Validation and Search of the Ideal Cut-Off of the Sysmex UF-1000i (R) Flow Cytometer for the Diagnosis of Urinary Tract Infection in a Tertiary Hospital in Spain

Urinary tract infections (UTI) are one of the most prevalent infections. A rapid and reliable screening method is useful to screen out negative samples. The objective of this study was to validate the Sysmex flow cytometer UF-1000i by evaluating its accuracy, linearity and carry-over; and define an optimal cut-off value to be used in routine practice in our hospital. For the validation of the UF-1000i cytometer, precision, linearity and carry-over were studied in samples with different counts of bacteria, leukocytes and erythrocytes. Between March and June 2016, urine samples were tested in the Clinical Microbiology Laboratory at University Miguel Servet Hospital, in Spain. Samples were analyzed with the Sysmex UF-1000i cytometer, and cultured. Growth of >= 10(5) CFUs/mL was considered positive. The validation study reveals that the precision in all the variables is acceptable; that there is a good linearity in the dilutions performed, obtaining values almost identical to those theoretically expected; and for the carry-over has practically null values. A total of 1, 220 urine specimens were included, of which 213 (17.4%) were culture positive. The optimal cut-off point of the bacteria-leukocyte combination was 138.8 bacteria or 119.8 leukocytes with an S and E of 95.3 and 70.4%, respectively. The UF-1000i cytometer is a valuable method to screen urine samples to effectively rule out UTI and, may contribute to the reduction of unnecessary urine cultures

Produzione, mercato e consumi della cerasicoltura spagnola

La crisi di sovrapproduzione di alcune specie tradizionali sta favorendo la crescita del ciliegio, che mostra un costante incremento delle superfici, favorito dal rinnovamento varietale, dall’aumento dei consumi, dal miglioramento delle tecnologie di produzione e dalla precocità di maturazione. Tutti fattori che garantiscono alla Spagna elevata competitività nelle esportazioni verso i Paesi dell’Ue.Colaboración en el blog: Rivista di frutticoltura e di ortofloricoltura. Disponible: http://www.rivistafrutticoltura.it

Dietary level of fibre and age at weaning affect the proliferation of Clostridium perfrigens in the caecum, the incidence of Epizootic Rabbit Enteropathy and the performance of fattening rabbits

An experiment was conducted to investigate the effects of dietary fibre content and weaning age on Clostridium perfringens proliferation in the caecum and fattening mortality in growing rabbits farmed in a facility having Epizootic Rabbit Enteropathy. The experiment consisted in a 2 × 2 factorial arrangement with two weaning ages (28 days vs. 42 days) and two levels of dietary neutral detergent fibre assayed with a heat stable amylase and expressed exclusive of residual ash (aNDFom; 330 g/kg vs. 425 g/kg). Controls were made during two consecutive experimental periods that differed in hygienic environmental conditions by modifying the intensity of cleaning and disinfection in the farm previous to the trial. An interaction (P<0.001) was detected among the independent variables studied on Cl. perfringens enumeration in the caecal contents, as minimal values for this trait were obtained in non-medicated animals reared in a clean environment, and especially when they were weaned at a later age and fed the diet with the lower fibre content. The treatments studied also led to a variation in fattening mortality (from 4.7% to 34.0%), which was highly and positively correlated (P<0.001) to the average Cl. perfringens caecal counts in each combination of treatments. The results of the current study indicate that high counts of Cl. perfringens in the caecum can be used as an indicator of Epizootic Rabbit Enteropathy, and suggest that strategies designed to control its proliferation in the caecum might help to limit fattening mortality in rabbit fed diets not-medicated with antibiotics

Comparing Two Automated Techniques for the Primary Screening-Out of Urine Culture

Urinary tract infection is the most common human infection with a high morbidity. In primary care and hospital services, conventional urine culture is a key part of infection diagnosis but results take at least 24 h. Therefore, a rapid and reliable screening method is still needed to discard negative samples as quickly as possible and to reduce the laboratory workload. In this aspect, this study aims to compare the diagnostic performance between Sysmex OF-1000i and FUS200 systems in comparison to urine culture as the gold standard. From March to June 2016, 1, 220 urine samples collected at the clinical microbiology laboratory of the "Miguel Servet" hospital were studied in parallel with both analysers, and some technical features were evaluated to select the ideal equipment. The most balanced cut-off values taking into account bacteria or leukocyte counts were 138 bacteria/mu L or 119.8 leukocyte/pl for the OF-1000i (95.3% SE and 70.4% SP), and 5.7 bacteria/mu L or 4.3 leukocyte/mu L for the FUS200 (95.8% SE and 44.4% SP). The reduction of cultured plates was 37.4% with the FUS200 and 58.3% with the UF-1000i. This study shows that both techniques improve the workflow in the laboratory, but the OF-1000i has the highest specificity at any sensitivity and the FUS200 needs a shorter processing time

Methylthioadenosine

5'-Methylthioadenosine (MTA) is a naturally occurring sulfur-containing nucleoside present in all mammalian tissues. MTA is produced from S-adenosylmethionine mainly through the polyamine biosynthetic pathway, where it behaves as a powerful inhibitory product. This compound is metabolized solely by MTA-phosphorylase, to yield 5-methylthioribose-1-phosphate and adenine, a crucial step in the methionine and purine salvage pathways, respectively. Abundant evidence has accumulated over time suggesting that MTA can affect cellular processes in many ways. MTA has been shown to influence numerous critical responses of the cell including regulation of gene expression, proliferation, differentiation and apoptosis. Although most of these responses have been observed at the pharmacological level, their specificity makes it tempting to speculate that endogenous MTA could play a regulatory role in the cell. Finally, observations carried out in models of liver damage and cancer demonstrate a therapeutic potential for MTA that deserves further consideration

NO sensitizes rat hepatocytes to proliferation by modifying S-adenosylmethionine levels

BACKGROUND & AIMS: Liver regeneration is a fundamental response of this organ to injury. Hepatocyte proliferation is triggered by growth factors, such as hepatocyte growth factor. However, hepatocytes need to be primed to react to mitogenic signals. It is known that nitrous oxide (NO), generated after partial hepatectomy, plays an important role in hepatocyte growth. Nevertheless, the molecular mechanisms behind this priming event are not completely known. S-adenosylmethionine (AdoMet) synthesis by methionine adenosyltransferase is the first step in methionine metabolism, and NO regulates hepatocyte S-adenosylmethionine levels through specific inhibition of this enzyme. We have studied the modulation of hepatocyte growth factor-induced proliferation by NO through the regulation of S-adenosylmethionine levels. METHODS: Studies were conducted in cultured rat hepatocytes isolated by collagenase perfusion, which triggers NO synthesis. RESULTS: The mitogenic response to hepatocyte growth factor was blunted when inducible NO synthase was inhibited; this process was overcome by the addition of an NO donor. This effect was dependent on methionine concentration in culture medium and intracellular S-adenosylmethionine levels. Accordingly, we found that S-adenosylmethionine inhibits hepatocyte growth factor-induced cyclin D1 and D2 expression, activator protein 1 induction, and hepatocyte proliferation. CONCLUSIONS: Together our findings indicate that NO may switch hepatocytes into a hepatocyte growth factor-responsive state through the down-regulation of S-adenosylmethionine levels

S-adenosylmethionine regulates MAT1A and MAT2A gene expression in cultured rat hepatocytes: a new role for S-adenosylmethionine in the maintenance of the differentiated status of the liver

Methionine metabolism starts with the formation of S-adenosylmethionine (AdoMet), the most important biological methyl donor. This reaction is catalyzed by methionine adenosyltransferase (MAT). MAT is the product of two different genes: MAT1A, which is expressed only in the adult liver, and MAT2A, which is widely distributed, expressed in the fetal liver, and replaces MAT1A in hepatocarcinoma. In the liver, preservation of high expression of MAT1A and low expression of MAT2A is critical for the maintenance of a functional and differentiated organ. Here we describe that in cultured rat hepatocytes MAT1A expression progressively decreased, as described for other liver-specific genes, and MAT2A expression was induced. We find that this switch in gene expression was prevented by adding AdoMet to the culture medium. We also show that in cultured hepatocytes with decreased MAT1A expression AdoMet addition markedly increased MAT1A transcription in a dose-dependent fashion. This effect of AdoMet was mimicked by methionine, and blocked by 3-deazaadenosine and L-ethionine, but not D-ethionine, indicating that the effect was specific and mediated probably by a methylation reaction. These findings identify AdoMet as a key molecule that differentially regulates MAT1A and MAT2A expression and helps to maintain the differentiated status of the hepatocyte

The importance of organizational variables in treatment time for patients with ST-elevation acute myocardial infarction improve delays in STEMI

Background: The time between arrival at the emergency department (ED) and balloon (D2B) in STEMI is one of the best indicators of the quality of care. Our aim is to describe treatment times and evaluate the causes of delay. Methods: This is an observational retrospective study, including all consecutive STEMI code patients ≥18 years old treated in the ED from 2013 to 2016.All the patients were stratified into two groups: delayed group with D2B > 70 min and non-delayed ≤70. The primary variable was D2B time. Findings: In total 327 patients were included, stratified according to their D2B as follows: 166 (67·48%) in the delayed group and 80 (32·52%) in the non-delayed group. The delayed group was older (p = 0·005), with more females (p = 0·060) and more atypical electrocardiogram (ECG) STEMI signs or symptoms (p = 0·058) (p = 0·087). Predictors of shorter D2B time were: typical STEMI ECG signs and short training sessions for nurses on identifying STEMI patients. Interpretation: There are delays particularly in specific groups with atypical clinical presentations. Short training sessions aimed at emergency nurses correlate with shorter delay. This suggests that continuing training for emergency nurses, along with organizational strategies, can contribute to increasing the quality of care. Clinical trial number: NCT0433338

Transformed but not normal hepatocytes express UCP2

Uncoupling protein 2 (UCP2) expression in liver is restricted to non-parenchymal cells. By means of differential display screening between normal rat liver and H4IIE hepatoma cells we have isolated a cDNA clone encompassing part of UCP2 cDNA. Northern blot analysis revealed that UCP2 is expressed in some hepatocarcinoma cell lines, while it is absent in adult hepatocytes. UCP2 mRNA in H4IIE cells was downregulated when cells were cultured for 36 h in 0.1% serum and its expression was restored upon addition of 10% serum or phorbol esters. Hypomethylation of UCP2 was observed in transformed UCP2 expressing cells. Our results indicate that UCP2 is expressed in some hepatocarcinoma cell lines and that serum components may participate in maintaining elevated UCP2 levels

Expression of Wilms' tumor suppressor in the liver with cirrhosis: relation to hepatocyte nuclear factor 4 and hepatocellular function

The Wilms' tumor suppressor WT1 is a transcriptional regulator present in the fetal but not in the mature liver. Its expression and functional role in liver diseases remains unexplored. In this study, we analyzed WT1 expression by reverse-transcription polymerase chain reaction (RT-PCR) and by immunohistochemistry in normal and diseased livers. In addition, we performed in vitro studies in isolated rat hepatocytes to investigate WT1 regulation and function. We detected WT1 messenger RNA (mRNA) in 18% of normal livers, 17% of chronic hepatitis with minimal fibrosis, 49% of chronic hepatitis with bridging fibrosis, and 71% of cirrhotic livers. In cirrhosis, WT1 immunoreactivity was localized to the nucleus of hepatocytes. WT1 mRNA abundance correlated inversely with prothrombin time (P =.04) and directly with serum bilirubin (P =.002) and with the MELD score (P =.001) of disease severity. In rats, WT1 expression was present in fetal hepatocytes and in the cirrhotic liver but not in normal hepatic tissue. In vitro studies showed that isolated primary hepatocytes express WT1 when stimulated with transforming growth factor beta (TGF-beta) or when the cells undergo dedifferentiation in culture. Moreover, we found that WT1 down-regulates hepatocyte nuclear factor 4 (HNF-4), a factor that is essential to maintain liver function and metabolic regulation in the mature organ. Hepatic expression of HNF-4 was impaired in advanced human cirrhosis and negatively correlated with WT1 mRNA levels (P =.001). In conclusion, we show that WT1 is induced by TGF-beta and down-regulates HNF-4 in liver cells. WT1 is reexpressed in the cirrhotic liver in relation to disease progression and may play a role in the development of hepatic insufficiency in cirrhosis