Funzionalita surrenale nel paziente canino in SIRS

Nei pazienti critici si può verificare una condizione di insufficienza corticosteroidea correlata allo stato di criticità denominata CIRCI, che è risultata associata a ipotensione e aumentata mortalità nell’uomo. In veterinaria pochi studi hanno finora valutato la presenza di tale sindrome e non ci sono studi pubblicati che valutino la risposta alla terapia con basse dosi di idrocortisone. L’obiettivo dello studio è di verificare la presenza di CIRCI nei pazienti canini affetti da sindrome da risposta infiammatoria sistemica (SIRS), valutarne l’eventuale correlazione con outcome e la risposta al trattamento con idrocortisone. Ventun pazienti in SIRS sono stati inclusi nello studio e per tutti è stata valutata la funzionalità surrenale tramite esecuzione di un test di stimolazione con ACTH, considerando cortisolo basale, cortisolo post ACTH, Δ cortisolo e aumento percentuale di cortisolo; sono stati inoltre valutati i parametri di pressione arteriosa media (MAP), glicemia, conta leucocitaria totale (WBC), percentuale di neutrofili banda, lattatemia, concentrazione di proteina c reattiva (CRP) e grading di SIRS. Quattordici pazienti sono stati trattati con idrocortisone a basse dosi, mentre i restanti 7 hanno costituito il gruppo di controllo. L’incidenza di CIRCI è risultata del 48 % basandosi su un Δ cortisolo ≤ 3 μg/dl. I pazienti CIRCI positivi avevano concentrazioni di CRP significativamente maggiori dei CIRCI negativi. L’aumentata mortalità non è risultata associata alla presenza di CIRCI, mentre è risultata significativamente associata a ipotensione, ipercortisolemia basale, leucocitosi e contemporanea presenza di ipercortisolemia basale e CIRCI. Sebbene la sopravvivenza sia maggiore nei trattati con idrocortisone, la differenza non è statisticamente significativa. A critical illness related corticosteroid insufficiency (CIRCI) occurs in critically ill human patients, and it’s associated with hypotension and increased mortality. A few veterinary studies have investigated CIRCI syndrome and no published studies have evaluated the benefit of low-dose hydrocortisone. The purpose of the current study is to verify the presence of CIRCI in dogs with systemic inflammatory response syndrome (SIRS), assess its relationship with outcome and the response to hydrocortisone therapy. Twenty-one SIRS dogs were included in the study and were all evaluated for adrenal function with ACTH stimulation test, considering baseline and ACTH stimulated serum cortisol concentrations, Δ cortisol and cortisol percentage increase. Mean arterial pressure (MAP), glycemia, white blood cell count (WBC), banda neutrophilia percentage, lactate concentration, c-reactive protein concentration (CRP) and SIRS grading were also assessed. Fourteen dogs were treated with low-dose hydrocortisone, the other seven dogs costituited the control group. CIRCI occurred in 48 % of the patients, basing on a Δ cortisolo ≤ 3 μg/dl. Patients with CIRCI had significantly higher CRP concentrations than patients without CIRCI. Increased mortality was not associated with CIRCI, but was significantly associated with hypotension, basal hypercortisolemia, leukocytosis and association of hypercortisolemia and CIRCI. Even if survival is greater in patients treated with hydrocortisone, the difference is not statistically significative

Indagine molecolare per la valutazione della qualita' e dell'amplificabilita' del DNA estratto da preparazioni a base di surimi: difficolta' e prospettive

Attualmente, a seguito dei cambiamenti dello stile di vita, i consumatori sono orientati verso prodotti ittici caratterizzati da praticità di preparazione (ready to eat e ready to cook products). Tra questi, le preparazioni a base di surimi, prodotti costituiti da pesce ricomposto addizionato con vari ingredienti. La perdita delle caratteristiche morfologiche che si verifica durante la produzione del surimi rende impossibile l’identificazione delle specie utilizzate tramite analisi morfologica: è quindi necessario ricorrere a metodiche di laboratorio basate sull’analisi del DNA. In questo lavoro, considerando le limitazioni associate all’uso di tecniche analitiche basate sul sequenziamento quando applicate a prodotti trasformati e multispecie, 40 preparazioni a base di surimi sono state analizzate al fine di valutare il livello di degradazione del DNA e la sua amplificabilità a mezzo di primers universali. In particolare, è stato testato un nuovo approccio (multi DNA barcoding) analizzando sia frammenti del gene mitocondriale che codifica per la subunità 16S ribosomiale, che frammenti del gene mitocondriale che codifica per la subunità I della Citocromo C Ossidasi, utilizzando 5 coppie di primers differenti (3 per il gene COI e 2 per il gene 16S) per l’amplificazione di target di lunghezza variabile (da 655 a 139 pb). Tali primers sono stati preventivamente testati su 132 campioni di DNA derivanti da 45 specie di riferimento (41 di pesci e 4 di cefalopodi) valutando sia la percentuale di amplificazione che la concentrazione dei prodotti di PCR. Successivamente gli stessi primers sono stati utilizzati per amplificare e sequenziare i DNA estratti dai 40 campioni commerciali. Tutte le sequenze ottenute sono state confrontate con i database pubblici (BOLD e GenBank). I livelli di degradazione del DNA estratto da tali campioni sono stati quindi valutati in funzione dei risultati dell’amplificazione e del sequenziamento, valutando anche la lunghezza delle sequenze ottenute rispetto a quella dell’amplicone atteso. Le etichette di tutti i prodotti sono state analizzate alla luce delle normative vigenti. Le analisi effettuate hanno evidenziato che il 35% dei campioni analizzati mostrava un pattern di degradazione del DNA totale con frammenti inferiori alle 500 pb. Nonostante questo, l’utilizzo di più coppie di primers per l’amplificazione di frammenti di lunghezza differente (lunghi e corti) ha permesso di sequenziare tutti i prodotti di PCR ottenuti. Nel complesso, il 76% delle sequenze ottenute sono state ritenute utilizzabili. Dai risultati ottenuti dal confronto con i database il 18,5% dei campioni è risultato non conforme. Nonostante la metodica applicata abbia permesso di verificare le informazioni riportate sull’etichetta di alcuni prodotti le sue limitazioni non hanno permesso di valutare in maniera esaustiva la composizione di tutti i prodotti analizzati. Il presente lavoro conferma la valutazione della degradazione del DNA e la selezione dei primers quali step essenziali per superare i limiti associati all’analisi di prodotti trasformati e multispecie. Currently, as a result of lifestyle changes, consumers moved towards seafood products characterized by a convenient preparation (ready to eat and ready to cook products). Among these, surimi-based products are preparations made of reassembled fish, supplemented with various ingredients. The loss of morphological characteristics of this processed product makes the species identification by morphological analysis impossible. Thus, it is necessary to use laboratory methods based on DNA analysis. In this work, considering the limitations associated with the use of analytical techniques based on sequencing when applied to processed and multispecies products, 40 commercial surimi-based products were analyzed in order to assess the level of degradation and the amplificability of DNA using universal primers. In particular, a new approach (multi DNA barcoding) was tested, analyzing both fragments belonging to the mitochondrial gene encoding the mitochondrial 16s ribosomal RNA gene (16s rRNA) and fragments belonging to the mitochondrial gene encoding the subunit I of Cytochrome C oxidase (COI), using 5 different couples of primers (3 for the COI gene and 2 for the 16s rRNA gene) to amplify target of different lengths (from 655 to 139 pb). These primers were tested on 132 samples of DNA extracted from 45 reference species (41 fishes and 4 cephalopods), in order to evaluate both the amplification rate and the concentration of PCR products. Then, the same primers have been used to amplify and sequence DNA extracts from 40 commercial samples. All the sequences obtained were compared with public database (BOLD and GenBank). The levels of DNA degradation of these samples were assessed in the light of the amplification and sequencing output, also comparing the lengths of the obtained sequences with those of the expected amplicons. The label of each product was analyzed according to the regulations in force. The performed analysis showed a pattern of degradation of total DNA with fragments less than 500 pb in 35% of the samples. Despite this, using multiple couples of primers to amplify fragments of different lengths (long and short), all the PCR products were successfully sequenced. Overall, 76% of the obtained sequences have been considered usable. The comparison with the databases highlighted that 18.5% of the samples was non-compliant. Despite the method applied has allowed to verify the information on the label of some products, limits associated to the technique did not allow to completely evaluate the composition of all the products. The present work confirms the evaluation of the DNA degradation and the selection of the primers as an essential step to overcome the limitations associated to the analysis of processed and multispecies products

The way to ultrafast, high-throughput enantioseparations of bioactive compounds in liquid and supercritical fluid chromatography

Until less than 10 years ago, chiral separations were carried out with columns packed with 5 or 3 μm fully porous particles (FPPs). Times to resolve enantiomeric mixtures were easily larger than 30 min, or so. Pushed especially by stringent requirements from medicinal and pharmaceutical industries, during the last years the field of chiral separations by liquid chromatography has undergone what can be defined a “true revolution”. With the purpose of developing ever faster and efficient method of separations, indeed, very efficient particle formats, such as superficially porous particles (SPPs) or sub-2 μm FPPs, have been functionalized with chiral selectors and employed in ultrafast applications. Thanks to the use of short column (1–2 cm long), packed with these extremely efficient chiral stationary phases (CSPs), operated at very high flow rates (5–8 mL/min), resolution of racemates could be accomplished in very short time, in many cases less than 1 s in normal-, reversed-phase and HILIC conditions. These CSPs have been found to be particularly promising also to carry out high-throughput separations under supercritical fluid chromatography (SFC) conditions. The most important results that have been recently achieved in terms of ultrafast, high-throughput enantioseparations both in liquid and supercritical fluid chromatography with particular attention to the very important field of bioactive chiral compounds will be reviewed in this manuscript. Attention will be focused not only on the latest introduced CSPs and their applications, but also on instrumental modifications which are required in some cases in order to fully exploit the intrinsic potential of new generation chiral columns

An unusual complication after endovascular aneurysm repair for giant abdominal aortic aneurysm with aortocaval fistula: High bilirubin levels

Abdominal aortic aneurysm has among its rare complications the aortocaval fistula. It is observed in less than 1% of all abdominal aortic aneurysms and represents 3%–7% of clinical presentation in case of rupture. A male patient was presented to the emergency department with pulsating mass with continuous vascular systo-diastolic bruit, located in the lower part of abdomen with the back pain radiating anteriorly in lower abdomen. After diagnosis of abdominal aortic aneurysm with aortocaval fistula, a trimodular Endurant endograft was placed. Migration of the endoprosthesis was treated with Endoanchor and endovascular aneurysm sealing device. In the postoperative course, the patient had jaundice due to high bilirubin levels, cholestasis and increased hepatocyte cytolysis: aspartate aminotransferase and alanine aminotransferase. The treatment with appropriate continuous filtration rapidly reduced bilirubin values and the patient gradually improved

Drug resistance of BRAF-mutant melanoma: Review of up-to-date mechanisms of action and promising targeted agents

Abstract Melanoma onset and progression are associated with a high variety of activating mutations in the MAPK-pathway, most frequently involving BRAF (35–45%) and NRAS (15–25%) genes, but also c-KIT and PTEN. Targeted therapies with BRAF and MEK inhibitors showed promising results over the past years, but it is known that most responses are temporary, and almost all of patients develop a tumor relapse within one year. Different drug-resistance mechanisms underlie the progression of disease and activation of both MAPK and PI3K/AKT/mTOR pathways. Therefore, in this article we reviewed the main studies about clinical effects of several target inhibitors, describing properly the most prominent mechanisms of both intrinsic and acquired resistance. Furthermore, suggestive strategies for overcoming drug resistance and the most recent alternative combination therapies to optimize the use of MAPK pathway inhibitors were also discussed

GTE. A new software for gravitational terrain effect computation: theory and performances

The computation of the vertical attraction due to the topographic masses, the so-called Terrain Correction, is a fundamental step in geodetic and geophysical applications: it is required in high-precision geoid estimation by means of the remove–restore technique and it is used to isolate the gravitational effect of anomalous masses in geophysical exploration. The increasing resolution of recently developed digital terrain models, the increasing number of observation points due to extensive use of airborne gravimetry in geophysical exploration and the increasing accuracy of gravity data represents nowadays major issues for the terrain correction computation. Classical methods such as prism or point masses approximations are indeed too slow while Fourier based techniques are usually too approximate for the required accuracy. In this work a new software, called Gravity Terrain Effects (GTE), developed to guarantee high accuracy and fast computation of terrain corrections is presented. GTE has been thought expressly for geophysical applications allowing the computation not only of the effect of topographic and bathymetric masses but also those due to sedimentary layers or to the Earth crust-mantle discontinuity (the so-called Moho). In the present contribution, after recalling the main classical algorithms for the computation of the terrain correction we summarize the basic theory of the software and its practical implementation. Some tests to prove its performances are also described showing GTE capability to compute high accurate terrain corrections in a very short time: results obtained for a real airborne survey with GTE ranges between few hours and few minutes, according to the GTE profile used, with differences with respect to both planar and spherical computations (performed by prism and tesseroid respectively) of the order of 0.02 mGal even when using fastest profiles

Characterisation of the antibiotic profile of Lysobacter capsici AZ78, an effective biological control agent of plant pathogenic microorganisms

Determining the mode of action of microbial biocontrol agents plays a key role in their development and registration as commercial biopesticides. The biocontrol rhizobacterium Lysobacter capsici AZ78 (AZ78) is able to inhibit a vast array of plant pathogenic oomycetes and Gram-positive bacteria due to the release of antimicrobial secondary metabolites. A combination of MALDI-qTOF-MSI and UHPLC-HRMS/M was applied to finely dissect the AZ78 metabolome and identify the main secondary metabolites involved in the inhibition of plant pathogenic microorganisms. Under nutritionally limited conditions, MALDI-qTOF-MSI revealed that AZ78 is able to release a relevant number of antimicrobial secondary metabolites belonging to the families of 2,5-diketopiperazines, cyclic lipodepsipeptides, macrolactones and macrolides. In vitro tests confirmed the presence of secondary metabolites toxic against Pythium ultimum and Rhodococcus fascians in AZ78 cell-free extracts. Subsequently, UHPLC-HRMS/MS was used to confirm the results achieved with MALDI-qTOF-MSI and investigate for further putative antimicrobial secondary metabolites known to be produced by Lysobacter spp. This technique confirmed the presence of several 2,5-diketopiperazines in AZ78 cell-free extracts and provided the first evidence of the production of the cyclic depsipeptide WAP-8294A2 in a member of L. capsici species. Moreover, UHPLC-HRMS/MS confirmed the presence of dihydromaltophilin/Heat Stable Antifungal Factor (HSAF) in AZ78 cell-free extracts. Due to the production of HSAF by AZ78, cell-free supernatants were effective in controlling Plasmopara viticola on grapevine leaf disks after exposure to high temperatures. Overall, our work determined the main secondary metabolites involved in the biocontrol activity of AZ78 against plant pathogenic oomycetes and Gram-positive bacteria. These results might be useful for the future development of this bacterial strain as the active ingredient of a microbial biopesticide that might contribute to a reduction in the chemical input in agricultur

Cervical manipulation and cerebrovascular ischemia. A rare but dangerous association. Monocentric experience in a high CEA volume center and literature review.

Abstract Background: A correlation between stroke and cervical manipulation in the neck is reported in the literature with increasing frequency, and each new publication seems to rekindle the debate. The manipulations in the cervical region is potentially dangerous for arterial vessels. The aim of this work is to analyze the incidence of these events in the experience of a center with high volume surgery, in which emergency interventions are performed for acute cerebrovascular Ischemia. Methods: Data were collected regarding patients undergoing carotid endarterectomy surgery for neurological symptoms of cerebrovascular insufficiency over a seven-year period. Results: The intervention was conducted urgently in 66 cases for acute symptoms (7.6%) with a median symptom interval / CEA: 39.4 ± 5.4 h) in 32 cases for hemispheric transient ischemic attack, while in 34 for minor stroke. Only in 1 case (1.5%) did a condition of severe spondyloarthrosis of the cervical spine occur in acute carotid plaque in C4-C5. In another case (1.5%) a cervical trauma caused by beatings is documented. Conclusions: Manipulations of the neck and cervical spine can rarely cause or contribute to worsening neurological symptoms, however they should always be considered

Catecholamine-induced Takotsubo syndrome: a case series

Background: Catecholamine-induced Takotsubo Syndrome (cat-TS) is a type of secondary Takotsubo syndrome, characterized by rapid onset of symptoms, high rate of complications during the acute phase, good short-term prognosis, and frequent apical sparing at echocardiogram. We present two clinical cases of cat-TS treated in our department. Case summary: Case one: 78-year-old man, admitted to Ear Nose and Throat Unit for surgical removal of oral squamous cellular carcinoma. During surgery, the occurrence of hypotensive episode was treated with catecholamines. After surgery, the occurrence of atrial fibrillation was followed by evidence of phasic increase of troponin levels and akinesia of midventricular segments. Angiography showed the absence of significant coronary stenoses, and during hospital stay, we observed rapid recovery of wall motion abnormalities. Case two: 64-year-old woman, admitted for hysteropexy surgery, during which cardiac arrest occurred, treated with epinephrine i.v.1 mg and DC shock. Two hours after resuscitation, the patient developed pulmonary oedema, troponin levels increased progressively, and the echocardiogram demonstrated hypokinesia in all midventricular segments with apical sparing. Afterwards, an urgent angiography highlighted normal coronary anatomy. Cardiac magnetic resonance imaging (MRI) revealed oedema corresponding to hypokinetic areas. On the seventh day, echocardiogram showed a complete remission of wall motion abnormalities. Discussion: These cases warn the physicians about the importance of routinely screening myocardial impairment through clinical assessment, electrocardiogram (ECG) monitoring, and serial cardiac troponin testing after catecholamine i.v. bolus administration. In case of alterations of these exams, performing a prompt echocardiogram allows early detection of cat-TS, to provide immediate suitable medical support and avoid complications

Successful conversion strategy in patient submitted to EVAR demanding open surgery: comparative analysis 1997-2011 vs 2012-2020

: Late open conversion in our center has been reviewed in the past 8 years, comparing 1997-2011 (first period group A) with 2012-2020 (second period group B). A retrospective analysis of patients treated at our centre by standard EVAR for infrarenal aortic aneurysm requiring late open conversion between January 1997 and February 2020 was performed. All stent grafts were implanted according to their current IFU all patients. The data concerning intra and postoperative complications were collected. Post-operative evaluated data include: ICU (Intensive Care Unit) stay, major peri-operative (<30 days) complications, in hospital mortality, length of hospital stay, 30-days mortality, and mid-term outcomes. Between January 2012 and February 2020 (group B), in our institution 8 patients previously treated by stent graft with endoleak underwent open surgery. The incidence of conversions and the 30-day mortality rate were compared with that of previous years, from January 1997 to December 2011 (group A). 481 patients submitted to EVAR in a second part of the analysis have been considered, 8 patients underwent late open conversion (1.7%) (Group B) due to endoleak. Among January 1997 and December 2011 overall 268 EVAR were performed; during this first study period, surgical conversion had been performed in 14 patients (5.2%) (Group A). The average time from EVAR to open conversion was four years (range 12-88 months) in Group B, and it was 30 months (range 1-82 months) in Group A. In most cases, in both group A and group B the proximal aortic cross-clamping were infrarenal. After the emergent procedure in Group B (12.5%), we have observed a death, whereas three patients died in Group A in urgent situations (21.4%). The more frequent indication for open surgery is the Endoleak type 1 and migration in the two considered periods. Adherence with current IFU and the technical progress in endoprosthesis design maintain lower rate incidence. In most cases, open surgery for prostheses that require explantation can be performed with infrarenal clamping. Partial removal of the endoprosthesis in selected cases makes open conversion easier and appears durable. The results are unfair by numerous comorbidities; in both periods, urgent graft removal seems to elevate both mortality and morbidity, compared to elective surgery