Subcellular Localizations of RIG-I, TRIM25, and MAVS Complexes.

The retinoic acid-inducible gene 1 (RIG-I) signaling pathway is essential for the recognition of viruses and the initiation of host interferon (IFN)-mediated antiviral responses. Once activated, RIG-I interacts with polyubiquitin chains generated by TRIM25 and binds mitochondrial antiviral signaling protein (MAVS), leading to the production of type I IFN. We now show specific interactions among these key partners in the RLR pathway through the use of bimolecular fluorescence complementation (BiFC) and super-resolution microscopy. Dimers of RIG-I, TRIM25, and MAVS localize into different compartments. Upon activation, we show that TRIM25 is redistributed into cytoplasmic dots associated with stress granules, while RIG-I associates with TRIM25/stress granules and with mitochondrial MAVS. In addition, MAVS competes with TRIM25 for RIG-I binding, and this suggests that upon TRIM25-mediated activation of RIG-I, RIG-I moves away from TRIM25 to interact with MAVS at the mitochondria. For the first time, the distribution of these three proteins was analyzed at the same time in virus-infected cells. We also investigated how specific viral proteins modify some of the protein complexes in the pathway. The protease NS3/4A from hepatitis C virus redistributes the complexes RIG-I/MAVS and MAVS/MAVS but not RIG-I/TRIM25. In contrast, the influenza A virus NS1 protein interacts with RIG-I and TRIM25 in specific areas in the cell cytoplasm and inhibits the formation of TRIM25 homocomplexes but not the formation of RIG-I/TRIM25 heterocomplexes, preventing the formation of RIG-I/MAVS complexes. Thus, we have localized spatially in the cell different complexes formed between RIG-I, TRIM25, and MAVS, in the presence or absence of two viral IFN antagonistic proteins. IMPORTANCE: The first line of defense against viral infections is the innate immune response. Viruses are recognized by pathogen recognition receptors, such as the RIG-I like receptor family, that activate a signaling cascade that induces IFN production. In the present study, we visualized, for the first time in cells, both in overexpression and endogenous levels, complexes formed among key proteins involved in this innate immune signaling pathway. Through different techniques we were able to analyze how these proteins are distributed and reorganized spatially within the cell in order to transmit the signal, leading to an efficient antiviral state. In addition, this work presents a new means by how, when, and where viral proteins can target these pathways and act against the host immune system in order to counteract the activation of the immune response.

Exploring a novel preparation method of 1D metal organic frameworks based on supercritical CO2.

The preparation of copper(II) one-dimensional MOFs using an eco-efficient method is reported here. This method is based exclusively on using supercritical CO2 as a solvent, without the addition of any other additive or co-solvent. Neutral acetylacetonate copper complexes and two linear linkers, namely, the bidentate 4,4'-bipyridine and 4,4'-trimethylenedipyridine molecules, were reacted under compressed CO2 at 60 °C and 20 MPa for periods of 4 or 24 h. The success achieved in the synthesis of the different studied 1D-MOFs was related to the solubility of the reagents in supercritical CO2. The reaction yield of the synthesized coordination polymers via the supercritical route was close to 100% because both the reactants were almost completely depleted in the performed experiments.

Negative impact of bone metastasis on outcome in clear-cell renal cell carcinoma treated with sunitinib.

BACKGROUND: The aim of our study was to determine whether the presence of bone metastases affects outcomes in patients with metastatic clear-cell renal cell carcinoma (m-ccRCC) receiving sunitinib. PATIENTS AND METHODS: We reviewed the charts of all patients in four academic centers in Belgium and France who started first-line sunitinib (50 mg/day; 4 weeks on and 2 weeks off) between January 2005 and December 2008. Data were collected on known prognostic factors for metastatic renal cell carcinoma and metastatic sites. Response and progression were evaluated by computed tomography scan (according to RECIST). RESULTS: Two hundred twenty-three patients were identified. With a median follow-up of 40 months, median progression-free survival (PFS) and median overall survival (OS) were significantly shorter in patients with bone metastases than in those without: respectively, 8.2 versus 19.1 months (P<0.0001) and 19.5 versus 38.5 months (P<0.0001). On multivariate analysis, taking on account platelet count, Eastern Cooperative Oncology Group performance status, number of metastatic sites, neutrophil count, corrected serum calcium, time from diagnosis to systemic treatment, and the presence of bone metastases, bone metastasis was the independent variable most significantly associated with poor PFS (P<0.0001) and OS (P=0.001). CONCLUSION: The presence of bone metastases in m-ccRCC patients has a significant and clinically relevant negative impact on outcome on sunitinib.

[Anti-angiogenic therapies and hemorrhagic risk]. / Risque hémorragique et anti-angiogéniques.

Anti-angiogenic therapies induce an ischemic necrosis of tumoral tissue, which exposes to a risk of hemorrhagic complications. The frequency and the intensity of the symptoms may differ according to molecules and according to studies. The risk seems to be maximal with tyrosin kinase inhibitors, particularly with sunitinib, with 26% of hemorrhagic complications. In most cases, events are grade 1 or 2, severe adverse effects (grade 3 or 4) appear rarely. Intra-cerebral bleeding remains rare and seems to be observed essentially in case of cerebral metastases. The hemorrhagic risk seems to be correlated with arterial high blood pressure and concomitant thrombopenia. The management of hemorrhagic risk is essentially based on precautionary measures. For any surgical procedure, it is advised to interrupt the treatment at least 4 weeks before and 4 weeks later, in order to avoid parietal complications.

[Angiogenesis inhibition: review of the activity of sorafenib, sunitinib and bevacizumab]. / Inhibiteurs de l'angiogenèse : revues de l'apport thérapeutique du sorafenib, du sunitinib et du bevacizumab dans le cancer du rein métastatique.

Renal cell carcinoma accounts for approximately 3% of all human malignancies. The use of cytokines in metastatic stage of disease has been the standard until last decades, presenting partial and short duration responses. Research on angiogenesis in renal carcinoma has brought important advances to understand tumor biology and to allow us development of new antiangiogenic drugs. Sunitinib (SUTENT), sorafenib (NEXAVAR) and bevacizumab (AVASTIN) are actually three molecules accepted to use in metastatic renal cell carcinoma (mRCC), with a good tolerability demonstrated in different studies. Clinical evidence shows sunitinib to be reference standard of care for the first-line treatment of mRCC. The use of bevacizumab in combination with interferon alfa (IFN alfa) can also be considered in this setting. Sorafenib is recommended for second-line treatment in cytokine-refractory patients, sunitinib being also accepted in this situation. Other combination of these molecules and their use as neo-adjuvant and adjuvant therapy is being evaluated and should change in the short term the management of the disease.

Low temperature N,N-dimethylformamide-assisted synthesis and characterization of anatase-rutile biphasic nanostructured titania.

Anatase and rutile biphasic nanostructured titania (TiO(2)) has been synthesized via hydrolysis of titanium tetraisopropoxide in an aqueous solution of hydrobromic acid (HBr) and N,N-dimethylformamide (DMF) at 80 degrees C for 16 h. The presence of DMF, which was partially hydrolyzed during the process, determined the formation of a biphasic material. Powder x-ray diffraction showed the presence of both anatase and rutile titania phases in a ratio of approx. 1:1. Transmission electron microscope analysis showed that rutile was present as radial flower-like nanorods, which were surrounded by anatase spherical nanoparticles of 5 nm diameter. Low temperature nitrogen adsorption-desorption analysis showed the characteristic hysteresis loop of a mesoporous material. Specific surface area reached a value of 120 m(2) g(-1) and the average pore diameter was 50 A. X-ray photoelectron spectroscopic analysis revealed that interstitial nitrogen was incorporated (0.35 at.%) during the annealing process. According to ultraviolet (UV)-visible diffuse reflectance spectroscope characterization, the N-doping caused a bandgap reduction from 3.0 to 2.9 eV. Photocatalytic activity of the material was tested for the degradation of methylene blue, methyl orange and 4-nitrophenol under near-UV and visible light radiation.

H2O2 formation from photocatalytic processes at the ZnO/water interface.

The paper focuses on the production of H2O2 by photocatalysis over ZnO in an aerated aqueous phase. The presence of different reductants that increase the H2O2 production in the aqueous phase is analysed; particular attention is paid to nitrite, which has been shown to be the reductant that produces the most significant increments in the H2O2 production. The photocatalytic anodic decomposition of ZnO in the presence of the different reductants is also investigated. From the results obtained, the relevance of the ZnO photocatalysis in the formation of environmental hydrogen peroxide is estimated.

Laparoscopic cholecystectomy vs open cholecystectomy in the treatment of acute cholecystitis: a prospective study.

OBJECTIVE: To compare the results of laparoscopic cholecystectomy (LC) with those of open cholecystectomy (OC) in the treatment of acute cholecystitis. DESIGN: A prospective, nonrandomized trial. SETTING: "Virgen de la Arrixaca" University Hospital, El Palmar (Murcia), Spain. PATIENTS: One hundred fourteen patients underwent LC, and 110 underwent OC. The patients underwent surgery within 72 hours of the onset of symptoms. The patients were selected for LC or OC depending on the surgeon's experience in laparoscopic surgery. MAIN OUTCOME MEASURES: Operating time, rate of conversion from LC to OC, complications, and length of hospital stay. RESULTS: Conversion from LC to OC was necessary in 15% of the patients. The mean operating time was 77 minutes for the OC group and 88 minutes for the LC group (P<.001). Complications occurred in 14% of the patients in the LC group and in 23% of the patients in the OC group, with no significant differences between the 2 groups (P=.06). The number of moderate or severe complications was similar in both groups, whereas mild complications were more common in the OC group (P<.02). The length of the hospital stay averaged 8.1 days for the OC group and 3.3 days for the LC group (P<.001). CONCLUSIONS: Laparoscopic cholecystectomy is a safe, valid alternative to OC in patients with acute cholecystitis. The technique has a low rate of complications, implies a shorter hospital stay, and offers the patient a more comfortable postoperative period than OC.

Laparoscopic cholecystectomy in the treatment of acute cholecystitis.

BACKGROUND: Laparoscopic cholecystectomy has several advantages over traditional cholecystectomy, which make it the treatment of choice for patients with uncomplicated biliary lithiasis. However, in patients with acute cholecystitis, the role of this technique remains controversial and some clinicians regard this condition as a contraindication to laparoscopic cholecystectomy. STUDY DESIGN: Between June, 1991 and July, 1993, a total of 259 patients with cholelithiasis underwent laparoscopic cholecystectomy at the "Virgen de la Arrixaca" University Hospital. Of these patients, 60 underwent laparoscopic cholecystectomy for acute cholecystitis. RESULTS: Conversion to laparotomy was necessary in eight patients (13 percent). Mean operating time was 83 minutes (range, 45 to 180 minutes). Overall mean hospital stay (laparoscopy and conversions) was 3.1 days (range, one to nine days). There was no mortality or injury to the common bile duct in our series. CONCLUSIONS: We believe that laparoscopic cholecystectomy in patients with acute cholecystitis is a safe and effective procedure, in which the patient can benefit from the advantages of laparoscopic surgery without an increase in mortality and morbidity rates.

Laparoscopic versus open appendicectomy: a prospective assessment.

A prospective study of 200 consecutive patients with suspected acute appendicitis was performed to compare open and laparoscopic appendicectomy. Formal randomization was precluded by instrument availability. Some 100 patients underwent laparoscopic appendicectomy (conversion to laparotomy was carried out in five) and 100 had conventional surgery. The groups were similar in sex ratio, age, degree of appendiceal inflammation and antibiotic treatment. The mean duration of open appendicectomy was 46 min and of the laparoscopic procedure 51 min (P not significant). Postoperative complications in patients who underwent laparoscopic appendicectomy included: intra-abdominal abscess (two patients), wound infection (one), early bowel obstruction (four; all resolved with medical treatment) and umbilical haematoma (two). There were no reoperations in the immediate or late postoperative period. Complications after open operation were: wound infection (seven patients) (P < 0.05), early bowel obstruction (five; three resolved with medical treatment, two required surgery) and haematoma of the surgical wound (one). The mean hospital stay was 4.8 days for laparoscopic appendicectomy and 6.0 days for the open operation (P < 0.05). There were no deaths.

Management of the perineal wound following abdominoperineal resection: prospective study of three methods.

A prospective study was made of three procedures for treating the perineal wound and presacral cavity in 102 patients undergoing abdominoperineal excision for cancer of the rectum: (1) packing of the presacral space after suture of the pelvic peritoneum; (2) suture of the pelvic peritoneum and perineal wound, leaving two drains through the perineum; and (3) no suture of the pelvic peritoneum, and primary closure of the perineal wound, leaving drains through the abdomen for physiological saline irrigation. The parameters analysed were incidence of infection, primary healing of the perineum, extraperineal complications and mean hospital stay. Primary healing of the perineum was best with method 3, and overall incidence of infection highest with method 2. There were no differences between the methods with regard to extraperineal complications. Hospital stay was shortest with method 3.

[Surgical treatment of gastric cancer. Evaluation of results in 313 consecutive cases]. / Tratamiento quirúrgico del cáncer gástrico. Valoración de resultados en 313 casos consecutivos.

The results of 313 gastric adenocarcinomas, treated by the same surgical team during 10 years have been evaluated retrospectively. The pathological characteristics, operability (88.8%) and resectability (69.8%) rates, surgical technique and morbility and mortality rates are analyzed. The causes of late mortality, total five-year survival, depending on the stage of the tumors, and survival in relation to treatment are reported.