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

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

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

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

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

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

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

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

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

Comparison of Different Self-Sampling Devices for Molecular Detection of Human Papillomavirus (HPV) and Other Sexually Transmitted Infections (STIs): A Pilot Study

Background: Cervical cancer is the fourth most common cancer in women, and it is well known that high-risk human papillomavirus (hrHPV) infections are the necessary carcinogenic factors for the development of cervical tumors. Moreover, the interaction between HPV and other sexually transmitted infections (STIs) may increase the risk of cancer progression. Self-sampling has been demonstrated to represent a valid and well-accepted alternative, favoring women’s participation in screening programs. This study aimed to investigate the use of FLOQSwabs® (FS) as compared to two other vaginal self-collection devices for the detection of hrHPV and other sexually transmitted infections. Methods: Cervical and vaginal self-samples were collected, using two different combinations of vaginal self-sampling devices, from 40 women referred to colposcopy for a documented abnormal Pap smear. All samples were tested for hrHPV and seven STI pathogens using two commercial molecular assays. Results: Data on hrHPV detection from the first group of women showed an almost perfect agreement (kappa: 0.89) between cervical vs. FS vaginal self-samples, and a substantial agreement (kappa: 0.79) between cervical and HerSwab™ (HS) samples. In the second group of women, an almost perfect agreement (kappa: 0.90) was demonstrated in the detection of hrHPV between cervical samples vs. FS, and a moderate agreement (kappa: 0.60) for cervical vs. Evalyn®Brush (EB) self-collected samples. STI detections showed a very good agreement (kappa: 0.89 and kappa: 1.00) both among FS vs. HS and FS vs. EB, respectively. There was no statistically significant difference between the different devices used. The most frequently detected hrHPV genotypes in the studied population were HPV 16, 31, 35, 51, and 56; whilst the most frequently identified STI pathogens were Ureaplasma parvum and Mycoplasma hominis. Overall, investigated women did not report any discomfort in using the different vaginal self-collection devices. Conclusion: Evaluation of the three different vaginal self-collection devices confirmed their overall good acceptability by the studied population, as well as a similar agreement for hrHPV detection as compared to cervical samples. Our study indicated that the use of self-collected samples offers an alternative strategy to improve women’s participation in cervical cancer screening programs, but also underlined the importance of evaluating the concordance in hrHPV detection of collection devices in combination with the molecular hrHPV assay

Human papillomavirus DNA detection in plasma and cervical samples of women with a recent history of low grade or precancerous cervical dysplasia.

Circulating HPV DNA has been previously described in women with advanced stages of cervical cancer and has been suggested to be a prognostic marker of disease recurrences and metastases. Only a few studies have reported the presence of HPV DNA in bloodstream of patients with low grade or precancerous cervical lesions. This study aimed to define if HPV DNA could be detected in plasma samples of 120 women referred for a recent history of cervical dysplasia who presented with lesions ranging from High Squamous Intraepithelial Lesion (H-SIL) to regressed normal cytology. HPV DNA detection was carried out in both plasma and cervical samples using type-specific real-time quantitative PCR assays identifying oncogenic HPV 16, 18, 31, 33, 45, 51 and 52. Overall, 34.2% (41/120) of plasma samples were shown to be positive for HPV DNA detection; HPV 45 (46.3%), HPV-51 (29.6%), and HPV 16 (18.5%) were the most frequently identified genotypes. The rate of HPV detection in paired cervical and plasma samples increased with advancing disease stage, ranging from 15.4% in women with regressed lesions to 38.9% in women with HSIL; HPV 16 resulted the most common genotype identified in women found to be HPV DNA positive in both cervical and plasma samples. Moreover, HPV 16 showed the highest median viral load value in both cervical and plasma samples, with 48,313 copies/104 cells and 1,099 copies/ml, respectively. Results obtained in this study confirm that HPV DNA can be detected and quantified in plasma samples of women with asymptomatic cervical infection. Further knowledge on HPV dissemination through the blood stream of women with cervical lesions would be very important in better understanding the natural history of HPV infection as well as its potential role in other distant tumors

Design, synthesis, and biological evaluation of 4-alkyliden-beta lactams: New products with promising antibiotic activity against resistant bacteria

The design, synthesis, and antibacterial activity of 4-alkyliden-azetidin-2-ones as new antimicrobial agents against multidrug-resistant pathogens is reported. 4-Alkyliden-azetidin-2-ones were easily obtained using an original protocol starting from 4-acetoxy-azetidinones and diazoesters. Parent compounds were further elaborated to obtain a small library of 4-alkylidene derivatives. A molecular modeling approach using GRID descriptors based on the concept of VRS identified attractive drug candidates and contributed to the rationalization of functional group effects in QSARs. The in vitro antibacterial activity of the new agents was evaluated against 43 recent clinical isolates of antibiotic-susceptible and -resistant Gram-positive and Gram-negative pathogens by determining their minimum inhibitory concentrations (MICs). The most active compound showed MIC values ranging from 0.25 to 32 mg/L against some of the bacterial species tested. Interestingly, some compounds demonstrated similar activity against methicillin-susceptible and -resistant strains of Staphylococcus aureus suggesting possible alternative mechanisms of action of these agents, supported by citotoxicity and preliminary scanning electron microscopy studies

HPV genotypes distribution in cervical and plasma samples.

<p>Correlation of HPV types identified in cervical and plasma samples with cervical cytology.</p