Epidemiological study of pathogens collected from blood for a period of a year (2008-2009)

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Unraveling Energy and Dynamics Determinants to Interpret Protein Functional Plasticity: The Limonene-1,2-epoxide-hydrolase Case Study

The balance between structural stability and functional plasticity in proteins that share common three-dimensional folds is the key factor that drives protein evolvability. The ability to distinguish the parts of homologous proteins that underlie common structural organization patterns from the parts acting as regulatory modules that can sustain modifications in response to evolutionary pressure may provide fundamental insights for understanding sequence–structure–dynamics relationships. In applicative terms, this would help develop rational protein design methods. Herein, we apply recently developed computational methods, validated by experimental tests, to address these questions in a set of homologous enzymes representative of the limonene-1,2-epoxide-hydrolase family (LEH) characterized by different stabilities, namely Rhodococcus erythropolis LEH (Re-LEH), Tomks-LEH, CH55-LEH, and the two thermostable Re-LEH variants Re-LEH-F1b and Re-LEH-P. Our results show that these enzymes, despite significant sequence variations, exploit a few highly conserved stabilization determinants to guarantee structural stability linked to biological functionality. Multiple sequence analysis shows that these key elements are also shared by a larger set of LEHs structural homologues, despite very low sequence identity and functional diversity. In this framework, stabilizing elements that we hypothesize to have an accessory role are characterized by a lower degree of sequence identity and higher mutability. We suggest that our approach can be successfully used to pinpoint the distinctive energy fingerprint of a class of proteins as well as to locate those modulators whose modification could be exploited to tune protein stability and dynamic properties

Epidemiological study of pathogens collected from blood for a period of a year (2008-2009)

Objectives. An epidemiological study, addressed to identify the pathogens isolated from blood, and their antibiotic susceptibility patterns, was conducted. Methods. 12 laboratories, homogeneously distributed in a Northern area of Italy, were required to collected all consecutive non-duplicated strains isolated from blood during February 2008 to February 2009 and sent them to the reference laboratory. Results. A total of 1092 microorganisms were collected, including 653 gram-positive, 385 gram-negative and 54 fungi. Escherichia coli 234, Staphylococcus epidermidis 205, S. aureus 142, S. hominis 87, Enterococcus faecalis 47, S. haemolyticus 33, Klebsiella pneumoniae 33, Pseudomonas aeruginosa 32, Candida albicans 28, Enterobacter cloacae 21 were the prevalent microrganisms found. Samples were collected mainly from medicine (255 strains), intensive care units (154), surgery (99), infectious diseases (93), paediatrics (62) and nephrology (62). Antibiotic resistance (in %) in staphylococci was 65.7 (methicillin), 33.5 (gentamicin), 61.8 (azithromycin), 59.6 (erythromycin), 45.2 (ciprofloxacin) 14.8 (chloramphenicol), 2.0 (teicoplanin), and 24.1 (trimethoprim-sulfamethoxazole) no vancomycin-resistant strain was found. Enterococci showed resistance to vancomycin (10.8), ampicillin (34.4), gentamycin (42.9), ciprofloxacin (42.2) teicoplanin (7.6), erythromycin (54.7) and chloramphenicol (17.5). Enterobacteriaceae exhibited resistance to ciprofloxacin(27.0), ampicillin (74.1), ceftazidime (15.8), cefoxitin (14.7), cefepime (13.3), ceftriaxone (15.0), both imipenem and amikacin (0.95), piperacillin-tazobactam (5.1) and trimethoprim-sulfamethoxazole (32.7). Non fermenting gram negative strains were found resistant to ciprofloxacin (27.3), ceftazidime (9.5), cefepime (14.6), ceftriaxone (81.6), both imipenem and amikacin (18.6), trimethoprim-sulfamethoxazole (65.2), and piperacillin-tazobactam (7.5). Conclusions.These data show a prevalent incidence of gram-positive (59.7 %) in comparison to gram-negative (35.3%) bacteria isolated from blood.A high percentage of methicillin-resistant staphylococci as well as ceftazidime-resistance among Enterobacteriaceae is also observed suggesting that this phenomenon requires periodically surveillance

The diagnosis of multiple sclerosis: pinpointing the concept of "no better explanation"

Background: Since the first appearance of McDonald’s diagnostic criteria for Multiple Sclerosis (MS), it was clear that beyond the demonstration of the disease dissemination in space and time, a crucial role in the diagnostic work up would have been played by the exclusion of other neurological diseases. Aim of this study is to provide an accurate picture of the main diseases that can mimic MS in the “real world” setting of a large number of specialist MS centres. Methods: Prospective, observational clinical and radiological study Including all patients presenting with symptoms suggestive of MS, in which a further examination was required in order to exclude or confirm MS diagnosis. Each patient after a regular diagnostic workup (including magnetic resonance imaging, visual evoked potentials, blood and cerebrospinal fluid examinations) was included in a 2 years clinical and radiological follow up. Results: In the first 2 months of recruitment, 300 patients were included and underwent clinical, imaging and paraclinical (blood and CSF) examinations. Among these patients, in 123 (41.3%) it has been possible to diagnose MS according to the most recently revised MS diagnostic criteria, in 55 (18.3%) a different diagnosis was made and in the remaining 121 (40.3%) patients a clinical and radiological follow up was required to clarify the diagnosis. Among the 55 patients in which a MS diagnosis was excluded and other diagnoses were already possible, the most frequent diagnoses were migraine (in 15 cases), vascular encephalopathy including PTO (in 13 cases), recurrent optic neuritis (in 2 cases), systemic erythematous lupus (in 2 cases), dizziness (in 2 cases), neuroBechet’s disease (in 2 cases), neuromyelitis optica (in 2 cases). Discussion: Our preliminary analysis confirm that according to the new diagnostic criteria, the MS diagnosis can be made at the end of diagnostic work-up after the first clinical evaluation in almost 40% of the patients presenting with symptoms suggestive of MS. Migraine and vascular encephalopathy are by far the most frequent alternative diagnosis, while, so far, no infective diseases have been observed. Results of our study will be helpful to recommend an appropriate diagnostic algorithm based on the most frequent conditions entering the differential diagnosis with MS

Can Baseline [18F]FDG PET/CT Predict Response to Immunotherapy After 6 Months and Overall Survival in Patients with Lung Cancer or Malignant Melanoma? A Multicenter Retrospective Study

: Aim: To assess the role of baseline 18F-fluorodeoxyglucose ([18F]FDG)-positron emission tomography/computed tomography (PET/CT) in predicting response to immunotherapy after 6 months and overall survival (OS) in patients with lung cancer (LC) or malignant melanoma (MM). Methods: Data from a multicenter, retrospective study conducted between March and November 2021 were analyzed. Patients >18 years old with a confirmed diagnosis of LC or MM, who underwent a baseline [18F]FDG-PET/CT within 1-2 months before starting immunotherapy and had a follow-up of at least 12 months were included. PET scans were examined visually and semiquantitatively by physicians at peripheral centers. The metabolic tumor burden (number of lesions with [18F]FDG-uptake) and other parameters were recorded. Clinical response was assessed at 3 and 6 months after starting immunotherapy, and OS was calculated as the time elapsing between the PET scan and death or latest follow-up. Results: The study concerned 177 patients with LC and 101 with MM. Baseline PET/CT was positive in primary or local recurrent lesions in 78.5% and 9.9% of cases, in local/distant lymph nodes in 71.8% and 36.6%, in distant metastases in 58.8% and 84%, respectively, in LC and in MM patients. Among patients with LC, [18F]FDG-uptake in primary/recurrent lung lesions was more often associated with no clinical response to immunotherapy after 6 months than in cases without any tracer uptake. After a mean 21 months, 46.5% of patients with LC and 37.1% with MM had died. A significant correlation emerged between the site/number of [18F]FDG foci and death among patients with LC, but not among those with MM. Conclusions: In patients with LC who are candidates for immunotherapy, baseline [18F]FDG-PET/CT can help to predict response to this therapy after 6 months, and to identify those with a poor prognosis based on their metabolic parameters. For patients with MM, there was only a weak correlation between baseline PET/CT parameters, response to therapy, and survival