Cryptogenic stroke as a working diagnosis: the need for an early and comprehensive diagnostic work-up

Atrial fibrillation; Cryptogenic stroke; Implantable cardiac monitorFibril·lació auricular; Ictus criptogènic; Monitor cardíac implantableFibrilación auricular; Ictus criptogénico; Monitor cardiaco implantableIn the Nordic Atrial Fibrillation and Stroke (NOR-FIB) study, the causes of ischemic stroke were identified in 43% of cryptogenic stroke patients monitored with implantable cardiac monitor (ICM), but one-third of these patients had non-cardioembolic causes. These results suggest the need for an early and comprehensive diagnostic work-up before inserting an ICM

Identification of the most abundant proteins in equine amniotic fluid by a proteomic approach

Characterisation of the physiologic equine amniotic fluid (AF) proteome is a prerequisite to study its changes during diseases and discover new biomarkers. The aim of this study was to identify by a proteomic approach the most abundant proteins of equine AF. AF samples were collected at parturition from 24 healthy mares that delivered healthy foals. All samples were subjected to sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) on 4\u201312% gels. A pool of the 24 samples, after SDS-PAGE, was cut in 25 slices, trypsin-digested and analysed by mass spectrometry (MS) for protein identification. Mean AF protein concentration was 1.96\ua0\ub1\ua01.12\ua0g/L. Thirty-four proteins were successfully identified by MS and subsequently categorised according to Gene Ontology (GO). Twelve proteins (e.g. fibronectin, lumican, thrombospondin and fibulin) belonged to or interacted with the extracellular matrix (ECM) playing an important role in the development of foetal tissues. Most of the remaining proteins were classified as transport (e.g. albumin, major allergen Equ c1 and alpha-fetoprotein) delivering nutrients, ions and lipids essential for foetal growth and development. Among these proteins, major allergen Equ c1 is widely studied in human medicine because it induces Ig-E mediated type I allergic reaction. The absence of immunoglobulins in equine AF was also confirmed

Measurement of cerebrospinal fluid albumin in healthy dogs

Background: Measurement of CSF albumin aids diagnosis in human medicine but technical difficulties related to its low CSF concentration prohibit its routine use in veterinary medicine. High-resolution electrophoresis (HRE) has been described but often results in non-interpretable integration profiles preventing albumin determination. Fraction quantification using HRE may be more precise after concentration (cHRE) using a membrane microconcentrator technique but has not been evaluated in CSF with total protein levels below 20mg/dL. Immunoturbidimetry is routinely used for human CSF albumin measurement and was recently applied on canine samples with encouraging results. Objective: The purpose of this study was to compare HRE (including the use of a concentration step) and immunoturbidimetric assay for the measurement of albumin levels in normal canine CSF. Methods: 30 CSF specimens from 15 healthy dogs were evaluated. CSF total protein was measured by the pyrogallol red methoda and CSF albumin was determined by HREb (n=15), cHREc (n=30) and immunoturbidimetric assayd (n=30). Validation of the human immunoturbidimetric assay was performed using a purified canine albumin standarde. Results: Mean CSF total protein was 17.5 (range 7-39) mg/dL. HRE integration profiles were non-interpretable in all unconcentrated specimens. However, clear distinction of the major protein fractions was achieved for all cHRE specimens. CSF albumin levels were measureable in 29/30 specimens using immunoturbidimetry. Excellent correlation (Pearson r=0.92, p<0.001) was found between the two techniques. Conclusion: Immunoturbidimetry and cHRE may be used for routine measurement of CSF albumin

Proteomic analysis of urine from healthy and CKD cats

In veterinary medicine there is an increasing interest in the application of proteomic techniques to investigate protein patterns in healthy and diseased animals; however, data on urine proteome are still limited. The aims of our study were to identify a urine protein profile in healthy cats and to compare it with those obtained in patients affected by chronic kidney disease (CKD). Urine samples were collected by cystocentesis from 23 healthy and 18 CKD cats. For all samples urinalysis and urine protein to creatinine ratio (UPC) were performed. Urine proteins were further separated by SDS-PAGE and the bands were reduced, alkylated and then digested by trypsin before ESI-Q-TOF mass spectrometry analysis. Protein identification was performed using MASCOT science search engine. Healthy cats had significantly (p<0.01) lower values of UPC than CKD cats. SDS-PAGE allowed to visualize an \u201chealthy profile\u201d with many different bands (median 32; range 26-47), including Albumin (70 kDa), Cauxin (carboxylesterase 5A, 61 kDa), Uromodulin (73 kDa), Transferrin (80 kDa), Angiotensin-converting enzyme 2 (93 kDa), Inter-alpha-trypsin inhibitor heavy chain H4 (103 kDa) and \u3b12Macroglobulin (170 kDa); at lower molecular weights, Albumin- (55kDa) and Cauxin- (40kDa) fragments, Haptoglobin (45kDa) and immunoglobulin light chains (24kDa) were present. Retinol binding protein 4 (23 kDa) and Cystatin-M(16 kDa) were identified only in urine of CKD patients. Proteomic techniques were successfully used to investigate proteinuria, revealing different protein patterns between healthy and CKD cats. Some of these proteins could be considered as promising biomarkers of chronic renal damage in feline patients

Urine proteome from healthy and CKD cats

In veterinary medicine there is an increasing interest in the application of proteomic techniques to investigate protein patterns in healthy and diseased animals; however, data on urine proteome are still limited (1,2,3). The aims of our study were to identify a urine protein profile in healthy cats and to compare it with those obtained in patients affected by chronic kidney disease (CKD). Urine samples were collected by cystocentesis from 23 healthy and 18 CKD cats. For all samples urinalysis and urine protein to creatinine ratio (UPC) were performed. Urine proteins were further separated by SDS-PAGE and the bands were reduced, alkylated and then digested by trypsin before ESI-Q-TOF mass spectrometry analysis. Protein identification was performed using MASCOT science search engine. Healthy cats had significantly (p<0.01) lower values of UPC than CKD cats. SDS-PAGE allowed to visualize an “healthy profile” with many different bands (median 32; range 26-47), including Albumin (70 kDa), Cauxin (carboxylesterase 5A, 61 kDa), Uromodulin (73 kDa), Transferrin (80 kDa), Angiotensin-converting enzyme 2 (93 kDa), Inter-alpha-trypsin inhibitor heavy chain H4 (103 kDa) and α2Macroglobulin (170 kDa); at lower molecular weights Albumin- (55kDa) and Cauxin- (40kDa) fragments, Haptoglobin (45kDa) and immunoglobulin light chains (24kDa) were present. Retinol binding protein 4 (23 kDa) and Cystatin-M(16 kDa) were identified only in urine of CKD patients. Proteomic techniques were successfully used to investigate proteinuria, indicating that two proteins are differentially expressed in urine of healthy and CKD cats. These proteins could be considered as promising biomarkers of chronic renal damage in feline patients. References 1. Lemberger SI, Deeg CA, Hauck SM, Amann B, Hirmer S, Hartmann K, Dorsh R. Comparison of urine protein profiles in cats without urinary tract disease and cats with idiopathic cystitis, bacterial urinary tract infection, or urolithiasis. Am J Vet Res. 2011;72(10):1407-15. 2. Schaefer H, Kohn B, Schweigert FJ, Raila J. Quantitative and qualitative urine protein excretion in dogs with severe inflammatory response syndrome. J Vet Intern Med. 2011;25(6):1292-7. 3. Nabity MB, Lees GE, Dangott LJ, Cianciolo R, Suchodolski JS, Steiner JM. Proteomic analysis of urine from male dogs during early stages of tubulointerstitial injury in a canine model of progressive glomerular disease. Vet Clin Pathol. 2011;40(2):222. Figures Figure 1. SDS-PAGE of urine sampples from healthy and CKD cats stained with silver staining. a) Electrophoretic profiles: 1 MW marker; 2-6 CKD urine samples; 7-8, pools of healthy urine samples female and male respectively; b) Pherograms obtained by ImageJ software

Analysis of online reports on the potential misuse of benzidamine

Benzydamine/Tantum Rosa is a drug for external use. It is typically available in Europe, without the need of a medical prescription, for the treatment of vaginal inflammatory processes. Between December 2009 and January 2010, the Milan and Pavia (I) Poison Centres have identified some 50 cases of inappropriate benzydamine ingestion. Reasons for this misuse have been attributed to an allegedly misleading television advert. However, the recreational misuse of benzydamine is a well-known phenomenon in Brazil and in some EU countries as well, notably in Poland and Romania. It is here suggested that the recent increase in benzydamine misuse reports in Italy may well be associated with a parallel increase in level of online information regarding the molecule potential for misuse. According to the online reports, benzydamine is typically taken at a dosage of 1-4 sachets, dissolved in water and ingested orally. Its intake may be associated with hallucinations (mostly visual), sleeping disorders and euphoria. Only future, prospective, studies will confirm and better describe the benzydamine misuse potentialPeer reviewe

Adenosine receptors expression in cardiac fibroblasts of patients with left ventricular dysfunction due to valvular disease

Context: Adenosine restores tissue homeostasis through the interaction with its membrane receptors (AR) expressed on fibroblasts, endothelial cells, smooth muscle cells and leukocytes, but their modulation is still not fully understood. Objective: To evaluate whether changes in the transcriptomic profiling of adenosine receptors (AR) occur in cardiac fibroblasts (CF) of patients (pts) with LV dysfunction due to valvular disease (V). The secondary aim was to compare in the same pts the results obtained at cardiac level with those found in circulating leukocytes. Materials and methods: Auricle fragments were excised from 13 pts during prosthetic implantation while blood samples were collected from pts (n = 9) and from healthy subjects (C, n = 7). In 7 pts cardiac biopsy and blood samples were taken simultaneously. A human CF atrial cell line (cc) was used as control. Results: AR higher levels of mRNA expression were observed with real-time PCR in Vpts compared to C, both at cardiac (overexpression A1R:98%, A2AR:63%, A2BR:87%, A3R:85%, CD39:92%, CD73:93%) and at peripheral level (A1R vs C: p = .0056; A2AR vs C: p = .0173; A2BR vs C: p = .0272; A3R vs C: p = .855; CD39 vs C: p = .0001; CD73 vs C: p = .0091). Conclusion: All AR subtypes were overexpressed in CF of Vpts. The same trends in AR expression at cardiac level was assessed on circulating leukocytes, thus opening a new road to minimally invasive studies of the adenosinergic system in cardiac patients

Ralstonia mannitolilytica infections in an oncologic day ward: description of a cluster among high-risk patients

Abstract Background Ralstonia spp, an environmental microorganism, has been occasionally associated with healthcare infections. The aim of this study was to investigate an outbreak caused by Ralstonia mannitolilytica in oncology patients. Methods Case definition: Oncology outpatients attending a day ward, with positive blood and/or central venous catheter (CVC) culture for Ralstonia spp from September 2013 – June 2014. We analysed medical records, procedures and environmental samples. R. mannitolilytica was identified by 16S rRNA sequencing, and typed by Pulsed Field Gel Electrophoresis (PFGE); resistance to carbapenemes was investigated by phenotypic and molecular methods. Results The patients (N = 22) had different malignancies and received different therapy; all had a CVC and 16 patients presented chills and/or fever. R. mannitolilytica was isolated from both blood and CVC (n = 12) or only blood (n = 6) or CVC tips (n = 4). The isolates had indistinguishable PFGE profile, and showed resistance to carbapenems. All the isolates were negative for carbapenemase genes while phenotypic tests suggests the presence of an AmpC β-lactamase activity,responsible for carbapenem resistance. All patients had had CVC flushed with saline to keep the venous access pervious or before receiving chemotherapy at various times before the onset of symptoms. After the first four cases occurred, the multi-dose saline bottles used for CVC flushing were replaced with single-dose vials; environmental samples were negative for R. mannitolilytica. Conclusions Although the source of R. mannitolilytica remains unidentified, CVC flushing with contaminated saline solution seems to be the most likely origin of R. mannitolilytica CVC colonization and subsequent infections. In order to prevent similar outbreaks we recommend removal of any CVC that is no longer necessary and the use of single-dose solutions for any parenteral treatment of oncology patients

Association Between PNPLA3 rs738409 C>G Variant and Liver-Related Outcomes in Patients with Non-alcoholic Fatty Liver Disease

Patients with nonalcoholic fatty liver disease (NAFLD) have an increased risk for liver-related complications, such as decompensation, hepatocellular carcinoma (HCC), and death; the severity of liver fibrosis and metabolic comorbidities are the main risk factors. A single nucleotide polymorphism in patatin-like phospholipase domain-containing-3 (PNPLA3) gene is associated with higher prevalence of liver damage and HCC, but there are no data from prospective studies of outcomes of patients with this polymorphism. We investigated whether the common rs738409 variant in PNPLA3 gene associates with the occurrence of liver-related events and death in a large cohort of patients with NAFLD

Phenomenon of new drugs on the Internet : the case of ketamine derivative methoxetamine

Copyright © 2012 John Wiley & Sons, Ltd.On the basis of the material available both in the scientific literature and on the web, this paper aims to provide a pharmacological, chemical and behavioural overview of the novel compound methoxetamine. This is a dissociative drug related to ketamine, with a much longer duration of action and intensity of effects. A critical discussion of the availability of information on the web of methoxetamine as a new recreational trend is here provided. Those methodological limitations, which are intrinsically associated with the analysis of online, non-peer reviewed, material, are here discussed as well. It is concluded that the online availability of information on novel psychoactive drugs, such as methoxethanine, may constitute a pressing public health challenge. Better international collaboration levels and novel forms of intervention are necessary to tackle this fast-growing phenomenon. Copyright © 2012 John Wiley & Sons, Ltd.Peer reviewe