A genome-wide search for promoters that respond to increased MYCN reveals both new oncogenic and tumor suppressor microRNAs associated with aggressive neuroblastoma

MYCN is a major driver of neuroblastoma tumorigenesis and MYCN amplification is the worst prognostic indicator of aggressive NB. To identify potentially therapeutic tumor suppressor microRNAs for aggressive NB, we utilized a conditional MYCN system to simulate MYCN-amplified and nonamplified tumor types and performed a genome-wide search for MYCN target microRNA promoters differentially repressed under high MYCN conditions. We identified 20 gene promoters hosting 30 microRNAs that were directly bound and differentially regulated by MYCN. Eleven of these genes showed significant clinical correlations for neuroblastoma with 4 genes linked with better survival and 7 genes linked with poor survival. Surprisingly, expression analysis of host genes and microRNAs demonstrated that 8 of 11 pairs were repressed by high levels of MYCN regardless of the clinical correlation of the host gene. We therefore predicted these intronic microRNAs would be tumor suppressors. In fact, detailed gain of function studies for two miRs, miR-591 and miR-558, confirmed potent tumor suppressive effects for miR-591 in orthotopic neuroblastoma xenografts. However, miR-558 markedly increased colony formation, proliferation, and tumor growth in vivo. Our data reveal host-gene independent functions of MYCN-target microRNAs and demonstrate that MYCN represses both tumor suppressive and proproliferative microRNAs.     

Interleukin (IL)-18, a biomarker of human ovarian carcinoma, is predominantly released as biologically inactive precursor

Interleukin (IL)-18 is a proinflammatory and immune-enhancing cytokine, which exerts antitumor effects in vivo, mediated by the induction of interferon (IFN)γ. We previously reported that IL-18 processing is defective in epithelial ovarian carcinoma (EOC) cells, which secrete an inactive precursor (pro-IL-18) in vitro. In addition, IL-18 was reported as a potential biomarker of EOC. Here, we further investigated its role as a serological marker in human EOC and addressed its possible biological activity in vivo. Our data indicate that immunoreactive IL-18 is increased in EOC patients' sera at diagnosis as compared with age-matched healthy women. IL-18 levels were higher in the ascitic fluids than in sera, suggesting a local production in the peritoneal cavity. Indeed, immunohistochemical analysis of tumors showed IL-18 expression in cytokeratine-positive neoplastic cells, although also scattered histiocytes and some lymphoid cells stained for IL-18. The detection of human IL-18 in sera and ascitic fluids of immunodeficient mice, orthotopically implanted with human EOC cells, further suggested that circulating IL-18 is tumor-derived. However, IL-18 is not an EOC specific biomarker, as increased serum levels were found also in some endometrial cancer patients. By means of a new monoclonal antibody, we characterized IL-18 present in the ascitic fluid as pro-IL-18, which is biologically inactive. Accordingly, IFNγ was not increased in EOC patients' sera and ascitic fluids and showed no correlation with IL-18 levels. Altogether these data indicate that IL-18 in EOC fluids is predominantly tumor-derived and that its lack of biological activity may represent a mechanism of tumor-escape.     

Spirulina did not ameliorate idiopathic chronic fatigue in four N-of-1 randomized controlled trials

Idiopathic chronic fatigue is an exclusion diagnosis established when no chronic disease is found. Spirulina platensis is an alga with a rich content of proteins, vitamins, minerals and amino acids and is considered as a bioactive additive with multiple effects, among them being effects against fatigue. However, despite the worldwide utilization of Spirulina, there are only a few quality studies with it and none concerning fatigue. The N-of-1 randomized trials are made on one patient, and by this kind of study the efficacy of a treatment on that particular patient can be assessed. A series of four N-of-1 double-blind, randomized trials were performed on four physicians who complained of chronic fatigue. Each patient was his own control and received three pairs of treatments comprising 4 weeks of spirulina and 4 weeks of placebo. Spirulina platensis was administered in a dose of 3 g/day. For each pair, the order of treatments was randomized. Outcome measures were severity of fatigue measured on a 10-point scale.The scores of fatigue were not significantly different between spirulina and placebo.Spirulina administered in a dose of 3 g/day did not ameliorate fatigue more than the placebo in any of the four subjects, and possibly it has no effect on chronic fatigue.     

First clinical experience of orally active epidermal growth factor receptor inhibitor combined with simplified FOLFOX6 as first-line treatment for metastatic colorectal cancer

BACKGROUND: Gefitinib, an orally active inhibitor of epidermal growth factor receptor (EGFR) tyrosine kinase, combined with chemotherapy, has shown efficacy as second-line treatment for advanced colorectal cancer (CRC). Gefitinib combined with FOLFOX6 (oxaliplatin plus folinic acid and 5-fluorouracil) was tested as a first-line therapy. METHODS: Patients with metastatic EGFR-positive CRC received gefitinib at a dose of 250 mg/day combined with simplified FOLFOX6. Gefitinib was continued as maintenance treatment in nonprogressing patients. Responses were assessed by Response Evaluation Criteria in Solid Tumors (RECIST) criteria and adverse events were assessed with the National Cancer Institute Common Toxicity Criteria (NCI-CTC) scale. RESULTS: A total of 56 patients were recruited. There were 26 men and 30 women, with a median age of 57.5 years. The Eastern Cooperative Oncology Group (ECOG) performance status was as follows: 0 in 39 patients, 1 in 12 patients, and 2 in 5 patients. Thirty-nine patients (69.6%) had stage IV disease at diagnosis, 92.9% had liver involvement, and 46.4% had > or =2 metastatic sites. All patients were evaluated for safety, and 53 were evaluated for response: 40 patients (71.4%; 95% confidence interval [95% CI], 57.8%-82.6%) had complete or partial responses, and 11 patients (19.6%) had stable disease. Median time to progression was 7 months (range, 2.1-33.0 months; 95% CI, 6.2-9.0 months). Radical surgery or thermoablation of metastatic sites was performed in 14 patients (25%). NCI-CTC grade 3-4 events occurred in 36 patients (64.3%): diarrhea in 9 patients (16.1%), and hematologic toxicity in 13 patients (23.2%). Four patients (7.1%) were withdrawn for drug-related adverse events. CONCLUSIONS: The regimen has shown promising efficacy with manageable toxicity as a first-line treatment for patients with advanced CRC.     

Leishmania OligoC-TesT as a Simple,Rapid, and Standardized Tool for Molecular Diagnosis of Cutaneous Leishmaniasis in Peru

Molecular methods such as PCR have become attractive tools for diagnosis of cutaneous leishmaniasis (CL), both for their high sensitivity and for their specificity. However, their practical use in routine diagnosis is limited due to the infrastructural requirements and the lack of any standardization. Recently, a simplified and standardized PCR format for molecular detection of Leishmania was developed. The Leishmania OligoC-TesT is based on simple and rapid detection using a dipstick with PCR-amplified Leishmania DNA. In this study, we estimated the diagnostic accuracy of the Leishmania OligoC-TesT for 61 specimens from 44 CL-suspected patients presenting at the leishmaniasis clinic of the Instituto de Medicina Tropical Alexander von Humboldt, Peru. On the basis of parasitological detection and the leishmanin skin test (LST), patients were classified as (i) confirmed CL cases, (ii) LST-positive cases, and (iii) LST-negative cases. The sensitivities of the Leishmania OligoC-TesT was 74% (95% confidence interval (CI), 60.5% to 84.1%) for lesion aspirates and 92% (95% CI, 81.2% to 96.9%) for scrapings. A significantly higher sensitivity was observed with a conventional PCR targeting the kinetoplast DNA on the aspirates (94%) (P = 0.001), while there was no significant difference in sensitivity for the lesion scrapings (88%) (P = 0.317). In addition, the Leishmania OligoC-TesT was evaluated for 13 CL-suspected patients in two different peripheral health centers in the central jungle of Peru. Our findings clearly indicate the high accuracy of the Leishmania OligoC-TesT for lesion scrapings for simple and rapid molecular diagnosis of CL in Peru.Leishmaniasis is a vector-borne disease caused by obligatory intracellular parasites of the genus Leishmania. Several clinical manifestations are classified under the term leishmaniasis, but three are the most prominent: visceral leishmaniasis (VL), cutaneous leishmaniasis (CL), and mucocutaneous leishmaniasis (MCL), which result from replication of parasites in macrophages in the internal organs, dermis, and naso-oropharyngeal mucosa, respectively (15).In Latin America, CL and MCL are important health problems, and Brazil and Peru are the two most affected countries (9). An increase of cases has been reported for Colombia, Ecuador, and Argentina (4, 25, 26). Human-made risk factors, such as migration, urbanization, and deforestation, likely contribute to the spread of the disease (10).Over 10,000 CL cases per year are reported to occur in Peru, and more than a million people are at risk for infection (11, 22). Furthermore, this disease is endemic in 70% of Peruvian territory, causing high morbidity, lifelong scars, and major health problems for many communities (19).Diagnosing CL is challenging because of its wide spectrum of clinical presentations. Lesions may vary in severity, clinical appearance, and duration (23). Moreover, differential diagnosis with other cutaneous diseases is often difficult (14, 15). In addition, CL can be caused by different Leishmania species. In Peru, the disease is mainly caused by Leishmania (Viannia) braziliensis, Leishmania (Viannia) peruviana, and Leishmania (Viannia) guyanensis, but Leishmania (Viannia) lainsoni and Leishmania (Leishmania) amazonensis infections have also been reported (1, 18).Routine diagnosis of CL is still based on demonstration of amastigotes in skin lesion scrapings through microscopic analysis of direct smears or prior in vitro culture of the parasite (15, 23). Both methods require skilled personnel, and their sensitivities tend to be low and variable (12, 21, 27). Furthermore, in vitro culture is cumbersome and time-consuming. The leishmanin skin test (LST) detects cell-mediated immunity and is frequently used in Peru to support clinical diagnosis of CL. However, it cannot distinguish between past and present infections (12).The PCR is a useful tool for detection of Leishmania parasites in clinical specimens, since high sensitivity and specificity have been reported. Attractive PCR targets are high-copy-number sequences, such as kinetoplast DNA (kDNA) (2, 8, 30) and the ribosomal small subunit (20, 28). Several PCR formats have been designed, but there is actually a demand for simplified and standardized approaches (24). Access to sophisticated equipment such as real-time PCR machines is often limited in Peru. Recently, a simple and rapid dipstick format for detection of amplified Leishmania DNA was developed (Leishmania OligoC-TesT) (7). The test is based on PCR amplification of a small sequence of the 18S rRNA gene followed by visualization of the PCR products on a dipstick by hybridization with a gold-conjugated probe. PCR product detection can be performed in 10 min, and no equipment other than a heating block and a pipette is needed. The test is a promising “low-tech” standardized PCR application for diagnosis of leishmaniasis and can be applied in a mid- to low-level-equipped laboratory (6).In this report, we estimated the sensitivity of the Leishmania OligoC-TesT for 61 skin lesion scrapings from 44 Peruvian patients suspected of having CL. To assess the performance and to demonstrate the applicability of the test in low-level-equipped laboratories, two trials in rural hospitals in the Peruvian jungle were conducted.     

Selective versus Standard Ligature of the Deep Venous Complex during Laparoscopic Radical Prostatectomy: Effects on Continence, Blood Loss, and Margin Status

Background

Continence after laparoscopic radical prostatectomy is critical to patients and to surgeons. In this setting, the management of deep venous complex (DVC) without involvement of the sphincter fibres could be an important step of the procedure.

Objective

To evaluate the effects of a personal selective suture of the plexus (selective ligature of the deep venous complex [SLDVC]) on continence, blood loss, and surgical margin status during laparoscopic radical prostatectomy (LRP).

Design, setting, and participants

We planned a prospective randomised study. Sixty consecutive patients with clinically localised prostate cancer were involved in the study and were divided into two groups: group A (30 patients) underwent LRP with extraperitoneoscopic approach with standard management of DVC; group B (30 patients) underwent LRP with SLDVC.

Intervention

In group A, a standard ligature of DVC was performed (ligature and subsequent section); in group B, a selective ligature of DVC after its section was performed.

Measurements

Continence was evaluated during follow-up visits at catheter removal, and after 1, 3, 6, and 12 mo, perioperative variables and pathologic features of specimens were recorded.

Results and limitations

The two groups were comparable in terms of age, body mass index (BMI), prostate-specific antigen (PSA) values, and Gleason score at biopsy. No differences were found between the two groups in terms of operative times, blood loss, catheterisation time, and postoperative stay or histologic status. As far as continence rate is concerned, a significant difference was recorded between the groups (53% in group A vs 80% in group B) after 3 mo.

Conclusions

This selective ligature of the DVC after its section can contribute to early recovery of continence. Our data suggest that SLDVC compromises neither the safety of the procedure nor its oncologic effectiveness.     

Determination of Pump Flow Rate During Cardiopulmonary Bypass in Obese Patients Avoiding Hemodilution

Abstract   Background and Aim: During cardiopulmonary bypass the pump flow is usually set on 2.4 L/min/m² of body surface area (BSA) to guarantee adequate tissue perfusion without differences for patient constitutional type. The present study attempts to evaluate the adequacy of pump flow rate in obese patients, considering the ideal weight instead of the real one, avoiding the overflow side effects and hemodilution. Methods: Obese patients with body mass index (BMI) > 30 presented for cardiac surgery were randomized in two groups: in one the cardiopulmonary bypass was led traditionally, in the other, pump flow rate was calculated on ideal BMI of 25. Results: Demographics, preoperative tests, and monitoring data were registered. Mortality at hospital discharge and 30 days after were analyzed. The pump flow rate between the groups was different (4.46 vs. 4.87; p = 0.004); there were no differences in organ perfusion (SvO₂; diuresis) and mortality, but the study group presented fewer complications and blood transfusions. Conclusions: The BSA is widely used as the biometric unit to normalize physiologic parameters included pump flow rate, but it is disputable if this practice is correct also in obese patients. The study group, in which pump flow rate was set on ideal BSA, presented no difference in diuresis and mixed venous saturation but fewer complications and fewer perioperative blood transfusions.     

Role of intravascular ultrasound in the management of intracoronary dislodged stent

We describe a nonagenarian patient in whom a paclitaxel-eluting stent was lost during an attempt of direct stent implantation at the distal right coronary artery after removal of the guide-wire. The potential usefulness of intravascular ultrasound in the management of this complication is illustrated. The dislodged stent could not be located by fluoroscopy. However, intravascular ultrasound allowed to find the undeployed stent at the proximal segment of the right coronary artery. It showed that the guide-wire was placed outside the lost stent lumen and this undeployed stent was crushed against a previously implanted stent by another stent with optimal intravascular ultrasound-guided implantation.     

Reliability and validity of the Older Americans Resources and Services (OARS) social resources scale in six European countries

OBJECTIVES: The purpose of this article is to examine data quality, reliability, and construct validity of the Older Americans Resources and Services social resources scale in six European countries (The Netherlands, Luxembourg, Italy, Austria, the United Kingdom, and Sweden). METHODS: A questionnaire was administered through face-to-face interviews in five countries, and postal interview in the sixth, to representative populations of adults aged 50 to 90 living independently (N = 12,478). This article examines missing values and distribution of items in the social resources scale, and consistency of skew and kurtosis across countries. We performed item-total correlations and ran confirmatory factor analyses to test a three-factor model obtained in previous U.S. and Spanish analyses. Cronbach's alpha determined the reliability of the factors. RESULTS: We observed a relatively large proportion of missing data for one item (have someone who would help you). All items correlated with a score equal to or greater than 0.20. Although the confirmatory factor analyses generally supported the acceptability of the three-factor structure in the European data, the reliability of two dimensions (dependability and affective) was unacceptably low. DISCUSSION: Differences across countries make it unlikely that researchers can develop a single social resources scale that would have item equivalence in multiple countries.