineate the differences in molecular constituents of adaptive response to the environment we performed the whole-blood transcriptome analysis in Italian Holstein (IH) and Italian Simmental (IS) breeds. For this, 663 IH and IS cows from six commercial farms were clustered according to the blood level of APPs. Ten extreme individuals (five APP+ and APP- variants) from each farm were selected for the RNA-seq using the Illumina sequencing technology. Differentially expressed (DE) genes were analyzed using dynamic impact approach (DIA) and DAVID annotation clustering. Milk production data were statistically elaborated to assess the association of APP+ and APP- gene expression patterns with variations in milk parameters. Results: The overall de novo assembly of cDNA sequence data generated 13,665 genes expressed in bovine blood cells. Comparative genomic analysis revealed 1,152 DE genes in the comparison of all APP+ vs. all APP- variants; 531 and 217 DE genes specific for IH and IS comparison respectively. In all comparisons overexpressed genes were more represented than underexpressed ones. DAVID analysis revealed 369 DE genes across breeds, 173 and 73 DE genes in IH and IS comparison respectively. Among the most impacted pathways for both breeds were vitamin B6 metabolism, folate biosynthesis, nitrogen metabolism and linoleic acid metabolism. Conclusions: Both DIA and DAVID approaches produced a high number of significantly impacted genes and pathways with a narrow connection to adaptive response in cows with high level of blood APPs. A similar variation in gene expression and impacted pathways between APP+ and APP- variants was found between two studied breeds. Such similarity was also confirmed by annotation clustering of the DE genes. However, IH breed showed higher and more differentiated impacts compared to IS breed and such particular features in the IH adaptive response could be explained by its higher metabolic activity. Variations of milk production data were significantly associated with APP+ and APP- gene expression patterns

Additional file 3: of Breed and adaptive response modulate bovine peripheral blood cells’ transcriptome

Complete list of genes included in DAVID Annotation Clusters. Each sheet of the.xlsx file includes one of the Annotation Clusters listed in Table 6 of the present article. In each Annotation Cluster the included genes are listed. The genes found to be common in each comparison are bolded. (XLSX 57 kb

Additional file 1: of Breed and adaptive response modulate bovine peripheral blood cells’ transcriptome

Characteristics of animals chosen for the transcriptome analysis in the selected commercial farms. F: farm, IH: Italian Holstein, IS: Italian Simmental, DIM: days in milk, BCS: body condition score, SCC: somatic cell count, values are expressed as mean ± SD. (XLSX 11 kb