Search

A real-world population-based longitudinal study, aimed at determining the magnitude and duration of immunity induced by different types of vaccines against COVID-19, started in 2021 by enrolling a cohort of 2,497 individuals at time of their first vaccination. The study cohort included both healthy adults aged ≤65 years and elderly subjects aged >65 years with two or more co-morbidities. Here, patterns of anti-SARS-CoV-2 humoral and cell-mediated specific immune response, assessed on 1,182 remaining subjects, at 6 (T6) and 12 months (T12) after the first vaccine dose, are described. At T12 median anti-Spike IgG antibody levels were increased compared to T6. The determinants of increased anti-Spike IgG were the receipt of a third vaccine dose between T6 and T12 and being positive for anti-Nucleocapside IgG at T12, a marker of recent infection, while age had no significant effect. The capacity of T12 sera to neutralize in vitro the ancestral B strain and the Omicron BA.5 variant was assessed in a subgroup of vaccinated subjects. A correlation between anti-S IgG levels and sera neutralizing capacity was identified and higher neutralizing capacity was evident in healthy adults compared to frail elderly subjects and in those who were positive for anti-Nucleocapside IgG at T12. Remarkably, one third of T12 sera from anti-Nucleocapside IgG negative older individuals were unable to neutralize the BA.5 variant strain. Finally, the evaluation of T-cell mediated immunity showed that most analysed subjects, independently from age and comorbidity, displayed Spike-specific responses with a high degree of polyfunctionality, especially in the CD8 compartment. In conclusion, vaccinated subjects had high levels of circulating antibodies against SARS-CoV-2 Spike protein 12 months after the primary vaccination, which increased as compared to T6. The enhancing effect could be attributable to the administration of a third vaccine dose but also to the occurrence of breakthrough infection. Older individuals, especially those who were anti-Nucleocapside IgG negative, displayed an impaired capacity to neutralize the BA.5 variant strain. Spike specific T-cell responses, able to sustain immunity and maintain the ability to fight the infection, were present in most of older and younger subjects assayed at T12., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2023 Fedele, Schiavoni, Trentini, Leone, Olivetta, Fallucca, Fiore, Di Martino, Abrignani, Baldo, Baldovin, Bandera, Clerici, De Paschale, Diaco, Domnich, Fortunato, Giberti, Gori, Grifantini, Lazzarotto, Lodi, Mastroianni, Prato, Restivo, Vitale, Brusaferro, Merler, Palamara, Stefanelli and the Study Group for the Immunological Monitoring post Covid19 vaccination.)

Objectives: Monitoring of SARS-CoV-2 spread and vaccination strategies have relied on antibody (Ab) status as a correlate of protection. We used QuantiFERON™ (QFN) and Activation-Induced Marker (AIM) assays to measure memory T-cell reactivity in unvaccinated individuals with prior documented symptomatic infection (late convalescents) and fully vaccinated asymptomatic donors (vaccinees)., Methods: Twenty-two convalescents and 13 vaccinees were enrolled. Serum anti-SARS-CoV-2 S1 and N Abs were measured using chemiluminescent immunoassays. QFN was performed following instructions and interferon-gamma (IFN-γ) measured by ELISA. AIM was performed on aliquots of antigen-stimulated samples from QFN tubes. SARS-CoV-2-specific memory CD4+CD25+CD134+, CD4+CD69+CD137+ and CD8+CD69+CD137+ T-cell frequencies were measured by flow cytometry., Results: In convalescents, substantial agreement was observed between QFN and AIM assays. IFN-γ concentrations and AIM+ (CD69+CD137+) CD4+ T-cell frequencies correlated with each other, with Ab levels and AIM+ CD8+ T-cell frequencies, whereas AIM+ (CD25+CD134+) CD4+ T-cell frequencies correlated with age. AIM+ CD4+ T-cell frequencies increased with time since infection, whereas AIM+ CD8+ T-cell expansion was greater after recent reinfection. QFN-reactivity and anti-S1 titers were lower, whereas anti-N titers were higher, and no statistical difference in AIM-reactivity and Ab positivity emerged compared to vaccinees., Conclusions: Albeit on a limited sample size, we confirm that coordinated, cellular and humoral responses are detectable in convalescents up to 2 years after prior infection. Combining QFN with AIM may enhance detection of naturally acquired memory responses and help stratify virus-exposed individuals in T helper 1-type (TH1)-reactive (QFNpos AIMpos Abshigh), non-TH1-reactive (QFNneg AIMpos Abshigh/low), and pauci-reactive (QFNneg AIMneg Abslow)., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Gatti et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Objectives: Parvovirus B19 (B19V) infection in pregnancy is generally asymptomatic, but in about 3% it can cause complications, including miscarriage, severe foetal anaemia and foetal hydrops. The seroprevalence in pregnancy ranges from 20% to 82% in Africa, but there are no data for Benin. We therefore retrospectively assessed the seroprevalence of B19V in pregnant women attending the Saint Jean de Dieu Hospital in Tanguiéta, a rural district of Atacora, in northern Benin., Methods: We searched for anti-B19V immunoglobulin M (IgM) and immunoglobulin G (IgG) antibodies in 227 sequential sera from as many women (mean age 26.3 years, range: 16-41) of whom 30 were in the first trimester, 66 in the second and 131 in the third. Samples that tested positive for IgM were analysed with an immunoblot test and the viral genome (DNA-B19V) was searched for using a polymerase chain reaction., Results: Of the 227 women, 153 (67.4%) were positive for IgG anti-B19V, 7 (3.1%) for IgM and 73 (32.2%) were non-immune. Six IgM-positive women were also IgG positive. The difference in IgG seroprevalence between trimesters or ages was not statistically significant. Of the seven IgM-positive samples, three were confirmed positive by immunoblot (of which two were DNA-B19V positive), three were indeterminate (DNA-B19V negative) and one was negative (DNA-B19V negative). Of the three women with confirmed positive IgM, two were in the third trimester and one in the second trimester of pregnancy., Conclusions: The seroprevalence of anti-B19V IgG among pregnant women in Benin is high and in line with those reported in some African countries. IgM seroprevalence is also similar to that described in some African countries in non-epidemic periods. The low viral load observed depicts non-acute infections, but it is difficult to establish the precise time of the infection, especially for women tested in the second or third trimester of pregnancy, when the observed viremia could be a sign of an acute infection that occurred in the previous trimester. Consequently, clinical follow-up and further investigations to highlight possible foetal consequences are indicated., (© 2023 John Wiley & Sons Ltd.)

Parvovirus B19 (B19V) infection in pregnancy is mostly asymptomatic, but can cause complications including abortion and fetal hydrops. Although its infection is ubiquitous, seroprevalence among pregnant women varies according to different geographical areas. Since seroprevalence data in Italy are limited, the prevalence of antibodies and DNA in pregnant women was evaluated retrospectively, correlating the clinical situation of mothers and newborns. One thousand eight hundred and ninety-three sequential sera were examined from pregnant women (60.8% in the first trimester, 16.6% in the second one, and 22.6% in the third one, respectively) for anti-B19V IgG and IgM (confirmed by immunoblot); 1402 (74.1%) were of Italian origin and 491 (25.9%) non-Italian women. Molecular tests were used to search for viral genome. One thousand three hundred and fifteen (69.5%) samples were IgG-positive, 21 (1.1%) IgM-positive, and 578 (30.5%) nonimmune. The difference in IgG seroprevalence between Italian (71.1%) and non-Italian women (64.8%) was statistically significant. Of the 21 IgM-positive women, 16 were confirmed positive also by immunoblot (prevalence: 0.8%), of which 11 were viraemic (prevalence: 0.6%; mean 1.3 × 10 4  geq/ml). Mothers were asymptomatic, and the newborns had no clinical signs of congenital infection. IgG seroprevalence in Italy is high, with differences between Italian women and non-Italian women from geographic areas with lower endemic levels of B19V. The consistent migratory flows in place could lead to an increase in the number of susceptible women. The prevalence of viremia is low, and has not been associated with evident fetal damage at birth., (© 2022 Wiley Periodicals LLC.)

To date there has been limited head-to-head evaluation of immune responses to different types of COVID-19 vaccines. A real-world population-based longitudinal study was designed with the aim to define the magnitude and duration of immunity induced by each of four different COVID-19 vaccines available in Italy at the time of this study. Overall, 2497 individuals were enrolled at time of their first vaccination (T0). Vaccine-specific antibody responses induced over time by Comirnaty, Spikevax, Vaxzevria, Janssen Ad26.COV2.S and heterologous vaccination were compared up to six months after immunization. On a subset of Comirnaty vaccinees, serology data were correlated with the ability to neutralize a reference SARS-CoV-2 B strain, as well as Delta AY.4 and Omicron BA.1. The frequency of SARS-CoV-2-specific CD4+ T cells, CD8+ T cells, and memory B cells induced by the four different vaccines was assessed six months after the immunization. We found that mRNA vaccines are stronger inducer of anti-Spike IgG and B-memory cell responses. Humoral immune responses are lower in frail elderly subjects. Neutralization of the Delta AY.4 and Omicron BA.1 variants is severely impaired, especially in older individuals. Most vaccinees display a vaccine-specific T-cell memory six months after the vaccination. By describing the immunological response during the first phase of COVID-19 vaccination campaign in different cohorts and considering several aspects of the immunological response, this study allowed to collect key information that could facilitate the implementation of effective prevention and control measures against SARS-CoV-2., Competing Interests: The authors declare that in this study the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Fedele, Trentini, Schiavoni, Abrignani, Antonelli, Baldo, Baldovin, Bandera, Bonura, Clerici, De Paschale, Fortunato, Gori, Grifantini, Icardi, Lazzarotto, Lodi, Mastroianni, Orsi, Prato, Restivo, Carsetti, Piano Mortari, Leone, Olivetta, Fiore, Di Martino, Brusaferro, Merler, Palamara and Stefanelli.)

Introduction: Most infants at risk for cytomegalovirus (CMV)-associated sensorineural hearing loss (SNHL) are unrecognized because of the absence of a universal neonatal CMV screening. The search of CMV-DNA by molecular methods in salivary swabs was demonstrated to be a reliable approach. This study describes the results obtained by carrying out a universal screening for congenital CMV (cCMV) infection including all live-born newborns in three Italian sites, as well as the therapeutic interventions and clinical outcome of the CMV-infected neonates. Moreover, CMV maternal infection's characteristics were evaluated., Methods: To confirm or exclude cCMV infection, a CMV-DNA-positive result on a first salivary swab was followed by repeated saliva and urine samples collected within 21 days of age. Breast milk samples were also collected. The search of CMV-DNA was performed with a single automated quantitative commercial real-time PCR assay, regardless of the type of samples used., Results: A total of 3,151 newborns were enrolled; 21 (0.66%) of them were congenitally infected (median saliva viral load at screening, 6.65 [range, 5.03-7.17] log 10 IU/ml). Very low/low viral load in screening saliva samples (median value, 1.87 [range, 1.14-2.59] log 10 IU/ml) was associated with false-positive results ( n = 54; 1.7%). CMV-DNA was detected in almost half of the breast milk samples of mother-infant pairs with a false-positive result, suggesting that contamination from breast milk may not be the only explanation in the study population. cCMV infection confirmation with the search of CMV-DNA in a urine sample proved to be the gold standard strategy, since false-positive results were observed in 4/54 (7.5%) of the repeated saliva samples. Symptomatic cCMV infection was observed in 3/21 (14.3%) infants; notably, one (4.7%) developed moderate unilateral SNHL at 5 months after birth. Finally, two symptomatic cCMV infections were associated with primary maternal infection acquired in the first trimester of gestation; one newborn with severe cCMV symptoms was born to a mother with no CMV checkups in pregnancy., Conclusion: Without universal neonatal CMV screening, some infected infants who develop late neurological sequelae may not be recognized and, consequently, they are not able to benefit early from instrumental and therapeutic interventions to limit and/or treat CMV disease., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict., (Copyright © 2022 Chiereghin, Pavia, Turello, Borgatti, Baiesi Pillastrini, Gabrielli, Gibertoni, Marsico, De Paschale, Manco, Ruscitto, Pogliani, Bellini, Porta, Parola, Scarasciulli, Calvario, Capozza, Capretti, Laforgia, Clerici and Lazzarotto.)

Not available.

Since the spreading of Sar-CoV-2 in March 2020, many serologic tests have been developed to identify antibody responses. Indeed, different commercial kits are directed against different antigens and could utilise different methods thereby triggering confusion and criticism. Here, we compared two Food and Drug Administration (FDA)-approved automatized assays that detect IgG responses against spike or nucleocapsid protein of Sars-Cov-2 virus in 127 subjects among healthcare workers of IRCCS Policlinico San Donato (MI), Italy. We observed different kinetics of IgG responses, demonstrating the importance of timing of sampling to correctly interpret the results both for infection diagnosis and for epidemiologic studies. We observed that Anti-N response starts earlier than Anti-S1/S2 response but also decreases earlier, affecting the sensitivity of the tests at different time points. Combining two different assays, designed against different antigens, could reduce false negative results. Finally, we observed a patient who produced anti-nucleocapsid IgG, but not anti-spike IgG. In conclusion, we investigated antibody responses in Covid-19 disease, aiming to direct clinicians and laboratory scientists to correctly interpret serologic results by always paying attention to clinical history correlation, timing of sampling, methods and antigens used, to avoid false negative results and obtain relevant epidemiologic data., (Copyright 2020 Biolife Sas. www.biolifesas.org.)

Background: Congenital cytomegalovirus (CMV) infection is the leading infectious cause of neurological impairment for which, currently, there are no approved antenatal treatment options., Objectives: The aim of this article was to summarize the available evidence on the use of valacyclovir during pregnancy to prevent and treat congenital CMV infection and disease., Sources: Two databases (PubMed and ClinicalTrial.gov) were reviewed., Content: Six relevant documents were identified, namely one observational study, three clinical trials, two case reports. Most relevant findings were those from two clinical trials. A phase 2/3 placebo-controlled study showed a decrease of 71% (5 of 45 vs 14 of 47) in rate of CMV vertical transmission in women treated with 8 g/day valacyclovir following primary CMV infection in pregnancy. A phase 2, single-arm clinical trial, showed that 8 g/day valacyclovir administered to mothers of symptomatic infected foetuses increased the portion of asymptomatic neonates to 82% (34 of 41), compared with 43% (20 of 47) in untreated pregnancies from a historical cohort., Implications: Studies in favour of using valacyclovir during pregnancy for prevention and treatment of congenital CMV infection are emerging but are still few. Randomized clinical trials on large cohorts of patients investigating the efficacy on prevention and treatment of congenital CMV are required. Unfortunately, this will be probably not be feasible at least in the short period. In the meantime, data on the 'off label' use of valacyclovir for CMV in pregnancy could be collected within a multicentre observational study., (Copyright © 2020 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.)

The efficacy of domestic laundering of healthcare staff clothing is still debated. This study aimed to compare the performance of decontamination of different domestic laundering with that of industrial laundering. Fourteen naturally contaminated white coats of healthcare workers (5 fabric squares from each coat) and fabric squares of artificially contaminated cotton cloth (30 fabric squares per each bacterial strain used) were included. Four domestic laundering procedures were performed; two different washing temperatures (40 °C and 90 °C) and drying (tumble dry and air dry) were used. All fabric squares were ironed. Presence of bacterial bioburden on the fabric squares after domestic and industrial laundering was investigated. None of the naturally contaminated fabric squares resulted completely decontaminated after any of the domestic washes. At 24, 48, and 72 h of incubation, bacterial growth was observed in all the laundered fabric squares. Besides environmental microorganisms, potentially pathogenic bacteria (i.e., Acinetobacter lwoffii, Micrococcus luteus, coagulase-negative staphylococci) were isolated. On the artificially contaminated fabric squares, the bioburden was reduced after the domestic laundries; nevertheless, both Gram-negative and -positive pathogenic bacteria were not completely removed. In addition, a contamination of the fabric squares by environmental Gram-negative bacteria was observed. In both the naturally and artificially contaminated fabric squares, no bacterial growth at all the time-points analyzed was observed after industrial laundering, which provided to be more effective in bacterial decontamination than domestic washes. For those areas requiring the highest level of decontamination, the use of specialized industrial laundry services should be preferred.

Anthelvencins A and B are pyrrolamide metabolites produced by Streptomyces venezuelae ATCC 14583 and 14585. Isolated in 1965, they were reported to exhibit anthelmintic and moderate antibacterial activities. In this study, we revise the structure of anthelvencin A and identify a third anthelvencin metabolite, bearing two N -methylated pyrrole groups, which we named anthelvencin C. We sequenced the genome of S. venezuelae ATCC 14583 and identified a gene cluster predicted to direct the biosynthesis of anthelvencins. Functional analysis of this gene cluster confirmed its involvement in anthelvencin biosynthesis and allowed us to propose a biosynthetic pathway for anthelvencins. In addition to a nonribosomal peptide synthetase (NRPS), the assembly of anthelvencins involves an enzyme from the ATP-grasp ligase family, Ant23. We propose that Ant23 uses a PCP-loaded 4-aminopyrrole-2-carboxylate as substrate. As observed for the biosynthesis of the other pyrrolamides congocidine (produced by Streptomyces ambofaciens ATCC 25877) and distamycin (produced by Streptomyces netropsis DSM 40846), the NRPS assembling anthelvencins is composed of stand-alone domains only. Such NRPSs, sometimes called type II NRPSs, are less studied than the classical multimodular NRPSs. Yet, they constitute an interesting model to study protein-protein interactions in NRPSs and are good candidates for combinatorial biosynthesis approaches.

SETTING: QuantiFERON TB assay (QFT) is used to screen tuberculosis (TB) infection, but it cannot distinguish active TB from latent TB infection (LTBI). OBJECTIVE: To evaluate the quantitative expression of the high-affinity FCgamma receptor I (CD64) on neutrophils (NE) and monocytes (MO) in peripheral blood using flow cytometry, measured in antibody binding capacity (ABC) units as a predictive biomarker of TB. DESIGN: Fifty-two patients were enrolled (45 QFT-positive and 7 QFT-indeterminate). Cultures and molecular analyses were performed. RESULTS: Of the 45 QFT-positive patients, 29 were culture-positive (active TB) and 16 were negative (LTBI). The median NE CD64 ABC and MO CD64 ABC expression was significantly higher ( P < 0.001) in culture-positive patients. The NE CD64 and MO CD64 area under the receiver operating characteristic curve values were respectively 0.948 (95%CI 0.838-0.992) and 0.989 (0.901-1.000). By setting the cut-off NE CD64 value at >2400 ABC or MO CD64 value >25 800 the assay sensitivity increased to 95.5% with 100% specificity and 100% positive predictive value. In the QFT-indeterminate group, five culture-positive cases had NE CD64 >2400 ABC or MO CD64 value >25 800; two culture-negative cases had lower values. CONCLUSION: The CD64 quantitative expression on peripheral blood cells may be used as a predictive biomarker for active TB.

Cytomegalovirus (CMV) remains a major cause of morbidity and mortality in allogeneic hematopoietic stem cell transplantation (allo-HSCT) and solid organ transplantation (SOT) recipients. In view of the uncertainties on the assessment and prevention of CMV infection in both transplant procedures, three Italian scientific societies for HSCT and SOT and for Clinical Microbiology appointed a panel of experts to compose a framework of recommendations. Recommendations were derived from a comprehensive analysis of the scientific literature and from a multidisciplinary consensus conference process. The lack of adequate clinical trials focused on certain diagnostic procedures, and antiviral intervention forced the panel to use the methods of consensus for shaping some recommendations. Recommendations concerning the two types of transplant were given for the following issues: assessment of pretransplant CMV serostatus, immunological monitoring after transplant, CMV prophylaxis with antivirals, CMV preemptive strategy, and CMV prophylaxis with immunoglobulin infusion and with adoptive immunotherapy. The questions raised by and the recommendations resulting from this consensus conference project may contribute to the improvement of certain crucial aspects of the management of CMV infections in allo-HSCT and in SOT populations., (© 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Introduction: A variety of national and international guidelines exist around the management of carbapenem resistant Enterobacteriaceae (CREs), but some of these are several years old and do not reflect current epidemiology and they also do not necessarily give pragmatic advice around active surveillance of CREs in countries with a high burden of cases and limited resources. This paper aims to provide a best practice position paper to guide active surveillance in a variety of scenarios in these settings, and discusses which patients should be screened, what methods could be used for screening, and how results might influence infection prevention interventions., Methods: This paper was developed as a result of a series of meetings of expert opinion leaders representing the major infectious disease and infection prevention societies in Italy and having the endorsement of AMCLI (Italian Association of Clinical Microbiology) and SITA (Italian Society for Anti-infective Therapy). There was no attempt to undertake a full systematic review of the evidence, as it was felt that this was inadequate to inform a pragmatic view on the best way forward based on current epidemiology and infection rates., Key Recommendations: Key recommendations focus on the urgent need to promote measures to prevent transmission and infection, focusing on high risk patients and clinical areas, as well as outbreak situations. Active surveillance leading to appropriate infection prevention precautions plays a major role in this., Conclusions: There are limited national or international guidelines giving pragmatic advice on the most appropriate measures for active surveillance and management of colonized patients in a high-burden setting such as Italy. While individual hospitals and regions will need to formulate their own policies based on local epidemiology, this position paper attempts to highlight current best practice in this area and provide pragmatic advice for clinicians, infection prevention staff, and healthcare managers., Competing Interests: Competing interestsFor activities outside of the submitted work, SA has received fees as speaker from ADA, Biomerieux, Cepheid, MB has received funding for scientific advisory boards, travel and speaker honoraria from Angelini, Astellas Pharma, AstraZeneca, Bayer, Biomerieux, Cepheid, Cidara, Pfizer, Menarini, MSD, Nabriva, Paratek, Roche, Shionogi, Tetraphase, Thermofisher and the Medicine Company, NP has received fees as speaker from MSD, Pfizer, Cepheid, Takeda, Shionogi, Angelini, Zambon, Becton & Dickinson, Johnson & Johnson, PV has served as a consultant for MSD, PFIZER, VENATORX, NABRIVA, GILEAD, THERMOFISHER and BIOMERIEUX and received payment for serving on the speaker’s bureau for MSD, PFIZER, CORREVIO and GILEAD, GMR has received honoraria for scientific advisory boards, travel and speaker’s bureau from Accelerate, Angelini, AstraZeneca, Basilea, Beckman Coulter, Biotest, Cepheid, Curetis, Elitech, Menarini, Merck, Nordic Pharma, Novartis, Pfizer, Qpex, Rempex, Roche, Thermo Fisher, VenatorX, Zambon; and research grants to the laboratory from from Accelerate, Alifax, Angelini, Arrow, AstraZeneca, Basilea, Becton-Dickinson, bioMérieux, Biotest, Cepheid, Checkpoints, Elitech, Estor, Liofilchem, Menarini, Merck, Nordic Pharma, Novartis, Pfizer, Rempex, Seegene, Shionogi, VenatorX, Zambon. PC and FT declare that they have no competing interests.

While implant-related infections continue to play a relevant role in failure of implantable biomaterials in orthopaedic and trauma there is a lack of standardised microbiological procedures to identify the pathogen(s). The microbiological diagnosis of implant-related infections is challenging due to the following factors: the presence of bacterial biofilm(s), often associated with slow-growing microorganisms, low bacterial loads, previous antibiotic treatments and, possible intra-operative contamination. Therefore, diagnosis requires a specific set of procedures. Based on the Guidelines of the Italian Association of the Clinical Microbiologists (AMCLI), the World Association against Infection in Orthopaedics and Trauma has drafted the present document. This document includes guidance on the basic principles for sampling and processing for implant-related infections based on the most relevant literature. These procedures outline the main microbiological approaches, including sampling and processing methodologies for diagnostic assessment and confirmation of implant-related infections. Biofilm dislodgement techniques, incubation time and the role of molecular approaches are addressed in specific sections. The aim of this paper is to ensure a standardised approach to the main microbiological methods for implant-related infections, as well as to promote multidisciplinary collaboration between clinicians and microbiologists.

Background: Simultaneous detection of HIV 1 and 2 antibodies and HIV-1 p-24 antigen in the 4th generation tests is particularly effective for the identification of early acute HIV infections while maintaining accurate detection of long-established infections., Objectives: The aim of this study was to evaluate the new 4th generation VITROS HIV Combo test from Ortho-Clinical Diagnostics by comparing its results with those obtained using a 3rd generation HIV 1/2 antibody test (VITROS Anti HIV 1 + 2 from Ortho-Clinical Diagnostics) and a 4th generation test (LIAISON XL HIV Ab/Ag, DiaSorin) currently used in the Microbiology Unit of Legnano Hospital., Study Design: One thousand and three samples of the normal daily routine (Group 1) were analyzed simultaneously with the three systems. The concordant and discordant sample results were further tested using Western blot and HIV-RNA assay (Roche). One hundred samples (Group 2) of known HIV positive subjects (63 of subtype B, 37 subtype non-B, and 51 with positive viraemia) and 50 samples (Group 3) with indeterminate Western blot were also examined using the three systems. From Group 3, 24 samples were collected from patients diagnosed with acute infection., Results: The overall agreement between the three systems was 99.4% (99.5% in group 1, 100% in group 2 and 96.6% in group 3) with a coefficient Fleiss Kappa of 0.9814. Notably, the VITROS HIV Combo test was positive in all known HIV positive samples of group 2 without any statistically significant difference in the values of the sample/cut off ratios between the B and non-B subtypes and between the positive and negative viraemia samples in established infections. The VITROS HIV Combo test was also positive in all samples of patients with acute infection in group 3., Conclusions: The VITROS HIV Combo test has shown comparable performance to the other two assays in use of 3rd and 4th generation tests and is able to correctly identify both acute and established HIV infections independently of viraemia and HIV subtype., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Objective: To develop a risk-scoring tool to predict multidrug-resistant (MDR) etiology in patients with bloodstream infections (BSI) caused by Gram-negative bacilli (GNB)., Methods: A prospective multicenter study analyzed patients with BSI hospitalized in 31 Internal Medicine wards in Italy from March 2012 to December 2012. Patients with BSI caused by MDR-GNB (non-susceptible to at least one agent in three antimicrobial categories) were compared to those with BSI due to susceptible GNB. A logistic regression to identify predictive factors of MDR-GNB was performed and the odds ratio (OR) were calculated. A score to predict the risk of MDR was developed., Results: Of 533 BSI episodes, 253 (47.5%) were caused by GNB. Among GNB-BSI, 122 (48.2%) were caused by MDR-GNB while 131 (51.8%) by non-MDR GNB. At multivariate analysis transfer from long-term care facility (OR 9.013, 95% CI 1.089-74.579, p = 0.041), hospitalization in the last 3 months (OR 2.882, 95% CI 1.580-5.259, p = 0.001), urinary catheter (OR 2.315, 95% CI 1.202-4.459, p = 0.012), antibiotic therapy in the last 3 months (OR 1.882, 95% CI 1.041-3.405, p = 0.036), age ≥ 75 years (OR 1.866, 95% CI 1.076-3.237, p = 0.026) were factors independently associated with MDR etiology. A score ranging from 0 to 10 was useful to recognize patients at lowest risk (0 points: Negative Likelihood Ratio 0.10) and those at highest risk (>6 points, Positive Likelihood Ratio 11.8) of GNB bacteremia due to a MDR strain., Conclusions: Specific predictors of MDR etiology are useful to calculate probabilities of MDR etiology among hospitalized patients with blood cultures positive for GNB., (Copyright © 2018 European Federation of Internal Medicine. Published by Elsevier B.V. All rights reserved.)

Background: Mycoplasmas are frequently isolated from the genital tract. New molecular PCR-based methods for the detection of mycoplasmas can better define the real epidemiology of these microorganisms. The aim of this study was to evaluate the prevalence of mycoplasmas in a population of childbearing age women by means of PCR., Methods: This 21-month multicentre observational study was conducted at four Italian clinical microbiology laboratories. Women reporting symptoms of vaginitis/cervicitis, or with history of infertility, pregnancy, miscarriage or preterm birth were included. Detection of Ureaplasma parvum, Ureaplasma urealyticum, Mycoplasma hominis, Mycoplasma genitalium was performed from cervical swabs by means of a commercially available multiplex real-time PCR., Results: a total of 1761 women fulfilled the inclusion criteria and were included in the study. The overall prevalence was: U. parvum 38.3%, U. urealyticum 9%, M. hominis 8.6% and M. genitalium 0.6%. The proportion of foreign patients positive for U. parvum was significantly higher compared to Italian patients (37% vs 30.1%, p = 0.007) and also for overall mycoplasma colonization (53.4% vs 45.8%, p = 0.011). The number of symptomatic patients positive for M. hominis was significantly higher than that of negative controls (2.9% vs 1%, p = 0.036). A significant positive trend in mycoplasma colonization was found in relation to the pregnancy week for U. urealyticum (p = 0.015), M. hominis (p = 0.044) and for overall mycoplasma colonization (p = 0.002)., Conclusion: multiplex RT-PCR can be a valuable tool to evaluate the real epidemiology of cervical mycoplasma colonization., (Copyright © 2017. Published by Elsevier B.V.)

Background: Double gallbladder is a rare biliary anomaly. Perinatal diagnosis of the disorder has been reported in only 6 cases, and in 5 of them the diagnosis was based on ultrasound imaging only. However, the ultrasound technique alone does not provide a sufficiently precise description of cystic ducts and biliary anatomy, an information that is crucial for a correct classification and for a possible future surgery., Case Presentation: At 21 weeks of gestational age of an uneventful pregnancy in a 38 year old primipara mother, a routine ultrasound screening detected a biliary anomaly in the fetus suggestive of a double gallbladder. A neonatal abdominal ultrasonography performed on postnatal day 2 confirmed the diagnosis. On day 12 the newborn underwent a Magnetic Resonance Cholangiopancreatography (MRCP) that clearly characterized the anatomy of the anomaly: both gallbladders had their own cystic duct and both had a separate insertion in the main biliary duct., Conclusions: We report a case of early prenatal suspected duplicate gallbladder that was confirmed by a neonatal precise diagnosis of a Type 2, H or ductular duplicate gallbladder, using for the first time 3D images of Magnetic resonance cholangiopancreatography in a newborn. An accurate anatomical diagnosis is mandatory in patients undergoing a possible future cholecystectomy, to avoid surgical complications or reoperations. Therefore, in case of a perinatal suspicion of a double gallbladder, neonates should undergo a Magnetic resonance cholangiopancreatography. A review of the Literature about this variant is included.

Background: There are no Italian data regarding the strategies for preventing neonatal group B streptococcal (GBS) infection. We conducted a national survey in order to explore obstetrical, neonatal and microbiological practices for the GBS prevention., Methods: Three distinct questionnaires were sent to obstetricians, neonatologists and microbiologists. Questionnaires included data on prenatal GBS screening, maternal risk factors, intrapartum antibiotic prophylaxis, microbiological information concerning specimen processing and GBS antimicrobial susceptibility., Results: All respondent obstetrical units used the culture-based screening approach to identify women who should receive intrapartum antibiotic prophylaxis, and more than half of the microbiological laboratories (58%) reported using specimen processing consistent with CDC guidelines. Most neonatal units (89 out of 107, 82%) reported using protocols for preventing GBS early-onset sepsis consistent with CDC guidelines., Conclusions: The screening-based strategy is largely prevalent in Italy, and most protocols for preventing GBS early-onset sepsis are consistent with CDC guidelines. However, we found discrepancies in practices among centers that may reflect the lack of Italian guidelines issued by public health organizations.

Some international guidelines recommend evaluating the need to confirm positive anti-hepatitis C virus (HCV) antibody screening results by means of a more specific antibody or molecular biology test on the basis of a screening threshold value (such as the sample signal/cut-off ratio) that can predict the positivity of additional antibody testing in at least 95% of cases. The aim of this study was to determine the threshold value of the DiaSorin LIAISON XL chemiluminescence test. Two hundred and twenty-eight routine laboratory samples that were chemiluminescence positive for anti-HCV antibodies but had different signal/cut-off ratios were assayed using immunoblotting, which indicated that 155 (68.0%) were positive, 40 (17.5%) were negative, and 33 (14.5%) were indeterminate. When the samples were divided on the basis of their signal/cut-off ratios, 95.5% of the samples with a ratio of ≥3.5 were positive as against 74.1% of the positive or indeterminate samples with a ratio of <3.5. Statistical analysis using Youden's index and a receiver operating characteristic curve showed that the optimum cut-off value was 3.65. These findings indicate that, when using the LIAISON XL system for anti-HCV antibody screening, a signal/cut-off ratio of ≥3.65 makes further confirmatory tests unnecessary., (© 2017 Wiley Periodicals, Inc.)

A sequence of oxidative cleavage/reductive amination/reductive cleavage enables the preparation of N-substituted cis 5-amino-3-hydroxypiperidines from a readily available bicyclic adduct. This new route provides straightforward and versatile access to drug-relevant scaffolds or fragments.

The aim of this study was to compare the data obtained using the new LIAISON® XL chemiluminescence system to search for HBsAg, anti-HCV, and anti-HIV1-2/p24 Ag with those obtained using the VITROS system currently adopted by the Microbiology Unit of the Hospital of Legnano. Routine samples of patients who were referred by practitioners for the determination of HBsAg (1,000 samples) and/or anti-HCV (1,002 samples) and/or anti-HIV1-2 (995 samples) were simultaneously analyzed using both systems. The concordant positive and discordant samples were re-examined for confirmation by means of an HBsAg neutralization assay, anti-HCV immunoblot, or anti-HIV1-2 Western blot; HBV-DNA, or HCV-RNA or HIV-RNA was also sought in the discordant samples. Samples of patients known to be positive were tested (100 HBsAg positive, 100 anti-HCV positive, and 100 HIV 1-2 positive) as well throughout treatment, with viremia levels becoming undetectable after treatment. The HBsAg, anti-HCV, and anti-HIV1-2 concordance between the two systems in routine series was respectively 99.8%, 98.5% and 99.7%, and 100% for all markers in samples known positive. The various molecular biology and confirmatory tests of the discordant samples were all negative (except for one anti-HCV positive sample). Measure of Cohen's kappa coefficient for HBsAg, anti-HCV, and anti-HIV gave K values of respectively 0.992, 0.946, and 0.980. In conclusion, the performance of the LIAISON® XL system in the routine laboratory determination for all three markers was comparable with that of the VITROS system. J. Med. Virol. 89:489-496, 2017. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)

Competing Interests: The Authors declare no conflicts of interest.

Varicella-zoster virus, which is responsible for varicella (chickenpox) and herpes zoster (shingles), is ubiquitous and causes an acute infection among children, especially those aged less than six years. As 90% of adults have had varicella in childhood, it is unusual to encounter an infected pregnant woman but, if the disease does appear, it can lead to complications for both the mother and fetus or newborn. The major maternal complications include pneumonia, which can lead to death if not treated. If the virus passes to the fetus, congenital varicella syndrome, neonatal varicella (particularly serious if maternal rash appears in the days immediately before or after childbirth) or herpes zoster in the early years of life may occur depending on the time of infection. A Microbiology laboratory can help in the diagnosis and management of mother-child infection at four main times: (1) when a pregnant woman has been exposed to varicella or herpes zoster, a prompt search for specific antibodies can determine whether she is susceptible to, or protected against infection; (2) when a pregnant woman develops clinical symptoms consistent with varicella, the diagnosis is usually clinical, but a laboratory can be crucial if the symptoms are doubtful or otherwise unclear (atypical patterns in immunocompromised subjects, patients with post-vaccination varicella, or subjects who have received immunoglobulins), or if there is a need for a differential diagnosis between varicella and other types of dermatoses with vesicle formation; (3) when a prenatal diagnosis of uterine infection is required in order to detect cases of congenital varicella syndrome after the onset of varicella in the mother; and (4) when the baby is born and it is necessary to confirm a diagnosis of varicella (and its complications), make a differential diagnosis between varicella and other diseases with similar symptoms, or confirm a causal relationship between maternal varicella and malformations in a newborn.

Only a few studies provided data on the clinical history of sepsis within internal Medicine units. The aim of the study was to assess the short-term mortality and to evaluate the prognostic risk factors in a large cohort of septic patients treated in internal medicine units. Thirty-one internal medicine units participated to the study. Within each participating unit, all admitted patients were screened for the presence of sepsis. A total of 533 patients were included; 78 patients (14.6%, 95%CI 11.9, 18.0%) died during hospitalization; mortality rate was 5.5% (95% CI 3.1, 9.6%) in patients with nonsevere sepsis and 20.1% (95%CI 16.2, 28.8%) in patients with severe sepsis or septic shock. Severe sepsis or septic shock (OR 4.41, 95%CI 1.93, 10.05), immune system weakening (OR 2.10, 95%CI 1.12, 3.94), active solid cancer (OR 2.14, 95% CI 1.16, 3.94), and age (OR 1.03 per year, 95% CI 1.01, 1.06) were significantly associated with an increased mortality risk, whereas blood culture positive for Escherichia coli was significantly associated with a reduced mortality risk (OR 0.46, 95%CI 0.24, 0.88). In-hospital mortality of septic patients treated in internal medicine units appeared similar to the mortality rate obtained in recent studies conducted in the ICU setting., Competing Interests: The authors have no conflicts of interest to disclose.

Objectives: Hepatitis E virus (HEV) is the cause of enterically transmitted non-A, non-C hepatitis (an infection that is particularly severe during pregnancy) in tropical and subtropical countries. As there are no published data concerning the prevalence of HEV antibodies in Benin, their presence was investigated in pregnant women undergoing routine HIV screening in a rural area in northern Benin and in pregnant women with acute non-A, non-C hepatitis., Methods: A total of 278 serum samples were collected from asymptomatic pregnant women in 2011 were tested for HEV and hepatitis A virus (HAV) antibodies, and the HEV IgM-positive samples were further tested for HEV-RNA. A further seven samples of pregnant women with acute non-A, non-C hepatitis collected during episodes of acute hepatitis in 2005 were also analysed., Results: Of the 278 samples collected in 2011, 16.19% were positive for HEV IgG and 1.44% for HEV IgM (none positive for HEV-RNA), and 99.64% were positive for total HAV antibodies (none positive for HAV IgM). Six of the seven samples collected in 2005 were positive for HEV IgG and IgM, and two were also positive for HEV-RNA., Conclusions: The circulation of HEV infection is significant among pregnant women in Benin, in whom the consequences may be fatal., (© 2015 John Wiley & Sons Ltd.)

Pregnant women are not screened for HBsAg and anti-HCV antibodies in many African countries. As there are few data concerning the prevalence of HBV, HDV, and HCV serological markers in Benin, the aim of this study was to evaluate their 2011 prevalence in pregnant women undergoing HIV screening in a rural area of north Benin, and compare the data with those reported for the same area in 1986. The sera of 283 women were examined for HBsAg, anti-HBs, anti-HBc, anti-HCV, and anti-HIV 1/2 antibodies. In the case of HBsAg positivity, a search was made for the HBeAg, anti-HDV, and HBV genotypes; in the case of anti-HCV positivity, a search was made for the HCV genotypes. HBsAg, anti-HBs, anti-HBc, anti-HCV, and anti-HIV 1/2 were positive in respectively 44 (15.5%), 82 (29.0%), 234 (82.7%), 21 (7.4%), and nine samples (3.2%). Of the HBsAg-positive samples, five (11.4%) were positive for HBeAg, five (11.4%) for anti-HDV, and 19 for HBV genotype E. Of the anti-HCV-positive samples, five were positive for genotype 2a/2c and one for genotype 1a. The prevalence of anti-HBc alone (HBsAg and anti-HBs negative) was very high (41.3%). In comparison with the 1986 data, the prevalence of HBsAg and anti-HBc remained unchanged, that of HBeAg and anti-HDV had decreased, and that of anti-HIV 1/2 had increased. As these data confirm that HBV and HCV are highly endemic in the study area, it may be appropriate to introduce HBsAg and anti-HCV screening for pregnant women. J. Med. Virol. 86:1281-1287, 2014. © 2014 Wiley Periodicals, Inc., (© 2014 Wiley Periodicals, Inc.)

Objectives: Toxoplasma gondii, cytomegalovirus (HCMV) and rubella virus infections are among the most serious of those contracted during pregnancy in terms of foetal consequences. Toxoplasma, HCMV and rubella antibody screening is unusual in Africa, and there are few published data. The aim of this study was to evaluate the prevalence of these markers among pregnant women in northern Benin on the occasion of routine syphilis screening., Methods: Toxoplasma, HCMV and rubella IgG and IgM antibodies were determined in the serum of 283 women attending Saint Jean de Dieu de Tanguiéta hospital, using an enzyme immunoassay, and IgM were confirmed using an enzyme-linked fluorescent assay (ELFA). In the case of IgM positivity, the avidity of anti-HCMV and anti-Toxoplasma IgG was measured. Total anti-Treponema pallidum antibodies were determined using an enzyme immunoassay and confirmed by immunoblotting. In the case of positivity, the Venereal Disease Research Laboratory (VDRL) test was used., Results: The prevalence of anti-Toxoplasma, anti-HCMV, anti-rubella IgG and total anti-Treponema antibodies was, respectively, 30.0%, 100%, 94% and 2.5%. The VDRL test was positive in 62.5% of the anti-Treponema-positive samples. The prevalence of anti-Toxoplasma, anti-HCMV and anti-rubella IgM was, respectively, 0.4%, 1.4% and 0%. There were no statistically significant differences in terms of age class or trimester of pregnancy. Anti-Toxoplasma and anti-HCMV IgG avidity was always high., Conclusions: The prevalence of HCMV and rubella antibodies is high in northern Benin, whereas that of Toxoplasma antibodies is lower. As nearly two-thirds of the pregnant women were anti-Toxoplasma seronegative, antibody screening should be introduced., (© 2014 John Wiley & Sons Ltd.)

The aim of this study was to determine short-term and long-term changes in motor function in patients with chronic hemiparesis who underwent robot training and to evaluate its long-term benefit after 6 months. This was a longitudinal study with a 6-month follow-up. The 15 patients included in this study underwent the Fugl-Meyer test, the Ashworth Scale test, the Frenchay Arm test, and the Box and Block test according to the following schedule: immediately before (T1, T3) and after each treatment (T2, T4), and 6 months after T4 (T5). There were statistically significant improvements in Fugl-Meyer test between T1 and T2 and between T1 and T4; the score increased in the Ashworth Scale test for Shoulder between T1 and T3 and between T1 and T5; a statistically significant decrease was found between T1 and T2 and between T1 and T4, in the Box and Block test between T1 and T4, and also between T1 and T5. This original rehabilitation treatment may contribute toward increasing upper limb motor recovery in stable chronic stroke patients.

Objectives: The introduction of vaccination against hepatitis B initially reduced the number of HBV (hepatitis B virus) and HDV (hepatitis delta virus) infections, but the decreasing trend of HDV infection seems to have stopped. The aim of this study was to assess the prevalence of HDV infection in the general population living in the catchment area of Legnano Hospital in northern Italy., Methods: Of the 22,758 subjects tested in 2007-2008, the 488 who were HBsAg (hepatitis B surface antigen)-positive [including 107 (21.9%) of non-Italian origin] were subsequently tested for anti-HDV antibodies., Results: Of the 488 subjects who tested positive for HBsAg, 24 (4.9%) were anti-HDV positive, all aged between 30 and 60 years. The difference in prevalence between males (7.1%) and females (1.9%) was statistically significant (p < 0.05), but not that between Italian (5.0%) and non-Italian patients (4.7%). The differences in anti-HDV seropositivity between the patients with acute (0%) and chronic infections (6.3%), and between the incident (2.5%) and prevalent cases (7.4%), were not statistically significant, but there was a significant difference (p < 0.01) between those with asymptomatic (2.1%) and clinically symptomatic infections (10.3%). Intravenous drug abuse was the main source of infection., Conclusions: In the catchment area of our hospital, the prevalence of HDV infection does not seem to be due to patients of non-Italian origin, but to Italian patients who are not vaccinated against HBV and who survived the HDV epidemic of the 1970s and 1980s. Nevertheless, the increase in the number of immigrants from non-EU countries in recent years is soon likely to lead to a change in the epidemiology of HDV.

Background: The implementation of mass vaccinations against hepatitis B virus (HBV) has significantly reduced the prevalence of HBsAg-positive subjects. At the same time, the prevalence of the other markers of infection has decreased, but there has been an increase in the percentage of subjects with markers of a successful vaccination. It has been suggested that increasing immigration from countries in which this virus is highly endemic is changing the epidemiology of HBV infection. The aim of this study was to assess the prevalence of the serological markers of HBV in Italian and non-Italian HBsAg-negative subjects., Materials and Methods: In the years 2007-2008, 8,018 samples from HBsAg-negative subjects (7,521 Italians and 497 non-Italians) were received for detection of anti-HBs and/or anti-HBc. The findings in the 1,358 samples from candidate blood donors were compared with those obtained in 1991 and 1999., Results: The rate of anti-HBc positivity was 18.3% in the Italian samples and 32.8% in the non-Italian samples; the corresponding percentages of anti-HBs/anti-HBc positive samples (indicating past infection), anti-HBs positive only samples (vaccination) and anti-HBc positive only were, 11.3% vs. 22.5%, 25.8% vs. 17.2%, and 6.9% vs. 9.9% in Italians and non-Italians, respectively. The differences were more marked when stratified by age. In relation to candidate blood donors, simultaneous positivity for anti-HBs and anti-HBc decreased from 11.0% in 1991 to 8.1% in 1999 and 3.9% in 2007-2008, whereas isolated anti-HBs positivity increased from 2.2% in 1991 to 21.4% in 1999 and 42.9% in 2007-2008., Conclusions: The frequency of markers of past infection among Italians has decreased over time as a result of mass vaccination and is significantly lower than that observed in non-Italians. The increasing number of immigrants from countries in which HBV is highly endemic is changing the epidemiology of HBV infection in Italy.

Various countries have implemented anti-rubella vaccination campaigns with the main aim of preventing congenital infection. In 2003, Italy joined the European WHO programme for the elimination of congenital rubella and issued a special healthcare plan, one of the objectives of which was to reduce the proportion of rubella-susceptible pregnant women to less than 5% by 2005. The aim of this study was to determine the percentage of seronegative pregnant women after the implementation of this plan. Anti-rubella IgG and IgM antibodies were sought in 2385 pregnant women who attended our hospital for serological screening between 1 July 2008 and 30 June 2010. They included 750 women of foreign origin (31.4%). Eight percent of the women were anti-rubella seronegative: 6.2% of the Italians and 11.7% of the non-Italians. Among the women of foreign origin, the percentage of seronegativity ranged from 5.6% of those coming from Eastern Europe to 17.7% of those coming from Latin America. The level of seropositivity among women of Italian origin is high, although the objective of ensuring less than 5% of susceptible pregnant women has not yet been quite reached in our area. However, particular attention needs to be given to women coming from geographical areas characterised by different epidemiologies and vaccination strategies because the percentage of seronegativity is in some cases double that of Italian women.

Serological tests for antibodies specific for Epstein-Barr virus (EBV) antigens are frequently used to define infection status and for the differential diagnosis of other pathogens responsible for mononucleosis syndrome. Using only three parameters [viral capsid antigen (VCA) IgG, VCA IgM and EBV nuclear antigen (EBNA)-1 IgG],it is normally possible to distinguish acute from past infection: the presence of VCA IgM and VCA IgG without EBNA-1 IgG indicates acute infection, whereas the presence of VCA IgG and EBNA-1 IgG without VCA IgM is typical of past infection. However, serological findings may sometimes be difficult to interpret as VCA IgG can be present without VCA IgM or EBNA-1 IgG in cases of acute or past infection, or all the three parameters may be detected simultaneously in the case of recent infection or during the course of reactivation. A profile of isolated EBNA-1 IgG may also create some doubts. In order to interpret these patterns correctly, it is necessary to determine IgG avidity, identify anti-EBV IgG and IgM antibodies by immunoblotting, and look for heterophile antibodies, anti-EA (D) antibodies or viral genome using molecular biology methods. These tests make it possible to define the status of the infection and solve any problems that may arise in routine laboratory practice.

Mono thiomalonates (MTMs) are introduced as thioester enolate equivalents. Asymmetric organocatalyzed conjugate addition reactions to nitroolefins proceed under mild conditions to afford synthetically useful γ-nitrothioesters with excellent yields and enantioselectivities.

The distribution of the different subtypes of HIV varies from one region of the world to another. Subtype B is predominant in Europe and the USA, but there has been a gradual increase in non-B subtypes as a result of migration from regions where they are endemic, and this may have important implications for the control of HIV-1. The aim of this study was to assess the prevalence of HIV-1 subtypes in an urban area of northern Italy in the period 1997-2008. Forty-nine (12.2%; 95% CI, 9.00-15.40) of 401 patients investigated carried a non-B subtype, the prevalence of which was 7.7% (95% CI, 4.96-10.44) among native Italians and 55.3% (95% CI, 39.49-71.11) among non-Italians, 1.6% (95% CI, 0.00-3.81) among ex-intravenous drug addicts, 7.6% (95% CI, 1.21-13.99) among homosexual/bisexual men and 20.5% (95% CI, 14.83-26.17) among heterosexuals, 6.8% (95% CI, 3.37-10.23) among Italians infected as a result of sexual contacts in Italy, and 55.0% (95% CI, 33.20-76.80) among Italians infected abroad or by foreign partners. Overall prevalence increased from 2.9% (95% CI, 0.00-6.11) before 1993 to 23.0% (95% CI, 16.31-29.69) in the period 2001-2008. The results demonstrate that there has been an increase in non-B subtypes (especially sexually transmitted infections), particularly among patients infected abroad or by foreign partners., (2010 The Authors. Clinical Microbiology and Infection; 2010 European Society of Clinical Microbiology and Infectious Diseases.)

This study was aimed at verifying the improvement on the motor impairment and functionality in 19 patients with chronic hemiparesis after stroke treated with a robot-aided rehabilitation protocol using the ReoGo™ system (Motorika Medical Ltd, Israel), and at evaluating the persistence of the effects after 1 month. The study also focused on the actual possibility of administering the robot-aided therapy with the ReoGo™ for the upper limbs and on the patients' degree of acceptance and compliance with the treatment. Subjects underwent an assessment prior to the start of the rehabilitation project (T-1), one at the start (T0), one at the end of the treatment (T1) and one after one month from the end of the treatment (T2). The following tests were administered: (i) Fugl-Meyer (FM) upper limb; Ashworth scale (AS); Functional Independence Measure (FIM™) (T-1 - T2); (ii) strength evaluation; Visual Analogue Scale (VAS) for pain; Frenchay Arm test (FAT); Box and Block test (BBT); Timed Up and Go (TUG) test (T0 - T2). Additionally, the Euro-QoL questionnaire and a VAS for the treatment satisfaction were administered to the subjects. Non-statistical difference of scores at T-1 and T0 on almost the entire battery of tasks suggested a stable patients' performance prior to the start of the rehabilitation. With the exception of the Medical Research Council (MRC) and the AS sub-scales measuring -as appropriate- strength and spasticity of the shoulder, triceps and wrist, all scores showed a significant increase between T0 and T1. The improvement on the pain could not be proved significant (p = 0.10). A significant increase between T0 and T2 was found for all assessment scores, with the exception of the MRC for external shoulder rotators (p = 0.05) and of the AS for shoulder (p = 0.32) and wrist (p = 0.08). Substantial stability was observed between T1 and T2. Patients were capable of completing the treatment and showed good participant satisfaction. This pilot study led to the finding of a clinical improvement and excellent patients compliance. It is possible that the learning process experienced by the patients was robot-dependent, especially in consideration of the general maintenance of the achievements observed on all activities.

A search for specific IgM antibodies was used for the detection of primary human cytomegalovirus (HCMV) infections, but the significance of the results is limited by the possible persistence of specific IgM over time, the fact that they are also produced during episodes of reactivation or reinfection, and possible cross-reactions with other viruses. Anti-HCMV antibody screening was carried out to assess the positive predictive value of detecting specific IgM antibodies using IgG and IgM enzyme-linked immunosorbent assays (ELISAs) in 6990 patients examined during the period 2005-2007. In comparison with IgG avidity, the positive predictive value of screening by IgM ELISA alone was 49.3%, which increased to 73% when the presence of IgM was confirmed by an enzyme-linked fluorescent assay (ELFA). The predictive values of highly or weakly positive IgM ELISA alone were respectively 68.8% and 16.4%, but increased to 83.1% and 39.1% if IgM was confirmed by ELFA. The positive predictive value of the IgM/IgG ratio ranged from 26.7% for a low ratio and the detection of IgM by ELISA alone, to 90.7% for a high ratio and ELFA-confirmed IgM detection. These findings indicate that a specimen in which highly positive IgM ELISA values are confirmed by ELFA, or which shows a high IgM/IgG ratio, is a good indicator of infection occurring within the previous three months., (Copyright 2010 Elsevier B.V. All rights reserved.)

The aim of this study was to characterise the AIDS presenters diagnosed between 2000 and 2008 in Legnano (Italy), and describe their initial response to highly active antiretroviral therapy (HAART) and trends over time. Seventy-six (48.7%) of 156 patients diagnosed as having AIDS in the period 2000-2008 were AIDS presenters. The proportion of AIDS presenters increased from 23.8% in 2000 to 70.6% in 2008 (p = 0.009). The major risk factors were heterosexual transmission and a foreign place of birth, and did not significantly change over time. The median CD4+ cell count at diagnosis was 30 cells/microl and the median level of HIV RNA was 5.38 log copies/ml, with no differences between the transmission risk groups. Fifteen AIDS presenters died of AIDS-defining diseases; the others started HAART (72% with 2 NRTIs + boosted PI), and 40% after a drug resistance test. The median duration of the initial HAART was 107 days. After three months, 34% of the patients had undetectable HIV-RNA levels and the median CD4+ cell count was 140 cells/microl; the corresponding figures after 12, 24 and 48 months were respectively 84%, 82.3% and 94.1%, and 310, 370 and 380 cells/microl. In conclusion, the AIDS presenters were mainly heterosexual men and immigrants. Their proportion increased significantly over time, and a substantial proportion maintained an immunovirological response to HAART.

Background: Since 1998, Italian law requires serological screening for toxoplasmosis by the thirteenth week of pregnancy, and seronegative women should undergo further checks every 30 - 40 days until delivery (a total of 5 - 7 screenings). This is an improvement of the previous law which foresaw three free tests (one by the end of the third month, one in the fifth, and one in the ninth month of pregnancy). The aim of this study was to assess the implementation, in an urban area of Northern Italy, of the 1998 law about 10 years after its entry into force., Methods: Of the 4,694 women who initiated and completed a pregnancy in the period 2006 - 2008, we recorded the trimester of pregnancy in which they underwent their first screening, the total and average number of screenings during pregnancy, and the trimester distribution of the screenings., Results: A total of 84.1% of the women underwent their first screening during the first trimester. The negative cases underwent an average of 3.7 screenings during pregnancy, with 34.9% undergoing five or more. Sixty percent of the women underwent at least one screening per trimester., Conclusions: Our data indicates active screening during the first trimester, but fewer screenings than required by law during pregnancy as a whole. Therefore further efforts are needed to improve screening implementation., Keywords: Anti-Toxoplasma antibodies; Congenital infection; Pregnancy trimester; Toxoplasma screening; Seroconversion; Seronegative women; Seropositive women; Screening protocol.

The proposed model is aimed at assessing work-related stress and consists of a preliminary phase during which the Organization is monitored, Indexes and stressors are defined, characterized and then weighted; existing symptoms (if any) are also identified. A 'Probability vs. Severity' Matrix is then built up as a result: these tasks can profitably be performed by a technical professional, typically the Responsible of the Safety and Health Committee. According to found evidences, a second phase, strictly based upon the application of psychosocial research tools, might be needed to investigate group of workers that resulted troublesome during the preliminary phase. The preliminary phase of investigation on organizational stressors and indexes can be easily and successfully integrated with the 'Safety Assessment' steps provided for the B-BS protocol, also aimed at monitoring the organizational wellbeing and consequently acting on the workers' behavior. The model has been specifically designed for Small and Medium Enterprises, with the global objective of preventing accidents at work due to misbehavior and distraction, by correctly and safely applying operational procedures and mutual relationships.

The presence of an "isolated viral capsid antigen (VCA) IgG" pattern in serum is not easy to interpret without the aid of further tests, such as specific immunoblotting or a virus genome search, that often give rise to organisational and economic problems. However, one alternative is to use an enzyme-linked immunosorbent assay (ELISA) to detect anti-early antigen (EA) antibodies, which can be found in about 85% of subjects with acute Epstein-Barr virus (EBV) infections. The purpose of this work was to search for anti-EA(D) antibodies in 130 samples with an isolated VCA IgG pattern at ELISA screening and classified as being indicative of past (102 cases) or acute (28 cases) infection on the basis of the immunoblotting results. Thirty-seven samples (28.5%) were positive for anti-EA(D), of which 25 (89.3%) had been classified by immunoblotting as indicating acute and 12 (11.8%) past EBV infection. This difference was statistically significant (P < .01). The results of our search for anti-EA(D) antibodies correctly identified nearly 90% of acute (presence) or past EBV infections (absence). When other tests are not available, the search for anti-EA antibodies may therefore be helpful in diagnosing patients with an isolated VCA IgG pattern at screening tests.

Objective: To verify the possibility of administering robot-aided therapy for the upper limbs in patients after stroke; to evaluate patients' degree of acceptance and compliance with the treatment; to establish if the treatment has an effect on motor impairment and functional outcome., Design: Quasi-experimental, uncontrolled study., Subjects: Fourteen patients with chronic hemiparesis after stroke., Methods: Patients were treated with a robotic system for the upper limbs (ReoGo; Motorika Medical Ltd, Israel). Subjects performed the following assessment, at the start (T0), at the end of treatment (T1), and at the follow-up performed one month after the end of treatment (T2): Fugl-Meyer test (FM) for upper limbs; strength evaluation; Ashworth scale; visual analogue scale (VAS) for pain; Frenchay Arm test (FAT); Box and Block test (B & B); Functional Independence Measure (FIM); ABILHAND Questionnaire; Timed Up and Go test (TUG); Euro-Quality of Life questionnaire and; a VAS for treatment satisfaction were administered to the subjects., Results: Total scores of FM, B&B, FAT and FIM showed a statistically significant improvement from T0 and T1 (FM p < 0.002, B&B p < 0.012, FAT p < 0.023, FIM p < 0.007) and from T0 and T2 (FM p < 0.003, B&B p < 0.011, FAT p < 0.024, FIM p < 0.027). No statistically significant differences were found between evaluations at T1 and T2 (FM p < 0.595, B&B p < 0.491, FAT p < 0.317, FIM p < 0.180)., Conclusion: The sample was capable of completing the treatment and demonstrated good participant satisfaction. This pilot study led to the finding of a clinical improvement and excellent patient compliance. It can be hypothesized that the results are robot-dependent and that they were learned and then maintained. However, the study is limited in that a control group was not used. As such, it is desirable to continue this study with a control group, as well as by designing a prospective longitudinal randomized controlled trial study.

Objective: To determine the genetic cause of primary amenorrhea in a 46,XY woman., Design: Case report., Setting: Centre of Gynecological Endocrinology and Cytogenetics and Molecular Genetics Laboratory of university medical school., Patient(s): A 19-year-old woman referred for primary amenorrhea., Intervention(s): Clinical, endocrinologic, and ultrasonographic investigation and SRY mutation analysis., Main Outcome Measure(s): Hormone profile (LH, FSH, PRL, leptin, E(2), 17alpha-hydroxyprogesterone, 3alpha-androstanediol glucuronide), ultrasonographic evaluation, clinical follow-up., Result(s): A new SRY sporadic mutation due to a single nucleotide insertion at codon 13 position 38 (38-39insA) was found in a 46,XY woman with sex reversal. This mutation determined a frameshift of the reading frame sequence and a protein truncation at codon 16. Clinical and endocrinologic data are reported., Conclusion(s): This is a new rare case of a single nucleotide insertion affecting the SRY gene in 46,XY females with sex reversal. This new mutation should be considered in genetic counseling.

The presence of VCA IgG in the absence of VCA IgM and EBNA-1 IgG antibodies makes classifying EBV infection more difficult as this serological picture can be seen in the case of past infection with EBNA-1 IgG loss or non-appearance, or acute infections with the early disappearance or delayed onset of VCA IgM. The aim of this study was to assess the prevalence of this pattern in 2,422 outpatients with suspected EBV infection examined in 2005-2006, and to interpret its significance by means of immunoblotting. One hundred and seventy-seven (7.3%) of the patients were VCA IgG-positive, VCA IgM-negative and EBNA-1 IgG-negative, 15 of whom (8.5%) presented with heterophile antibodies. Analysis by age class showed that the prevalence of isolated VCA IgG ranged from 4.5% in the subjects aged 1-10 years to 9% in those aged >60 years. Immunoblotting allowed 18.9% of the cases to be classified as acute and 81.1% as past infections, the latter being observed in about 37% of the patients aged less than 10 years and in 100% of those aged >30 years. Therefore, in our case series, the presence of isolated VCA IgG was associated usually with past infection, particularly among adults. In children aged less than 10 years, it was associated mainly with acute infection but as past infection may be present in about one-third of such children, this possibility should not be overlooked., ((c) 2008 Wiley-Liss, Inc.)

The fetal consequences of CMV infection make it one of the most serious infections contracted during pregnancy, but the scientific community is divided over the proposed implementation of preventive screening for anti-CMV antibodies. The aim of this study was to assess the incidence and risk of infection during pregnancy in 2817 women who underwent anti-CMV IgG and IgM antibody screening during the period 2005-2007. The prevalence of anti-CMV IgG antibodies was 68.3% (95% CI: 66.6-70.0); the seroconversion rate in the 892 seronegative women was 0.32%; the results of IgG avidity testing revealed an cumulative incidence of 1.4% (95% CI: 0.97-1.83), density incidence of 0.8% (as cases/pregnant woman-trimester) (95% CI: 0.47-1.13), and a risk of infection of 0.5% (95% CI: 0.24-0.76). The screening identified 13 cases of primary infection (84.6% of which occurred in the first trimester of pregnancy). The possibility to identify these cases and consequently to plan appropriate interventions, supports the use of screening during pregnancy, especially in the first trimester when the risk of infection is greater.

Background: Data on self-reported symptoms and/or functional impairments are important in research on work-related musculoskeletal disorders (WRMSDs). In such cases the availability of international standardized questionnaires is extremely important since they permit comparison of studies performed in different Countries., Objectives: Translation into Italian and validation of the Nordic Musculoskeletal Questionnaire (NMQ), a tool which is widely used in studies on WRMSDs in the international scientific literature., Methods: The extended Canadian version of the NMQ was translated into Italian. As per the current guidelines, the cross-cultural adaptation was performed by translation of the items from French, back-translation by independent mother-tongue translators and committee review. The resulting version of the questionnaire underwent pre-testing in 3 independent groups of subjects. The comprehensibility, reliability (internal consistency and reproducibility) and sensitivity were evaluated., Results: After translation/back-translation and review of the items the comprehensibility of the Italian version of the questionnaire was judged good in a group of 40 workers. The internal consistency was evaluated using the Cronbach's Alpha test in the same group and in another 98 engineering workers: the results were on the whole acceptable. The reproducibility, which was tested with Cohen's Kappa test in the 40 workers, was good/excellent. In a preliminary evaluation, performed in 30 outpatients of a of Rehabilitation Service, sensitivity was very good., Conclusions: The results show that the Italian version of the Nordic Musculoskeletal Questionnaire is valid for self-administration and can be applied in 'field" studies on self-reported musculoskeletal symptoms and functional impairments in group of workers.