Search

The members of the formyl peptide receptor (FPR) family are involved in the sensing of chemoattractant substances, including bacteria-derived N-formylated peptides and host-derived peptides and proteins. We have recently described two chemoattractant receptor inhibitors from Staphylococcus aureus. Chemotaxis inhibitory protein of S. aureus (CHIPS) blocks the formyl peptide receptor (FPR) and the receptor for complement C5a (C5aR), while FPR-like 1 (FPRL1) inhibitory protein (FLIPr) blocks the FPRL1. Here, we describe another staphylococcal chemoattractant-inhibiting protein with 73% overall homology to FLIPr and identical first 25 aa, which we termed FLIPr-like. This protein inhibits neutrophil calcium mobilization and chemotaxis induced by the FPRL1-ligand MMK-1 and FPR-ligand fMLP. While its FPRL1-inhibitory activity lies in the comparable nanomolar range of FLIPr, its antagonism of the FPR is ∼100-fold more potent than that of FLIPr and comparable to that of CHIPS. The second N-terminal phenylalanine was required for its inhibition of the FPR, but it was dispensable for the FPRL1. Furthermore, the deletion of the first seven amino acids reduced its antagonism of the FPRL1, and the exchange of the first six amino acids with that of CHIPS-conferred receptor specificity. Finally, studies with cells transfected with several chemoattractant receptors confirmed that FLIPr-like specifically binds to the FPR and FPRL1. In conclusion, the newly described excreted protein from S. aureus, FLIPr-like, is a potent inhibitor of the FPR- and FPRL1-mediated neutrophil responses and may be used to selectively modulate these chemoattractant receptors.

The aim of the study was to assess a new latex agglutination (LA) stool antigen assay (PYLOGEN; CerTest Biotec, Zaragoza, Spain) in the diagnosis of Helicobacter pylori infection and to monitor its eradication after treatment. The LA test has been approved for sale in Europe, and its approval from the US Food and Drug Administration is still pending. The individuals enrolled were classified into 3 groups of patients: Group 1 consisted of 38 patients who are H. pylori positive. The diagnosis of H. pylori infection was established if there was concordance between 2 test results (urea breath test [UBT], rapid urease test, and histopathologic study) or if the culture alone was positive. Patients with only 1 positive test were considered indeterminate and were excluded from the study. Group 2 comprised 9 patients without positive tests and who were considered to be H. pylori negative. Group 3 consisted of 57 patients who received eradication treatment. The sensitivity and specificity of the test were 78.9% and 100%, respectively. The results of the UBT of the patients were studied 6 weeks after eradication therapy. The sensitivity and specificity of the LA test relative to UBT for patients after treatment were 75% and 93.3%, respectively.

SUMMARYIn vitro tests have been developed for the diagnosis of tuberculosis (TB) infection. The objective was to analyse latent TB infection in drug and alcohol abusers through two interferon-γ techniques. One hundred and thirty-nine patients were admitted between February 2006 and May 2007. Mean age was 39·8 years [31% HIV positive]. The enzyme immunoassay (EIA) and enzyme-linked immunospot (ELISPOT) interferon-γ assays were positive in 34% of patients with an agreement of 83% (κ=0·63). Tuberculin skin test (TST) was positive in 29% of patients and the agreement of TST with EIA and ELISPOT interferon-γ assays was 85% (κ=0·62) and 83% (κ=0·57), respectively. Almost 50% of patients with history of TB had a positive in vitro test. In conclusion, we observed a high prevalence of latent TB and good agreement between the new in vitro tests that otherwise may continue to be positive long after developing TB disease.

Background. The aim of the present research was to study the possible interference of hemosiderin deposits with the histological detection of dextran-coated, iron-labeled, mesenchymal stem cells after intracoronary administration in a porcine model of myocardial infarction. Materials and methods. A myocardial infarction was induced in six animals that received intracoronary iron-labeled autologous mesenchymal stem cells (group 1; n 2) or placebo (group 2; n 4). Six control animals without myocardial infarction underwent direct intramyocardial injections of iron-labeled autologous mesenchymal stem cells (group 3; n 2) or placebo (group 4; n 4). Histological sections from explanted hearts were stained with Prussian blue to identify dextran-coated, iron-labeled, mesenchymal stem cells. Results. After Prussian blue staining, granular blue labeling in the tissue was observed in both groups of animals with infarcts. Similar granular blue labeling was detected in hearts from control animals without infarction that had received iron-labeled mesenchymal stem cells. However, hearts from control animals without infarction that received placebo did not have any granular blue labeling in the tissue. Conclusions. Hemosiderin from infarction hemorrhage interferes with detection of dextran-coated iron-labeled mesenchymal stem cells after intracoronary administration, suggesting that this marker is not useful to detect mesenchymal stem cells in a porcine model of myocardial infarction.

The purpose of this study was to evaluate the GenoType MTBDR plus assay (Hain Lifescience GmbH, Nehren, Germany) for its ability to detect resistance to rifampin (RIF) and isoniazid (INH) in Mycobacterium tuberculosis clinical strains and directly in clinical samples. A total of 62 clinical strains characterized with the Bactec 460TB system were included. For the INH-resistant strains, the MIC was measured and sequencing was performed. Sixty-five clinical samples from 28 patients (39 smear-positive samples and 26 smear-negative samples) were also tested directly. The corresponding isolates of the clinical specimens were studied with the Bactec 460TB system. The overall rates of concordance of the MTBDR plus assay and the Bactec 460TB system for the detection of RIF and INH susceptibility in clinical strains were 98.3% (61/62) and 79% (49/62), respectively. The rate of concordance between the Bactec 460TB system and the MTBDR plus test for the detection of INH resistance in the group of 27 strains with low-level resistance was 62.9% (17/27), and that for the detection of INH resistance in the group of 21 strains with high-level resistance was 85.71% (18/21). Valid test results were obtained for 78.45% (51/65) of the clinical samples tested. The rates of concordance between both assays for the detection of drug resistance in these samples were 98% (50/51) for RIF and 96.2% (49/51) for INH. Taking into account only one sample per patient, the overall rate of concordance between both tests was 92.85% (26/28). The GenoType MTBDR plus assay is easy to perform and is a useful tool for the management of tuberculosis, as it allows the detection of resistance to RIF and INH in M. tuberculosis strains and also in clinical samples.

Las micobacterias constituyen un grupo de bacterias de gran interes en medicina, ya que, junto a especies teluricas y oportunistas, se hallan 2 especies (Mycobacterium tuberculosis y Mycobacterium leprae) de gran importancia en salud publica. A pesar de los esfuerzos realizados para su control, la tuberculosis (TB) sigue siendo en la actualidad uno de los problemas sanitarios de mas trascendencia mundial. En los ultimos anos, la micobacteriologia ha experimentado importantes avances tecnologicos. A pesar de ello, el diagnostico temprano de la infeccion por micobacterias y, especialmente de la TB, sigue recayendo en el examen microscopico de las muestras tenidas de manera adecuada. En la actualidad, este sigue siendo el procedimiento mas simple, de mejor coste-efectividad y rapidez para proporcionar al clinico una orientacion diagnostica preliminar. El control efectivo de la TB se basa en la deteccion rapida de M. tuberculosis, seguido por la inmediata implementacion del tratamiento antituberculoso adecuado. La emergencia de cepas resistentes a los farmacos antituberculosos agudiza la necesidad de disponer de metodos rapidos de deteccion de M. tuberculosis y de resistencias. La disponibilidad de metodos de epidemiologia molecular de facil implementacion y estandarizacion, que nos permitan identificar casos relacionados, es fundamental para identificar brotes epidemicos que ayuden a controlar la propagacion de la TB. Aun reconociendo los evidentes progresos realizados en el diagnostico molecular de las infecciones micobacterianas, las tecnicas disponibles son todavia insuficientes. En esta revision, describimos el estado actual de las principales tecnicas moleculares para la deteccion directa de micobacterias en muestras clinicas, para su identificacion, deteccion de resistencias a los principales farmacos antituberculosos y de epidemiologia molecular. En cada caso, destacamos las ventajas y las limitaciones de ellas. En un proximo futuro la micobacteriologia clinica evolucionara, con bastante probabilidad, hacia la universalizacion de las tecnicas geneticas aplicadas al diagnostico directo y la deteccion de resistencias. La epidemiologia molecular de la TB se realizara, en sus diferentes aplicaciones, con tecnicas mas rapidas y automatizadas que las actuales.

Despite advances in medium formulations and pretreatment techniques, recovery of Legionella from water samples can still be quite low, difficult and time consuming. The aim of this study was to evaluate the utility of a Legionella urinary antigen enzyme immunoassay (Bartels ELISA, Trinity Biotech, Ireland) for the detection of Legionella in water samples. Reference ATCC Legionella strains were used to spike water samples to a final concentration of 10 4 –10 5 cfu/ml. The lower detection limit of the test for all Legionella pneumophila serogroups was assessed by serial dilutions of spiked water samples. Legionella antigen was detected in all filtered samples except for those spiked with L. bozemanii and L. longbeachae . The lower detection limit for soluble L. pneumophila serogroup 1 antigen was 780 cfu/ml. Bartels ELISA could be a useful method for antigen detection in water samples when a high recovery of L. pneumophila is suspected. The test could be used as a rapid screening method for the detection of Legionella in a large number of samples. However, the low sensitivity of the test requires to keep on performing conventional culture for isolation and for further studies on isolated bacteria.

We evaluated the T-SPOT.TB and Quantiferon-TB Gold In tube (QFN-G-IT) tests for diagnosing Mycobacterium tuberculosis infection. T-SPOT.TB was more sensitive than QFN-G-IT in diagnosing both active and latent infection. Both gamma interferon tests were unaffected by prior Mycobacterium bovis BCG vaccination. Among children who were not BCG vaccinated but had a positive tuberculin skin test, QFN-G-IT was negative in 53.3% of cases, and T-SPOT.TB was negative in 50% of cases.

Background Umbilical cord blood (UCB) has been widely used for hematopoietic stem cell transplantation. The UCB-derived stem cells (UCBSCs) have been proposed as an alternative to bone marrow (BM)–derived mesenchymal stem cells (MSCs) for cardiac cell-based therapy. Herein we studied whether UCBSCs spontaneously exhibit cardiac-specific markers in vitro. Methods Human UCBSCs were isolated, expanded, and phenotyped by flow cytometry, quantitative RT-PCR, and immunofluorescence. Cell pluripotency and proliferation were also assessed by adipogenic and osteogenic media and in growth assays. Results Among 25 analyzed UCB, 16% of cases afforded primary culture satisfactory generation of UCBSCs. Duplication time (Td) of cultures was 2.16 ± 0.06 days. The cells were strongly positive for CD105 (18.5 ± 0.14), CD44 (27 ± 2.8), CD166 (13 ± 9), CD29 (59 ± 9.4), CD90 (60 ± 11) and consistently negative for CD117 (1.2 ± 0.1), CD106 (1.1 ± 0), CD34 (1.2 ± 0.2), CD14 (1 ± 0), and CD45 (1 ± 0), consistent with a mesenchymal lineage. Adipogenesis and osteogenesis of cells resulted in low accumulation of intracellular lipid droplets and high deposition of calcium. The UCBSCs showed gene transcripts for α-actinin, connexin (Cx)-43, SERCA-2, and stromal cell-derived factor (SDF)-1α. At the protein level, the cells abundantly expressed α-actinin, Cx-43, SERCA-2 and SDF-1α. In contrast, these cells did not express the cardiac transcription factors GATA-4, Tbx5, and Nkx2.5, nor the sarcomeric proteins β-myosin heavy chain (β-MyHC) or cardiac troponin I (cTnI). Conclusions Human UCBSCs may represent an alternative source of stem cells for myocardial-cell replacement. These cells can be highly expanded. They spontaneously express proteins of paramount importance for cardiovascular regeneration, such as Cx-43, SERCA-2, and SDF-1α.

Human bone marrow-derived mesenchymal stem cells (MSCs) exhibit limited in vitro growth. Fibroblast growth factors (FGFs) elicit a variety of biological responses, such as cell proliferation, differentiation and migration. FGF-4 represents one of the FGFs with the highest cell mitogenic activity. We studied the effect of FGF-4 on MSCs growth and pluripotency. MSCs duplication time (Td) was significantly reduced with FGF-4 compared to controls (2.2 +/- 0.2 vs. 4.1 +/- 0.2 days, respectively; p = 0.03) while BMP-2 and SCF-1 did not exert a significant growth effect. MSC expression of surface markers, differentiation into adipogenic and osteogenic lineages, and baseline expression of cardiomyogenic genes were unaffected by FGF-4. In summary, exogenous FGF-4 increases the rate at which MSC proliferate and has no significant effect on MSC pluripotency.

Objective To validate high serum procalcitonin (PCT) as a predictor of vesicoureteral reflux (VUR) in children with a first febrile urinary tract infection (UTI). Study design This secondary analysis of prospective hospital-based cohort studies included children ages 1 month to 4 years with a first febrile UTI. Results Of the 398 patients included in 8 centers in 7 European countries, 25% had VUR. The median PCT concentration,vas significantly higher in children with VUR than in those without: 1.6 versus 0.7 ng/ml, (P = 10(-4)). High PCT (>= 0.5 ng/ml) was' associated with VUR (OR: 2.3. 95% Cl, 1.3 to 3.9; P = 10(-3)). After adjustment for all cofactors, the association remained significant (OR: 2.5: 95% Cl, 1.4 to 4.4; P = 10(-3)). The strength of the relation increased with tire grade of reflux (P = 10(-5).). The sensitivity of procalcitonin was 75% (95% Cl, 66 to 83) for all-grade VUR and 100% (95% Cl, 81 to 100) for grade >= 4 VUR. both with 43% specificity (95% Cl, 37 to 48). Conclusions High PCT is a strong, independent and now validated predictor of VUR that can be used to identify low-risk patients and thus avoid one third of the unnecessary cystourethrographies in children with a first febrile UTI.

Treating a myocardial infarction (MI), the most frequent cause of death worldwide, remains one of the most exciting medical challenges in the 21st century. Cardiac tissue engineering, a novel emerging treatment, involves the use of therapeutic cells supported by a scaffold for regenerating the infarcted area. It is essential to select the appropriate scaffold material; the ideal one should provide a suitable cellular microenvironment, mimic the native myocardium, and allow mechanical and electrical coupling with host tissues. Among available scaffold materials, natural scaffolds are preferable for achieving these purposes because they possess myocardial extracellular matrix properties and structures. Here, we review several natural scaffolds for applications in MI management, with a focus on pre-clinical studies and clinical trials performed to date. We also evaluate scaffolds combined with different cell types and proteins for their ability to promote improved heart function, contractility and neovascularization, and attenuate adverse ventricular remodeling. Although further refinement is necessary in the coming years, promising results indicate that natural scaffolds may be a valuable translational therapeutic option with clinical impact in MI repair.

Introduction: Staphylococcus aureus is frequently isolated in respiratory samples both as colonizer and causing lower respiratory tract infection (LRTI) and may persist despite treatment with risk of developing resistance. Objective: To investigate the correlation of persistent isolation of S.aureus with clinical and microbiological variables in patients undergoing mechanical ventilation treated as LRTI. Methods: Clinico-epidemiological data were collected from patients with isolation of S.aureus in tracheal aspirate. Persistence was considered when lasted more than 3 days despite treatment adjusted to susceptibility profile. Strains were phenotypically and genotypically characterized. Results: During the study period (3 years) S.aureus was isolated from 122 patients (82,3% nosocomial), stratified by CPIS as 28 pneumonia,43 tracheobronchitis and 51 bronchial colonization. Among the 91 patients that received antimicrobial treatment after first isolation, persistence was documented in 32 cases (35,2%)(8,9 days).None of the clinical variables correlated with persistence except ictus of ischemic origin (but only 5 cases).Regarding strains characteristics, there was no correlation with cloxacillin resistance (18%MRSA), clonal complex and presence of selected virulence factors. However, agrI allele (p=0,002) and absence of TSST-1 gene (p=0,025) were more frequent in persistent strains. Conclusion: Persistent isolation of S.aureus is frequent, without correlation to clinical variables, prognosis,nor resistance pattern.However, some genotypical features of S.aureus might be related, suggesting its adaptation to the lower respiratory tract allowing its survival.SEPAR054/2011;FIS PI13/01418.

Introduction: Staphylococcus aureus can colonize the respiratory tract or progress as etiologic agent of lower respiratory tract infection (LRTI). Activation of accessory gene regulator ( agr) operon leads to the expression of haemolysins and downregulation of surface proteins and its dysfunction has been associated with clinical failure in bacteremia. Objective: To study the clinical role of agr functionality in the respiratory tract. Methods: Strains isolated from nasal swabs and tracheal aspirates were phenotypically and genotypically characterized. agr activity was assessed by δ-haemolysin CAMP assay and Vesicle Lysis Test (VLT). Results: Patients were stratified as nasal carriers (n=13), bronchial colonization(n=27) and LRTI (28 tracheobronchitis, 25 pneumonia). agr was functional in 79,1% by CAMP and 80,2%, by VLT. Concordance between both tests was 85,22%(κ:0,516;SE:0,117). VLT was positive in 84,2% of pneumonia strains and 76,9% of nasal carriers but no correlation was observed with cloxacillin resistance (23%MRSA) nor persistent isolation (47,3% despite treatment adjusted to susceptibility profile). A high diversity of clonal complexes was observed being CC5 the most dominant. Prevalence of agr allele I and II were 41,8% and 42,9%, respectively. Presence of virulence factors (CHIPS,75.8%;SCIN,89%;TSST-1,18.7%;PVL one strain) did not correlate to study group nor to agr functionality. Conclusion. High agr activity and δ-toxin expression was observed in strains isolated from respiratory tract and did not correlate to clinical features nor genotype. Activity of agr could be important as an adaptation of S.aureus to the respiratory niche. SEPAR 054/2011; FIS PI13/01418.

The aim of this work was to study the diagnostic accuracy of pyrosequencing to detect resistance to fluoroquinolones, kanamycin, amikacin, capreomycin, and ethambutol (EMB) in Mycobacterium tuberculosis clinical strains. One hundred four clinical isolates previously characterized by BACTEC 460TB/MGIT 960 were included. Specific mutations were targeted in gyrA, rrs, eis promoter, and embB. When there was a discordant result between BACTEC and pyrosequencing, Genotype MTBDRsl (Hain Lifescience, Nehren, Germany) was performed. Sensitivity and specificity of pyrosequencing were 70.6% and 100%, respectively, for fluoroquinolones; 93.3% and 81.7%, respectively, for kanamycin; 94.1% and 95.9%, respectively, for amikacin; 90.0% and 100%, respectively, for capreomycin; and 64.8% and 87.8%, respectively, for EMB. This study shows that pyrosequencing may be a useful tool for making early decisions regarding second-line drugs and EMB resistance. However, for a correct management of patients with suspected extensively drug-resistant tuberculosis, susceptibility results obtained by molecular methods should be confirmed by a phenotypic method.

Bacteria have developed mechanisms to escape the first line of host defense, which is constituted by the recruitment of phagocytes to the sites of bacterial invasion. We previously described the chemotaxis inhibitory protein of Staphylococcus aureus, a protein that blocks the activation of neutrophils via the formyl peptide receptor (FPR) and C5aR. We now describe a new protein from S. aureus that impaired the neutrophil responses to FPR-like1 (FPRL1) agonists. FPRL1 inhibitory protein (FLIPr) inhibited the calcium mobilization in neutrophils stimulated with MMK-1, WKYMVM, prion-protein fragment PrP106–126, and amyloid β1–42. Stimulation with low concentrations of fMLP was partly inhibited. Directed migration was also completely prevented toward MMK-1 and partly toward fMLP. Fluorescence-labeled FLIPr efficiently bound to neutrophils, monocytes, B cells, and NK cells. HEK293 cells transfected with human C5aR, FPR, FPRL1, and FPRL2 clearly showed that FLIPr directly bound to FPRL1 and, at higher concentrations, also to FPR but not to C5aR and FPRL2. FLIPr can reveal unknown inflammatory ligands crucial during S. aureus infections. As a novel described FPRL1 antagonist, it might lead to the development of therapeutic agents in FPRL1-mediated inflammatory components of diseases such as systemic amyloidosis, Alzheimer’s, and prion disease.

Spontaneous calcium release from the sarcoplasmic reticulum in cardiac myocytes plays a central role in cardiac arrhythmogenesis. Compounds intended for therapeutical use that interfere with intracellular calcium handling may therefore have an undesired proarrhythmic potential. Here we have used isolated human atrial myocytes to compare the effect of the proarrhythmic antihistaminic drug terfenadine with the non-proarrhythmic antihistaminic drugs fexofenadine and rupatadine on intracellular calcium homeostasis. Perforated patch-clamp technique was used to measure ionic currents and to detect spontaneous calcium release from the sarcoplasmic reticulum. Our results show that the compound terfenadine, with known arrhythmogenic effects, inhibits L-type calcium current (I(Ca)) with an IC(50) of 185 nM when cells are stimulated at 1.0 Hz. The inhibitory effect of 0.3 muM terfenadine increased from 19+/-4% at stimulation frequency of 0.2 Hz to 63+/-6% at 2.0 Hz. Moreover, terfenadine also increased spontaneous calcium release from the sarcoplasmic reticulum. At a concentration of 1 muM, terfenadine significantly increased the spontaneous Na-Ca exchange current (I(NCX)) frequency from 0.48+/-0.25 to 1.93+/-0.67 s(-1). In contrast, fexofenadine and rupatadine did not change I(Ca) or the frequency of spontaneous I(NCX). We conclude that the proarrhythmic antihistaminic drug terfenadine alters intracellular calcium handling in isolated human atrial myocytes. This experimental model may be suitable to screen for potential arrhythmogenic side-effects of compounds intended for therapeutical use.

Summary Objectives The aim of the study is to assess the usefulness of C polysaccharide and polysaccharide capsular antigen detection by immunochromatography (ICT) and enzyme immunoassay (EIA), respectively, in serum samples for diagnosing pneumococcal pneumonia. Methods Adult patients included in the study were classified in the following groups: In group 1 we studied 101 serum samples from patients with pneumonia due to Streptococcus pneumoniae . In 53 cases the pneumonia was bacteremic. The second group contained 113 serum samples from patients with no pneumococcal pneumonia. Group 3 was made up of 40 serum samples from healthy subjects with no clinical or radiological signs of pneumonia. Results Using ICT, antigen was detected in 50% of patients with pneumococcal pneumonia, in 64.3 and 40.9% of patients with bacteremic and non-bacteremic pneumococcal pneumonia, respectively. Using EIA, antigens were detected in 35.8% of patients with pneumococcal pneumonia, in 45 and 22.2% of patients with bacteremic and non-bacteremic pneumococcal pneumonia, respectively. Conclusions In conclusion, the sensitivity of the tests is low. However, in special situations, where obtaining large volume of urine is difficult, they could be a complementary method in the rapid diagnosis of pneumococcal pneumonia.

Summary Study objectives To evaluate the rapid urine antigen tests, including a new rapid immunochromatographic test (ICT) for the detection of the Streptococcus pneumoniae antigen and an enzyme immunoassay (EIA) for the detection of the Legionella antigen, in order to improve the diagnosis of community-acquired pneumonia (CAP) in adults. Design Prospective study. Setting A tertiary hospital in Spain. Patients We consecutively recruited 107 adults with CAP evaluated at our hospital. Interventions The analyses included blood and sputum cultures, pleural fluid culture (if present) and serologic studies. The detection of the Legionella pneumophila urinary antigen was performed by EIA, and the detection of S. pneumoniae antigen in urine samples was performed by counterimmunoelectrophoresis (CIE) and a rapid ICT. Results Using conventional microbiologic tests we succeeded in performing the etiologic diagnosis of 39 out of the 107 cases (36.4%). The inclusion of rapid antigen detection techniques increased the percentage of diagnosis to 54.2%, which represents a total increase of 17.8% \( P = 0.034 \) . Conclusions The data obtained in this study indicate that rapid urine antigen tests are very useful to determine CAP etiology in adults and, consequently, to quickly identify a group of patients in whom narrow spectrum antibiotics may be used.

Summary Objective Clinical outcome of pneumonia depends on a multifaceted treatment approach. Not only diagnostic methods but also early indicators of the degree of inflammatory response can aid in therapeutic decisions. The objective was to evaluate the usefulness of procalcitonin and neopterin in distinguishing among aetiologies as well as severity in patients with pneumonia. Patients A total of one hundred sixteen patients with clinical, radiographic and microbiological diagnosis of pneumonia were grouped by aetiology, pneumonia severity index, and by the presence of unilobar or multilobar radiographic pulmonary infiltrates. Procalcitonin and neopterin were measured by immunoassays. Results Patients with pneumococcal pneumonia presented elevated procalcitonin and neopterin levels, being higher in bacteraemic than in non-bacteraemic pneumonia. Patients with Legionella pneumonia presented elevated neopterin levels and slightly elevated procalcitonin levels. Patients with tuberculosis and Pneumocystis jirovecii pneumonia presented elevated neopterin and low or not detectable procalcitonin. Procalcitonin and neopterin levels were increased in high-risk classes of pneumonia severity index. Both parameters yielded significant correlation to the radiographic extent and also to young age. Conclusions Procalcitonin and neopterin levels vary depending on age, aetiology and severity of pneumonia. Together with clinical and microbiological data, combined measurement can help to identify patients who might benefit from additional therapies.

Urinary tract infection (UTI) in young children carries the risk of parenchymal damage and sequelae. The location of the infection within the urinary tract influences decisions regarding both therapeutics and follow-up. Because clinical features and laboratory markers of infection at an early age are not specific, it is difficult to make a distinction between lower UTI and acute pyelonephritis. Procalcitonin (PCT) has been studied as a marker of severe bacterial infection. The aim of this study was to test the usefulness of PCT concentration in serum to distinguish between uncomplicated UTI and severe acute pyelonephritis with renal scars.PCT was measured by immunoluminometric assay in serum samples from children with microbiologically documented infection. Severe renal involvement was assessed by 99mTc-dimercaptosuccinic acid gammagraphy done 5 to 6 months after the episode to check for the presence of parenchymal scars. C-reactive protein (CRP) and leukocyte count were also measured.PCT at presentation showed a significant correlation (P0.001) with the presence of renal scars in children with UTI. Using a cutoff of 1 ng/ml for PCT and 20 mg/l for CRP, sensitivity and specificity in distinguishing between urinary tract infection with and without renal damage were 92.3 and 61.9%, respectively, for PCT and 92.3 and 34.4% for CRP. Positive and negative predictive values were 32 and 97.5%, respectively, for PCT and 23 and 95%, respectively, for CRP.A low PCT value at admission indicates a low risk of long term renal scarring. Increased PCT values at admission correlate with the presence of scars. PCT values have proved to be more specific than CRP and leukocyte count for identifying patients who might develop renal damage.

Objective This study was performed to determine the ability of the VITAL system to detect and allow recovery of microorganisms that are difficult to grow, such as Brucella spp., yeasts, or anaerobes, as well as to determine the need for blind subcultures after the incubation period. Methods A prospective evaluation of the system was performed, and 8247 blood culture bottles were processed. The standard was blind subculture from all the bottles after 5 days of incubation. Results There were 3.2% false-positive and 0.6% false-negative results (72% of clinical importance). The system sensitivity for yeasts was 41%. The mean time for detection of Neisseria meningitidis was 31.9 ± 2.8 h, for Brucella spp. 119.7 ± 2 h, and for yeast 51.5 ± 27.8 h. Conclusions The VITAL system poses has serious difficulties in the detection of N. meningitidis, Brucella spp., yeast and methicillin- and aminoglycoside-resistant Staphylococcus aureus (MARSA). The low system sensitivity for yeast detection makes the blind subculture necessary after the incubation period.

The aim of our work here was to evaluate the immunogenicity of 60 mycobacterial antigens, some of which have not been previously assessed, notably a novel series of in vivo-expressed Mycobacterium tuberculosis (IVE-TB) antigens. We enrolled 505 subjects and separated them in individuals with and without latent tuberculosis infection (LTBI) vs. patients with active tuberculosis (TB). Following an overnight and 7 days stimulation of whole blood with purified recombinant M. tuberculosis antigens, interferon-γ (IFN-γ) levels were determined by ELISA. Several antigens could statistically significantly differentiate the groups of individuals. We obtained promising antigens from all studied antigen groups [dormancy survival regulon (DosR regulon) encoded antigens; resuscitation-promoting factors (Rpf) antigens; IVE-TB antigens; reactivation associated antigens]. Rv1733, which is a probable conserved transmembrane protein encoded in DosR regulon, turned out to be very immunogenic and able to discriminate between the three defined TB status, thus considered a candidate biomarker. Rv2389 and Rv2435n, belonging to Rpf family and IVE-TB group of antigens, respectively, also stood out as LTBI biomarkers. Although more studies are needed to support our findings, the combined use of these antigens would be an interesting approach to TB immunodiagnosis candidates.

Background/objectives The aim of this study was to develop a myocardial bioprosthesis for cardiac repair with an integrated online monitoring system. Myocardial infarction (MI) causes irreversible myocyte loss and scar formation. Tissue engineering to reduce myocardial scar size has been tested with variable success, yet scar formation and modulation by an engineered graft is incompletely characterized. Methods Decellularized human pericardium was embedded using self-assembling peptide RAD16-I with or without GFP-labeled mediastinal adipose tissue-derived progenitor cells (MATPCs). Resulting bioprostheses were implanted over the ischemic myocardium in the swine model of MI ( n =8 treated and n =5 control animals). For in vivo electrical impedance spectroscopy (EIS) monitoring, two electrodes were anchored to construct edges, covered by NanoGold particles and connected to an impedance-based implantable device. Histological evaluation was performed to identify and characterize GFP cells on post mortem myocardial sections. Results Pluripotency, cardiomyogenic and endothelial potential and migratory capacity of porcine-derived MATPCs were demonstrated in vitro . Decellularization protocol efficiency, biodegradability, as well as in vitro biocompatibility after recellularization were also verified. One month after myocardial bioprosthesis implantation, morphometry revealed a 36% reduction in infarct area, Ki67 + -GFP + -MATPCs were found at infarct core and border zones, and bioprosthesis vascularization was confirmed by presence of Griffonia simplicifolia lectin I (GSLI) B4 isolectin + -GFP + -MATPCs. Electrical impedance measurement at low and high frequencies (10kHz–100kHz) allowed online monitoring of scar maturation. Conclusions With clinical translation as ultimate goal, this myocardial bioprosthesis holds promise to be a viable candidate for human cardiac repair.

BACKGROUND: Despite recent advances, myocardial infarction (MI) remains the leading cause of death worldwide. Pre-clinical animal models that closely mimic human MI are pivotal for a quick translation of research and swine have similarities in anatomy and physiology. Here, we compared coronary surgical ligation versus coil embolization MI models in swine.METHODS: Fifteen animals were randomly distributed to undergo surgical ligation (n=7) or coil embolization (n=8). We evaluated infarct size, scar fibrosis, inflammation, myocardial vascularization, and cardiac function by magnetic resonance imaging (MRI).RESULTS: Thirty-five days after MI, there were no differences between the models in infarct size (P=0.53), left ventricular (LV) ejection fraction (P=0.19), LV end systolic volume (P=0.22), LV end diastolic volume (P=0.84), and cardiac output (P=0.89). Histologically, cardiac scars did not differ and the collagen content, collagen type I (I), collagen type III (III), and the I/III ratio were similar in both groups. Inflammation was assessed using specific anti-CD3 and anti-CD25 antibodies. There was similar activation of inflammation throughout the heart after coil embolization (P=0.78); while, there were more activated lymphocytes in the infarcted myocardium in the surgical occlusion model (P=0.02). Less myocardial vascularization in the infarction areas compared with the border and remote zones only in coil embolization animals was observed (P=0.004 and P=0.014, respectively).CONCLUSIONS: Our results support that surgical occlusion and coil embolization MI models generate similar infarct size, cardiac function impairment, and myocardial fibrosis; although, inflammation and myocardial vascularization levels were closer to those found in humans when coil embolization was performed.

Summary Objective Performance of IFN-γ assays in children is compromised. Therefore, we investigated the utility of IP-10 for the detection of active tuberculosis (TB) and latent tuberculosis infection (LTBI) diagnosis in children; comparing its positivity with QuantiFERON-TB Gold In-Tube (QFN-G-IT) and T-SPOT.TB. Methods We studied 230 children from three groups: active TB, screening (healthy children without known exposure to active TB patient screened at school or by their paediatrician) and contact-tracing studies. IFN-γ release was determined by QFN-G-IT and T-SPOT.TB. IP-10 was detected in QFN-G-IT supernatants by ELISA. Results When combining QFN-G-IT and IP-10 assays, positive results improved significantly from 38.3% in QFN-G-IT and 33.9% in IP-10 to 41.3%. Age and type of contact were significant risk factors associated with positive QFN-G-IT and IP-10 results. IP-10 levels after antigen-specific stimulation were significantly higher in comparison to IFN-γ levels. Correlation between the three assays was good ( κ = 0.717–0.783). Conclusions IP-10 cytokine is expressed in response to TB specific-antigens used in QFN-G-IT. In conclusion, the use of IFN-γ T-cell based assays in combination with an additional IP-10 assay detection could be useful for diagnosing active TB and LTBI in children.

The optimal cell lineage for cardiac-regeneration approaches remains mysterious. Additionally, electrical stimulation promotes cardiomyogenic differentiation of stimulated cells. Therefore, we hypothesized that electrical conditioning of cardiomyocyte progenitor cells (CMPCs) might enrich their cardiovascular potential. CMPCs were isolated from human adult atrial appendages, characterized, and electrically stimulated for 7 and 14 days. Electrical stimulation modulated CMPCs gene and protein expression, increasing all cardiac markers. GATA-binding protein 4 (GATA4) early transcription factor was significantly overexpressed (P = 0.008), but also its coactivator myocyte enhancer factor 2A (MEF2A) was upregulated (P = 0.073) under electrical stimulation. Moreover, important structural proteins and calcium handling-related genes were enhanced. The cardioregeneration capability of CMPCs is improved by electrical field stimulation. Consequently, short-term electrical stimulation should be a valid biophysical approach to modify cardiac progenitor cells toward a cardiogenic phenotype, and can be incorporated into transdifferentiation protocols. Electrostimulated CMPCs may be best-equipped cells for myocardial integration after implantation. Electronic supplementary material The online version of this article (doi:10.1186/scrt482) contains supplementary material, which is available to authorized users.

Study objectives: Evaluation of a newly available rapid(15 min) immunochromatographic membrane test (ICT) to detect Streptococcus pneumoniae in urine samples, in order toassess its utility in the diagnosis of bacteremic and nonbacteremicpneumococcal pneumonia. Design: Retrospectivestudy. Setting: We studied urine samples from 51patients with bacteremic and nonbacteremic pneumonia due to Spneumoniae diagnosed by blood culture and pneumococcalpolysaccharide capsular antigen detection bycounterimmunoelectrophoresis in urine samples, 16 patients withprobable pneumococcal pneumonia, 71 patients with nonpneumococcalpneumonia, and 16 patients with pneumonia but no pathogen identified. Urine samples were collected and frozen at − 20°C until used. The ICT test was performed following the instructions of themanufacturer. Measurements and results: Spneumoniae antigen was detected in 41 of 51 patients withpneumococcal pneumonia (80.4%); results were positive in 23 of 28bacteremic cases (82.1%) and in 18 of 23 nonbacteremic cases (78.3%). From patients with a diagnosis of presumptive pneumococcal pneumonia, antigen was detected in seven urine samples (43.7%) and also in onecase of the 16 patients with pneumonia but no pathogen identified. Thespecificity of the ICT test was 97.2%. Conclusion: The ICT assay is a valuable tool for the diagnosis of pneumococcalpneumonia, especially for the nonbacteremic cases.

Nasopharyngeal aspirates, collected during outbreaks, of the novel influenza A (H1N1) virus in Barcelona, were used to compare the accuracy of a rapid antigen-based test (Binax) with the real-time RT-PCR assay developed by the CDC. The sensitivity, specificity and positive predictive value of the rapid test are higher in patients less than 18 years old and during the acute stage of the epidemic than in adult patients.

Sabado 1 Diciembre 2007. Volumen 129 - Numero 20 p. 770 - 772 Texto completo Descargar PDF Buscar en medline articulos de: Pere Tudela Josep Maria Modol Montserrat Gimenez Cristina Prat Blanca Vilaseca Jordi Tor Buscar: en Medicina Clinica en Vol. 129, Num. 20 Busqueda avanzada Originales breves Bacteriemia en pacientes no hospitalizados: tendencias evolutivas en los ultimos 10 anos Pere Tudelaa Josep Maria Modola Montserrat Gimenezb Cristina Pratb Blanca Vilasecac Jordi Tord aUnidad de Corta Estancia-Servicio de Urgencias. Hospital Germans Trias i Pujol. Badalona. Barcelona. Espana. bServicio de Microbiologia. Hospital Germans Trias i Pujol. Badalona. Barcelona. Espana. cUnidad de Corta Estancia-Servicio de Urgencias. Hospital Germans Trias i Pujol. Badalona. Barcelona. Espana. dServicio de Medicina Interna. Hospital Germans Trias i Pujol. Badalona. Barcelona. Espana. Fundamento y objetivo: Conocer la incidencia anual de bacteriemia en pacientes no hospitalizados (BPNH), analizar sus caracteristicas clinicas y epidemiologicas, asi como sus tendencias evolutivas. Pacientes y metodo: Se analizo un periodo de 10 anos, durante el cual se contabilizaron el porcentaje de hemocultivos positivos y, de estos, el porcentaje de BPNH. Se recogieron los siguientes parametros: ano, mes, edad, sexo, diagnostico inicial, factores de riesgo para la bacteriemia, microorganismo, diagnostico final, concordancia entre ambos y necesidad de ingreso. Se clasificaron en: infeccion urinaria, respiratoria, abdominal, de cateter venoso (ICV), cutanea, endocarditis, bacteriemia sin focalidad aparente (BSFA) y un grupo miscelanea. Se compararon las variables edad, sexo, ano y mes entre los grupos con y sin diagnostico concordante. Resultados: Se registraron 283 episodios. El porcentaje de hemocultivos positivos se mantuvo constante, mientras que el de BPNH tendio a descender. El microorganismo mas frecuente fue Escherichia coli (56,5%), y los principales focos, el urinario (59,7%) y la BSFA (19,7%). Los diagnosticos inicial y final no eran concordantes en el 37,1% de los casos. Preciso ingreso el 30,3%. Se detecto infeccion urinaria en el 93,5% de los casos, ICV solo en el 6,2% y ninguna BSFA. La presencia de los principales factores de riesgo para la bacteriemia a lo largo de los anos mostro un descenso de la infeccion por el virus de la inmunodeficiencia humana, mientras que se advertia un ascenso de las enfermedades neoplasicas. Conclusiones: La BPNH, en nuestra experiencia, tiende a un descenso progresivo. La de origen urinario no parece preocupante, mientras que es mejorable la sospecha de ICV. La BSFA constituye el grupo mas preocupante y supone un reto diagnostico para el futuro

This study investigated the utility of an immunochromatographic test (ICT) for the detection of Streptococcus pneumoniae antigens in pleural fluid. Antigen was detected in 15 of 19 (79%) patients with pneumococcal pneumonia. The ICT was always negative in patients with non-pneumococcal pneumonia, but was positive in three cases with a non-infectious aetiology. In patients with pneumonia for which no pathogen was identified, antigen was detected in one of 24 pleural fluids tested. The ICT can be a valuable tool for the management of pneumonia because it can detect pneumococcal antigen in pleural effusion samples.

Molecular mechanisms associated with pathophysiological changes in ventricular remodelling due to myocardial infarction (MI) remain poorly understood. We analyzed changes in gene expression by microarray technology in porcine myocardial tissue at 1, 4, and 6 weeks post-MI. MI was induced by coronary artery ligation in 9 female pigs (30–40 kg). Animals were randomly sacrificed at 1, 4, or 6 weeks post-MI (n = 3 per group) and 3 healthy animals were also included as control group. Total RNA from myocardial samples was hybridized to GeneChip® Porcine Genome Arrays. Functional analysis was obtained with the Ingenuity Pathway Analysis (IPA) online tool. Validation of microarray data was performed by quantitative real-time PCR (qRT-PCR). More than 8,000 different probe sets showed altered expression in the remodelling myocardium at 1, 4, or 6 weeks post-MI. Ninety-seven percent of altered transcripts were detected in the infarct core and 255 probe sets were differentially expressed in the remote myocardium. Functional analysis revealed 28 genes de-regulated in the remote myocardial region in at least one of the three temporal analyzed stages, including genes associated with heart failure (HF), systemic sclerosis and coronary artery disease. In the infarct core tissue, eight major time-dependent gene expression patterns were recognized among 4,221 probe sets commonly altered over time. Altered gene expression of ACVR2B, BID, BMP2, BMPR1A, LMNA, NFKBIA, SMAD1, TGFB3, TNFRSF1A, and TP53 were further validated. The clustering of similar expression patterns for gene products with related function revealed molecular footprints, some of them described for the first time, which elucidate changes in biological processes at different stages after MI.

During infancy and childhood, clinical signs of infection and conventional laboratory markers are not specific in the early phase of disease [1]. The availability of a parameter that more rapidly identifies children suspected to have bacterial sepsis before microbiological results are available would minimize unnecessary treatments and hospitalization. Since its original description, the importance of procalcitonin (PCT) as an indicator of systemic bacterial infection has been demonstrated in many reports [2, 3, 4, 5]. The aim of our study was to evaluate the reliability of PCT measurement by quantitative luminometric immunoassay (LIA) in distinguishing between systemic bacterial infection (sepsis and/or meningitis), localized bacterial infection and aseptic meningitis in children, compared to leukocyte count and C-reactive protein (CRP) levels. We also evaluated the correlation of a rapid immunochromatographic test (ICT) for semiquantitative PCT measurement in comparison with quantitative test results. The study was carried out on selected children aged between 1 month and 12 years who were admitted to the pediatric emergency department of our hospital after presenting with fever of less than 12-h duration. At the time of admission, blood samples were collected for leukocyte count, CRP and PCT measurement. Clinical specimens were collected for microbiological testing in order to correctly establish the etiology. Serial serum samples for PCT and CRP assays were collected daily when possible. Patients were grouped retrospectively according to the type of illness. Group 1 included 25 children diagnosed with bacterial sepsis and/or meningitis by culture of blood and/or cerebrospinal fluid (CSF) samples: Neisseria meningitidis was isolated in 18 cases, Streptococcus pneumoniae in 6 and Haemophilus influenzae in 1. Bacteria were isolated from CSF culture only in 8 patients, blood culture only in 10 and both culture types in 7. Group 2 included 18 children diagnosed with aseptic meningitis by compatible clinical presentation, negative Gram stain, cultures and antigen tests for bacterial infection using blood and CSF samples, compatible CSF analysis (pleocytosis with a predominance of mononuclear cells) and successful recovery without antibiotic therapy. Viral cultures were not performed. Group 3 included 22 children presenting with localized bacterial infection such as purulent conjunctivitis, acute otitis media, streptococcal pharyngitis, cellulitis or sinusitis, confirmed by culture of the site of infection and negative blood cultures. The bacteria isolated were Staphylococcus aureus, Streptococcus pneumoniae and Streptococcus pyogenes. In order to establish normal PCT levels for comparison, serum samples were also collected from a control group of 25 similarly aged healthy children admitted to the hospital for elective surgery. PCT was measured using LIA (Lumitest PCT; Brahms Diagnostica, Germany) for quantitative detection and ICT (PCT-Q; Brahms Diagnostica) for semiquantitative detection following the manufacturer’s instructions. LIA requires 20 l of serum and 90–120 min to be performed. ICT requires 200 l of serum and results can be obtained in 30 min; samples can be measured individually. CRP was measured using a turbidimetric assay (C-Reactive Protein FlexT reagent cartridge, Dimension; Dade Behring, USA) and leukocyte count was measured with Coulter Counter Maxm (Coulter, USA). Comparison between groups for quantitative parameters was performed using the non-parametric MannWhitney U test for CRP, leukocyte count and PCT. Diagnostic accuracy and optimum cut-off points were determined using a receiver operating characteristic curve. Comparisons between the two methods were made C. Prat ()) · J. Dom nguez · M. Gim nez · S. Blanco · V. Ausina Servei de Microbiologia, Hospital Universitari Germans Trias i Pujol, C/Canyet s/n, 08916 Badalona, Spain e-mail: crisprat@ns.hugtip.scs.es Tel.: +34-93-4978894 Fax: +34-93-4978895

Fundamento y objetivo La tuberculosis en el personalsanitario se considera una enfermedad profesional.El objetivo del estudio es describir loscasos de tuberculosis en personal sanitario observadosen nuestro hospital y comparar su incidenciaanual de tuberculosis con respecto a lade la poblacion general. Pacientes y metodo Estudio retrospectivo de loscasos de tuberculosis en el colectivo sanitario deun hospital universitario, durante el periodo comprendidoentre 1988 y 2002, y determinacion dela incidencia anual acumulada. Resultados Se detectaron 21 casos de tuberculosis,cuya forma mas frecuente fue la pulmonar(61,9%), seguida del derrame pleural (28%). Lacategoria profesional mas afectada fue la de medicosresidentes (38%) y el lugar de trabajo conmas riesgo fue el Servicio de Urgencias (48%). Apesar de la variabilidad entre poblaciones, destacamosincidencias acumuladas muy superioresen nuestro personal sanitario respecto a las de lapoblacion general. Conclusiones Existe riesgo de transmision de latuberculosis en personal sanitario, especialmenteen los profesionales que trabajan en el area de urgenciasy en los laboratorios de anatomia patologicay microbiologia. Los esfuerzos han de dirigirsea mantener unas medidas de prevencion eficacesen las areas donde existe mas riesgo de contagio.

Volume 54, no. 4, p. [1160–1163][1], 2016. Page 1160: The byline should appear as shown above. [1]: /lookup/doi/10.1128/JCM.03341-15

A rapid in-house mycobacteriophage-based assay to identify multidrug resistance by detecting the rifampin susceptibility of Mycobacterium tuberculosis in a microtiter plate format was evaluated. The sensitivity, specificity, and overall accuracy of the assay were 100%. This test is rapid to perform and suitable for widespread implementation.

Heart failure is the end-stage of many cardiovascular diseases-such as acute myocardial infarction-and remains one of the most appealing challenges for regenerative medicine because of its high incidence and prevalence. Over the last 20 years, cardiomyoplasty, based on the isolated administration of cells with regenerative capacity, has been the focal point of most studies aimed at regenerating the heart. Although this therapy has proved feasible in the clinical setting, the degree of infarcted myocardium regenerated and of improved cardiac function are at best modest. Hence, tissue engineering has emerged as a novel technology using cells with regenerative capacity, biological and/or synthetic materials, growth, proangiogenic and differentiation factors, and online registry systems, to induce the regeneration of whole organs or locally damaged tissue. The next step, seen recently in pioneering animal studies, is de novo generation of bioartificial hearts by decellularization and preservation of supporting structures for their subsequent repopulation with new contractile, vascular muscle tissue. Ultimately, this new approach would entail transplantation of the "rebuilt" heart, reestablishing cardiac function in the recipient.

Purpose: Urinary tract infection leads to a diagnosis of moderate or high grade (III or higher) vesicoureteral reflux in approximately 15% of children. Predicting reflux grade III or higher would make it possible to restrict cystography to high risk cases. We aimed to derive a clinical decision rule to predict vesicoureteral reflux grade III or higher in children with a first febrile urinary tract infection. Materials and Methods: We conducted a secondary analysis of prospective series including all children with a first febrile urinary tract infection from the 8 European participating university hospitals. Results: A total of 494 patients (197 boys, reflux grade III or higher in 11%) were included. Procalcitonin and ureteral dilatation on ultrasound were significantly associated with reflux grade III or higher and then combined into a prediction model with an ROC AUC of 0.75 (95% CI 0.690.81). Given the prespecified constraint of achieving at least 85% sensitivity, our model led to the clinical decision rule, for children with a first febrile urinary tract infection cystography should be performed in cases with ureteral dilatation and serum procalcitonin level 0.17 ng/ml or higher, or without ureteral dilatation (ie ureter not visible) when serum procalcitonin level is 0.63 ng/ml or higher. The rule had 86% sensitivity (95% CI 7493) with 47% specificity (95% CI 4251). Internal cross-validation produced 86% sensitivity (95% CI 7993) and 43% specificity (95% CI 3947). Conclusions: A clinical decision rule was derived to enable a selective approach to cystography in children with urinary tract infection. The rule predicts high grade vesicoureteral reflux with approximately 85% sensitivity and avoids half of the cystograms that do not find reflux grade III or higher. Further validation is needed before its widespread use. © 2012 American Urological Association Education and Research, Inc.

Recently, a selection of HLA class II-restricted epitopes of ESAT-6 and CFP-10 Mycobacterium tuberculosis proteins from the region of difference (RD) 1 have been described. We have evaluated the host interferon-gamma (IFN-γ) T cell response to these RD1 selected peptides at the beginning and during anti-tuberculosis therapy.We studied 29 pulmonary TB patients enrolled at the beginning of treatment and 24 enrolled during treatment. We performed T-SPOT.TB and ELISPOT with RD1 selected peptides.Patients included at the beginning of treatment responded producing IFN-γ after antigen stimulation in 89.7% by means of T-SPOT.TB and 79.3% by means of RD1 selected ELISPOT. In contrast, for patients included during treatment the percentages were 87.5% and 25%, respectively. Differences in sensitivities between patients evaluated at the beginning and during treatment were only significant for RD1 selected ELISPOT (p0.0001).The host immune response to RD1 selected peptides is lower than to T-SPOT.TB during therapy. Immunological assays based on RD1 selected peptides may be useful tools for studying the immune response during anti-tuberculosis therapy.

BACKGROUND: Myocardial salvage after coverage with a fat flap was recently demonstrated in acute coronary occlusion. The effect of this novel therapeutic strategy on a chronic myocardial scar is unknown.METHODS: Myocardial infarction (MI) was induced by coil deployment in the mid circumflex artery in the swine model. Two weeks after infarction, a pericardial-derived adipose flap was transposed, fully covering the scar, in the treated group. Infarct size and histopathology were analyzed on post mortem sections. To assess cell migration, adenoviral eGFP vectors were injected in the adipose flap and expression was evaluated upon sacrifice both at the flap and myocardium. Magnetic resonance imaging (MRI) was used to measure left ventricular (LV) ejection fraction and ventricular volumes at baseline, 2 weeks post-MI, and at 6 weeks.RESULTS: One month after flap transposition, histopathology confirmed a 34% reduction in infarct size (8.7% vs. 5.7%; P=0.04) and the presence of vascular connections at the flap-myocardium interface. High eGFP expression was detected at the infarct core both at the gene and protein level (negligible signal was detected at the flap on sacrifice). At the functional level, changes in LV ejection fraction and volumes (end-systolic and end-diastolic) were not significantly different between groups (all P values>0.1).CONCLUSIONS: Our data support the use of post-infarction scar coverage with a pericardial-derived fat flap to reduce infarct size, due partly to neovascular connections and cell trafficking at the flap-myocardium interface. Further studies are needed to validate the functional and clinical relevance of this intervention.

The aim of this study was to investigate whether procalcitonin (PCT), neopterin, C-reactive protein (CRP), and mid regional pro-atrial natriuretic peptide (MR-proANP) levels at admission and during the clinical course can be useful for the management of patients with pneumonia. The study population consisted of 75 patients with clinical and radiological diagnosis of pneumonia. Serum samples were collected at admission and during hospitalization. Complications were defined as intensive care unit (ICU) admission or death. The levels of PCT were significantly higher in pneumonia of definite bacterial origin in comparison to probable bacterial or unknown origin. The PCT levels were higher in pneumococcal pneumonia. The PCT and MR-proANP levels increased significantly according to the Pneumonia Severity Index (PSI). All biomarkers levels are higher in patients developing complications and who were dying. The serial levels of MR-proANP remain significantly elevated in patients developing complications and in patients classified in PSI and CURB-65 risk groups. In patients not developing complications, there is a significant decrease in the PCT levels. PCT can be useful for identifying pneumonia etiology. PCT and MR-proANP levels correlate with pneumonia severity rules. PCT and MR-proANP serial measurements can be useful for predicting short-term prognosis. Systemic biomarkers can provide additional information regarding clinical evolution, because these are dynamic and can be measured daily.

Aims Atrial fibrillation (AF) is associated with abnormal sarcoplasmic reticulum (SR) calcium release, which is promoted by adenosine A2A receptor (A2AR) activation. Here, we tested the hypothesis that abnormal calcium release in AF is linked to A2AR remodelling. Methods and results Western blotting and quantitative real-time PCR were used to determine A2AR mRNA and protein levels in right atrial samples from patients with and without AF. Effects of A2AR activation on calcium handling were assessed with patch-clamp technique and confocal calcium imaging. A2AR mRNA levels and functional A2ARs were moderately up-regulated in patients with atrial dilation and markedly up-regulated in those with AF. Accordingly, A2AR stimulation significantly increased ryanodine receptor phosphorylation in AF patients, and spontaneous calcium waves increased moderately in myocytes from patients with atrial dilation and strongly in patients with AF (2.2 ± 2.1 to 14.3 ± 8.8 min−1, n = 6, P = 0.01). Moreover, the high baseline level of calcium waves in AF was reduced by A2AR antagonists (3.5 ± 2.0 to 1.3 ± 1.3 min−1, n = 6, P = 0.007) or adenosine deaminase (1.7 ± 1.5 to 0.5 ± 0.6 min−1, n = 10, P = 0.02) suggesting that A2ARs are activated by endogenous adenosine. Indeed, intracellular perfusion with adenosine significantly increased the calcium wave frequency (1.1 ± 0.8 to 8.2 ± 3.3 min−1, n = 8), whereas adenosine removal from the cytosol decreased it (2.1 ± 0.9 to 0.3 ± 0.3 min−1, n = 8, P = 0.04). Conclusions Atrial fibrillation patients show increased A2AR expression that may account for the high baseline level of spontaneous SR calcium release seen in myocytes from these patients, and the ability of A2AR antagonists to reduce this abnormal calcium release points to the A2AR as a novel molecular target in AF.

Background Individuals infected with human immunodeficiency virus (HIV) have an increased risk of progression to active tuberculosis following Mycobacterium tuberculosis infection. The objective of the study was to determine IFN-γ responses for the detection of latent tuberculosis infection (LTBI) with QuantiFERON-TB GOLD In Tube (QFT-G-IT) and T-SPOT.TB in HIV patients, and evaluate the influence of CD4 cell count on tests performance. Methods We studied 75 HIV patients enrolled for ongoing studies of LTBI with T-SPOT.TB, QFN-G-IT and TST. Mean CD4 cell counts ± standard deviation was 461.29 ± 307.49 cells/μl. Eight patients had a BCG scar. Results T-SPOT.TB, QFN-G-IT and TST were positive in 7 (9.3%), 5 (6.7%) and 9 (12%) cases, respectively. Global agreement between QFN-G-IT and T-SPOT.TB was 89% (κ = 0.275). The overall agreement of T-SPOT.TB and QFN-G-IT with TST was 80.8% (κ = 0.019) and 89% (κ = 0.373), respectively. We have found negative IFN-γ assays results among 2 BCG-vaccinated HIV-infected individuals with a positive TST. In non BCG-vaccinated patients, QFN-G-IT and TST were positive in 5 cases (7.5%) and T-SPOT.TB in 7 (10.4%). In contrast, in BCG-vaccinated patients, only TST was positive in 4/8 (50%) of the cases. The differences obtained in the number of positive results between TST and both IFN-γ assays in BCG vaccinated patients were significant (95% CI 3-97%, p = 0.046), however, the confidence interval is very wide given the small number of patients. In patients with CD4< 200, we obtained only one (5%) positive result with T-SPOT.TB; however, QFN-G-IT and TST were negative in all cases. On the contrary, percentages of positive results in patients with CD4> 200 were 10.9% (6/55), 9.1% (5/55) and 16.4% (9/55) with T-SPOT.TB, QFN-G-IT and TST, respectively. Conclusions IFN-γ tests have the benefit over TST that are less influenced by BCG vaccination, consequently they are more specific than TST. Although our number of patients with advance immunosuppression is limited, our study suggests that IFN-γ assays are influenced with level of immunosuppression. The use of IFN-γ assays could be a helpful method for diagnosing LTBI in HIV population.

Chronic obstructive pulmonary disease (COPD) is still a leading cause of morbidity and mortality worldwide, with a huge socioeconomic impact. New strategies for the management of COPD are required, not only for identifying the origin of the exacerbation episodes, but also to assess an individual risk for each patient. A promising approach is to measure systemic biomarkers and correlate their levels with exacerbation characteristics and clinical prognosis of the disease. Several biomarkers have clearly correlated with the aetiology of lower respiratory tract infections and the response to antibiotic treatment, indicating a potential utility in COPD exacerbation. Nevertheless, the results available at the moment, together with the absence of a gold standard for identifying the aetiological origin of an exacerbation, impedes establishing the real utility of these biomarkers for this concrete task. Regarding the clinical evolution and prognosis, several clinical characteristics have been correlated to biomarker levels. The potential influence of many factors (severity of the disease, presence of comorbidities and treatment) leads to the conclusion that, in the future, the best option would be to monitor levels individually, rather than establishing cut-off points for the general COPD population.