STAT3 phosphorylation at serine 727 activates specific genetic programs and promotes clear cell renal cell carcinoma (ccRCC) aggressiveness.

The signal transducer and activator of transcription 3 (STAT3) is a transcription factor mainly activated by phosphorylation in either tyrosine 705 (Y705) or serine 727 (S727) residues that regulates essential processes such as cell differentiation, apoptosis inhibition, or cell survival. Aberrant activation of STAT3 has been related to development of nearly 50% of human cancers including clear cell renal cell carcinoma (ccRCC). In fact, phosho-S727 (pS727) levels correlate with overall survival of ccRCC patients. With the aim to elucidate the contribution of STAT3 phosphorylation in ccRCC development and progression, we have generated human-derived ccRCC cell lines carrying STAT3 Y705 and S727 phosphomutants. Our data show that the phosphomimetic substitution Ser727Asp facilitates a pro-tumoral phenotype in vitro, in a Y705-phosphorylation-independent manner. Moreover, we describe that STAT3 phosphorylation state determines the expression of different subsets of target genes associated with distinct biological processes, being pS727-dependent genes the most related to cellular hallmarks of cancer. In summary, the present study constitutes the first analysis on the role of overall STAT3 phosphorylation state in ccRCC and demonstrates that pS727 promotes the expression of a specific subset of target genes that might be clinically relevant as novel biomarkers and potential therapeutic targets for ccRCC.

Establishment of urinary exosome-like vesicles isolation protocol for FHHNC patients and evaluation of different exosomal RNA extraction methods.

BACKGROUND: Molecular and cellular pathophysiological events occurring in the majority of rare kidney diseases remain to be elucidated. Familial hypomagnesemia with hypercalciuria and nephrocalcinosis (FHHNC) is a rare autosomal recessive disorder caused by mutations in either CLDN16 or CLDN19 genes. This disease is characterized by massive urinary wasting of magnesium and calcium, osmosis deregulation and polyuria. Patients with p.G20D homozygous mutation in CLDN19 gene exhibit different progression to kidney failure suggesting that beyond the pathogenic mutation itself, other molecular events are favoring disease progression. Due to the fact that biopsy is not clinically indicated in these patients, urinary exosome-like vesicles (uEVs) can be envisioned as a valuable non-invasive source of information of events occurring in the kidney. Exosome research has increased notably to identify novel disease biomarkers but there is no consensus standardized protocols for uEVs isolation in patients with polyuria. For this reason, this work was aimed to evaluate and refine different uEVs isolation methods based on differential centrifugation, the gold standard method. RESULTS: Characterization by NTA, cryo-TEM and immunoblotting techniques identified the most appropriate protocol to obtain the highest yield and purest uEVs enriched fraction possible from urine control samples and FHHNC patients. Moreover, we tested five different RNA extraction methods and evaluated the miRNA expression pattern by qRT-PCR. CONCLUSIONS: In summary, we have standardized the conditions to proceed with the identification of differentially expressed miRNAs in uEVs of FHHNC patients, or other renal diseases characterized by polyuria.

Immune checkpoint inhibitor-related hypophysitis and endocrine dysfunction: clinical review.

Immune checkpoint inhibitors are a new and effective class of cancer therapy, with ipilimumab being the most established drug in this category. The drugs' mechanism of action includes promoting the effector T cell response to tumours and therefore increased autoimmunity is a predictable side effect. The endocrine effects of these drugs include hypophysitis and thyroid dysfunction, with rare reports of adrenalitis. The overall incidence of hypophysitis with these medications is up to 9%. Primary thyroid dysfunction occurs in up to 15% of patients, with adrenalitis reported in approximately 1%. The mean onset of endocrine side effects is 9 weeks after initiation (range 5-36 weeks). Investigation and/or screening for hypophysitis requires biochemical and radiological assessment. Hypopituitarism is treated with replacement doses of deficient hormones. Since the endocrine effects of immune checkpoint inhibitors are classed as toxic adverse events, most authors recommend both discontinuation of the immune checkpoint inhibiting medication and 'high-dose' glucocorticoid treatment. However, this has been challenged by some authors, particularly if the endocrine effects can be managed (e.g. pituitary hormone deficiency), and the therapy is proving effective as an anticancer agent. This review describes the mechanism of action of immune checkpoint inhibitors and details the key clinical endocrine-related consequences of this novel class of immunotherapies.

[Comparative analysis of patients admitted to Spanish Intensive Care Units due to medical and surgical disease]. / Análisis comparativo de pacientes ingresados en Unidades de Cuidados Intensivos españolas por causa médica y quirúrgica.

OBJECTIVE: To describe the characteristics of the patients case-mix admitted to ICUs due to medical and surgical disease, and to compare both groups. DESIGN: Analysis of data covering the period 2006-2011 in the ENVIN-HELICS registry. An observational, prospective, multicenter and voluntary participation study. SETTING: A total of 188 Spanish ICUs. PATIENTS: All patients admitted for more than 24 hours. MAIN VARIABLES: Demographic data, cause of admission, severity scores, length of stay, mortality. RESULTS: A total of 138,999 patients were analyzed. Of these, 65,467 (47.1%) were admitted due to a non-coronary medical cause, 27,785 (20,0%) due to coronary-related illness, 28,044 (20,2%) after elective surgery and 17,613 (12.7%) after urgent surgery. Use of devices, nosocomial infections and isolation of multirresistant organisms were more prevalent in urgent surgery patients. Longer length of stay (median 5 days; interquartile range 2-11) as well as higher severity scale values (APACHE II and SAPS II) corresponded to this same group of patients. Mortality was higher in non-coronay medical patients. On categorizing the patients according to the APACHE II score, mortality was seen to be higher in urgent surgery cases than in elective surgery patients in all groups. The largest difference was observed in the APACHE II score 6-10 group (3% vs. 0.9%) (OR: 2.14, 95% CI 1.825-2.513; p<0.001). CONCLUSIONS: The mortality rate is higher in non-coronary medical patients, though resource use per patient is greater in the urgent surgery cases. The APACHE II scale underestimates mortality in emergency surgery patients.

Guidelines for the prevention of ventilator-associated pneumonia and their implementation. The Spanish "Zero-VAP" bundle

"BACKGROUND: ""Zero-VAP"" is a proposal for the implementation of a simultaneous multimodal intervention in Spanish intensive care units (ICU) consisting of a bundle of ventilator-associated pneumonia (VAP) prevention measures. METHODS/DESIGN: An initiative of the Spanish Societies of Intensive Care Medicine and of Intensive Care Nurses, the project is supported by the Spanish Ministry of Health, and participation is voluntary. In addition to guidelines for VAP prevention, the ""Zero-VAP"" Project incorporates an integral patient safety program and continuous online validation of the application of the bundle. For the latter, VAP episodes and participation indices are entered into the web-based Spanish ICU Infection Surveillance Program ""ENVIN-HELICS"" database, which provides continuous information about local, regional and national VAP incidence rates. Implementation of the guidelines aims at the reduction of VAP to less than 9 episodes per 1000 days of mechanical ventilation. A total of 35 preventive measures were initially selected. A task force of experts used the Grading of Recommendations, Assessment, Development and Evaluation Working Group methodology to generate a list of 7 basic ""mandatory"" recommendations (education and training in airway management, strict hand hygiene for airway management, cuff pressure control, oral hygiene with chlorhexidine, semi-recumbent positioning, promoting measures that safely avoid or reduce time on ventilator, and discouraging scheduled changes of ventilator circuits, humidifiers and endotracheal tubes) and 3 additional ""highly recommended"" measures (selective decontamination of the digestive tract, aspiration of subglottic secretions, and a short course of iv antibiotic). DISCUSSION: We present the Spanish VAP prevention guidelines and describe the methodology used for the selection and implementation of the recommendations and the organizational structure of the project. Compared to conventional guideline documents, the associated safety assurance program, the online data recording and compliance control systems, as well as the existence of a pre-defined objective are the distinct features of ""Zero VAP"""

Invasive device-related infections after heart surgery.

OBJECTIVE: To analyze postoperative infections in critically ill patients undergoing heart surgery. SETTING: Intensive care units (ICUs). DESIGN: An observational, prospective, multicenter study was carried out. PATIENTS: Patients in the postoperative period of heart surgery admitted to the ICU and included in the ENVIN-HELICS registry between 2005 and 2011. MAIN OUTCOME VARIABLES: Mechanical ventilation associated pneumonia (MVP), urinary catheter-related infection (UCI), primary bacteremia (PB), PB related to vascular catheters (PB-VC) and secondary bacteremia. RESULTS: Of a total of 97,692 patients included in the study, 9089 (9.3%) had undergone heart surgery. In 440 patients (4.8%), one or more infections were recorded. Infection rates were 9.94 episodes of MVP per 1000 days of mechanical ventilation, 3.4 episodes of UCI per 1000 days of urinary catheterization, 3.10 episodes of BP-VC per 1000 days of central venous catheter, and 1.84 episodes of secondary bacteremia per 1000 days of ICU stay. Statistically significant risk factors for infection were ICU stay (odds ratio [OR] 1.18, 95%CI 1.16-1.20), APACHE II upon admission to the ICU (OR 1.05, 95%CI 1.03-1.07), emergency surgery (OR 1.67, 95%CI 1.13-2.47), previous antibiotic treatment (OR 1.38, 95%CI 1.04-1.83), and previous colonization by Pseudomonas aeruginosa (OR 18.25, 95%CI 3.74-89.06) or extended spectrum beta-lactamase producing enterobacteria (OR 16.97, 95%CI 5.4-53.2). The overall ICU mortality rate was 4.1% (32.2% in patients who developed one or more infections and 2.9% in uninfected patients) (P < .001). CONCLUSIONS: Of the patients included in the ENVIN-HELICS registry, 9.3% were postoperative heart surgery patients. The overall mortality was low but increased significantly in patients who developed one or more infection episodes.

Insulin-like growth factor-II and insulin-like growth factor-binding proteins in bovine cystic ovarian disease.

Cystic ovarian disease (COD) is one of the most common reproductive disorders of cattle and is considered to have multifactorial aetiology. An accepted hypothesis involves neuroendocrinological dysfunction of the hypothalamic-pituitary-gonadal axis; however, the role of growth factors in COD has not been extensively investigated. The present study examines the potential role of members of the insulin-like growth factor (IGF) family in COD. Expression of genes encoding IGF-II and insulin-like growth factor-binding proteins (IGFBPs) was examined and the distribution of IGF-II within the follicular wall was assessed immunohistochemically. Finally, the concentration of IGF-II protein was determined in follicular fluid. There was increased IGF-II mRNA in the wall of cystic follicles, mainly associated with granulosa cells. Additionally, there was significantly more IGF-II protein in granulosa and theca cells in cystic follicles, but no change in the concentration of IGF-II in follicular fluid. Total IGFBPs, assessed by western blotting, were similar in different structures. However, by discriminating each IGFBP a decrease was detected in IGFBP-2 expression in cystic follicles that may be related to the observed higher expression of IGF-II. In summary, the present study provides evidence to suggest that COD in cattle is associated with modifications in the IGF-II system.

Fungal colonization and/or infection in non-neutropenic critically ill patients: results of the EPCAN observational study.

The purpose of this paper is to determine the incidence of fungal colonization and infection in non-neutropenic critically ill patients and to identify factors favoring infection by Candida spp. A total of 1,655 consecutive patients (>18 years of age) admitted for > or = 7 days to 73 medical-surgical Spanish intensive care units (ICUs) participated in an observational prospective cohort study. Surveillance samples were obtained once a week. One or more fungi were isolated in different samples in 59.2% of patients, 94.2% of which were Candida spp. There were 864 (52.2%) patients with Candida spp. colonization and 92 (5.5%) with proven Candida infection. In the logistic regression analysis risk factors independently associated with Candida spp. infection were sepsis (odds ratio [OR] = 8.29, 95% confidence interval [CI] 5.07-13.6), multifocal colonization (OR = 3.49, 95% CI 1.74-7.00), surgery (OR = 2.04, 95% CI 1.27-3.30), and the use of total parenteral nutrition (OR = 4.37, 95% CI 2.16-8.33). Patients with Candida spp. infection showed significantly higher in-hospital and intra-ICU mortality rates than those colonized or non-colonized non-infected (P < 0.001). Fungal colonization, mainly due to Candida spp., was documented in nearly 60% of non-neutropenic critically ill patients admitted to the ICU for more than 7 days. Proven candidal infection was diagnosed in 5.5% of cases. Risk factors independently associated with Candida spp. infection were sepsis, multifocal colonization, surgery, and the use of total parenteral nutrition.

Has double-phase MIBI scintigraphy usefulness in the diagnosis of hyperparathyroidism?

The usefulness of double-phase parathyroid technetium-99m-MIBI scintigraphy for the detection of hyperplastic parathyroid tissue has been described. The aim of the present study was to establish the effectiveness of this new technique in the morphological and functional assessment of parathyroid glands in patients with different types of hyperparathyroidism. We performed 99mTc-MIBI scintigraphy (MIBI) and neck ultrasonography in 38 patients with primary (n=16) or secondary (n=22) hyperparathyroidism. All patients underwent surgical neck exploration, removing a total of 84 parathyroid glands. Before and after surgery, blood intact parathyroid hormone (iPTH) was measured peripherally and in both the right and left internal jugular veins. In patients with primary hyperparathyroidism, ultrasonography showed one enlarged gland in 11 cases (69%), while MIBI was positive in 15 (94%) (including two ectopic glands). The sensitivity of MIBI (93%) was greater than that of ultrasonography (68%), with a similar specificity (100 and 97%, respectively). In patients with secondary hyperparathyroidism, there was a discrepancy between both imaging modalities in 29 glands (33%). The sensitivity of both techniques was similar (41 and 54%, respectively), with the same specificity (89%). There were more difficulties in detecting the upper than the lower pathological glands. MIBI reflected more accurately the functionality of the glands, and ultrasonography has a better correlation with the volume and weight. In conclusion, Tc-99m-MIBI scintigraphy is a good technique to identify parathyroid hyperfunctioning tissue in cases of primary hyperparathyroidism and to detect ectopic glands, but it does not give significantly better results than conventional ultrasonography in patients with secondary hyperparathyroidism.

[Postoperative infections in critically ill patients]. / Infección posquirúrgica en pacientes graves.

Patients subjected to surgery often develop nosocomial infections, among which the intra-abdominal ones stand out as being a common cause of septicemia, multi-organ failure, and death of the critical patients. Advances have been made in the study of the physiopathology by studying the mediators which are responsible for the systemic inflammatory response, the microbiology (changes in the pathogen type and in the antimicrobial sensitivity), and for the clinical picture (cholecystitis, tertiary peritonitis). Abdominal ultra-sound and computerized axial tomography have contributed greatly to the diagnosis of these infections. The new treatment techniques are discussed, both of the drainage of the septic focus (percutaneous or surgical), as of the antimicrobial treatment and the supportive measures. The diagnostic and therapeutic advances have modified the prognosis of these patients, although this continues to be poor when there is development of the multi-organ failure syndrome.

Endoscopic evolution of laryngeal injuries caused by translaryngeal intubation.

Despite improvements in tube design and materials, the longer survival rates of critically ill patients make laryngeal and tracheal lesions still common following prolonged translaryngeal intubation. The time of intubation is almost the only factor employed in deciding whether or not tracheotomy has to be performed. Some patients will not develop laryngeal lesions afer long intubation periods, whereas some already have clinical symptoms after short periods of time. If the conditions of the larynx and trachea could be assessed before irreversible complications take place, then timing of tracheotomy could be individualized to avoid laryngeal stenosis as well as unnecessary tracheostomies. We present the preliminary results of an endoscopic study of the early laryngeal changes that take place during translaryngeal intubation. The method of exploration is explained and tissue changes seen and their evolution after extubation are described, emphasizing those that could have a predictive value.

Fatal chemical pneumonitis due to cadmium fumes.

Acute exposure to high concentrations of cadmium fumes may cause acute chemical pneumonitis with a possibly fatal outcome. The etiologic diagnosis of acute cadmium intoxication from inhaled fumes may be difficult and can be confused with other forms of acute respiratory failure. We report on a case of a fit 53 year-old man who was exposed to cadmium fumes after flame-cutting an alloy containing around 10% of cadmium for a period of 60-75 minutes. He developed severe chemical pneumonitis and died 19 days after exposure.