Search

Background: Acute Infectious Diarrhea (AID) and the short- and long-term complications associated with it are major causes of hospitalization worldwide. In Italy, due to a lack of robust surveillance programs, only limited data has been collected on their prevalence and circulation. This study aims to evaluate the resistance pattern of enteric pathogens and their epidemiological trends over a six-year period., Methods: This cross-sectional retrospective study was conducted from January 2018 to December 2023. Stool samples were analyzed during routine diagnosis with culture methods, syndromic molecular tests, and enzyme immunoassay., Results: Bacteria were the most isolated enteric pathogens (62.2%), followed by fungi (29.0%), viruses (8.2%), and parasites (0.6%). Most bacteria were isolated from outpatients (29.5%) and from patients in the Oncology ward (26.2%). The most prevalent target was EPEC (11.1%), followed by C. difficile toxin A/B-producing strains (8.3%), C. jejuni (2.5%), and S. enterica , (1%.). Norovirus and Candida spp. were the most prevalent in pediatric patients (6.5% and 39.6%, respectively). In the last years, enteric pathogens have been a frequent cause of infections characterized by a problematic resistance to common antimicrobials. In our study, S. enterica showed resistance to amikacin, gentamicin, ampicillin, levofloxacin, and ciprofloxacin. C. jejuni was susceptible to all tested drugs., Conclusion: Timely notification of gastroenteric infections is crucial in identifying potential outbreak sources and ensuring strict adherence to food safety and hygiene practices, so as to protect the most vulnerable populations. The present study offers insights into the epidemiological characteristics and the antibiotic susceptibility of the main enteric AID pathogens in order to implement infection control measures in health care settings.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) persistence in COVID-19 patients could play a key role in the emergence of variants of concern. The rapid intra-host evolution of SARS-CoV-2 may result in an increased transmissibility, immune and therapeutic escape which could be a direct consequence of COVID-19 epidemic currents. In this context, a longitudinal retrospective study on eight consecutive COVID-19 patients with persistent SARS-CoV-2 infection, from January 2022 to March 2023, was conducted. To characterize the intra- and inter-host viral evolution, whole genome sequencing and phylogenetic analysis were performed on nasopharyngeal samples collected at different time points. Phylogenetic reconstruction revealed an accelerated SARS-CoV-2 intra-host evolution and emergence of antigenically divergent variants. The Bayesian inference and principal coordinate analysis analysis showed a host-based genomic structuring among antigenically divergent variants, that might reflect the positive effect of containment practices, within the critical hospital area. All longitudinal antigenically divergent isolates shared a wide range of amino acidic (aa) changes, particularly in the Spike (S) glycoprotein, that increased viral transmissibility (K417N, S477N, N501Y and Q498R), enhanced infectivity (R346T, S373P, R408S, T478K, Q498R, Y505H, D614G, H655Y, N679K and P681H), caused host immune escape (S371L, S375F, T376A, K417N, and K444T/R) and displayed partial or complete resistance to treatments (G339D, R346K/T, S371F/L, S375F, T376A, D405N, N440K, G446S, N460K, E484A, F486V, Q493R, G496S and Q498R). These results suggest that multiple novel variants which emerge in the patient during persistent infection, might spread to another individual and continue to evolve. A pro-active genomic surveillance of persistent SARS-CoV-2 infected patients is recommended to identify genetically divergent lineages before their diffusion., (© 2024 Wiley Periodicals LLC.)

Data on cellular immunity mediators in the early phase of human leishmaniasis are still limited and controversial. In order to mimic the changes of humoral mediators during the early phase of human natural infection, some Th1, Th2, Treg, and Breg cytokines, MCP-1, and the nitric oxide (NO) from human PBMC, stimulated by Leishmania infantum, Leishmania major, Leishmania donovani and Leishmania tropica infective metacyclic promastigotes, were determined. After 4 h of L. major, L. donovani, and L. tropica challenge, TNFα, IL-1β, IL-6 levels were significantly higher than negative control cultures with saline (SF) instead of Leishmania promastigotes, unlike L. infantum-stimulated TNFα and L. major-stimulated IL-1β. We obtained higher levels of IL-4 and IL-10 cytokines after stimulation of human PBMCs by L. infantum and L. donovani, compared to those observed after the challenge of PBMCs by L. major and L. tropica. Regarding IL-35, such cytokine levels were significantly increased following infection with L. infantum and L. donovani, in contrast to L. major and L. tropica. Up to our knowledge, we are the first to study the effect of four different species of Leishmania on IL-35 levels in human cells. Our study highlights how several Leishmania species can up-regulate different groups of cytokines (Th1, Th2, Treg and Breg) and modulate NO release in a different way. This original aspect can be explained by different Leishmania cell products, such as LPG, obtained from different strains/species of live parasites. Our findings would contribute to the development of new therapeutics or vaccination strategies., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2023. Published by Elsevier B.V.)

Background: A prompt set of suitable biomarkers is needed in suspected COVID-19 patients. This study aims to assess patients positive for one or more gene associated with the C reactive protein (CRP) and procalcitonin (PCT) as non-specific pro-inflammatory markers and IgG and IgM kinetic as specific diagnostic and prognostic tools in SARS-CoV-2 RT-PCR positive patients., Methods: We enrolled 101 patients within a two month time span (March 26 th , 2020 to May 31 st , 2020). A reverse transcription-Real-Time PCR assay on nasopharyngeal/oropharyngeal swabs was used for SARS-CoV-2 identification. Serum anti-SARS-CoV-2 IgM and IgG were measured by enzyme immunoassay, PCT levels by Enzyme linked fluorescent assay (ELFA)and CRP by nephelometry., Results: We found that older patients were significantly associated with a worse prognosis. Serum IgM levels were significantly lower during the late stage of the disease, regardless of the presence of one or three genes and patients' outcome. On the contrary, IgG levels exhibited a higher concentration in the late phases of the illness, regardless of the gene found or patients' prognosis. With the exception of the very first sample tested, an increase in CRP in surviving patients (both one and three genes) and a time-dependent decrease of deceased patients CRP was found. PCT levels were always within the normal reference range. The difference between one gene and three genes patients was significant during late disease stages regarding IgG levels and also between three genes survivors versus three genes deceased, where the IgG levels were progressively increasing over time., Conclusions: The relevant finding of the present study is the significant and consistent increase of IgG and IgM in deceased patients. The associated evaluation of antibody kinetics and non specific inflammatory markers (CRP and PCT) in positive patients stratified according to the presence of one gene or three genes could help the clinician in both the diagnosis and prognosis of COVID-19 patients.

The host response to helminth infections is characterized by systemic and tissue-related immune responses that play a crucial role in pathological diseases. Recently, experimental studies have highlighted the role of regulatory T (Tregs) and B (Bregs) cells with secreted cytokines as important markers in anti-schistosomiasis immunity. We investigated the serical levels of five cytokines (TNFα, IFN-γ, IL-4, IL-10 and IL-35) in pre- and post-treatment samples from chronic Schistosoma infected patients to identify potential serological markers during follow-up therapy. Interestingly, we highlighted an increased serum level of IL-35 in the pre-therapy samples (median 439 pg/mL for Schistosoma haematobium and 100.5 pg/mL for Schistsoma mansoni infected patients) compared to a control group (median 62 pg/mL and 58 pg/mL, respectively, p ≤ 0.05), and a significantly lower concentration in post-therapy samples (181 pg/mL for S. haematobium and 49.5 pg/mL for S. mansoni infected patients, p ≤ 0.05). The present study suggests the possible role of IL-35 as a novel serological biomarker in the evaluation of Schistosoma therapy follow-up.

Leishmaniasis is a vector-borne disease caused by obligate intracellular protozoan parasites that can infect humans and other mammals. Pro- and anti-inflammatory cytokines are important regulators of innate and specific responses in Leishmania infection. Resistance to leishmaniasis is related to T helper 1 (Th1) response with the production of pro-inflammatory cytokines: IL-12, IL-1β, IFN-γ, TNF-α, IL-2 leading to activation of macrophages and parasite killing. Instead, a more intense Th2 (IL-4, IL-5, IL-13), Treg (IL-10 and TGF-β) and Breg response (IL-10 and IL-35) are related to parasite persistence through the inhibition of macrophage activation, which promotes the escape from host immune system. Interestingly, a cytokine involved in the parasite killing in one form of leishmaniasis may be "pathogen friendly" in another form of the disease. To date, few studies are focusing on the role of Treg and Breg cytokines in human models of leishmaniasis; therefore, further investigations are needed to clarify their potential role in the diagnosis and prognosis of such protozoan infections, as well as in the development of vaccines against leishmaniasis. This review summarizes the current knowledge about the role of cytokines produced by Th1, Th2, Treg, and Breg cells involved in Leishmania disease progression and host protection. Some cytokines might play a role as diagnostic and prognostic clinical markers, or they could represent a novel approach leading to new anti-leishmaniasis therapies. Overall, advances in knowledge of the complex network of cytokines secreted by immune cells could help to better understand signaling pathways and host immune responses during Leishmania infection. This approach would allow these mediators to be used as therapeutic strategies against leishmaniasis., Competing Interests: Declaration of Competing Interest The authors have no conflicts of interest to disclose., (Copyright © 2023. Published by Elsevier B.V.)

Background: General population data on hepatitis C virus (HCV) prevalence in Italy come mostly from studies conducted in small towns. The highest rates have consistently been found in southern regions, especially in Calabria. Herein, we aimed to determine HCV prevalence, awareness, and risk factors in the general population of Catanzaro, the capital city of Calabria, Italy., Methods: A stratified probability-based random sample of adult population was drawn from the Census. Anti-HCV and HCV-RNA were assayed. Data on sociodemographycs, risk factors and awareness of infection status were also collected. Crude and age and sex directly standardized rates (DSR), using Catanzaro's general population as standard, were calculated. Log binomial regressions with sampling weights was used to identify independent predictors of infection., Results: The final study population consisted of 1003 people. Of them 27 (2.69%; 95% confidence interval, [CI] 1.78-3.89) (DSR: 2.34%; 95% CI: 1.37-3.30) and 9 (0.9%; 95% CI: 0.41-1.70) (DSR: 0.79%; 95% CI: 0.21-1.37) were anti-HCV and HCV RNA positive, respectively. Most HCV-positive participants were older people. Age ≥65 and past use of illicit drugs were both positive independent predictors of anti-HCV positivity, while female sex was an independent protective predictor of infection. Only 9 (33.3%) of anti-HCV positive participants had awareness of their status., Conclusions: We detected a much lower anti-HCV prevalence than those previously found in Calabria, along with a substantial change in HCV transmission modes. Infected people were almost only elderly and mostly unaware of their infection. Improving diagnosis and linkage to care for these infected persons would be needed.

Reverse Transcription Polymerase Chain Reaction (RT-PCR) conducted on nasopharyngeal swabs is the gold standard in the diagnosis of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2). In Italy, recent guidelines indicate that rapid antigen tests (RATs) can be used for the isolation of positive patients or for the interruption of quarantine, but they are often less sensitive to detect positive subjects. Indeed, the performance of these RATs depends on the timing and the population on which they are evaluated. Herein, we evaluated the performance of BIOCREDIT COVID-19 Ag and Fluorecare ® SARS-CoV-2 Spike Protein Test during a population screening in the Calabria Region, Southern Italy. We report that both antigen test shows low sensitivity in contrast to the high sensitivity declared by manufacturer (90% and 92%, respectively) and that the area under the curve (AUC) was good for Fluorecare ® SARS-CoV- 2 Spike Protein Test but very poor for BIOCREDIT COVID-19 Ag. We suggest that these RATs should be re-evaluated in the current pandemic era., Competing Interests: Conflict interest All authors declared no conflict of interest., (Copyright © 2016 - 2022 InfezMed.)

In this study, we report on the results of SARS-CoV-2 surveillance performed in an area of Southern Italy for 12 months (from March 2021 to February 2022). To this study, we have sequenced RNA from 609 isolates. We have identified circulating VOCs by Sanger sequencing of the S gene and defined their genotypes by whole-genome NGS sequencing of 157 representative isolates. Our results indicated that B.1 and Alpha were the only circulating lineages in Calabria in March 2021; while Alpha remained the most common variant between April 2021 and May 2021 (90 and 73%, respectively), we observed a concomitant decrease in B.1 cases and appearance of Gamma cases (6 and 21%, respectively); C.36.3 and Delta appeared in June 2021 (6 and 3%, respectively); Delta became dominant in July 2021 while Alpha continued to reduce (46 and 48%, respectively). In August 2021, Delta became the only circulating variant until the end of December 2021. As of January 2022, Omicron emerged and took over Delta (72 and 28%, respectively). No patient carrying Beta, Iota, Mu, or Eta variants was identified in this survey. Among the genomes identified in this study, some were distributed all over Europe (B1_S477N, Alpha_L5F, Delta_T95, Delta_G181V, and Delta_A222V), some were distributed in the majority of Italian regions (B1_S477N, B1_Q675H, Delta_T95I and Delta_A222V), and some were present mainly in Calabria (B1_S477N_T29I, B1_S477N_T29I_E484Q, Alpha_A67S, Alpha_A701S, and Alpha_T724I). Prediction analysis of the effects of mutations on the immune response (i.e., binding to class I MHC and/or recognition of T cells) indicated that T29I in B.1 variant; A701S in Alpha variant; and T19R in Delta variant were predicted to impair binding to class I MHC whereas the mutations A67S identified in Alpha; E484K identified in Gamma; and E156G and ΔF157/R158 identified in Delta were predicted to impair recognition by T cells. In conclusion, we report on the results of SARS-CoV-2 surveillance in Regione Calabria in the period between March 2021 and February 2022, identified variants that were enriched mainly in Calabria, and predicted the effects of identified mutations on host immune response., 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., (Copyright © 2022 De Marco, Veneziano, Massacci, Pallocca, Marascio, Quirino, Barreca, Giancotti, Gallo, Lamberti, Quaresima, Santamaria, Biamonte, Scicchitano, Trecarichi, Russo, Torella, Quattrone, Torti, Matera, De Filippo, Costanzo and Viglietto.)

Antimicrobial Susceptibility Testing is mandatory for Bloodstream Infections management in order to establish appropriate antimicrobial therapy. Herein we evaluated new approach based on AST results directly from positive blood cultures, using Microscan WA to carry out rapid phenotypical profile of antibiotic resistance. Our investigations allow to reduce time versus traditional results., Competing Interests: Declarations of competing interest There are no conflicts of interest., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Hepatitis C virus subtype 1b (HCV1b) is still the most prevalent subtype worldwide, with massive expansion due to poor health care standards, such as blood transfusion and iatrogenic procedures. Despite safe and effective new direct antiviral agents (DAA), treatment success can depend on resistance-associated substitutions (RASs) carried in target genomic regions. Herein we investigated transmission clusters and RASs among isolates from HCV1b positive subjects in the Calabria Region. Forty-one NS5B and twenty-two NS5A sequences were obtained by Sanger sequencing. Phylogenetic analysis was performed using the maximum likelihood method and resistance substitutions were analyzed with the Geno2pheno tool. Phylogenetic analysis showed sixteen statistically supported clusters, with twelve containing Italian sequences mixed with foreign HCV1b isolates and four monophyletic clusters including only sequences from Calabria. Interestingly, HCV1b spread has been maintained by sporadic infections in geographically limited areas and by dental treatment or surgical intervention in the metropolitan area. The L159F NS5B RAS was found in 15 isolates and in particular 8/15 also showed the C316N substitution. The Y93H and L31M NS5A RASs were detected in three and one isolates, respectively. The A92T NS5A RAS was found in one isolate. Overall, frequencies of detected NS5B and NS5A RASs were 36.6% and 22.7%, respectively. For the eradication of infection, improved screening policies should be considered and the prevalence of natural RASs carried on viral strains.

The activity of the SARS-CoV-2 virus has not yet been studied in a post-mortem setting. The absence of these data has led to the prohibition of exposure of infected corpses during burial procedures. Our aim was to assess the virus's persistence and the possibility of transmission in the post-mortem phase including autopsy staff. The sample group included 29 patients who were admitted to our Covid-19 Centre who died during hospitalisation and the autopsy staff. All the swabs were subjected to a one-step real-time reverse transcription polymerase chain reaction with cycle threshold (Ct) values. Swab collection was performed at 2 h, 4 h, 6 h, 12 h, over 24 since death. The following were the analysis of patients' swabs: 10 cases were positive 2 h after death; 10 cases positive 4 h after death; 9 cases were found positive 6 h after death; 7 cases positive 12 h after death; 9 cases remained positive 24 h after death. The swabs performed on all the forensic pathologist staff on duty who performed the autopsies were negative. The choice to avoid rituals and the display of corpses before and at the burial procedures given appears cautiously valid due to the persistence of the SARS-CoV-2 virus in the post-mortem period. Although the caution in choosing whether or not to perform an autopsy on infected corpses is acceptable, not to perform autopsies is not biologically supported.

Background: In December 2019, a new coronavirus (named severe acute respiratory syndrome coronavirus 2, SARS-CoV-2) spread from China, causing a pandemic in a very short time. The main clinical presentation of SARS-CoV-2 infection (COVID-19, coronavirus disease-2019) is pneumonia, but several cardiovascular complications may also occur (e.g., acute coronary syndromes, pulmonary embolism, stroke, arrhythmias, heart failure and cardiogenic shock). Direct or indirect mechanisms induced by SARS-CoV-2 could be implicated in the pathogenesis of these events., Case Presentation: We report herein the third case of COVID-19 autoimmune haemolytic anaemia (AIHA) reported so far, which occurredwithout any other possible explanations in a Caucasian patient. The patient also suffered from ST-elevation myocardial injury., Conclusions: Both complications occurred quite late after COVID-19 diagnosis and were probably precipitated by systemic inflammation, as indicated by a significant delayed increase in inflammatory markers, including interleukin-6 (IL-6).

Aims: Patients with acute coronary syndrome (ACS) often arrive in the catheterization (cath) lab directly from the field or an emergency department without an accurate triage for Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection.Although in the pandemic period the treatment in the cath laboratory of high-risk ACS should not be delayed because the operators wear special protection systems, the subsequent risk of contagion in a non-Covid coronary care unit could be high in the case of patients positive for SARS-CoV-2., Methods: We tested the possibility of a fast-track protocol in 51 consecutive patients (mean age 65 ± 12 years) transferred from spokes centres or from the field to our HUB centre and admitted to our coronary care unit (CCU). Once the patient had arrived in the cath lab, the nasopharyngeal swab was performed. The real-time PCR to extract RNA for SARS-CoV-2 detection was performed with an automated rapid molecular Xpert Xpress test. Meanwhile, coronary angiography or percutaneous coronary intervention was performed if necessary., Results: In this fast-track protocol, the time to perform nasopharyngeal swab was 11 ± 11 min; time spent to transport nasopharyngeal swab to the laboratory was 29 ± 20 min; time to detect viral nucleic acid was 68 ± 16 min. The overall time from the execution of nasopharyngeal swab to the result was 109 ± 26 min. The results were immediately put into the hospital computer system and made readily available. Depending on the test result, patients were then transferred to the regular CCU or Covid area., Conclusion: This study demonstrates that 0-1.5 h fast-track triage for coronavirus disease 2019 (COVID 19) is feasible in patients with ACS. The execution of nasopharyngeal swab in the cath lab and its analysis with a rapid molecular test allows rapid stratification of SARS-CoV-2 infection.

In patients with influenza-related myocarditis, prompt diagnosis and treatment are important. Intravenous zanamivir can be an alternative to oral oseltamivir, especially in severe cases and when drug intestinal malabsorption is suspected or proven., Competing Interests: The authors declare that they have no competing interests., (© 2019 The Authors. Clinical Case Reports published by John Wiley & Sons Ltd.)

Background: Intravenous (IV) zanamivir could be a suitable alternative for the treatment of severe influenza A(H1N1)pdm09 infection in patients who are unable to take oral or inhaled medication, for example, those on mechanical ventilation and extracorporeal membrane oxygenation (ECMO). However, data on the clinical outcomes of such patients is limited., Case Presentation: We report the clinical outcomes of four patients who were admitted at the intensive care unit during the 2017-2018 influenza season with severe sepsis (SOFA score > 11) and acute respiratory distress syndrome requiring ECMO and mechanical ventilation. Two patients were immune-compromised. The A(H1N1)pdm09 genome was confirmed by polymerase chain reaction (PCR) on nasopharyngeal specimen swabs prior to administration of IV zanamivir at a dose of 600 mg twice daily. Weekly qualitative PCR analysis was done to monitor viral clearance, with zanamivir treatment being discontinued upon receipt of negative results. In addition, the patients were managed for concomitant multidrug-resistant bacterial infections, with infection resolution confirmed with blood cultures. The median time for zanamivir treatment was 10 days (IQR 10-17). The clinical outcome was favourable with all four patients surviving and improving clinically. All four patients achieved viral clearance of A(H1N1)pdm09 genome, and resolution of multidrug-resistant bacterial infections., Conclusions: IV zanamivir could be a good therapeutic option in patients with severe influenza A(H1N1)pdm09 infection who are unable to take oral or aerosolised antiviral medication. We recommend prospective randomized control trials to support this hypothesis.

Direct-acting antiviral drugs to cure infections with Hepatitis C virus (HCV) achieve a sustained virological response (SVR) in more than 90% of adult patients. At present, clinical trials are ongoing and real-life data are still limited in children. Herein, we report two cases of pediatric patients treated with fixed-dose combination of sofosbuvir/ledipasvir, already approved to treat HCV4 genotype. Both young girls achieved SVR even though HCV4 isolates carried L28M and M31L NS5A resistance-associated substitutions (RASs). Therefore, possible effects of these RASs merit further study, especially in children.

Sequence analysis of HPV16 isolates reveals the presence of genome variants with characteristic mutations. The HPV16 variants have different geographical distribution and diverge into four phylogenetic lineages (A, B, C and D) and 16 sub-lineages: A1, A2, A3 (previously known as European variants), A4 (Asian variant), B1, B2, B3, B4, C1, C2, C3, and C4 (African variants), D1 (North-American variant), D2, D3 (Asian-American variants) and D4. Population studies showed that infections with viruses belonging to specific HPV16 sublineages confer different risks of viral persistence and cancer. In this study, 39 HPV16-positive cervical smears from European women living in Calabria (Italy) were analyzed for the presence of HPV16 variants. Cervical DNA extracts were processed by PCR to amplify L1, the Long Control Region (LCR), E6 and E7, which were sequenced. The sequences were concatenated and the 3169 nucleotides long fragments were characterized by BLAST and phylogenetic analysis. A total of 96 Single Nucleotide Polymorphism (SNPs) were detected, 29 of which mapping in the L1, 45 in the LCR, 15 in the E6 and 7 in the E7. The most common SNP was the T350G (29/39 samples, 74.4%), causing the L83 V amino acid change in the E6. Most of the HPV16 isolates (89.7%) had 99% of nucleotide (nt) identity to members of the A1 and A2 sublineages, while 4 isolates had 99% nt identity to members of the B2, B4, C1 and D4 sublineages. In conclusion, viruses belonging to the A1, A2, B2, B4, C1 and D4 HPV16 sublineages were found to circulate in the Calabria region., (Copyright © 2018. Published by Elsevier B.V.)

Hepatitis C virus (HCV) infects around 71 million people worldwide and in 2018 it is still a major health problem. Since 2011, anti-HCV therapy with availability of direct-acting antiviral drugs has revolutionized the clinical response and paved the way to eradication strategies. However, despite the high rate of sustained virological response, treatment failure may occur in a limited percentage of patients, possibly due to resistance-associated substitutions (RASs), either emergent or pre-existent even in minority viral populations. Clearly this problem may impair success of eradication strategies. With this background, several questions marks still exist around HCV treatment, including whether pan-genotypic treatments with complete effectiveness in any clinical conditions really exist outside clinical trials, the actual cost-effectiveness of genotyping testing, and utility of RAS detection in viral quasispecies by next generation sequencing approach. In this review, we describe these critical points by discussing recent literature data and our research experience.

In recent years, migration has become a significant challenge in Western countries. Migrant populations, coming from hyper-endemic areas, may present parasitic infections that remain latent and asymptomatic even for years, eventually leading to severe complications. Italian guidelines have been established to perform screening guided by the presence of symptoms and/or hypereosinophilia. Parasitological screening was conducted in a migrant population to carry out preventative measures. All migrants were asked to report any symptoms suggesting parasitic infections and list any previous treatment received. Travel data were recorded. Parasitological examination of stools and urine were conducted in all patients regardless of symptoms. In all, 208 consecutive patients were enrolled in our outpatient clinic from November 2016 to August 2017. Thirty-four patients were excluded due to the previous assumption of albendazole or because they did not exhibit suitable samples. Prevalence of parasitic infections was 33/174 (18.9%). A statistically significant difference for the prevalence of parasitic infections was not found between patients who were asymptomatic and without hypereosinophilia compared to those who presented symptoms and/or hypereosinophilia (27/151 [17.9%] vs. 6/23 [26.0%]; p=0.39). By contrast, a statistically significant difference was found for the length of time between arrival in Italy and parasitological examinations (4/51 [7.8%] migrants who arrived in Italy more than six months prior to screening vs. 29/123 [23.6%] migrants who arrived within six months; p= 0.016). Our results did not demonstrate any significant differences in prevalence of parasitic infections between symptomatic or hypereosinophilic and asymptomatic migrants. Thus we feel it inappropriate to support recent guidelines recommending parasitological examinations only in migrants with symptoms and/or hypereosinophilia. By contrast, it would appear important to perform parasitological screening in migrants as soon as possible after their arrival. Since such infestations, if untreated, could result in chronic diseases and complications, and could be transmitted in the host countries, our results have potential implications for public health.

Skin leishmaniasis includes lesions of different appearance, shape, and severity, spanning from alarming diffuse lesions to an asymptomatic course. Moreover, aspecific presentation, as well as challenging differential diagnosis of cutaneous leishmaniasis, may request more in-depth investigations on the intriguing and complex pathogenesis of such infection. A 7-year case of worsening cutaneous leishmaniasis in the left frontoparietal region of the scalp, achieving omolateral eyebrow, in a 68-year-old male patient prompted us to address the immunity profile of peripheral blood lymphocytes. An increase of regulatory CD19 + /CD38 bright /CD24 bright B cell lymphocytes was observed at the front of normal levels of other lymphocytes subpopulations, including CD4 + /CD25 bright T cells. The total IgG and IgM, as well as proinflammatory subclasses of IgG, were below the normal range. However, IgG4 subclass was found normal. In conclusion, our data may indicate inhibition of humoral immunity associated with an increase of lymphocyte B-regulatory subpopulation., (Copyright © 2018 Elsevier Inc. All rights reserved.)

We report a real-life 3D therapy failure in a patient treated with ombitasvir (OMV)/paritaprevir/ritonavir and dasabuvir without ribavirin (3D-R). He had therapy failure at week 12 after the end of treatment. We detected resistance-associated substitutions (RASs) plus polymorphisms on NS3, NS5A, and NS5B target regions by population sequencing (15% cut-off) at baseline, at relapse and during follow-up. About this, NS5A RASs generally persist longer than resistances in the other target genes and may impact treatment outcome. Therefore, to evaluate OMV drug-resistance mechanism, we studied the acquired RAS plus polymorphisms on NS5A phosphoprotein by computational studies. OMV showed a higher affinity towards baseline and 93H/108 K mutant structure (follow-up) with respect to 93H/R108 mutant structure (relapse) on phosphoprotein. By Molecular Dynamics simulations (MDs), structural information about the protein stability in presence of OMV were observed. According to our data, molecular modeling approach has proved to be a powerful method to evaluate the impact of these RASs plus specific amino acid (AA) changes on phosphoprotein., (© 2018 Wiley Periodicals, Inc.)

Background/aims: Correct renal function evaluation is based on estimated glomerular filtration rates (eGFR) and complementary renal damage biomarkers, such as neutrophil gelatinase associated lipocalin (NGAL). The aim of this study was to evaluate eGFR and NGAL modifications and renal impairment during treatment with a direct acting antiviral (DAA) for chronic hepatitis C virus (HCV) infection., Methods: A retrospective cohort study evaluated eGFR modification during treatment with DAA. Subgroup analysis on serum NGAL was conducted in those receiving sofosbuvir/ledipasvir, with complete follow-up until week 12 after the end of treatment (FU-12)., Results: In the 102 enrolled patients, eGFR reduction was observed (from 86.22 mL/min at baseline to 84.43 mL/min at FU-12, P=0.049). Mean NGAL increased in 18 patients (from 121.89 ng/mL at baseline to 204.13 ng/mL at FU-12, P=0.014). At FU-12, 38.8% (7/18) of patients had a plasmatic NGAL value higher than the normal range (36-203 ng/mL) compared with 11.1% (2/18) at baseline (χ 2 =3,704; P=0.054). In contrast, eGFR did not change significantly over the follow-up in this subgroup., Conclusions: In conclusion, compared to a negligible eGFR decline observed in the entire cohort analyzed, a significant NGAL increase was observed after HCV treatment with DAA in a small subgroup. This could reflect tubular damage during DAA treatment rather than glomerular injury.

Background: Bartonella species are intracellular bacteria capable of producing several diseases in humans. The three most common and wellknown diseases are cat scratch disease (CSD), caused by B. henselae, trench fever, caused by B. quintana and Carrion's Disease, caused by B. bacilliformis . Signs and symptoms are very different and aspecific: Fatigue, fever, headache, lymphadenopathy, malaise, loss of weight. No data exist to support guidelines' recommendations to decide which drugs should be optimally used and how long they should be administered. Therefore, a marker of treatment response is needed to guide treatment strategies., Methods: We report herein three cases in which a species specific Reverse-Transcriptase Polymerase-Chain-Reaction (RT PCR) developed in-house was performed and compared to serology in order to make diagnosis and to evaluate treatment response., Results: Our species-specific RT PCR seemed to play a fundamental role both in diagnosis and treatment. Moreover, a discrepancy with the serology results was found., Conclusion: Further studies are necessary to validate these results and elucidate what is the best treatment for this pleomorphic disease. However, in absence of clear guidelines, RT PCR may be useful to orientate kind of treatment ad its duration.

We aim to investigate some of the pathogenetic mediators of the human echinococcosis and to obtain updated epidemiological findings on cases of echinococcosis in Calabria, Southern Italy. Echinococcosis diagnosis was based on imaging, serological investigations, and molecular assay. Indeed, real-time PCR indicated the presence of G2/G3 genotypes of Echinococcus granulosus complex. Regarding pathogenesis, a relevant novel tool of immune depression should be deemed the reduced level of serum MCP-1. Also, we found a previously unreported VEGF, possibly associated with neovascularization requested by the parasite cyst metabolism. Cytokine profiles suggest a bias of the immunity toward Th2 and Treg responses. Nitric oxide levels exhibited a significant decrease one week after therapy versus basal level measured before surgery and/or chemotherapy. An increase of serum total IgE class and IgG4 subclass was found in Echinococcus -positive patients versus controls. Our data demonstrated an endemic spreading, at least in the province of Catanzaro and neighboring Calabria territories, for such parasitosis with the novel issue of the number of female overcoming male cases. In conclusion, the novel findings of this study were the increased VEGF and the reduced serum MCP-1 in the studied cases, as well as the number of Echinococcus -infected females overcoming the infected males.

Background: Although analysis of the Human papillomavirus (HPV) genotype spread in a particular area has a crucial impact on public health and prevention programmes, there is a lack of epidemiological data regarding HPV in the Calabria region of Italy. We therefore update information on HPV age/genotype distribution by retrospectively analysing a cohort of women, with and without cervical lesions, living in Calabria, who underwent HPV DNA testing; moreover, we also evaluated HPV age/genotype distribution in a subset of patients with cervical lesions., Methods: Cervical scrape specimens obtained from 9590 women (age range 20-75 years) from January 2010 to December 2015 were tested for HPV DNA. Viral types were genotyped by Linear Array HPV Genotyping® test (Roche, USA) at the Clinical Microbiology Operative Unit of six hospitals located in four provinces of the Calabria region. Cervical scrape specimens were also used to perform Pap smears for cytological analysis in a subset of 405 women; cytological classification of the samples was performed according to the Bethesda classification system., Results: A total of 2974 women (31%) (C.I. 95% 30.09-31.94) were found to be HPV DNA positive for at least one (57.3%) or several (42.7%) HPV genotypes. Of single genotype HPV infections, 46.5% and 36.4 % were classed as high-risk (HR, Group 1) and low-risk (LR, Group 3) respectively, while 16.9% were classed as probably/possibly carcinogenic and 0.2% undetermined risk. Stratified by age, total HPV distribution, showed the highest prevalence within the range 30-39 years (37.2%), while single genotype infection distribution displayed a peak in women from the age range 20-29 years (37.5%). The most common high-risk HPV type was HPV 16 (19.1%), followed by HPV 31 (9.1%)., Conclusions: We provide epidemiological data on HPV age/genotype distribution in women living in the Calabria region with or without cytological abnormalities, further to the enhancement of HPV screening/prevention programmes for the local population.

Background: We evaluated the role of presepsin (soluble CD14 subtype, sCD14-ST) in predicting the outcome of critically ill septic patients in parallel with procalcitonin and C-reactive protein., Methods: This study was an observational, prospective study that enrolled 58 surgical and medical intensive care unit patients with suspected sepsis. All studied subjects were retrospectively stratified into survivors and nonsurvivors based on 28 days survival and according to microbiological results in blood culture positive and negative groups. Plasma and serum samples from each patient were collected at admission (T-0), after 24-48 hours (T-1) and after 7 days (T-2). Statistics were obtained using Student׳s t test and ANOVA, as well as Bonferroni post hoc test. Receiver-operating characteristic (ROC) analysis was also performed., Results: Presepsin levels were significantly higher at T-0 (P = 0.0007), at T-1 (P < 0.0001) and at T-2 (P < 0.0001) in nonsurvivors versus survivors at the same time point. Presepsin concentrations were significantly increased at T-0 (P = 0.0073), T1 (P = 0.0111) and T2 (P = 0.0167) in patients with positive blood cultures in comparison to patients with negative cultures at the same time. For all time periods evaluated, presepsin data from nonsurviving and surviving individuals were subjected to ROC analysis that demonstrated an excellent accuracy and significant area under the ROC curve (P < 0.0001). Results of multivariate analysis indicated presepsin as a predictive independent variable among prognosis markers at T-0 (P = 0.016)., Conclusions: Presepsin revealed an optimal prognostic performance in patients with severe sepsis and provided interesting diagnostic value. Prediction of outcome in critically ill patients is crucial to optimize management decisions and level of treatment., (Copyright © 2017 Southern Society for Clinical Investigation. Published by Elsevier Inc. All rights reserved.)

Naturally occurring resistance-associated substitutions (RASs) can negatively impact the response to direct-acting antivirals (DAAs) agents-based therapies for hepatitis C virus (HCV) infection. Herein, we set out to characterize the RASs in the HCV1b genome from serum samples of DAA-naïve patients in the context of the SINERGIE (South Italian Network for Rational Guidelines and International Epidemiology, 2014) project. We deep-sequenced the NS3/4A protease region of the viral population using the Ion Torrent Personal Genome Machine, and patient-specific majority rule consensus sequence summaries were constructed with a combination of freely available next generation sequencing data analysis software. We detected NS3/4A protease major and minor variants associated with resistance to boceprevir (V36L), telaprevir (V36L, I132V), simeprevir (V36L), and grazoprevir (V36L, V170I). Furthermore, we sequenced part of HCV NS5B polymerase using Sanger-sequencing and detected a natural RAS for dasabuvir (C316N). This mutation could be important for treatment strategies in cases of previous therapy failure., Competing Interests: The authors declare no conflict of interest.

Aim: To evaluate neutrophil gelatinase associated lipocalin (NGAL) in patients infected by hepatitis C virus (HCV) before and during treatment with directly acting antivirals (DAAs)., Methods: NGAL was measured in a group of patients with chronic HCV infection ranked, at baseline, by age, gender, anti-hypertensive therapy, HCV viral load, liver fibrosis stage and, either at baseline or after 1 year, estimated glomerular filtration rate (eGFR). Then, NGAL and eGFR evolutions were monitored in a subgroup of patients who started antiviral therapy with DAAs. Differences of median NGAL levels were evaluated through Wilcoxon-Mann-Whitney test for non-parametric data. Differences in dichotomous variables were evaluated through χ (2) test. At baseline, a univariate regression analysis was conducted to verify if NGAL values correlated with other quantitative variables [age, fibrosis four (FIB-4), AST to platelet ratio index (APRI), and eGFR]., Results: Overall, 48 patients were enrolled, 8 of them starting HCV treatment. At baseline, statistically significant differences were found in median NGAL values only between patients with eGFR < 60 mL/min vs patients with eGFR ≥ 90 mL/min. Differences in NGAL were not significant among patients ranked by HCV viral load, FIB-4 score and APRI, when patients with NGAL > 118.11 ng/dL were compared with those of NGAL ≤ 118.11 ng/dL, not statistically significant differences were present for age, gender, chronic kidney disease classification and liver fibrosis (P > 0.05). Linear correlation was found between NGAL and both age (P = 0.0475) and eGFR (P = 0.0282) values. Not statistically significant predictions of NGAL at baseline were demonstrated for eGFR evolution 1 year later. Interestingly, in the 8 patients treated with DAAs, median NGAL significantly increased at week 12 compared to baseline (P = 0.0239)., Conclusion: Our results suggest that NGAL should be further evaluated as an adjunct marker of kidney function in these patients.

Nasopharyngeal carcinoma (NPC) is a rare neoplasm which is associated with Epstein-Barr virus (EBV). First of all, we reviewed the literature on NPC treatment. Radio/chemotherapy is currently the gold standard but unfortunately is affected by rates of failure ranging from 7% up to 58%. Because NPC development is promoted by the EBV latent life cycle, EBV-targeted treatments were investigated. Firstly, forcing cytolytic virus activation through administration of gemcitabine and/or valproic acid before administration of a nucleoside analogue showed anti-tumoral activity in vitro as well as in murine model and it was also well tolerated in humans. Secondly, the association of autologous EBV-specific cytotoxic T lymphocytes with chemotherapy correlated with an improved median survival and was safe but not effective versus metastatic lesions. Thirdly, suppression of late membrane protein-1 in the clinic proved controversial because it gave resistance to chemotherapy and, on the other hand, increased radiosensitivity. Finally, we suggest future perspectives for clinical research which should include both prospective and observational cohort studies to assess the role of different risk factors in the development of NPC and the effectiveness of new investigational treatments.

We report herein the first case of a coinfection with Brucella spp., M. bovis, and Enterobacter cloacae in a butcher who moved from Bulgaria to Italy. Molecular typing suggested professional acquisition of M. bovis in Italy. So, surveillance and preventive measures need to be implemented.

Circulation of HCV genotype 2 has been described in European Countries where numerous subtypes and unclassified HCV 2 lineages have been reported. In Italy, subtype 1b is the most prevalent, followed by genotype 2. In the present study, phylogeny of HCV 2c was investigated. The phylogeny of HCV 2c isolated from 54 Italian patients in the Calabria region (Southern Italy) was investigated by analyzing a fragment of the NS5B gene. Patients came from 5 metropolitan areas and a small village (Sersale). These areas were geographically dispersed throughout the entire region. A Bayesian coalescent-based framework was used to estimate origin and spreading of HCV 2c in this region. Phylogenetic analysis showed that 28 Italian sequences were intermixed with foreign HCV 2c reference sequences and grouped into 3 major clades: A, B, and C. Nineteen inter-clade sequences were associated uniquely with surgery as risk factor for HCV acquisition. By contrast, a sub-cluster within clade B was associated with blood transfusion. Moreover, sequences from Sersale village grouped in the Italian sub-cluster and were intermixed with 10 sequences from metropolitan areas. The three isolates with the longest branch came from Sersale and belonged to patients who had glass syringes as risk factor. HCV 2c isolates from the Calabria region shared a common ancestor whose origin was traced back to 1889. Our results suggest that, after its introduction - possibly as a result of population movements between Italy and African Countries during Italian colonialism - HCV 2c spread through multiple risk factors, not including intravenous drug use. So, transmission chains followed a pathway different from other European Countries. Although HCV incidence is decreasing, these ways are still ongoing, possibly justifying stability in the relative prevalence of HCV 2c., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Late/chronic Lyme neuroborreliosis (LNB) represents a challenging entity whose diagnosis requires a combination of clinical and laboratory findings, surrounded by much controversy. Here we describe a patient who had a peculiar form of late LNB with CNS lesions shown by magnetic resonance imaging (MRI), and epileptic seizures, etiologically diagnosed by conventional and molecular methods. The current case provides evidence that patients presenting with epileptic seizures and MRI-detected multifocal lesions, particularly when a facial palsy has also occurred, should raise the suspicion of LNB, as this diagnosis has important implications for treatment and prognosis.

Background: In Italy, anti-HCV drugs are provided free of charge by the National Health System. Since 2011, three drug regimens including a directly acting antiviral (DAA) are considered the gold standard for HCV treatment. However, these drugs add a significant cost (roughly €26,000) to the combination of pegylated-interferon-α/ribavirin (PEG-IFN/RBV), which before DAA represented the unique treatment. To provide the National Health System potential useful information, we estimated costs to provide anti-HCV drugs to treat a population experienced for PEG-INF/RBV., Methods: Genotype 1 HCV mono-infected or HIV/HCV co-infected individuals who were treated with PEG-IFN/RBV between 2008 and 2013 were included. The cost to treat these patients with PEG-IFN/RBV was calculated (cost 1). We also estimated costs if we had to treat these patients with a lead-in period of PEG-INF/RBV followed by PEG-IFN/RBV and a DAA in naïves (cost 2), in addition to cost 1 plus the estimated cost to re-treat with PEG-IFN/RBV and a DAA patients who had a relapse or a non response (cost 3). Moreover, all costs were normalized by SVR. Rates of foreseen response with DAA were obtained from literature data., Results: The overall study population consisted of 104 patients. The rate of sustained virological response (SVR) was 55%, while it was estimated that SVR would be obtained in 75% of patients with a lead-in period with PEG-IFN/RBV followed by a DAA combination, and in 78% if this treatment is used to re-treat experienced patients with a DAA. Drug costs associated with these treatments were: €1,214,283 for cost 1, €3,474,977 for cost 2 and €3,002,095 for cost 3. Costs per SVR achieved were: €22,284 for cost 1, €44,643 for cost 2 and €38,322 for cost 3., Conclusions: Treatments including DAAs achieve a SVR in more patients than PEG-IFN/RBV but they cost around three times more than PEG-IFN/RBV alone regimens. Also, cost per SVR is almost twofold greater than PEG-IFN/RBV regimens. Therefore, it is mandatory to implement use of DAA in clinical practice, but the National Health System should allocate adequate resources to provide drugs, which challenges sustainability. Cost reduction for anti-HCV drugs should be pursued.

Systemic Bartonella henselae infections are unusual in immunocompetent adults. However, here we report one such case of bartonellosis in a 34-year-old patient, who presented with fever and multinodular splenomegaly. We also describe a novel method of identifying Bartonella henselae by real-time quantitative polymerase chain reaction and sequencing of amplified products. This could prevent splenic bartonellosis being mistaken for lymphoma and thereby avert unnecessary splenectomy.

Background: Hepatitis B virus infection has decreased in Italy. The aims of this study were to identify changes, if any, in the epidemiological pattern of HBV infection in a southern Italian town first surveyed in 1996 and to assess the effectiveness of vaccination campaign against hepatitis B., Methods: In 2010, subjects were selected from the census by a systematic 1:4 random sampling procedure. Hepatitis B surface antigen (HBsAg) and antibodies to hepatitis B core antigen (anti-HBc) were detected by ELISA. Associations (odds ratios) linking exposure to hepatitis B virus infection to potential risk factors were estimated by univariate and multivariate analyses., Results: Of the 1100 eligible subjects, 1020 (92.0%) agreed to participate. The prevalences of HBsAg (0.6%) and anti-HBc (15.2%) were significantly lower than in 1996 (0.8% and 21.5%) (p<0.01). No subject below 30 years of age (those that had been targeted for compulsory immunization) had been exposed to HBV infection. At multiple logistic regression analysis, age>45 years (OR=9.8; 95% CI=5.1-18.7) and past use of glass syringes (OR=1.9; 95% CI=1.2-3.1) independently predicted the likelihood of anti-HBc positivity., Conclusions: These results, albeit obtained in a small town and thus not generalizable, confirm the continuous decreasing trend of HBV infection and demonstrate the effectiveness of the Italian hepatitis B vaccination program., (Copyright © 2012 European Federation of Internal Medicine. Published by Elsevier B.V. All rights reserved.)

Purpose: Universal anti-hepatitis B vaccination of infants and of 12-year-old children became mandatory in Italy in 1991. The purpose of this study was to evaluate the persistence of anti-hepatitis B surface (HBs) antibodies several years after a primary course of vaccination., Methods: In 2010, anti-HBs titers were measured in all subjects aged between 5 and 25 years residing in a southern Italian town. Individuals with an anti-hepatitis B antibody concentration of 10 IU/ml or more were considered to be protected., Results: Of the 671 subjects evaluated, 149 (30%) lacked protective antibodies. Fifty-three (29.4%) of the subjects had been vaccinated ≤10 years earlier and 96 (30.3%) more than 10 years earlier (P = not significant). Subjects vaccinated in infancy were more likely to lack protective anti-HBs antibodies than subjects vaccinated at 12 years of age, regardless of the years elapsed since immunization., Conclusions: Most subjects maintained protective antibodies for a considerable number of years after vaccination. Vaccination in adolescence results in more prolonged immunogenicity than vaccination in infancy.

Analysis of the Hepatitis C Virus (HCV) genotype spread in a particular area has a crucial impact on public health. In this study, we update information on the distribution of HCV genotypes, by evaluating a hospital-based cohort of 2,153 chronic hepatitis C patients, collected prospectively among subjects attending University Hospital of Catanzaro, within an area of Southern Italy. We assessed the rates (%) of HCV genotypes during two consecutive periods, from 2001 to 2005 and from 2006 to 2011, according to age and gender. Considering overall observation time, subtype 1b was predominant followed by subtypes 2a/2c, genotype 3 and 4. Statistical evaluation of the age of HCV patients stratified by genotypes, revealed a slight but significant increase in the median age of 1b, 2a/2c and 3 HCV genotype-infected subjects, during the 2006-2011 period, whilst genotype 4 patients exhibited a decrease in the median age during the same period studied. Moreover genotype 4 increased between 2002 and 2003 as well as between 2010 and 2011. Due to the peculiar diagnostic/clinical/therapeutic features of HCV-4, our findings warrant a deeper investigation to better control infections caused by such genotype.

We describe a SYBR Green I-based real-time PCR targeting Bartonella quintana transcriptional regulatory protein (bqtR) gene, recently found as invariant gene among different B. quintana strains. Melting curve analysis allowed us to discriminate between B. quintana and Bartonella henselae amplified products. We also show its usefulness in the management of a blood culture-negative patient affected by enlarged cervical lymphonodes and long-lasting fever. B. quintana DNA detection in patient whole blood samples and blood culture bottles was confirmed by sequencing and analyzing amplified products., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Twenty-nine staphylococcal isolates from different clinical samples were tested for slime production: phenotypic characterization was carried out using Christensen test (tube test) and Congo red agar plate test (CRA plate test), while the presence and expression of icaA and icaD genes were evaluated by real-time PCR. In 79.3% of studied strains there was a concordance between slime production and presence of icaA and icaD genes, and between lack of slime production and absence of both or only one of the tested genes. In four of five strains where positive phenotype was not associated with the presence of ica genes, gene co-expression (evaluated by mRNA determination) was lacking, while in only a case out of five, there was the presence of transcripts without phenotype. Our study, for the first time, introduces real-time PCR for the detection of both icaA and icaD genes and their mRNA and, furthermore, addresses the relationship between slime phenotype absence and mRNA expression, in clinical isolates of coagulase-negative staphylococci.

We evaluated the applicability of the LightCycler Staphylococcus M(GRADE0 assay on artificially infected blood samples from healthy donors and on clinical specimens of 31 hospitalized patients. The sensitivity and specificity of the assay for detecting Staphylococcus aureus was 100% in blood samples, and 100% in blood culture bottles, when data from the BACTEC 9120 blood culture system were taken as gold standard. The same specificity and sensitivity was found during the search for CoNS (Coagulase Negative Staphylococci) in blood culture bottles, whereas a 93.33% sensitivity and 100% specificity was observed for detecting CoNS directly in blood clinical specimens.

Stenotrophomonas maltophilia is an emerging pathogen implicated in an increasing number of severe pulmonary infections and nosocomial sepsis. We evaluated the influence of four different antibiotics on the bacterial count and LPS activity found in broth cultures of S. maltophilia. After 4 h ceftazidime (CTZ) decreased live bacteria but increased endotoxin activity, whilst isepamicin (ISE), tobramycin (TB), and polymyxin B (PB) reduced both of them. We also investigated the influence of the above mentioned antibiotics on the ability of S. maltophilia culture filtrates and S. maltophilia LPS, extracted in our laboratory, to stimulate sepsis mediators such as tumor necrosis factor a (TNF-a), interleukin 8 (IL-8), interleukin 10 (IL-10), Nitric Oxide (NO) and as bactericidal/permeability-increasing protein (BPI) in human whole blood. Our results demonstrated that both single polycationic antibiotics and the combination of two molecules are able to kill bacteria and neutralize released S. maltophilia LPS. Combination between beta-lactams and aminoglycosides is often able to reduce the pro-inflammatory effects of S. maltophilia culture filtrates.

Previous studies demonstrated that live Bartonella quintana often induces angioproliferative lesions in humans. It modulates endothelial cell apoptotic and inflammatory patterns, thus inducing a very early overexpression of caspase 8 and Apaf-1 and increasing mRNA production of TNF-alpha, interleukin-8, and E-selectin. However, starting at 10 hours postinfection, the bacteria provoke antiapoptotic effects that induce an increase of bcl-2 gene transcription. To gain further insight into the cellular mechanisms that regulate apoptosis, survival and proliferation, we studied the modulation of mitogen-activated protein kinase (MAPK) and the activation state of cdc2 kinase, which regulates progression into mitosis. Confocal microscopy findings indicated a maximum rate of Bartonella entry into host cells between postinfection hours 6 and 10. Live bacteria caused substantially higher apoptosis of human umbilical vein endothelial cells-cryopreserved (HUVEC-C) than heat- and trypsin-inactivated microorganisms. During the first 6 hours postinfection, B. quintana triggered a peak of apoptosis, induced activation of p38 MAPK and stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK), with bacterial clusters appearing at the cellular surface of the HUVEC-C. However, at 8 to 24 hours postinfection, B. quintana was internalized and inhibited proapoptotic signals such as p38 MAPK and SAPK/JNK while inducing antiapoptotic signals. Indeed, expression of the bcl-2 gene and the increase of the bcl-2 kinase active form was concomitant to activation of mitosis, as shown by cdc2 protein activation. These data thus suggest that mechanisms that induce mitotic activity and inhibit apoptotic signals may contribute to the ability of B. quintana to cause vascular proliferation.

Geographical position, an increasing flow of immigrants and refugees coming from regions where malaria is endemic might further increase those cases of malaria imported to Calabria due to travel on military missions, visiting relatives, business and leisure. However, few reports have been published regarding malaria imported into the southern Italian region of Calabria. Based on data from our laboratory, official reports received from the Italian Ministry of Health and Regional Health Offices, an epidemiological analysis of malaria cases registered in Calabria from January 1988 to December 2001 is reported. The epidemiological and clinical features concerning the cases are discussed. A total of 34 slide-confirmed malaria cases were observed in Calabria during the period in question. Infections were mostly acquired in Africa (84.8%), while remaining infections came from Asia (9.1%) and South America and Europe (6.0%). Length of stay in the endemic area did not increase the infection risk. Etiological diagnosis indicated Plasmodium falciparum as the species most often involved (60.6%), followed by Plasmodium vivax (36.3%) and P. vivax/Plasmodium malariae mixed infection (3.0%). The mortality rate was about 3.0%. The number of cases during the second seven-year period of this study was almost double that of the first seven-year period. Correct chemoprophylaxis was performed by only 27.3% of our studied subjects. Delay of malaria diagnosis ranged between 4 days and 1 month. In conclusion, increases in malaria cases, mostly due to P. falciparum, delay in diagnosis and reporting to the Regional Health Office, as well as the increasing arrival of refugees from endemic areas, are epidemiological concerns in Calabria, the southernmost region of continental Italy.

Bartonella quintana, an emerging gram-negative pathogen, may cause trench fever, endocarditis, cerebral abscess and bacillary angiomatosis usually with the absence of septic shock in humans. B. quintana lipopolysaccharide (LPS), a deep rough endotoxin with strong reactivity in the limulus amebocyte lysate (LAL)-assay, was studied in human whole blood and in a rat model. A significant (P<0.05) increase of interleukin-8 (IL-8) concentration, comparable to the level induced by enterobacterial LPS, was stimulated in the human whole blood by B. quintana LPS. Isolated human neutrophils delayed their apoptotic behavior in the presence of B. quintana LPS. In the rat, B. quintana LPS induced a significant (P<0.001) increase in white blood cell count, both 30 and 60 min after intravenous injection. Such leukocytosis was inhibited by pretreatment with prazosin, an alpha-adrenergic antagonist. B. quintana LPS did not significantly change heart rate (HR), hematocrit (HCT) and platelet count in the above reported in vivo model, and regarding mean blood pressure (MAP) only a very early (5 min after LPS) and mild (yet significant) hypotension was observed. In contrast, a long-lasting decrease of MAP was found in Salmonella minnesota R595 LPS-treated animals. Blood TNFalpha levels did not change significantly from the baseline in rats injected with either saline or with B. quintana LPS, on the contrary S. minnesota R595 LPS-injected animals showed substantial increase of TNFalpha levels up to 2924 pg/ml at 60 min after LPS injection. B. quintana LPS as well as Salmonella LPS-injected rats exhibited an increase of the blood levels of GRO/CINC-1, particularly at 240 min after LPS administration. Apical part of rat gut villi showed several TUNEL-positive cells in tissue sections from B. quintana LPS-treated animals. Taken together, our data demonstrates that B. quintana LPS is able to selectively stimulate some inflammatory mediators. B. quintana LPS-induced leukocytosis appears mediated by an alpha-adrenergic receptor. The delayed apoptotic process of leukocytes and the chemokine increase may explain the apoptotic cells found in the rat gut and the inflammatory reactions in some human Bartonella diseases. This peculiar inflammatory pattern induced by B. quintana LPS, may partially account for the lack of severe septic shock, observed in human B. quintana infections.