Improved diagnosis of central venous catheter-related bloodstream infections using the HB&L UROQUATTRO™ system.

The diagnosis of catheter-related bloodstream infections (CRBSIs) in febrile patients with indwelling central venous catheters (CVCs) needs improvement. To diagnose CRBSIs more efficiently, we have developed a novel culture approach using the catheter tips removed from febrile patients. CVCs and blood cultures from 1,070 patients with only CVC-related infections were obtained over a period of 3 years (January 2009 to December 2011). The CVCs were evaluated by a semi-quantitative catheter culture method according to Maki's method and by our novel method, which is based on the use of the HB&L UROQUATTRO™ system (Alifax, Padova, Italy). Using our new method, 571 (571/1,070) of the infections were confirmed as CRBSIs. The remaining 487 patients had infections that were associated with hematologic malignancies, neutropenia, prior exposure to antibiotics, and a decreased CVC removal rate. Twelve samples were identified as false-positives. The percentage of patients with CRBSIs confirmed using the HB&L UROQUATTRO™ system was 53.36 % versus 34.95 % (p-value 0.004) using Maki's method (374/1,070 CVC Maki-positive samples). Our results indicate that our new culture method allows for an improved CRBSI diagnosis rate. A significant number of tip cultures (18.41 %) tested positive for CRBSIs using our system but were negative when tested using Maki's method. Moreover, the use of the HB&L UROQUATTRO™ system allowed us to significantly reduce diagnosis time; a negative CRBSI diagnosis could be made within 6 h and a positive diagnosis could be made within 22-28 h.

Role of the quantiferon-TB test in ruling out pleural tuberculosis: a multi-centre study.

Diagnosing pleural tuberculosis (plTB) might be difficult due to limited sensitivity of conventional microbiology tools. As M. tuberculosis (MTB)-specific T cells are recruited into pleural space in plTB, their detection may provide useful clinical information. To this aim, in addition to standard diagnostic tests, we used the QuantiFERON-TB Gold In-Tube (QFT-IT) test in blood and pleural effusion (PE) samples from 48 patients with clinical suspicion of plTB, 18 (37.5%) of whom had confirmed plTB. Four of them (22.2%) tested positive with a nucleic acid amplification test for MTB. The tuberculin skin test was positive in most confirmed plTB cases (88.9%). Positive QFT-IT tests were significantly more frequent in patients with confirmed plTB, as compared to patients with an alternative diagnosis, both in blood (77.7 vs 36.6%, p=0.006) and in PE samples (83.3% vs 46.6%, p=0.02). In addition, both blood and PE MTB-stimulated IFN-gamma levels were significantly higher in plTB patients (p=0.03 and p=0.0049 vs non-plTB, respectively). In blood samples, QFT-IT had 77.8% sensitivity and 63.3% specificity, resulting in 56.0% positive (PPV) and 82.6% negative (NPV) predictive values. On PE, QFT-IT sensitivity was 83.3% and specificity 53.3% (PPV 51.7% and NPV 84.2%). The optimal AUC-derived cut-off for MTB-stimulated pleural IFN-gamma level was 3.01 IU/mL (77.8% sensitivity, 80% specificity, PPV 68.4% and NPV 82.8%). These data suggest that QFT-IT might have a role in ruling out plTB in clinical practice.

Pulmonary sarcoidosis in a patient with psoriatic arthritis during infliximab therapy.

The occurrence of sarcoidosis during anti-TNF-alpha therapy has occasionally been published. We report the case of a psoriasis patient who developed pulmonary sarcoidosis during a cycle of therapy with infliximab.

Percutaneous management of complications of tuberculous spondylodiscitis: short- to medium-term results.

PURPOSE: Psoas abscesses are the most frequent complication of tuberculosis with skeletal involvement. The aim of this paper is to report our experience with the systematic application of percutaneous drainage to tuberculous psoas abscesses. MATERIALS AND METHODS: Between January 1997 and December 2005, 23 patients (14 men and nine women; age range 21-48 years), after a previous study with computed tomography (CT) and/or magnetic resonance (MR) imaging, underwent percutaneous drainage of a tuberculous fluid collection in the psoas muscles. Follow-up consisted of monthly clinical and laboratory assessment, and plain chest radiography and spinal CT every 6-12 months. RESULTS: Spondylodiscitis involved the thoracolumbar spine. Fluid collections were bilateral in 14 cases and communicating in ten of these. Maximum transverse diameter was 7 cm, whereas longitudinal diameter was 14 cm. Placement of the drainage catheter was successful in all cases, and the catheter was left in place for 5-36 (mean 18.4) days. Symptom regression occurred immediately after drainage of the fluid collection. The drainage procedure was curative in 100% of cases. Dislodgement of the drainage catheter occurred in two cases as a result of excessive traction during dressing removal. CONCLUSIONS: A serious complication of bone tuberculosis, psoas abscesses, can be effectively treated by percutaneous drainage, leading to immediate pain resolution. The drainage catheter requires daily monitoring to identify when it can be safely removed without risk of recurrence.

Polymorphisms in the genes of the urokinase plasminogen activation system in relation to colorectal cancer.

BACKGROUND: Extracellular matrix degradation, mediated by the urokinase plasminogen activation (uPA) system, is a critical step in tumor invasion and metastasis. High tumor levels of uPA and its inhibitor PAI-1 have been correlated with poor cancer prognosis. We examined four single nucleotide polymorphisms (SNPs) with a potential effect on expression of genes in the uPA system for their role in colorectal cancer susceptibility and prognosis. PATIENTS AND METHODS: We genotyped the SNPs in 308 Swedish incident colorectal cancer patients with up to 16 years of follow-up and in 585 age- and sex-matched controls. We evaluated the associations between genotypes and colorectal cancer and Dukes' stage. Survival probabilities were compared between different subgroups. RESULTS: Patients with PAI-1 -675 5G/5G genotype had better survival than patients with 4G/4G or 4G/5G genotypes when they had Dukes' stage A or B tumors (P = 0.023 and P = 0.015, respectively). No statistically significant association was observed between the SNPs and the risk of colorectal cancer or Dukes' stage. CONCLUSIONS: Our results suggest a role for the PAI-1 genotype in colorectal cancer prognosis, but further studies are needed to evaluate the impact of our finding in the clinic.

The combined use of VIGI@ct (bioMérieux) and fluorescent amplified length fragment polymorphisms in the investigation of potential outbreaks.

Even with good surveillance programmes, hospital-acquired infections (HAIs) are not always recognized and this may lead to an outbreak. In order to reduce this risk, we propose a model for prompt detection of HAIs, based on the use of a real-time epidemiological information system called VIGI@ct (bioMèrieux, Las Balmas, France) and on the rapid confirmation or exclusion of the genetic relationship among pathogens using fluorescent amplified length fragment polymorphism (f-AFLP) microbial fingerprinting. We present the results of one year's experience with the system, which identified a total of 306 suspicious HAIs. Of these, 281 (92%) were 'confirmed' by clinical evidence, 16 (5%) were considered to be simple colonization and the latter nine (3%) were archived as 'not answered' because of the absence of the physician's cooperation. There were seven suspected outbreaks; of these, f-AFLP analysis confirmed the clonal relationship among the isolates in four cases: outbreak 1 (four isolates of Pseudomonas aeruginosa), outbreak 2 (three Escherichia coli isolates), outbreak 6 (two Candida parapsilosis isolates) and outbreak 7 (30 ESbetaL-producing Klebsiella pneumoniae subsp. pneumoniae). Based on our results, we conclude that the combination of VIGI@ct and f-AFLP is useful in the rapid assessment of an outbreak due to Gram-positive or Gram-negative bacteria and yeasts.

NONO regulates the intra-S-phase checkpoint in response to UV radiation.

The main risk factor for skin cancer is ultraviolet (UV) exposure, which causes DNA damage. Cells respond to UV-induced DNA damage by activating the intra-S-phase checkpoint, which prevents replication fork collapse, late origin firing and stabilizes fragile sites. Recently, the 54-kDa multifunctional protein NONO was found to be involved in the non-homologous end-joining DNA repair process and in poly ADP-ribose polymerase 1 activation. Interestingly, NONO is mutated in several tumour types and emerged as a crucial factor underlying both melanoma development and progression. Therefore, we set out to evaluate whether NONO could be involved in the DNA-damage response to UV radiations. We generated NONO-silenced HeLa cell clones and found that lack of NONO decreased cell growth rate. Then, we challenged NONO-silenced cells with exposure to UV radiations and found that NONO-silenced cells, compared with control cells, continued to synthesize DNA, failed to block new origin firing and impaired CHK1S345 phosphorylation showing a defective checkpoint activation. Consistently, NONO is present at the sites of UV-induced DNA damage where it localizes to RAD9 foci. To position NONO in the DNA-damage response cascade, we analysed the loading onto chromatin of various intra-S-phase checkpoint mediators and found that NONO favours the loading of topoisomerase II-binding protein 1 acting upstream of the ATM and Rad3-related kinase activity. Strikingly, re-expression of NONO, through an sh-resistant mRNA, rescued CHK1S345 phosphorylation in NONO-silenced cells. Interestingly, NONO silencing affected cell response to UV radiations also in a melanoma cell line. Overall, our data uncover a new role for NONO in mediating the cellular response to UV-induced DNA damage.

Emergency room technetium-99m sestamibi imaging to rule out acute myocardial ischemic events in patients with nondiagnostic electrocardiograms.

OBJECTIVES: The goal of this study was to determine the role of nuclear imaging in patients with chest pain. BACKGROUND: The diagnosis of myocardial ischemic events in patients with chest pain and a nondiagnostic electrocardiogram (ECG) is problematic. METHODS: Rest tomographic technetium-99m sestamibi imaging (740 MBq intravenously) was performed in 64 patients presenting to the emergency room with chest pain of suspected cardiac origin and a nondiagnostic ECG. Patients were admitted to the coronary care unit on the basis of clinical criteria only and were strictly monitored. RESULTS: Thirty patients showed a perfusion defect on admission. Of these, 13 developed myocardial infarction within 12 h. Coronary artery disease was diagnosed in 14 patients and the remaining 3 patients were classified as having false positive findings. Normal perfusion scans were seen in 34 patients, none of whom were ultimately diagnosed as having coronary artery disease. A 100% sensitivity was demonstrated versus the final diagnosis of acute cardiac ischemia (kappa 0.91, 95% confidence interval 0.8 to 1.0). A follow-up period of up to 18 months (mean 11 +/- 3) was also carried out for major cardiac events (death, myocardial infarction, coronary angioplasty and coronary artery bypass grafting). Six events (two coronary bypass procedures, three angioplasty procedures and one death) were observed at follow-up in the group of patients with a technetium-99m sestamibi perfusion defect. Patients with normal perfusion scans on admission had no major cardiac events at follow-up study. CONCLUSIONS: Technetium-99m sestamibi perfusion imaging is a promising technique for ruling out acute myocardial ischemia in the emergency room. More efficient utilization of intensive therapy beds may be expected with this approach.

Solution structure, domain features, and structural implications of mutants of the chromo domain from the fission yeast histone methyltransferase Clr4.

The encapsulation of otherwise transcribable loci within transcriptionally inactive heterochromatin is rapidly gaining recognition as an important mechanism of epigenetic gene regulation. In the fission yeast Schizosaccharomyces pombe, heterochromatinization of the mat2/mat3 loci silences the mating-type information encoded within these loci. Here, we present the solution structure of the chromo domain from the cryptic loci regulator protein Clr4. Clr4 is known to regulate silencing and switching at the mating-type loci and to affect chromatin structure at centromeres. Clr4 and its human and Drosophila homologs have been identified as histone H3-specific methyltransferases, further implicating this family of proteins in chromatin remodeling. Our structure highlights a conserved surface that may be involved in chromo domain-ligand interactions. We have also analyzed two chromo domain mutants (W31G and W41G) that previously were shown to affect silencing and switching in full-length Clr4. Both mutants are significantly destabilized relative to wild-type.

Solution structure of interleukin-13 and insights into receptor engagement.

The complex and interrelated function of the interleukin cytokines relies on a range of pro-inflammatory and anti-inflammatory immune responses mediated by an array of receptors, and there is considerable cross-reactivity for related cytokines. Recent findings continue to elucidate the expression patterns of interleukin receptors associated with a range of diseases, including cancer. We report here the first experimentally determined high-resolution structure of human interleukin-13 (IL-13). The experimental structure is significantly different from an earlier homology model, which could have led to improper estimation of receptor interaction surfaces and design of mutational experiments. Similarities between the presented IL-13 structure and the homologous interleukin-4 (IL-4) are discussed. Additionally, mutation data for IL-4 and IL-13 are analyzed and combined with a detailed structural analysis of the IL-4/IL4Ralpha interface that leads us to postulate interactions at the IL-13/receptor interface. The structural comparison is used to interpret the different affinities for various receptors and establishes the basis for further mutational experiments and antagonist design.

Solution structure of the human Hck SH3 domain and identification of its ligand binding site.

SH3 domains are protein binding domains that occur widely among signal transduction proteins. Here, we present the NMR-determined solution structure of the SH3 domain from the cytoplasmic protein tyrosine kinase, Hck. Hck is involved in a number of cell signal transduction pathways, frequently in pathways associated with immune response. SH3 domains bind proteins via a left-handed polyproline type II helix on the target protein. We have assessed the structural impact of binding to a ligand through addition of a peptide corresponding to a proline-rich region of a Hck target, the GTPase activating protein of the Ras pathway. Ligand binding effects small structural changes and stabilizes the SH3 domain structure. Also, we have compared the solution structure of the Hck SH3 domain to the crystal structure of Hck, in which the SH3 domain exhibits an intramolecular binding to an interdomain linker region. These structures are interpreted as the apo- and holo- forms of the Hck SH3 domain.

Radiometric characterisation of more representative natural building materials in the province of Rome.

Natural building materials, characterised by middle-low-activity concentrations of primordial radionuclides ((40)K, (232)Th and (238)U series) are widely used in Italy. Since natural materials reflect the geological variability of their sites of origin, a systematic study was carried out in the province of Rome and the results are reported in this paper. In the present work, in order to evaluate average, minimum and maximum contents of primordial radionuclides, more representative lithologies outcropping on the territory of the province of Rome were identified and around 150 samples were collected. Also, these lithologies were characterised from a radioprotection point of view, by means of the evaluation of the index, I, when they are used as building materials. The results confirm the high-primordial radionuclide content within some materials used in Latium (central Italy). Although the study was carried out in a limited area, the results confirm considerable variation in the primordial radionuclide content depending on the sites of origin.

[Evaluation of central venous catheters by endocavitary ECG]. / Registrazione elettrocardiografica intratoracica (EC-ECG) per valutare il catetere venoso centrale.

Jugular vein catheterization (JVC) is adopted for blood access in patients with acute renal failure, chronic renal failure and when patients demonstrate traditional vascular access (VA) failure. We believe that the best technique, first described by Serafini et al, to establish the position of a central venous catheter (CVC) is endocavitary electrocardiography (EC-ECG) and its use is recommended in all uremic patients requiring hemodialysis (HD). This technique uses the tip of the CVC as a reference point in standard electrocardiography. From 2001 to March 2004, we successfully applied this CVC technique in 33 patients requiring HD. The EC-ECG technique is a method that complies with the Food and Drug Administration guidelines regarding catheter tip location in uremic patients.

Sequential assignments and secondary structure of the RNA-binding transcriptional regulator NusB.

The NusB protein is involved in transcriptional regulation in bacteriophage lambda. NusB binds to the RNA form of the nut site and along with N, NusA, NusE and NusG, stabilizes the RNA polymerase transcription complex and allows stable, persistent antitermination. NusB contains a 10 residue Arg-rich RNA-binding motif (ARM) at the N-terminus but is not sequentially homologous to any other proteins. In contrast to other known ARM-containing proteins, NusB forms a stable structure in solution in the absence of RNA. NMR spectroscopy was used to determine that NusB contains six alpha-helices: R10-Q21, 127-F34, V45-L65, Q79-S93, Y100-F114 and D118-L127. The structure of NusB makes it a member of a newly emerging class of alpha-helical RNA-binding proteins.

Oral contraceptives and cancer: an update.

Most up-to-date information on oral contraceptives (OCs) and breast cancer risk comes from a collaborative re-analysis of individual data on 53297 cases and 100239 controls. It is now established that there is a moderately increased breast cancer risk among current OC users, which tends to level off in the few years after stopping use. With regard to cervical cancer, OC use has been found to be associated with increased risk in human papilloma virus-positive women. With reference to the well known protective effects of OCs against endometrial carcinogenesis, additional information has suggested a consistent protection across types of OCs used. Further data on ovarian cancer confirm that the protection of OCs is long lasting, and may well be observed 15 to 20 years after stopping use. Several studies have suggested an inverse relationship between use of OCs and risk of colorectal cancer, and in a meta-analysis of published data the pooled relative risk of colorectal cancer for DC ever-use was 0.82 (95% confidence interval 0.74 to 0.97). There was no association with duration of use. The increased risk for hepatocellular carcinoma in the absence of hepatitis B viruses is the only established evidence of a direct association between OC use and cancer risk, which led an International Agency for Research on Cancer Working Group to classify OCs as carcinogenic to humans in 1998.

Cigarette smoking and risk of Hodgkin's disease.

The role of cigarette smoking on the risk of Hodgkin's disease remains controversial. To provide further information on the issue, we analysed data of a case-control study from northern Italy. The cases were 158 patients with incident, histologically confirmed Hodgkin's disease, and the controls were 316 patients, frequency-matched to the cases by age, sex and study centre, and admitted to the same network of hospitals for acute, non-neoplastic, non-alcohol- or non-tobacco-related conditions. Compared with those who had never smoked, the multivariate odds ratio was 0.54 for former and 0.85 for current smokers. No trend in risk was found for either the number of cigarettes smoked or the duration of consumption. None of the estimates, or the corresponding trends in risk, was statistically significant. Our results are consistent with those of several studies indicating no direct association between cigarette smoking and risk of Hodgkin's disease.

Hormone replacement therapy and risk of lymphomas and myelomas.

The relation between post-menopausal hormone replacement therapy (HRT) and lymphoid neoplasms was analysed using data from a case-control study conducted in northern Italy between 1983 and 1992. Cases included 26 incident, histologically confirmed, post-menopausal female patients of Hodgkin's Disease (HD), 145 non-Hodgkin's lymphomas (NHL) and 65 multiple myelomas (MM), between 45 and 79 years of age. The control group comprised 361 women (age range 45-79 years) admitted to the same network of hospitals for acute, non-neoplastic conditions. The multivariate odds ratio (OR) for ever HRT users was 0.7 [95% confidence interval (CI) 0.2-2.8] for HD, 0.7 (95% CI 0.3-1.4) for NHL, and 0.2 (95% CI 0.1-1.0) for MM. No clear pattern of association was found for duration or recently of use. Despite the relatively small number of cases, due to the rarity of the disease, and the low frequency of HRT use in this population, these findings provide additional information for any global risk-benefit assessment of HRT use.

Aerosolized interferon-alpha treatment in patients with multi-drug-resistant pulmonary tuberculosis.

Multi-drug-resistant tuberculosis (MDR-TB) has emerged as an obstacle to the control of tuberculosis. Recent data however, suggest that interferon-(IFN)-gamma and IFN-alpha may improve disease evolution in subjects affected with pulmonary tuberculosis caused by multi-resistant (IFN-gamma) and sensitive (IFN-alpha) strains. The mechanisms involved are not known, even though it has been reported that IFN-gamma-secreting CD4+ Th cells may possess antitubercular effects. In addition, IFN-alpha can induce IFN-gamma secretion by CD4+ Th cells, and both types of IFN may stimulate macrophage activities. The aim of this study was to explore the possibility that aerosolized IFN-alpha, administered concomitantly with conventional antitubercular chemotherapy, may improve the course of pulmonary tuberculosis. After six months of directly observed therapy (DOT), seven patients who were non-responders to a second line antitubercular therapy were given an IFN-alpha aerosol (3 MU, three times a week) for two months as adjunctive therapy. All strains were resistant to at least two first-line drugs. After IFN-alpha administration, the patients were followed up for a further six months with the same DOT. Sputum samples were collected monthly during the study period, with the exception of the IFN-alpha administration period, when the observations were performed weekly. High resolution computed tomography (HRCT) chest scans were performed before and after IFN-alpha inhalations. The analysis of the results showed that the mean number of Mycobacterium tuberculosis (Mt) had remained statistically unchanged (p = 0.80) during the first 6 months of DOT. During the following 2 months of IFN-alpha administration, 5 patients became negative (p = 0.02). After the end of treatment a progressive increase in Mt number was observed (p = 0. 02). Sputum cultures remained positive for all patients throughout the study period, although a significant decrease (p = 0.02) in the colony number per culture was observed after adjunctive treatment with IFN-alpha. After stopping administration of IFN-alpha, a significant increase (p = 0.03) in the colony number per culture was noted as well as in Mt numbers. HRCT scans were slightly improved in all patients. These preliminary data suggest that aerosolized IFN-alpha may be a promising adjunctive therapy for patients with MDR-TB. Optimal doses and schedules however, require further studies.

Fluid intake and risk of bladder and other cancers.

There are appreciable differences in total fluid intake at the individual and population level, and substantial difficulties in obtaining valid measures of fluid intake. Epidemiological studies have examined the association between fluid intake and different types of cancer. For bladder cancer, fluid consumption has been associated with a moderate increase of risk in some studies, including a multicentric case-control study from the United States, based on about 3000 cases, with a decrease in others, including the Health Professional Follow-up study, or with no material association. The evidence, therefore, is far from consistent. Sources and components of fluids were also different across different types studies. From a biological point of view, a decreased fluid intake could result in a greater concentration of carcinogens in the urine or in a prolonged time of contact with the bladder mucosa because of less frequent micturition. Carcinogenic or anticarcinogenic components of various beverages excreted in the urine may also play a role in the process. It has been suggested that fluid consumption has a favorable effect on colorectal cancer risk. Fluid intake may reduce colon cancer risk by decreasing bowel transit time and reducing mucosal contact with carcinogens. Low fluid intake may also compromise cellular concentration, affect enzyme activity in metabolic regulation, and inhibit carcinogen removal. However, epidemiological data are inadequate for evaluation. Data are sparse and inconsistent for other neoplasms, including breast cancer. The fluid constituent of foods, confounding, interactions and possible influences of specific types of beverages should be investigated further. In conclusion therefore the association between total fluid intake and cancer risk remains still open to debate.

Microbiological indoor air quality in healthy buildings.

There is a growing interest in indoor air quality for a better quality environment both at home and at work because many people spend at least 80% of their time indoors. The aim of our study was to evaluate the indoor concentration of airborne bacteria and fungi in a University auditorium, in an office of public buildings and in an apartment in the presence and in absence of building's occupants, building materials and furnishings. The concentrations of airborne bacteria and fungi were determined using a Surface Air System (SAS). In presence of people and furnishings the average air concentrations of bacteria (University auditorium: 925-1225 CFU m(-3); office: 493 CFU m(-3); apartment: 92-182 CFU m(-3)) were higher than in absence (respectively: 190-315 CFU m(-3); 126 CFU m(-3); 66-80 CFU m(-3)). The average air concentrations fungal were higher in presence of people and furnishings (University auditorium: 1256-1769 CFU m(-3); office: 858 CFU m(-3); apartment: 147-297 CFU m(-3)) than in absence (respectively: 301-431 CFU m(-3); 224 CFU m(-3); 102-132 CFU m(-3)). The obtained data can be considered as a step to identify acceptable levels for bioaerosols in common indoor environments.