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SUMMARYThe objective of this study was to assess the prevalence of Ureaplasma urealyticum and Mycoplasma hominis infections and to investigate associations between their presence in the lower female genital tract and lifestyle characteristics. The study was performed on a population of 3115 women, comparing the demographic and behavioural characteristics of 872 women with U. urealyticum infection and 142 women with M. hominis with uninfected women, using univariate and multiple logistic regression analysis. The prevalence of infection with U. urealyticum was 28% and M. hominis was 4·6%. In multivariate logistic regression analysis, intrauterine device, number of sexual partners and age (U. urealyticum while previous induced abortion, condom use and young age at first intercourse (M. hominis infection. U. urealyticum infection presents the same demographic and behavioural characteristics of a sexually transmitted disease. The unprotective role of condom use suggests a non-sexual mode of transmission of M. hominis infection.

Objectives/Hypothesis: This study was undertaken in two phases. In the first phase, we considered patients affected by chronic external otitis treated either by chemical ear peeling (CEP) or by antibiotic/steroid treatment to compare the clinical and microbiological outcomes. In the second phase, we compared the microscopic findings observed in the CEP samples of patients affected by chronic otitis externa's acute exacerbation or by acute otitis externa to demonstrate the role of biofilm in the pathogenesis of chronic otitis externa. Study Design: Prospective, double-blind, controlled study. Methods: In phase 1 we compared clinical and microbiological data collected from two groups of 25 patients with chronic otitis externa treated by CEP or by conventional antibiotic/steroid treatment. In phase 2 we compared the results of the optical and electron microscopic analysis of specimens obtained by performing CEP in two groups of patients (25 with chronic otitis externa exacerbation and 15 with acute otitis externa). Results: In phase 1 the disease control rate yielded markedly better results when treated with CEP. In phase 2 biofilms were identified in 23 of the 25 patients with chronic otitis externa exacerbation (92%) and in only three acute external otitis cases (20%). Conclusions: CEP is a simple and effective method for the treatment of chronic external otitis. The removal of the bacterial biofilm has a high correlation with a long-term clinical remission. Laryngoscope, 121:2626–2633, 2011

Brevundimonas vesicularis is a non-fermenting gram-negative bacillus, aerobic and motile. This microrganism is ubiquitous in the environment and has rarely been implicated in human infections. We present the second case of cutaneous infection caused by B. vesicularis in an immunocompetent patient.

The possible role of human papillomavirus (HPV) in the aetiology of papillary tumor of the bladder has been evaluated and a review of the literature concerning this issue was made. A group of 17 patients affected by bladder papillary tumor was analysed. Surgical specimens were collected for virological and histological analysis. The DNA of the following viruses was searched by polymerase chain reaction (PCR): Adenovirus, Herpes simplex virus type 1 (HSV-1), Herpes simplex virus type 2 (HSV-2), Human Papillomaviruses (HPV), Polyomaviruses (BKV and JCV). The results showed that 15/17 (88 %) patients with papillary bladder tumor were found negative for each viral-searched DNA; only one sample was positive for HPV (6 %) genotype 6, which is considered to convey a low risk for cancer development and only one was positive for BKV (6 %). From the results obtained there seems to be no relationship between viral infection and the presence of bladder papillary tumor. Moreover, in the examined population the association bladder carcinoma-HPV, found by others, has not been confirmed. The homogeneity of the specimens studied was such that it would not be affected by the temporal factor, as were cases of more or less advanced cancers. Nonetheless specimens from patients with advanced cancers (G III) were negative to HPV infection. The data do not appear indicative for a correlation between viral DNA presence and histological parameters. Thus, in the light of the data emerging from this investigation, no causal relationship can be established between HPV infection and papillary bladder tumors.

We describe a 79-year-old female with a chronic venous ulceration infected by Staphylococcus aureus and Enterococcus faecalis and not responsive to conventional treatments. The patient was treated with Methyl-Aminolaevulinate Photodynamic Therapy (MAL-PDT). After four weeks the cutaneous swabs become negative and we observed a significant clinical improvement. Therefore we suppose that MAL-PDT could represent a valid therapeutic option in the treatment of infected chronic ulcers.

(2008). A Dialysis Patient with Bacteremia Caused by a Community-Acquired Methicillin-Resistant Staphylococcus aureus (CA-MRSA) Carrying the Staphylococcal Chromosome Cassette (SCC) mec type V. Journal of Chemotherapy: Vol. 20, No. 3, pp. 402-404.

The effect of different natural and semi-synthetic polysaccharides on Chlamydia trachomatis multiplication in Hela 229 cells was evaluated. Some neutral, negatively and positively charged carbohydrates were able, in a dose-dependent fashion, to inhibit chlamydial infection by interfering mainly with the adsorption process. The inhibiting compounds, whose effect was shown within the concentration range of 8-200 micrograms/ml, were in order of action: dextran sulphate > glyloid sulphate 4327 > glycogen sulphate 4427 > arabic gum = glyloid > chitosan > glycogen. Data obtained suggested that antichlamydial activity was not only related to the electric charge of these molecules but could also be attributed to other features of their polymeric backbone. Since carbohydrate polymers have also been shown to inhibit the early stages of infection by viral agents causing sexually transmitted diseases, the employment of these molecules for prevention or treatment of mixed viral-C. trachomatis infections can be hypothesized.

Patients in end-stage renal disease undergoing renal replacement treatment (ESRD-RRT) are considered immunocompromised. The hemodialysis (HD) or peritoneal dialysis (PD) procedures seem to produce alterations of the immune status. Interest in immunosuppression has increased due to the poliomavirus BK (BKV) infection. Our study evaluated the prevalence of BKV infection in ESRD-RRT patients and viral replication on HD or PD. From 2006 to 2011 we selected 58 patients (34 males) in ESRD-RRT for inclusion in our study. BKV replication was evaluated by qualitative real-time polymerase chain reaction. In ESRD-RRT patients, the prevalence of BKV replication on plasma was 21%. We identified two groups of patients according to the dialysis procedure: 36 patients on HD (HD group) and 22 on PD (PD group). BKV replication in the HD group was 33% (12 of 36) versus 0% (0 of 22) in the PD group. Different age, number of months on RRT, and preserved diuresis was observed in the HD versus PD groups. With our results we can speculate that BKV infection in ESRD-RRT patients is linked to factors involved in the uremia-related immune dysfunction but also to specific mechanisms related to the different RRTs. PD is an option that could be associated with a better transplant outcome for patients undergoing kidney transplantation.

As reliable data on Chlamydia trachomatis infection in Italy are lacking and as there is no Italian screening policy, epidemiological analyses are needed to optimise effective strategies for surveillance of the infection in the country. We collected data from 6,969 sexually active women aged 15 to 55 years who underwent testing for endocervical C. trachomatis infection at the Cervico-Vaginal Pathology Unit in the Department of Gynaecology and Obstetrics of Sapienza University in Rome between 2000 and 2009. The mean prevalence of C. trachomatis endocervical infection during this period was 5.2%. Prevalence over time did not show a linear trend. Univariate analysis demonstrated a significant association of infection with multiple lifetime sexual partners, younger age (

Psoriasis vulgaris (PsV) and psoriatic arthritis (PSA) are inter-related heritable inflammatory skin diseases. Psoriatic lesions develop as a result of abnormal immune responses, hyperproliferation and altered differentiation of keratinocytes, and a notable subset of psoriatic patients develops PsA, characterized by joints inflammation. Recently, biological drugs were introduced to treat these diseases. However, this therapy has already been associated with the development of serious life-threatening infections, such as the reactivation of human polyomavirus JC (JCV), responsible for the progressive multifocal leukoencephalopathy (PML), a lethal demyelinating disease caused by oligodendrocytes lytic infection. Therefore, the aims of our study were the investigation of the possible JCV reactivation in PsV and PsA patients treated with adalimumab, etanercept, and methotrexate, performing quantitative real-time PCR in sera and skin biopsies at the time of recruitment (T0) and after 3 (T3) and 6 (T6) months of treatment, and the sequencing analysis of the JCV non-coding control region (NCCR). We found JCV DNA in 5/15 PsV patients and in 2/15 PsA patients and JCV NCCR sequence analysis always showed a structure similar to non-pathogenic CY archetype, with random occurrence of a few irrelevant point mutations. Nevertheless the poor number of patients analyzed, our preliminary data can pave the way for taking into account that the follow-up of JCV DNA detection and the JCV NCCR sequence analysis in psoriatic patients may be important to evaluate the risk of PML onset, considering that patients affected by autoimmune diseases and treated with biologics continue to rise.

Progressive multifocal leukoencephalopathy (PML) is a demyelinating disease of the central nervous system caused by the neurotropic human polyomavirus JC (JCV) lytic infection of oligodendrocytes. PML was first described as a complication of lymphoproliferative disorders more than 50 years ago and emerged as a major complication of human immunodeficiency virus (HIV) infection in the 1980s. Despite the ubiquity of this virus, PML is rare and always seen in association with underlying immunosuppressive condition, such as HIV infection, autoimmune diseases, cancer, and organ transplantation. JCV remains quiescent in the kidneys, where it displays a stable archetypal non-coding control region (NCCR). Conversely, rearranged JCV NCCR, including tandem repeat patterns found in the brain of PML patients, have been associated with neurovirulence. The specific site and mechanism of JCV NCCR transformation is unknown. According to one model, during the course of immunosuppression, JCV departs from its latent state and after entering the brain, productively infects and destroys oligodendrocytes. Although the majority of PML cases occur in severely immunesuppressed individuals, PML has been increasingly diagnosed in patients treated with biological therapies such as monoclonal antibodies (mAbs) that modulate immune system functions: in fact, CD4+ and CD8+ T lymphopenia, resulting from this immunomodulatory therapy, are the primary risk factor. Furthermore, JCV reactivation in nonpermissive cells after treatment with mAbs, such as intestinal epithelial cells in Crohn's disease patients, in association with other host tumor-inducing factors, could provide valid information on the role of JCV in several malignancies, such as colorectal cancer.

Polyomavirus-associated nephropathy (PVAN) has a predilection for kidney rather than for other solid organ transplants such as the liver. Immunosuppression is widely recognized to be a major risk factor for PVAN development. Since end-stage liver disease (ESLD) patients are immunocompromised and immunosuppression is a major cause of BK virus reactivation, we sought to evaluate BK virus replication in patients listed for liver transplantation. From April to October 2010, we enrolled 20 patients listed for liver transplantation. BK virus load was measured by quantitative real-time polymerase chain reaction on plasma and urine samples. Viremia occurred in only 1 among 20 patients. We hypothesized that in ESLD patients, the low prevalence of BK virus infection may be related to the prevalent impairment of antibacterial immunity rather than to the viral-specific one. In BK virus reactivation, not only the immunodepressive state itself, but also the specific immunologic mechanisms involved may have a role.

Background Nowadays, better immunosuppressors have decreased the rates of acute rejection in kidney transplantation, but have also led to the emergence of BKV-associated nephropathy (BKVAN). Therefore, we prospectively investigated BKV load in plasma and urine samples in a cohort of kidney transplants, receiving basiliximab combined with a mycophenolate mofetil-based triple immunotherapy, to evaluate the difference between BKV replication during the first 3 months post-transplantation, characterized by the non-depleting action of basiliximab, versus the second 3 months, in which the maintenance therapy acts alone. We also performed sequencing analysis to assess whether a particular BKV subtype/subgroup or transcriptional control region (TCR) variants were present. Methods We monitored BK viruria and viremia by quantitative polymerase chain reaction (Q-PCR) at 12 hours (Tx), 1 (T1), 3 (T2) and 6 (T3) months post-transplantation among 60 kidney transplant patients. Sequencing analysis was performed by nested-PCR with specific primers for TCR and VP1 regions. Data were statistically analyzed using χ2 test and Student's t-test. Results BKV was detected at Tx in 4/60 urine and in 16/60 plasma, with median viral loads of 3,70 log GEq/mL and 3,79 log GEq/mL, respectively, followed by a significant increase of both BKV-positive transplants (32/60) and median values of viruria (5,78 log GEq/mL) and viremia (4,52 log GEq/mL) at T2. Conversely, a significantly decrease of patients with viruria and viremia (17/60) was observed at T3, together with a reduction of the median urinary and plasma viral loads (4,09 log GEq/mL and 4,00 log GEq/mL, respectively). BKV TCR sequence analysis always showed the presence of archetypal sequences, with a few single-nucleotide substitutions and one nucleotide insertion that, interestingly, were all representative of the particular subtypes/subgroups we identified by VP1 sequencing analysis: I/b-2 and IV/c-2. Conclusions Our results confirm previous studies indicating that BKV replication may occur during the early hours after kidney transplantation, reaches the highest incidence in the third post-transplantation month and then decreases within the sixth month, maybe due to induction therapy. Moreover, it might become clinically useful whether specific BKV subtypes or rearrangements could be linked to a particular disease state in order to detect them before BKVAN onset.

The methicillin-resistance is increasing all over the world in the last decade. It is more frequent among coagulase-negative staphylococci (MRCoNS); infact the 52% of S. epidermidis strains results to be resistant to methicillin.The methicillin-resistant strains also show a reduced sensitivity towards the first-line agents such as glycopeptides and other antibiotics commonly used in therapy such as trimethoprim-sulphamethoxazole, imipenem, gentamycin, fosfomycin and chlarytromicin. Unlike MRSA (Methicillin-resistant S. aureus), MRCoNS resistance to glycopeptides generally concerns teicoplanin. Although vancomycin resistance is rare in Staphylococcus isolates, the detected shift towards higher values of MICs might affect patient’s clinical outcome.

The recent introduction of monoclonal antibodies in Crohn's disease (CD) management has been associated with the development of serious complications, such as the progressive multifocal leukoencephalopathy (PML), caused by JC polyomavirus (JCV) reactivation. Therefore, the aims of our study have been the investigation of the possible JCV reactivation in pediatric CD patients treated or not with infliximab, performing quantitative PCR in urine, plasma, and intestinal biopsies at the time of recruitment (t0) and every 4 months in 1 year of follow-up (t1, t2, and t3), and the analysis of the JCV noncoding control region (NCCR) to detect cellular transcription factors binding site mutations. Results obtained showed that, in urine and ileal specimens, JCV load significantly increased in infliximab-treated patients after 1 year of treatment (t3), while viremia was significantly higher at t1. JCV NCCR sequence analysis showed a structure similar to CY archetype in 65/80 analyzed sequences, but the remaining 15/80, obtained exclusively from plasma and biopsies, evidenced a CY NCCR organization with two recurrent nucleotide changes, the 37-T to G transversion in box A Spi-B binding site and the 217-G to A transition in box F, and a box D deletion. These rearrangements were always found at t3 within seven infliximab-treated CD patients, who presented a very severe disease at t0. We can conclude that our rearranged NCCR sequences could be considered a marker of JCV virulence during mAb treatment, although none of our examined patients developed PML, and further studies on a larger cohort of patients should be performed.

Polyomavirus BK (BKV) infection is ubiquitous in the human population. Under immunosuppression, BKV can undergo reactivation resulting in viral replication. What really happens in the early hours posttransplantation is not clearly defined; the meaning of early viremia and viruria is not clear. BKV viremia is considered a marker of infection. The aim of our study was to investigate the prevalence of early BKV infection in kidney transplant patients, to evaluate the relationship to infections at 3 and 6 months and the association with recipient, donor, and graft features. We enrolled 36 kidney transplanted patients from May 2006 to April 2007. BKV load was measured on plasma and urine samples by Q-PCR at 12 hours (T 0 /early) as well as 3 (T 3 ) and 6 (T 6 ) months thereafter. A high percentage of BKV infections were detectable in the first hours after transplantation (33.3%), which remained unchanged to month 6 post transplantation. Moreover, patients who were positive at T 0 had a high probability of remaining positive thereafter. The number of copies in plasma samples tended to increase at 3 months and to decrease thereafter, whereas the urine viral load tended to steadily increase. Among BKV-positive patients, we identified 2 groups according to viremic state at T 0 : 9 patients (group A); who were already positive and remained so to T 6 5 and 3 patients who turned positive at 3 or at 6 months, respectively (group B). Group A included 75% of positive patients at T 0 and 90% of positive patients at either T 3 or T 6 ( P = .007). The most important contribution of our study was to highlight the presence of BKV infection in renal transplant recipients from the first hours posttransplantation. This condition seemed to be the most important risk factor for persistent infection in the first 6 months.

Although the remarkable efficacy of biological therapy has resulted in significant success in inflammatory bowel disease (IBD) management,susceptibility to infections remains a concern. The biological agents include the tumor necrosis factor-a (TNF-a) inhibitors, for instanceinfliximab,andotherimmunomodulatingagents,suchasnatalizumab.Progressivemultifocalleukoencephalopathy(PML),ararebutmostlyfatalopportunistic brain infection caused by reactivation of the human polyomavirus JC virus (JCV), has been found in two patients with multiplesclerosis and one patient with Crohn’s disease (CD), linked to treatment with natalizumab. After these cases of PML, the commercial andinvestigational use of natalizumab was suspended in February 2005 but was subsequently resumed for multiple sclerosis and for CD, onlythroughaspecialrestricteddistributionprogram.Thisreview,startingfromanextensiveliteraturesearchbythePubMeddatabase,resumestheclinicalaspectsandpathophysiologyofCDandfocusesonthebiologicsincurrentuseinCD(infliximab,adalimumab,andnatalizumab),inordertoprovideareferenceandgatewaytoprevention,recognition,andmanagementofJCV,intheearlyyearsofbiologicalagentstherapy.Italso proposed to provide an overview on the hypothetical mechanism of reactivation of JC virus related to the use of these drugs.J. Cell. Physiol. 224: 316–326, 2010. 2010 Wiley-Liss, Inc.Crohn’s disease (CD) is a chronic inflammatory bowel disease(IBD)ofunknownetiologythatresultsinsignificantmorbidityandhealth carecosts. Morethan400,000peoplein theUnitedStateshave CD. The prevalence continues to increase because of itsrising incidence and improved survival (Loftus and Sandborn,2002;ComerfordandBickston,2004).Thechronicnatureoftheillness causes frequent hospitalizations; most patients eventuallyrequire surgery secondary to complications, such as strictures,abscesses, fistula, or refractory disease. The disease seems tooccur when the intestinal immune cascade is triggered by anantigeningeneticallysusceptibleindividuals.Overactivationoftheenteric immune and inflammatory pathways causes mucosaldamage resulting in the clinical signs and symptoms. Variousmedications, including 5-aminosalicylates, antibiotics,corticosteroids, and immunomodulators, such as purineantimetabolites and methotrexate, have traditionally been usedtocontrolinflammation.Theiruseisintendedtopreventsurgeryandimprovethepatient’squalityoflife.Nonecurethediseaseandunfortunately, many patients require steroids to control theirsymptoms. A wide range of dose-related adverse effects makesthis an unappealing strategy. Immunomodulators are effectivemaintenance drugs, but have a slow onset of action with clinicalremission rates of approximately 40%. The limitations ofconventionalagentsleavetheclinicianinneedofamedicationthatissteroid-sparing,quickacting,andmaintainsremission(Hanaeuret al., 2002).Recently, biological therapy has brought a paradigm shift inthemanagementofIBD.Theintroductionofbiologicagentshasproduced an astounding transformation by halting or slowingthe progression of IBD, rheumatoid arthritis (RA), psoriaticarthritis and multiple sclerosis (MS) resulting in markeddecrease of disability and improvement in quality of life andhealth outcomes (Saketkoo and Espinoza, 2006). Along withgreatreliefandhope,biologicagentshavebroughttheneedforexpert vigilance requiring judicious selection of candidates,close observation, and careful attention to patient education.Theearlyyearsofbiologicalagentstherapyhasbeenassociatedwith the development of serious life-threatening infections inaddition to other well documented hematologic, immunologic,cardiovascular, and malignant adverse effects. Furthercomplications include the concomitant use of otherimmunosuppressive therapies, such as prednisone andmethotrexate, coexistent morbidities (Hyrich et al., 2006) andunderlying immune dysfunction associated with autoimmunediseases (Askling et al., 2005).Biologic therapies encompass agents with diverse modesof action. They include: (a) native biologic preparations; (b)recombinant peptides or proteins; (c) antibody-based therapy;(d) nucleic acid based therapies; and (e) cell and genetherapy (Sands, 1997; Bosani et al., 2009). At this time, thebiotechnology therapies that are being used in clinical practiceor investigated for the treatment of IBD are predominantlyrecombinant human proteins with immunoregulatory effects,monoclonalantibodies (chimeric, humanized, andfullyhuman)

This is a report concerning human polyomavirus JC (JCV) reactivation in a pediatric patient with Crohn's disease (CD) during the treatment with 5-aminosalicylic acid (5-ASA), a non-steroidal anti-inflammatory drug (NSAID). We examined 9 bioptic samples from three different bowel districts (ileum, cecum, rectum) of this child. These samples were analyzed by Quantitative PCR (Q-PCR) to investigate the presence of JCV DNA. JCV DNA was detected in one rectum biopsy taken two months after 5-ASA treatment. Although our result must be validated in a larger group of subjects and with a longer follow-up period, it underlines the importance of JCV monitoring in CD patients.

International audience; Haemorrhagic cystitis is characterized by haematuria due to inflammation of the bladder. In bone marrow transplants, this disease is linked to the infection by human polyomavirus BK, whereas the role of the human polyomavirus JC is unclear. The transcriptional control regions of both viruses contain important cellular transcription factor binding sites that undergo rearrangement process generating suitable variants that could be more active for viral replication and for the onset of haemorrhagic cystitis. In this study urine obtained from seven patients with bone marrow transplant were examined. Polyomavirus genomes were quantified by PCR and viral loads were compared. The transcriptional regions of both viruses were amplified and sequenced to determine the presence of variants. Subtypes of polyomaviruses were determined by amplification and sequencing of the viral protein 1 region. The results showed that 4 of 7 patients were positive for BK DNA, 2 of 7 patients had BK and JC DNA and 1 of 7 had JC DNA. Positive samples were amplified and sequenced successively for transcriptional regions. The viral archetype was always found in both viruses. Finally, typing showed that BK virus subtype I infected patients with BK, whereas JC virus genotype IA and genotype 1B were found in patients infected with JC. The data suggest that new and different approaches are required to improve the morbidity and mortality caused by polyoma-associated haemorrhagic cystitis, since it known that BK virus is involved in the onset of haemorrhagic cystitis, whereas the role of JC virus should be investigated further.

Introduction IFN-γ is a pivotal cytokine in the immune response to Myc. tuberculosis, infact this is the key cytokine produced in response to antigens specific following tuberculosis exposure causing either active or latent tuberculosis (TB) and this observation forms the basis of interferon gamma release assay (IGRA), but there are alternative or additional cytokines and chemokines that could be used to improve detection of Myc. tuberculosis infection.The aim of this study was to evaluate the diagnostic utility of chemokine CXCL10/IP-10 as biomarker of active TB and to compare the results with classical QuantiFERON-Gold assay . Methods CXCL10/IP-10 and IFN-γ responses to stimulation with ESAT-6 and CFP-10 were evaluated in 21 patients with active tuberculosis and in 6 healthy unexposed subjects with no history of TB or TB contact were used as controls healthy controls. QuantiFERON-TB Gold (QFT-G, Cellestis) was used for the measurement of IFN-γ levels; CXCL10/IP-10 was detected by ELISA (R&D Systems ). Results Of the 21 TB patients included, 11 had a QFT-G positive and 10 had negative QFT-G results.All QFT-G positive patients had increased levels of CXCL10/IP-10 (median, pg/ml) in both ESAT-6 and CFP-10 stimulated samples patients compared to healthy controls (1807 and 1111 vs 251 and 188 of controls, respectively) (p

Background High grade HPV infections and persistence are the strongest risk factors for cervical cancer. Nevertheless other genital microorganisms may be involved in the progression of HPV associated lesions. Methods Cervical samples were collected to search for human Papillomavirus (HPV), bacteria and yeast infections in gynaecologic outpatients. HPV typing was carried out by PCR and sequencing on cervical brush specimens. Chlamydia trachomatis was identified by strand displacement amplification (SDA) and the other microorganisms were detected by conventional methods. Results In this cross-sectional study on 857 enrolled outpatients, statistical analyses revealed a significant association of HPV with C. trachomatis and Ureaplasma urealyticum (at high density) detection, whereas no correlation was found between HPV infection and bacterial vaginosis, Streptococcus agalactiae, yeasts, Trichomonas vaginalis and U. urealyticum. Mycoplasma hominis was isolated only in a few cases both in HPV positive and negative women and no patient was infected with Neisseria gonorrhoeae. Conclusion Although bacterial vaginosis was not significantly associated with HPV, it was more common among the HPV positive women. A significant association between HPV and C. trachomatis was found and interestingly also with U. urealyticum but only at a high colonization rate. These data suggest that it may be important to screen for the simultaneous presence of different microorganisms which may have synergistic pathological effects.

Background: Studies performed in adults with inflammatory bowel disease (IBD) have suggested that mucosa-associated Escherichia coli strains may be involved in its pathogenesis. The aim of this study was to characterize E. coli strains from the intestinal mucosa of pediatric IBD patients to investigate whether a particular subset of strains could be associated with the disease. Methods: We analyzed the genomic and phenotypic traits of 60 E. coli strains isolated from biopsies of pediatric patients with Crohn's disease (CD), ulcerative colitis (UC), and from age-matched controls. Results: No noteworthy differences were found in the distribution of phylogroups. The percentage of adhesive E. coli strains was similar in biopsies from patients and controls. However, the adhesion ability of E. coli strains differed between ileal and colonic or rectal areas, only in the strains from CD and UC patients. The percentage of E. coli possessing more than 1 of the adhesive/virulence determinants was significantly higher in strains from UC than from CD and controls. Interestingly, the genetic profile examination revealed 2 large clusters of genetically linked E. coli strains from IBD patients. Ninety-two percent of the strains isolated from CD patients were in the first cluster (A) and were distributed between 2 genetic subclusters (A1 and A2), while a second cluster (B) contained most of the strains isolated from UC (78%; subcluster B1), and control strains (77%; subcluster B2). Conclusions: Genomic analysis of mucosa-associated E. coli strains found a close genetic association between strains isolated from CD and UC patients. (Inflamm Bowel Dis 2008)

Newborn babies admitted into the neonatal intensive care unit (NICU) often require many supportive invasive devices and frequently receive antimicrobial therapy. We investigated the microbial flora in NICU patients reporting the distribution of infections in different catheter sites. Results showed that 97% of samples were positive; in particular 11% were positive for two or more microbial agents. Coagulase negative Staphylococci were the most commonly isolated. The detection of Gram-negative bacteria and yeasts suggested that these microorganisms are also involved in infections of hospitalized infants. Finally, no correlation between a specific microbial agent and a particular catheter type was found.

Herpes simplex virus (HSV) infection is one of the most common sexually transmitted viral diseases worldwide. HSV type 2 causes most genital herpes and HSV type 1 is usually transmitted via non-sexual contacts. We studied 109 pregnant women between January 2007 and December 2008, in relation to their age, condom use, number of sexual partners, age at first intercourse, parity and smoking habits. The aim of this study is to evaluate the prevalence of HSV cervical infection and HSV co-infection with other genital microorganisms associated with poor neonatal outcome. Our results show that of the 109 outpatients enrolled, 30% were HSV1 and/or HSV2 positive, of whom 30% were infected with both HSV1 and HSV2, 18% were infected with HSV1 alone and 52% with HSV2 alone. A significant association between HSV1 and HSV2 infection was found, and the prevalence of HSV2 infection in women infected with HSV1 was 63%. The prevalence of HSV1/2 varied in the presence of other vaginal microorganisms but a statistical significant association was not found. This pilot study is probably too small to obtain statistically significant results. Nevertheless, using these observed results, we calculated that about 530 patients with comparable features should be enrolled to detect an increase of 50% in HSV infection due to the presence of other genital infections and potential risk factors.

Hospital-acquired infections (HAIs) represent an important cause of morbidity and mortality in neonatal intensive care units (NICUs).All neonates admitted for48 hours between January 2003 and December 2006 in the NICU of the teaching hospital Umberto I of Rome, Italy were considered.Of the 575 neonates evaluated, 76 (13.2%) developed a total of 100 HAIs, including 36 bloodstream infections (BSIs), 33 pneumonias, 19 urinary tract infections, 8 conjunctivitis, and 4 onphalitis. There were 7.8 HAIs/1000 patient-days and 12.5 BSIs/1000 days of umbilical catheterization. Logistic analysis identified an association with mechanical ventilation (odds ratio [OR] = 3.05; 95% confidence interval [CI] = 1.75 to 5.31; P.01) and birth weightor= 1500 g (OR = 2.34; 95% CI = 1.36 to 4.03; P.01). Thirty-five neonates (6.1%) died. Klebsiella pneumoniae (37.7%) and coagulase-negative staphylococci (28.6%) were the most frequently isolated microorganisms. Only 3 Candida spp determined BSIs (8.3%). BSI mortality was higher in infections with gram-negative pathogens (36.4%) than in infections with gram-positive pathogens (4.5%).Although we found a low infection and mortality rate, attention should be directed toward antibiotic-resistant gram-negative pathogens.

This study aims to characterize phenotypic and genotypic virulence traits in Escherichia coli strains, isolated from outpatients with urinary tract infections, comparing with those obtained from inpatients. Information on the pathogenic behavior of the uropathogenic strains was obtained by monitoring different biological properties, such as autoagglutination, hemagglutination, adhesiveness to and invasion of human bladder (HT1376) cells, biofilm formation, phylogenetic grouping, and virulence-related genes. The results show similar behavior in the two groups concerning autoagglutination, hemagglutination, and biofilm formation. None of the strains examined was invasive. However, in strains from outpatients there was an increased adhesion to HT1376 cells compared with clinical strains, a significant higher presence of genes codifying for adhesins and cell protection factors, and a lower proportion of strains belonging to B1 group. These findings add further information on the pathogenic traits of community E. coli, since strains isolated from the outpatients' group were differently “armed” in comparison with those of clinical cases, and more suitable to infect healthy individuals.

Clinical diagnosis of kidney transplants related illnesses is not a simple task. Several studies were conducted to define diseases and complications after renal transplantation, but there are no comprehensive guidelines about diagnostic tools for their prevention and detection. The Authors of this review looked for the medical literature and pertinent publications in particular to understand the role of Human Polyomavirus BK (BKV) in renal failure and to recognize analytical techniques for BK virus associated nephropathy (BKVAN) detection.

Introduction: The identification and characterization of two M. tuberculosis-specific antigens (ESAT-6 and CFP- 10) has led to the development of a whole blood new generation of M. tuberculosis specific diagnostic tests, that have several advantages over tuberculin skin test (TST), in terms of higher specificity, better correlation with surrogate measures of exposure to M. tuberculosis in low-incidence setting, and less cross-reactivity with M. bovis (BCG) vaccine and environmental mycobacteria.The role of these new tests in evaluating post-therapy tuberculosis eradication has not been investigated yet. Aim of this longitudinal study was to determinate changes of response to M. tuberculosis-specific antigens in patients during the standard tuberculosis treatment and to investigate the in vitro effects of tuberculosis drugs on the IFN-γ release. Methods: 23 individuals with active tuberculosis were enrolled and followed over time.They were tested with QuantiFERON TB-Gold (QFT-Gold) at four time points: at diagnosis (t0), after 3 and 6 months of treatment (t1- t2) and at the end of the specific treatment (t3). Results: At baseline all patients were positive by QFT-Gold.At second time-point 17 out of 23 (74%) were positive, at third time-point 11 of 23 (47%) were positive, at treatment completion 3/23 (13%) were positive.The conversion to negative response to M. tuberculosis-specific antigens was found in 87% patients analyzed after successful therapy. Longitudinal QFT-Gold testing shown a significant decrease (p

Aims: The ability of Listeria monocytogenes to survive and grow at high salt concentrations and low pH makes it a potential hazard after the consumption of milk and dairy products, often implicated in severe outbreaks of listeriosis. This study was designed to evaluate the behaviour of L. monocytogenes in traditional acid and salted Italian-style soft cheeses and to investigate whether Listeria occurrence and growth in these environments may represent a potential increase of hazard. Methods and Results: A first approach was addressed to in vitro evaluate survival, acid tolerance response, ability to produce biofilm, and capability to invade intestinal-like cells of a L. monocytogenes strain grown under experimental conditions mimicking environmental features that this pathogen encounters in soft cheeses (such as acid pH and high NaCl content). A second set of experiments was performed to monitor, during the storage at 4°C, the survival of acid-adapted and nonadapted Listeriae in artificially contamined soft cheeses. Both acid tolerance response and invasion efficiency of acid-adapted bacteria resulted in an increase, even when bacteria were simultaneously pre-exposed to increasing salt stress. The contamination of cheeses with acid-adapted and nonadapted bacteria evidenced in all products a good survival. A significant increased survival, the recovery of bacterial cells highly resistant to lethal pH exposure, and the prevalence of filamentous structures were observed in crescenza cheese during the storage. Conclusions: The Listeria survival and acid pH tolerance observed during refrigerated storage are probably related to the intrinsic acid and saline features of soft cheeses analysed. Significance and Impact of the Study: Italian soft cheeses tested may represent a potential hazard for the recovery of acid-adapted L. monocytogenes cells with enhanced ability to adhere to inert surfaces and/or to penetrate host cells.

Results.Forty-six single-nucleotide polymorphisms (SNPs) were identied. The SNPs indicated below were significantly (p< 0.05) more frequently dentified in IBD affected children than in healthy controls: hBD1-113T/C, BD1-361A/G, hBD2-319+26A/G, hBD2-319+53G/A, hBD3-87A/G, BD4-73+29G/T, hBD4-73+192T/C, hBD4-73+158A/G, hBD43+114C/A, hBD4-73+77C/T, hBD4-285+26C/T, hBD4-285+99C/T, BD4-285+13C/T and hBD-285+26A/G. No associations between enotype and type of disease were identified. Conclusions. This is the first investigation on hBD genotyping in chilren with IBD. Some hBDs genetic variants are significantly associated with BD. Our preliminary data support the hypothesis of a role of hBDs in the evelopment of paediatric onset IBD.

Background: Clinical and experimental observations in animal models indicate that intestinal commensal bacteria are involved in the initiation and amplification of inflammatory bowel disease (IBD). No paediatric reports are available on intestinal endogenous microflora in IBD. Aims: To investigate and characterise the predominant composition of the mucosa-associated intestinal microflora in colonoscopic biopsy specimens of paediatric patients with newly diagnosed IBD. Methods: Mucosa-associated bacteria were quantified and isolated from biopsy specimens of the ileum, caecum and rectum obtained at colonoscopy in 12 patients with Crohn’s disease, 7 with ulcerative colitis, 6 with indeterminate colitis, 10 with lymphonodular hyperplasia of the distal ileum and in 7 controls. Isolation and characterisation were carried out by conventional culture techniques for aerobic and facultative-anaerobic microorganisms, and molecular analysis (16S rRNA-based amplification and real-time polymerase chain reaction assays) for the detection of anaerobic bacterial groups or species. Results: A higher number of mucosa-associated aerobic and facultative-anaerobic bacteria were found in biopsy specimens of children with IBD than in controls. An overall decrease in some bacterial species or groups belonging to the normal anaerobic intestinal flora was suggested by molecular approaches; in particular, occurrence of Bacteroides vulgatus was low in Crohn’s disease, ulcerative colitis and indeterminate colitis specimens. Conclusion: This is the first paediatric report investigating the intestinal mucosa-associated microflora in patients of the IBD spectrum. These results, although limited by the sample size, allow a better understanding of changes in mucosa-associated bacterial flora in these patients, showing either a predominance of some potentially harmful bacterial groups or a decrease in beneficial bacterial species. These data underline the central role of mucosa-adherent bacteria in IBD.

In spite of the adoption of third generation cephalosporin restriction policies, two independent outbreaks by Extended Spectrum Beta-Lactamase (ESBL)-producing Klebsiella pneumoniae occurred in two different wards (Neonatal Intensive Care Unit, NICU and Neurosurgery) of a teaching hospital in Rome, Italy. In the former 19 infected neonates were reported, whereas in the latter there were 10 infected patients. In both wards no differences were observed in the mortality rates in periods of outbreak and those with no outbreak. Molecular typing on a total of 19 isolated strains was carried out and restriction patterns were compared. The PFGE showed that nine isolates responsible for infection in the NICU were all included in three closely related clusters. In Neurosurgery nine strains out of ten were strictly related and part of an outbreak occurring between August-December 2003, while one isolate was temporarily (February 2003) and genetically (seven band differences) unrelated to the outbreak strains. When ESBL producing K. pneumoniae clusters from the two wards (NICU vs Neurosurgery) were compared, they appeared to be completely different both for their genotype pattern and plasmid type or presence, thus demonstrating cross transmission by two different genotypes.

Bladder cancer is the second most commonly occurring genitourinary cancer in adults. The interaction of different carcinogenic and co-carcinogenic agents are responsible for bladder urothelial carcinoma: alcohol and smoking habits, Schistosoma haematobium infection, exposition to chemicals, analgesic and antineoplastic drugs prolonged use. Recently also viral infections have been associated to this pathology. In this study the correlation between viral infections and bladder carcinoma has been evaluated. A group of 32 patients affected by primary bladder neoplasia has been analysed. A control group of 20 autoptic samples of healthy bladder was analysed. The DNA of the following viruses has been searched by Polymerase chain reaction (PCR): Adenovirus, Herpes simplex virus type 1 (HSV-1), Herpes simplex virus type 2 (HSV-2), Human Papillomaviruses (HPV), Polyomaviruses (BKV and JCV). In the examined population the association bladder carcinoma-HPV, found by others, has not been confirmed. The high percentage of human polyomaviruses present in the samples is a statistically significant data (p=0.0087) and allows to presume that BKV and JCV may play a role in the aetiology of bladder tumor. In particular the polyomavirus BK, which is found in significative percentage both in single infection (p=0.0036) and in co-infections with other viral species (p=0.035), may be an important co-factor in the pathogenesis of bladder carcinoma.

Human papillomaviruses (HPVs) and herpes simplex virus type 2 (HSV-2) can establish latent or persistent infections in the host, and are involved in the aetiology of benign and/or malignant lesions of the urogenital tract. To investigate the putative interaction between these DNA viruses when a double infection occurs, we have studied the effect of HSV-2 infection in HeLa 229 cells containing 10–50 copies of HPV type 18 genomic DNA. Twenty hours post HSV-2 infection, the analysis of mRNA transcripts from El, E2, E6 early and L1 late HPV18 genes was performed in HeLa cells by a semi-quantitative RT-PCR assay. A modulation of HPV18 E1 and E6 early genes was observed, resulting in a 9-fold and 3-fold increased transcription respectively.

The present study analyses the susceptibility of human bladder-derived cells (HT-1376) to the infection by herpes simplex virus type 2 (HSV-2) and Chlamydia trachomatis, as well as to the adhesiveness of uropathogenic bacteria. HT-1376 cells were efficiently infected by HSV-2 strain 333, as demonstrated by immunofluorescence staining of viral antigens, titration of cytopathic effect, and visualisation by transmission electron microscopy. This cell model was also prone to C. trachomatis (serovar E, Bour strain) replication and to the adherence of clinical uropathogenic isolates of Escherichia coli, Pseudomonas aeruginosa, Proteus vulgaris and Enterococcus faecalis. The pre-infection of HT-1376 cells with HSV-2 caused a tenfold increased adherence of an E. coli strain (U1), isolated from a patient affected by severe haemorrhagic cystitis, whereas in HSV-2 pre-infected cells the number of C. trachomatis inclusion bodies was significantly reduced. Our findings indicate that these cells are a suitable in vitro model for studying infection and super-infection of the lower urinary tract by viruses and bacteria.

Chlamydia trachomatis (C.t.) cell culture represents a sensitive method for the diagnosis of chlamydial infection and the only one which makes it possible to determine the susceptibility of an isolate to antibiotics so that an appropriate drug can be selected for individual treatment. In 11 patients, affected by urethroprostatitis and suspected of treatment failure with standard drug regimens, either due to lack of compliance with therapy or antibiotic resistance, C.t. was isolated in McCoy cell culture from urethral swabs, after prostatic massage. The in vitro activity of azithromycin against these isolates and the in vivo efficacy of the drug in the patients treated with a single 1 g dose have been evaluated. All the C.t. strains tested were susceptible to the action of azithromycin (MIC range 0.125-1.0 microgram/ml). Bactericidal values were one dilution higher (MBC range 0.25-2.0 microgram/ml). These in vitro results are consistent with clinical observations as all the patients treated had negative culture at a 4-week follow-up visit.

A peptide fraction was isolated from the skin of Bombina variegata that showed antimicrobial activity. This fraction contained several molecular species, all of them consisting of 27 amino acid residues, with a constant C-terminal region (from residues 14-27), including an amidated carboxyl end and a variable N-terminal segment. These peptides are related but not identical to bombinin [Csordas, A. & Michl, H. (1970) Monatsh. Chem. 101, 182-189]. By using synthetic oligonucleotides corresponding to the C-terminal region of the peptides, a cDNA library from the skin of B. variegata was screened and several positive clones coding for the corresponding peptide precursors were isolated and sequenced. Each clone contained the genetic information for a different bombinin-like peptide. The antimicrobial activity towards different bacterial species of a synthetic peptide corresponding to one of the variants deduced from cDNA sequences was tested. This peptide was found to be mainly active against different isolates of Staphylococci and Escherichia coli.

Among Listeria genus, only two species, Listeria ivanovii and Listeria monocytogenes, are pathogenic. L. ivanovii is almost only associated with infections in animals, mainly sheep and cattle, and has rarely been associated with human infections, whereas L. monocytogenes causes severe illnesses in both humans and animals. To further investigate the pathogenetic features of L. ivanovii in humans, we undertook a study in which the intracellular behaviour of this pathogen was analysed in WISH cells, a cell line derived from human amniotic tissue, and compared to that of L. monocytogenes. Using microbiological, biochemical, and ultrastructural approaches, we demonstrate that L. ivanovii can adhere to and invade human amniotic cells, lyse the phagosomal membrane, polymerize host cell actin, and spread from cell to cell more efficiently than L. monocytogenes. However, although L. ivanovii is capable of specifically infecting and replicating in human amnion cells, its survival in cytoplasm is limited compared to that of L. monocytogenes.

The basic molecular mechanisms regulating prostate cancer (PCA) development and progression are very poorly understood. Different tumor suppressor genes are implicated in PCA. In particular, since the mutation rate of the p53 gene is also low, researchers have speculated that an infectious agent might play an important role in PCA. Polyomaviruses are candidates for this agent. We selected a patient with a diagnosis of PCA and underwent radical prostatectomy, to investigate the presence of Polyomavirus BK (BKV) sequences (urine and neoplastic tissues) and the mutation pattern of p53 gene. The results obtained showed the presence of BKV DNA and of p53 gene mutations in exons 6, 8 and 9. We speculate that BKV might contribute to cellular transformation process, triggered possibly by p53 gene mutations.

Interstitial cystitis (IC) is a syndrome consisting of severe refractory bladder symptoms of unknown etiology. The disease tends to affect Caucasian women with a mean age of 40 years, with 25% of patients under the age of 30. Few population based epidemiological studies of IC have been performed. We analyzed a case of interstitial cystitis in a 42-year-old non-smoker woman. In two biopsy samples the presence of viral DNA of human polyomavirus BK (BKV), human herpes virus type 1 and type 2 (HHV-1 and HHV-2), adenovirus, human papillomavirus (HPV) and bacterial DNA (Chlamydia trachomatis and Mycoplasma genitalium) were evaluated by means of polymerase chain reaction (PCR). Both samples resulted positive only for BKV and HPV DNA. HPV genotyping revealed the presence of HPV-66 that is associated with a high risk of cancer development. Thus the finding of a viral co-infection could support the hypothesis of the multi-factorial origin of this pathology.

Encrusted cystitis is a severe chronic inflammatory disease of the bladder characterized by excessively alkaline urine and calcifications within the bladder wall. A case of a 60 year-old man affected by systemic lupus erythematosus (SLE), which developed encrusted cystitis due to Corynebacterium urealyticum with E. coli coinfection, shows that the treatment of encrusted cystitis with a endoscopic debulking of the encrusted stones and an antimicrobial therapy specific for C. urealyticum often is not sufficient for the complete resolution of symptoms.