FDG-PET parameters predicting mediastinal malignancy in lung cancer.

BACKGROUND: Staging of mediastinal lymph nodes in non-small cell lung cancer (NSCLC) is mandatory. The maximum Standard Uptake Value (SUVmax) obtained using F-18 fluorodeoxyglucose positron emission tomography (FDG-PET) is the best non-invasive technique available for this evaluation, but its performance varies from center to center. The aim of the present study was to identify FDG-PET predictors of mediastinal malignancy that are able to minimize intercenter variability and improve the selection of subsequent staging procedures. METHOD: A multicenter study of NSCLC patients staged through FDG-PET and endobronchial ultrasonography with needle aspiration (EBUS-NA) was performed using therapeutic surgery with systematic nodal dissection as gold standard. Intercenter variability and predictive power for mediastinal malignancy of different FDG-PET measures were assessed, as well as the role of these measures for selecting additional staging procedures. RESULTS: One hundred and twenty-one NSCLC patients, of whom 94 (72%) had ≥1 hypermetabolic spots in the mediastinum, were included in the study. Mean SUVmax of the primary tumor was 12.3 (SD 6.3), and median SUVmax of the highest hypermetabolic spots in the mediastinum was 3.9 (IQR 2.4-7). Variability of FDG-PET measures between hospitals was statistically significant (p = 0.016 and p < 0.001 respectively), but lost significance when SUVmax in the mediastinum was expressed as a ratio or a subtraction from the primary tumor (SUVmax mediastinum/tumor, p = 0.083; and SUVmax mediastinum - tumor, p = 0.428 respectively). SUVmax mediastinum/tumor showed higher accuracy in the ROC analysis (AUC 0.77 CI 0.68-0.85, p < 0.001), and showed predictive power for mediastinal malignancy when using a 0.4 cutoff (OR 6.62, 95%CI 2.98-14.69). Sensitivities and negative predictive values of clinical staging through EBUS-NA attained values ranging between 57% and 92% after FDG-PET, which improved with additional techniques when the tumor had a diameter >3 cm and/or a SUVmax mediastinum/tumor ratio >0.4. CONCLUSION: The SUVmax mediastinum/tumor ratio is a good predictor of regional tumor extension in NSCLC. This measure is not influenced by intercenter variability and has an accuracy of over 70% for the identification of malignancy when using a 0.4 cutoff.

Bronchial microbiome of severe COPD patients colonised by Pseudomonas aeruginosa.

The bronchial microbiome in severe COPD during stability and exacerbation in patients chronically colonised by Pseudomonas aeruginosa (PA), has not been defined. Our objective was to determine the characteristics of the bronchial microbiome of severe COPD patients colonised and not colonised by P. aeruginosa and its changes during exacerbation. COPD patients with severe disease and frequent exacerbations were categorised according to chronic colonisation by P. aeruginosa. Sputum samples were obtained in stability and exacerbation, cultured, and analysed by 16S rRNA gene amplification and pyrosequencing. Sixteen patients were included, 5 of them showing chronic colonisation by P. aeruginosa. Pseudomonas genus had significantly higher relative abundance in stable colonised patients (p = 0.019), but no significant differences in biodiversity parameters were found between the two groups (Shannon, 3 (2-4) vs 3 (2-3), p = 0.699; Chao1, 124 (77-159) vs 140 (115-163), p = 0.364). In PA-colonised patients bronchial microbiome changed to a microbiome similar to non-PA-colonised patients during exacerbations. An increase in the relative abundance over 20 % during exacerbation was found for Streptococcus, Pseudomonas, Moraxella, Haemophilus, Neisseria, Achromobacter and Corynebacterium genera, which include recognised potentially pathogenic microorganisms, in 13 patients colonised and not colonised by P. aeruginosa with paired samples. These increases were not identified by culture in 5 out of 13 participants (38.5 %). Stable COPD patients with severe disease and PA-colonised showed a similar biodiversity to non-PA-colonised patients, with a higher relative abundance of Pseudomonas genus in bronchial secretions. Exacerbation in severe COPD patients showed the same microbial pattern, independently of previous colonisation by P. aeruginosa.

Electronic structure differences between H(2)-, Fe-, Co-, and Cu-phthalocyanine highly oriented thin films observed using NEXAFS spectroscopy.

Phthalocyanines, a class of macrocyclic, square planar molecules, are extensively studied as semiconductor materials for chemical sensors, dye-sensitized solar cells, and other applications. In this study, we use angular dependent near-edge x-ray absorption fine structure (NEXAFS) spectroscopy as a quantitative probe of the orientation and electronic structure of H2-, Fe-, Co-, and Cu-phthalocyanine molecular thin films. NEXAFS measurements at both the carbon and nitrogen K-edges reveal that phthalocyanine films deposited on sapphire have upright molecular orientations, while films up to 50 nm thick deposited on gold substrates contain prostrate molecules. Although great similarity is observed in the carbon and nitrogen K-edge NEXAFS spectra recorded for the films composed of prostrate molecules, the H2-phthalocyanine exhibits the cleanest angular dependence due to its purely out-of-plane π* resonances at the absorption onset. In contrast, organometallic-phthalocyanine nitrogen K-edges have a small in-plane resonance superimposed on this π* region that is due to a transition into molecular orbitals interacting with the 3dx(2)-y(2) empty state. NEXAFS spectra recorded at the metal L-edges for the prostrate films reveal dramatic variations in the angular dependence of specific resonances for the Cu-phthalocyanines compared with the Fe-, and Co-phthalocyanines. The Cu L3,2 edge exhibits a strong in-plane resonance, attributed to its b1g empty state with dx(2)-y(2) character at the Cu center. Conversely, the Fe- and Co- phthalocyanine L3,2 edges have strong out-of-plane resonances; these are attributed to transitions into not only b1g (dz(2)) but also eg states with dxz and dyz character at the metal center.

First Report of Damping-Off Caused by Cylindrocarpon pauciseptatum on Pinus radiata in Spain.

In the fall of 2009, damping-off of Pinus radiata seedlings was observed in a pine nursery in Sant Feliu de Buixalleu, Girona Province, northeastern Spain. Plants exhibited needle blight, extensive root necrosis, and root death. Root sections of symptomatic plants were cut, washed under running tap water, surface disinfected for 1 min in a 1.5% sodium hypochlorite solution, and washed twice with sterile distilled water. Small fragments of discolored tissues were plated onto potato dextrose agar (PDA) supplemented with 0.5 g liter-1 of streptomycin sulfate. Plates were placed at 25°C in the dark for 10 to 14 days, and all fungal colonies were transferred to PDA. A Cylindrocarpon sp. was consistently isolated from necrotic root tissues. Single-conidial isolates were obtained and grown on PDA and Spezieller Nährstoffarmer agar (SNA) (2) at 25°C for 10 days with a 12-h photoperiod. On PDA, the isolates developed abundant mycelium, which varied from white-to-grayish brown or golden brown. On SNA, all isolates produced two-septate, (35-) 39.4 (-40) × (7.5-) 7.7 (-8.75) µm, and three-septate, (32.5-) 40.9 (-52.5) × (7.5-) 7.7 (-8.75) µm, macroconidia. Microconidia, one-septate macroconidia, and chlamydospores were not observed. Identity of these isolates was determined by a multiplex PCR system using a set of three pair of specific primers (Mac1/MaPa2, Lir1/Lir2, and Pau1/MaPa2) (1), which generated a 117-bp product that was characteristic of Cylindrocarpon pauciseptatum Schroers & Crous. Morphological characteristics also supported this identification (4). Internal transcribed spacers regions (ITS1 and ITS4) of rDNA were obtained for isolate 1052 and deposited in GenBank (Accession No. HQ441248). This sequence was identical (100%) with the sequence of C. pauciseptatum (GenBank Accession No. HM036590). Pathogenicity tests were conducted with inoculum produced on wheat kernels that were soaked in distilled water in flasks for 12 h. Each flask contained 200 ml of kernels that were subsequently autoclaved three times after excess water was drained. Two fungal disks from a 2-week-old culture of C. pauciseptatum (isolate 1052) grown on PDA were placed aseptically in each flask. Cultures in flasks were incubated at 25°C for 4 weeks and shaken once a week. A plastic pot (220 cm3) was filled with a mixture of sterilized peat moss and 10 g of inoculum. A 1-month-old seedling of P. radiata was planted in plastic pots and placed in a greenhouse at 25 to 30°C in a completely randomized design with six replications. Controls contained sterile wheat kernels. The experiment was repeated. Symptoms developed 20 days after inoculation and consisted of root lesions, a reduction in root biomass, needle blight, and the death of all seedlings. The fungus was reisolated from affected seedlings. Damping-off was not observed on the control plants. C. pauciseptatum causing black foot disease of grapevine (3) was first found in Spain in 2008, but to our knowledge, this is the first report of C. pauciseptatum causing damping-off of P. radiata in Spain. References: (1) S. Alaniz et al. Plant Dis. 93:821, 2009. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Oxford, 2006. (3) M. T. Martin et al. Plant Dis. 95:361, 2011. (4) H. J. Schroers et al. Mycol. Res. 112:82, 2008.

Comparison of routinely used intestinal biopsy forceps in dogs: an ex vivo histopathological approach.

OBJECTIVES: To compare the quality of duodenal and ileal samples obtained with different biopsy forceps. METHODS: Fifteen dogs were included in a prospective ex vivo study. After euthanasia, the duodenum and the ileum were sampled with four different forceps and evaluated according to a standardised scoring system. The biopsy forceps evaluated had alligator jaws or cups with smooth edge with or without a needle. RESULTS: The global quality of the biopsies was better in the ileum that in the duodenum regardless of the biopsy forceps. Biopsy forceps with smooth edge including a needle resulted in fewer artefacts than biopsy forceps with smooth edge but no needle in both sites and those with alligator jaws without a needle provided deeper biopsies than those with smooth edge without a needle only in the duodenum. There was no effect of the biopsy forceps type on the size of the biopsies. CLINICAL SIGNIFICANCE: Our findings may aid in choosing the appropriate type of forceps for intestinal biopsy.

Competition between Superconductor - Ferromagnetic stray magnetic fields in YBa2Cu3O7-x films pierced with Co nano-rods.

Superconductivity and ferromagnetism are two antagonistic phenomena that combined can lead to a rich phenomenology of interactions, resulting in novel physical properties and unique functionalities. Here we propose an original hybrid system formed by a high-temperature superconducting film, patterned with antidots, and with ferromagnetic nano-rods grown inside them. This particular structure exhibits the synergic influence of superconductor (SC) - ferromagnetic (FM) stray fields, in both the superconducting behaviour of the film and the three-dimensional (3D) magnetic structure of nano-rods. We show that FM stray fields directly influence the critical current density of the superconducting film. Additional functionalities appear due to the interaction of SC stray fields, associated to supercurrent loops, with the non-trivial 3D remanent magnetic structure of FM nano-rods. This work unravels the importance of addressing quantitatively the effect of stray magnetic fields from both, the superconductor and the ferromagnet in hybrid magnetic nano-devices based on high temperature superconductors.

Electrical stimulation of cardiac adipose tissue-derived progenitor cells modulates cell phenotype and genetic machinery.

A major challenge of cardiac tissue engineering is directing cells to establish the physiological structure and function of the myocardium being replaced. Our aim was to examine the effect of electrical stimulation on the cardiodifferentiation potential of cardiac adipose tissue-derived progenitor cells (cardiac ATDPCs). Three different electrical stimulation protocols were tested; the selected protocol consisted of 2 ms monophasic square-wave pulses of 50 mV/cm at 1 Hz over 14 days. Cardiac and subcutaneous ATDPCs were grown on biocompatible patterned surfaces. Cardiomyogenic differentiation was examined by real-time PCR and immunocytofluorescence. In cardiac ATDPCs, MEF2A and GATA-4 were significantly upregulated at day 14 after stimulation, while subcutaneous ATDPCs only exhibited increased Cx43 expression. In response to electrical stimulation, cardiac ATDPCs elongated, and both cardiac and subcutaneous ATDPCs became aligned following the linear surface pattern of the construct. Cardiac ATDPC length increased by 11.3%, while subcutaneous ATDPC length diminished by 11.2% (p = 0.013 and p = 0.030 vs unstimulated controls, respectively). Compared to controls, electrostimulated cells became aligned better to the patterned surfaces when the pattern was perpendicular to the electric field (89.71 ± 28.47º for cardiac ATDPCs and 92.15 ± 15.21º for subcutaneous ATDPCs). Electrical stimulation of cardiac ATDPCs caused changes in cell phenotype and genetic machinery, making them more suitable for cardiac regeneration approaches. Thus, it seems advisable to use electrical cell training before delivery as a cell suspension or within engineered tissue.

Induced sputum in the diagnosis of peripheral lung cancer not visible endoscopically.

The diagnosis of small peripheral lung cancer is difficult to achieve by non-invasive methods. We hypothesized that in these patients induced sputum might ncrease the diagnostic yield over spontaneous sputum, representing a good diagnostic alternative in selected patients. We prospectively evaluated 60 patients with peripheral lung lesions and normal bronchoscopic evaluation. Six samples of sputum (three spontaneous and three induced with nebulization of hypertonic saline) before bronchoscopy and six samples of sputum after bronchoscopy (three spontaneous and three induced) were obtained in each subject. Forty-two out of the 60 patients included were finally diagnosed with lung cancer. Eighteen patients were diagnosed with different benign conditions of the lung. Overall, malignant cells in sputum were observed in 21 patients and in all but one, the final diagnosis of lung cancer was achieved. Only one patient with a pseudoinflammatory tumour of the lung had a false-positive result in one spontaneous sputum sample. The diagnosis of lung cancer was obtained in 18 patients with the induced sputum (43%) and in 14 patients with spontaneous sputum (31%) (P=NS). Samples of induced sputum were more adequate for cytological analysis than samples of spontaneous sputum (P < 0.001). Of 13 patients with peripheral lung neoplasms of 2 cm or less in diameter, five were diagnosed using induced sputum (38%) and only one using spontaneous sputum (8%) (P<0.05). In conclusion, induced sputum is a valuable technique for the diagnosis of peripheral lung cancer. Induced sputum gives better quality specimens and better diagnostic yield in small lesions than the spontaneous sputum and may be indicated in selected patients with disseminated disease, inoperability or severe co-morbities.

Relationship between ventilator-associated pneumonia and intramucosal gastric pHi: a case-control study.

PURPOSE: Prior investigations have suggested a clear relationship between nosocomial pneumonia and intramucosal gastric pH (pHi), a probable marker of bacterial translocation. METHODS: We studied 33 patients (18 with pneumonia and 15 without) admitted to an intensive care unit and hospitalized longer than 72 hours with the aim of assessing the relationship between nosocomial pneumonia, pHi, and outcome. pHi was estimated at the time of inclusion of patients into the study. Arterial pH (pHa) and bicarbonate and stomach pH and tonometer PtCO2 were also recorded. Values of < 7.32 or delta pHa-pHi of > +0.06 were used to differentiate between normal and low pHi. Quantitative cultures of pharyngeal swabs, gastric lumen, and protected specimen brush from lower airways were also done. RESULTS: The mean pHi values were 7.397 +/- 0.105 (range, 7.14 to 7.53) and 7.452 +/- 0.059 (range, 7.37 to 7.56) for patients with and without pneumonia, respectively (P = .073). Five patients, all with pneumonia, had pHi < 7.32. No patients without pneumonia had pHi < 7.32 (P = .04). The mean delta pHa-pHi was 0.04 +/- 0.07 (range, -0.11 to 0.13) and 0.05 +/- 0.09 (range, -0.09 to 0.28; P = .72) for patients with and without pneumonia, respectively. However, there were significant differences when tonometer PtCO2 values of both groups were compared (38.9 +/- 8.3 and 30.6 +/- 4.7 mm Hg, respectively; P = .025). Patients with pneumonia had higher alkaline gastric lumen pH (5.2 +/- 1.0) than those without pneumonia (3.8 +/- 1.4; P = .006). Nonsurvivors (n = 7) had more acidic pHi (7.33 +/- 0.11) than survivors (7.44 +/- 0.06; P = .045). The mean gastric lumen bacterial concentration was 4.14 +/- 1.01 Log10 CFU/mL in patients with pneumonia and 4.28 +/- 1.22 Log10 CFU/mL in patients without pneumonia (P = NS). When patients with and without intramucosal gastric acidosis (pHi < 7.32) were compared, the gastric bacterial burden was 4.42 +/- 0.82 Log10CFU/mL and 4.32 +/- 1.03 Log10 CFU/mL (P = .08), respectively. CONCLUSIONS: Most patients with nosocomial pneumonia had no associated intramucosal gastric acidosis. However, low pHi was associated with increased mortality.

[A decrease in the level of CPAP required after prolonged treatment in patients with the obstructive sleep apnea syndrome]. / Disminución del requerimiento del nivel de CPAP tras un tratamiento prolongado en pacientes con síndrome de apneas obstructivas durante el sueño.

Application of continuous positive upper airway pressure (CPAP) is the treatment of choice in obstructive sleep apnea syndrome (OSAS). CPAP keeps the upper airway open by providing a "pneumatic splint" on the pharyngeal walls. However, it has been suggested that prolonged nasal CPAP treatment may also improve the physiopathological mechanisms responsible for OSAS by a variety of mechanisms such as the reduction of edema of the pharyngeal mucosa, modification of upper airway muscle dynamics; there may even be a reduction in the intensity of phenomena responsible for brief awakening and ending of apneic episodes. The objective of this study was to determine whether prolonged nasal CPAP is associated to a reduction of CPAP pressure required for eliminating apnea in patients with OSAS. We studied 22 patients (4 women) with OSAS diagnosed by conventional polysomnography, including study of neurological variables during sleep (electroencephalogram, ocular movement, submental electromyogram), respiratory variables (nasal air flow, thoracic-abdominal movement, O2 saturation in the blood), and others (electrocardiogram, leg movement). Treatment was exclusively with nasal CPAP. All 22 patients were middle-aged (52.3 +/- 10 years), presented a mean apnea/hypoapnea index of 65.2 +/- 24 per hour. CPAP levels needed to eliminate episodes of apnea, blood O2 desaturation and snoring at the time of diagnosis and after long-term treatment (8.5 +/- 4.6 months) were applied. Weight was recorder at both measurement times. Eighteen of the 22 patients (1 woman, mean age 53.1 +/- 11.4 years and apnea/hypopnea index 64.8 +/- 23.4 per hour) experienced no weight change during treatment.(ABSTRACT TRUNCATED AT 250 WORDS)

[Epidemic Kaposi's sarcoma in women: presentation of 4 cases]. / Sarcoma de Kaposi epidémico en mujeres: presentación de 4 casos.

Epidemic Kaposi's sarcoma is the neoplasm most frequently manifested in infection by the human immunodeficiency virus. Its prevalence is considerably higher among homosexual males than among intravenous drug users with practically exclusive infection in this sex. Four cases of Kaposi's sarcoma in women with the human immunodeficiency virus are described. Two of these subjects were intravenous drug users and the other two were heterosexually promiscuous as the only conduct of risk for acquiring infection by the human immunodeficiency virus. Kaposi's sarcoma was the form of presentation of the syndrome of acquired immunodeficiency syndrome in these four patients.

Lung inflammatory response in pneumonia.

In normal conditions, alveolar macrophages (AMs) are the main cells that respond to bacteria that reach lower airways. However, if the microbial inoculum is too high or too virulent to be stopped by AM alone, these cells recruit polymorphonuclear neutrophils (PMN) into the alveoli from the vascular compartment. Cytokines, such as tumour necrosis factor-alpha (TNF-alpha), interleukin-1-beta (IL-beta), interleukin-6 (IL-6), and interleukin-8 (IL-8), secreted by the AM are able to attract PMN enhanced for phagocytosis, ready to destroy the invading pathogens. However, excessive cytokine production has deleterious effects, with a systemic inflammatory response (sepsis) that can lead to multiorganic failure and death. Other cytokines, such as interleukin-10 (IL-10) balance this response, attenuating several inflammatory mechanisms. The inflammatory lung response in pneumonia has been well studied in animals, and more recently in humans, using bronchoalveolar lavage to measure some inflammatory mediators (TNF-alpha, IL-1 beta, IL-6, IL-8). From these studies, it seems that: 1) the inflammatory response to pneumonia is compartmentalized for most cytokines (in contrast to adult respiratory distress syndrome (ARDS)), except for IL-6 which is a general marker of inflammation. On the other hand, C-reactive-protein is an acute-phase protein synthesized by the liver through the stimulus of IL-6 that may also be an easy-to-measure marker of inflammation that is directly related to IL-6; 2) some of these cytokines may be useful as prognostic indices; 3) there is no clear relationship between the local lung bacterial burden and the intensity of the inflammatory response; and 4) the administration of granulocyte colony-stimulating factor (G-CSF) is a promising therapeutic approach that is still under clinical investigation. In the future, it is probable that the therapeutic goal in severe pneumonia will be to find the exact point at which inflammation is beneficial but not deleterious. The measurement of the inflammatory response may serve for this purpose.

Simultaneous surface plasmon resonance and x-ray absorption spectroscopy.

We present an experimental setup for the simultaneous measurement of surface plasmon resonance (SPR) and x-ray absorption spectroscopy (XAS) on metallic thin films at a synchrotron beamline. The system allows measuring in situ and in real time the effect of x-ray irradiation on the SPR curves to explore the interaction of x-rays with matter. It is also possible to record XAS spectra while exciting SPR in order to study changes in the films induced by the excitation of surface plasmons. Combined experiments recording simultaneously SPR and XAS curves while scanning different parameters can be also carried out. The relative variations in the SPR and XAS spectra that can be detected with this setup range from 10(-3) to 10(-5), depending on the particular experiment.

Effect of photodiode angular response on surface plasmon resonance measurements in the Kretschmann-Raether configuration.

We study the effect of photodiode angular response on the measurement of surface plasmon resonance (SPR) in metallic thin films using the Kretschmann-Raether configuration. The photodiode signal depends not only on the light intensity but also on the incidence angle. This implies that the photodiode sensitivity changes along the SPR curve. Consequently, the measured SPR spectrum is distorted, thus affecting fits and numerical analyses of SPR curves. We analyze the magnitude of this change, determine when it is significant, and develop a calibration method of the experimental setup which corrects for this type of spectral shape distortions.

Markers of ventilator-associated pneumonia.

The diagnosis of ventilator-associated pneumonia (VAP) is difficult for several reasons. Firstly, clinical markers show a large percentage of false-positive and false-negative results. Secondly, microbiological diagnosis based on quantitative cultures of protected specimen brush (PSB), bronchoalveolar lavage (BAL), and endotracheal aspirates also present false-positive and false-negative results. Furthermore, definite results are delayed for 48-72 hours. For all these reasons it would be an advantage to have a biological marker of ventilator-associated pneumonia in clinical practice. Since clinical features of pneumonia in mechanically ventilated patients are neither specific nor sensitive, rapid markers of pneumonia might be of great assistance to the clinician in deciding whether to start an empiric antibiotic regimen. A marker of ventilator-associated pneumonia could be a rapid alternative diagnostic method which permits the definite diagnosis of pneumonia. Accordingly, specific markers of VAP, namely the presence of intracellular microorganisms, the detection of elastin fibres, the antibody-coated bacteria test, the level of endotoxin in bronchoalveolar lavage fluid, the local production of interleukin-8, the levels of lactate dehydrogenase, and decreased surfactant protein A, may be important as they can provide a rapid diagnosis of VAP. Among the markers alluded to above, the search for intracellular bacteria in polymorphonuclear leukocytes or macrophages is the most widely validated technique with an excellent specificity, provided that prior antibiotics are not given. However, this technique has its own limitations; it requires a considerable time effort for the microbiologist, and also compels the performance of BAL, a technique not always harmless to the patient.