58 research outputs found

    Design and Synthesis of 4-Alkylidene-b-lactams: Benzyland Phenethyl-carbamates as Key Fragments to Switch on Antibacterial Activity

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    The emergence of multidrug-resistant bacterial strains is particularly important in chronic pathologies such as cystic fibrosis (CF), in which persistent colonization and selection of resistant strains is favored by the frequent and repeated use of antibacterial agents. Staphylococcus aureus is a common pathogen in CF patients that has an associated increased multidrug resistance. In previous studies we demonstrated that the presence of a 4-alkylidene side chain directly linked to a beta-lactam appeared to strengthen the potency against S. aureus, especially against methicillin-resistant S. aureus (MRSA) strains. In the present study, 21 new 4-alkylidene-beta-lactams were synthesized and evaluated for antibacterial activity. We designed the new compounds to have aryl, benzyl, or phenethyl-carbamate groups on the C3 hydroxyethyl side chain. We found a correlation between biological activity and the nitrogen substituent of the carbamate group, and two phenethyl-carbamate b-lactams were shown to be valuable antibacterial agents against selected linezolid-resistant strains, with a minimum inhibitory concentrations of 2–4 mgL-1

    Gender and Age-Dependent Etiology of Community-Acquired Urinary Tract Infections

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    Urinary tract infections (UTIs) are among the most frequent community-acquired infections worldwide. Escherichia coli is the most common UTI pathogen although underlying host factors such as patients' age and gender may influence prevalence of causative agents. In this study, 61 273 consecutive urine samples received over a 22-month period from outpatients clinics of an urban area of north Italy underwent microbiological culture with subsequent bacterial identification and antimicrobial susceptibility testing of positive samples. A total of 13 820 uropathogens were isolated and their prevalence analyzed according to patient's gender and age group. Overall Escherichia coli accounted for 67.6% of all isolates, followed by Klebsiella pneumoniae (8.8%), Enterococcus faecalis (6.3%), Proteus mirabilis (5.2%), and Pseudomonas aeruginosa (2.5%). Data stratification according to both age and gender showed E. coli isolation rates to be lower in both males aged ≥60 years (52.2%), E. faecalis and P. aeruginosa being more prevalent in this group (11.6% and 7.8%, resp.), as well as in those aged ≤14 years (51.3%) in whom P. mirabilis prevalence was found to be as high as 21.2%. Streptococcus agalactiae overall prevalence was found to be 2.3% although it was shown to occur most frequently in women aged between 15 and 59 years (4.1%). Susceptibility of E. coli to oral antimicrobial agents was demonstrated to be as follows: fosfomycin (72.9%), trimethoprim/sulfamethoxazole (72.9%), ciprofloxacin (76.8%), ampicillin (48.0%), and amoxicillin/clavulanate (77.5%). In conclusion, both patients' age and gender are significant factors in determining UTIs etiology; they can increase accuracy in defining the causative uropathogen as well as providing useful guidance to empiric treatment

    The cientificWorldJOURNAL Research Article Gender and Age-Dependent Etiology of Community-Acquired Urinary Tract Infections

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    Urinary tract infections (UTIs) are among the most frequent community-acquired infections worldwide. Escherichia coli is the most common UTI pathogen although underlying host factors such as patients' age and gender may influence prevalence of causative agents. In this study, 61 273 consecutive urine samples received over a 22-month period from outpatients clinics of an urban area of north Italy underwent microbiological culture with subsequent bacterial identification and antimicrobial susceptibility testing of positive samples. A total of 13 820 uropathogens were isolated and their prevalence analyzed according to patient's gender and age group. Overall Escherichia coli accounted for 67.6% of all isolates, followed by Klebsiella pneumoniae (8.8%), Enterococcus faecalis (6.3%), Proteus mirabilis (5.2%), and Pseudomonas aeruginosa (2.5%). Data stratification according to both age and gender showed E. coli isolation rates to be lower in both males aged ≥60 years (52.2%), E. faecalis and P. aeruginosa being more prevalent in this group (11.6% and 7.8%, resp.), as well as in those aged ≤14 years (51.3%) in whom P. mirabilis prevalence was found to be as high as 21.2%. Streptococcus agalactiae overall prevalence was found to be 2.3% although it was shown to occur most frequently in women aged between 15 and 59 years (4.1%). Susceptibility of E. coli to oral antimicrobial agents was demonstrated to be as follows: fosfomycin (72.9%), trimethoprim/sulfamethoxazole (72.9%), ciprofloxacin (76.8%), ampicillin (48.0%), and amoxicillin/clavulanate (77.5%). In conclusion, both patients' age and gender are significant factors in determining UTIs etiology; they can increase accuracy in defining the causative uropathogen as well as providing useful guidance to empiric treatment. Background Urinary tract infections (UTIs) are among the most frequent bacterial infections worldwide Although women, particularly those aged 16-64 years, are significantly more likely to experience UTIs than men Microbial etiology of UTIs has been regarded as well established, with E. coli being the causative pathogen in 50-80% of cases Methods A retrospective study was performed at the Bacteriological Laboratory of the "Centro Diagnostico Italiano" (CDI), based in Milan (Italy), on all bacterial strains isolated from consecutive urine samples received from outpatients clinics of a high-populated urban area of North Italy, between March 2008 and December 2009. Urine samples, accompanied by microbiology request forms, were delivered either directly to the CDI laboratory or through 7 collaborating laboratories. All sample processing and patients' data collection were carried out centrally by the CDI laboratory. CDI Laboratory follows Internal Quality Control procedures and participates to an External Program for Quality Assessment with positive evaluations. As part of the routine procedure, patients received indications to avoid antimicrobials assumption during the previous 7 days and instructions on urine sampling (including cleaning of the genital area prior to midstream specimen collection) and its transport to the laboratories (within 2 hours of collection). Specimens from collaborating laboratories were transported in Vacutainer tubes containing boric acid at 1-2% as preservative. All samples were plated as soon as possible and no later than 24 hours on Chromagar Orientation-BD plates and incubated for 18-24 hours at 37 • C. Criterion for defining significant bacteriuria (positive samples) was the presence of ≥10 5 colony-forming units (CFU)/mL of urine The statistical analysis was performed by the SAS System version 9.2. The difference between females and males in the frequency of positive samples of each agent was analyzed by the chi-square test. The frequency of the positive samples of each agent as a function of gender and age groups (≤14; 15-29; 30-59; ≥60 years) was analyzed by the logistic regression, including in the model the terms gender, age, and the gender by age interaction. Results A total of 61 273 urine cultures were performed over a 22-month period; of these 13 820 (22.6%) were found to be positive for bacterial infection. Nearly 80% of all isolates were from women (female to male ratio (F/M) = 3.8) and 58% from subjects aged 60 years or more. Cumulatively, the two younger age groups accounted for 11.2% of total isolates. Female to male ratio was highest in age group 15-29 years (F/M = 13.5) and lowest in the youngest age group (F/M = 1.4). Overall the most frequently encountered pathogen was Escherichia coli (67.6%), followed by Klebsiella pneumoniae (8.8%), Enterococcus faecalis (6.3%), Proteus mirabilis (5.2%), Pseudomonas aeruginosa (2.5%), and Streptococcus agalactiae (2.3%) Frequency of isolation of all six main species was found to be statistically different between females and males ( in females, whereas E. faecalis, P. mirabilis, and P. aeruginosa were more common in men. All the six most prevalent bacterial species, with the exception of K. pneumoniae, revealed statistically significant differences in isolation rates within the four chosen age groups Data stratification according to both gender and age showed that differences in frequency of isolation between females and males of E. faecalis and S. agalactiae were not consistent across all age groups Susceptibility to antimicrobials of main isolated uropathogens is shown in K. pneumoniae susceptibility to quinolones (93.0%-95.3%) and to TMP-SMX (89.8%) was higher in comparison to E. coli while fosfomycin activity resulted to be lower (81.0%). E. faecalis susceptibility to ampicillin and fosfomycin was high (96.1% and 100%, resp.), superior with respect to susceptibility to quinolones (71.9%-82.3%). S. agalactiae susceptibility to levofloxacin was found to be 91.1% (data not shown). Discussion As urinary tract infection is a very common disease, its diagnosis and treatment have important implications for patients' health, development of antibiotic resistance, and health care costs The present retrospective study describes the distribution and antimicrobial susceptibility of bacterial species isolated from a large number of urinary samples collected over a 22-month period, as part of routine analyses, from unselected community patients (male and female of any age and clinical condition) living in a urban area in the north of Italy. The high number of available isolates allowed to stratify data according to patients' gender and age and so to evaluate the association of such variables to UTI etiology. As expected E. coli was the most frequently encountered species in our study. Percentage of E. coli isolation (67.6%) well compares with those reported from other outpatients surveys conducted in north (64.6%) Other frequent isolates found in this study included K. pneumoniae, P. mirabilis, and E. faecalis, all having been reported to be highly prevalent species in UTIs Our study, however, showed that prevalence of urinary pathogens following data stratification was not consistent across all age groups further divided by gender. Escherichia coli, for example, was found to be less prevalent in the youngest and oldest male subjects (51.3% and 52.2%, resp.) and more frequent in female patients aged 15 years or older 4 The Scientific World Journal (approximately 71%), Proteus mirabilis prevalence was found to be highest (21.2%) in young males aged ≤14 years, whilst S. agalactiae was mostly found in women aged between 15 and 59 years (approximately 4.0%). Kiffer et al. [21] conducted a study comparable to ours, in terms of patient's population (both males and females of any age), number of isolates The Scientific World Journal 5 (0-13 years), accounting for 22.1% of isolates (15.6% in our study) with a females to males ratio of 0.45 (0.55 in our study). Already in 1972, Bergström [22] and, more recently, Modarres and Oskoii P. mirabilis has been described to be present in the preputial sac of boys, having been isolated in 22.6% of uncircumcised males of up to 14 years of age Susceptibility of uropathogenic bacteria to antimicrobials agents is also known to vary among countries and over time In our survey, 72.9% of E. coli isolates were susceptible to TMP/SMX. Susceptibility to oral antimicrobials of P. mirabilis strains isolated in our study, was generally lower than that reported both in Italy and other countries. Susceptibility to ciprofloxacin and TMP/SMX of P. mirabilis isolates in our study was of 62.9% and 51.5%, respectively, as compared to rates demonstrated by other authors ranging from 75.5% to 97.9% for ciprofloxacin and from 52.0% to 84.9% for TMP-SMX In conclusion, besides providing further data on the etiology of community-acquired UTIs and antimicrobial susceptibility of uropathogens in Italy, our results confirm that stratification of isolates from unselected patients on the basis of age and gender can improve the assessment of causative pathogens, providing guidance for empiric treatment and interesting clues to the understanding of UTIs etiopathology. In particular, P. mirabilis prevalence was found to be high both in boys (21.2%) and girls (11.8%) suggesting, as previously reporte

    The Effectiveness of 0.6% Povidone Iodine Eye Drops in Reducing the Conjunctival Bacterial Load and Needle Contamination in Patients Undergoing Anti-VEGF Intravitreal Injection: A Prospective, Randomized Study

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    The study purpose was to assess the efficacy of a preservative-free 0.6% povidone iodine eye drops as perioperative prophylactic treatment for reducing conjunctival bacterial load and the rate of needle contamination in patients undergoing intravitreal anti-vascular endothelial growth factor injection. Enrolled patients were randomized to either the study group (0.6% povidone iodine, three day-prophylactic treatment before the injection) or to the control group (placebo, three day-prophylactic treatment). Conjunctival swabs were obtained before and after the prophylactic treatment in both groups. Intravitreal injections were performed in a sterile fashion. The injection needle and a control needle were collected for microbiological culture. Data from 254 and 253 eyes in the study group and control group, respectively, were analyzed. Bacterial growth from conjunctival swab cultures was significantly lower after 0.6% povidone iodine prophylaxis compared to baseline and to placebo prophylaxis (p < 0.001), showing an 82% eradication rate in the study group. No injection needle showed bacterial contamination in the study group, whereas six needles were culture-positive in the control group (p = 0.015). No serious ocular and non-ocular adverse events were recorded. The 0.6% povidone iodine solution proved an effective treatment in reducing conjunctival bacterial load and risk of needle contamination

    Prevalence and type distribution of human papillomavirus infection in women from North Sardinia, Italy

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    <p>Abstract</p> <p>Background</p> <p>Human papillomavirus (HPV) has been associated with several disorders of the genital tract, skin and oropharynx. The aims of our study were to evaluate the prevalence of HPV infection in women between 15 and 54 years of age in North Sardinia, Italy, to identify the prevalence of High Risk - Human papillomaviruses (HR-HPV) genotypes and to establish a correlation between molecular and cytological results.</p> <p>Methods</p> <p>From 2007 to 2009 we consecutively enrolled women aged 15-54 years admitted to public and private outpatient settings. All the participants filled in a questionnaire about the socio-cultural state, sexual activity and awareness about HPV. 323 cervical specimens were tested for HPV-DNA and HPV genotypes with INNO-LiPA HPV Genotyping CE Amp kit. Samples showing positivity to some HPV genotypes were re-tested using "in house" quantitative Real-Time PCR assays.</p> <p>Results</p> <p>Overall HPV-DNA positivity was detected in 35.9% of the women. The prevalence of HR-HPV infection among HPV positive samples was 93.1% with a specific prevalence of HPV 16, 51, 31, 53 and 18 of 54.3%, 37.9%, 10.3%, 6.9% and 5.2%, respectively. Co-infection with any HPV, HR-HPV, LR-HPV and HR/LR-HPV type was 18.3%, 14.9%, 0.9% and 2.5%, respectively; HPV 16/51 co-infection was detected in 64.6% of the HR-HPV co-infection group. The most frequent HPV-genotypes detected were 16 (32.5%) and 51 (22.7%). Among the 57 patients harboring mono-infection the most prevalent HPV genotypes were 16 (38.6%) and 31(10.5%). A multivariate analysis identified a statistical significant association between HPV infection and age and between HPV infection and previous sexual transmitted diseases. A statistically significant association between cytological cervical lesions and generic HPV exposure was identified.</p> <p>Conclusions</p> <p>To our knowledge, this is the first survey evaluating the prevalence of HPV infection in Northern Sardinia and drawing attention to the unusual high proportion of genotype HPV 51. Given the recent implementation of a widespread immunization program with vaccines not containing HPV 51, it has been relevant to prove the high prevalence of this HPV genotype from the start of the vaccination campaign, in order to avoid in the future attributing to the vaccination program a possible selection effect (HPV replacement).</p

    The rapid spread of SARS-COV-2 Omicron variant in Italy reflected early through wastewater surveillance

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    The SARS-CoV-2 Omicron variant emerged in South Africa in November 2021, and has later been identified worldwide, raising serious concerns. A real-time RT-PCR assay was designed for the rapid screening of the Omicron variant, targeting characteristic mutations of the spike gene. The assay was used to test 737 sewage samples collected throughout Italy (19/21 Regions) between 11 November and 25 December 2021, with the aim of assessing the spread of the Omicron variant in the country. Positive samples were also tested with a real-time RT-PCR developed by the European Commission, Joint Research Centre (JRC), and through nested RT-PCR followed by Sanger sequencing. Overall, 115 samples tested positive for Omicron SARS-CoV-2 variant. The first occurrence was detected on 7 December, in Veneto, North Italy. Later on, the variant spread extremely fast in three weeks, with prevalence of positive wastewater samples rising from 1.0% (1/104 samples) in the week 5–11 December, to 17.5% (25/143 samples) in the week 12–18, to 65.9% (89/135 samples) in the week 19–25, in line with the increase in cases of infection with the Omicron variant observed during December in Italy. Similarly, the number of Regions/Autonomous Provinces in which the variant was detected increased fromone in the first week, to 11 in the second, and to 17 in the last one. The presence of the Omicron variant was confirmed by the JRC real-time RT-PCR in 79.1% (91/115) of the positive samples, and by Sanger sequencing in 66% (64/97) of PCR amplicons

    The rapid spread of SARS-COV-2 Omicron variant in Italy reflected early through wastewater surveillance

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    The SARS-CoV-2 Omicron variant emerged in South Africa in November 2021, and has later been identified worldwide, raising serious concerns. A real-time RT-PCR assay was designed for the rapid screening of the Omicron variant, targeting characteristic mutations of the spike gene. The assay was used to test 737 sewage samples collected throughout Italy (19/21 Regions) between 11 November and 25 December 2021, with the aim of assessing the spread of the Omicron variant in the country. Positive samples were also tested with a real-time RT-PCR developed by the European Commission, Joint Research Centre (JRC), and through nested RT-PCR followed by Sanger sequencing. Overall, 115 samples tested positive for Omicron SARS-CoV-2 variant. The first occurrence was detected on 7 December, in Veneto, North Italy. Later on, the variant spread extremely fast in three weeks, with prevalence of positive wastewater samples rising from 1.0% (1/104 samples) in the week 5-11 December, to 17.5% (25/143 samples) in the week 12-18, to 65.9% (89/135 samples) in the week 19-25, in line with the increase in cases of infection with the Omicron variant observed during December in Italy. Similarly, the number of Regions/Autonomous Provinces in which the variant was detected increased from one in the first week, to 11 in the second, and to 17 in the last one. The presence of the Omicron variant was confirmed by the JRC real-time RT-PCR in 79.1% (91/115) of the positive samples, and by Sanger sequencing in 66% (64/97) of PCR amplicons. In conclusion, we designed an RT-qPCR assay capable to detect the Omicron variant, which can be successfully used for the purpose of wastewater-based epidemiology. We also described the history of the introduction and diffusion of the Omicron variant in the Italian population and territory, confirming the effectiveness of sewage monitoring as a powerful surveillance tool

    Identification and quantification of oncogenic HPV nucleic acids by means of real-time PCR assays

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    Method for the identification and quantification of oncogenic HPV nucleic acids comprising: a) first line screening by means of 5 independent SYBR Green I Real-time PCR assays to determine the total viral load and to identify the presence of one or more of 13 high risk HPV genotypes in the sample; b) second line assays to be applied to samples positives for: - 5 independent TaqMan Real-time PCR assays to determine the presence and the viral load of the most common oncogenic HPV types: HPV types: 16, 18, 31, 45, 33 group (including 33, 52, 58, 67 genotypes). - 6 independent SYBR Green I RT Real-time PCR assays to determine the presence in the sample of the oncogenic transcripts E6/E7 of HPV types 16, 18, 31, 33, 45, 5
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