Intestinal spirochaetosis associated with hyperplastic and adenomatous colonic polyps.

Intestinal spirochaetosis is a human colonic infection due to Brachyspira aalborgi and Brachyspira pilosicoli causing various abdominal complaints. Although the presence of healthy epithelial cells was hypothesized to be essential for the adhesion of spirochaetes to the colonic mucosa, their adhesion to hyperplastic and adenomatous colonic polyps has been observed recently. We report a case of a woman with long-standing abdominal symptoms, in whom spirochaetes were found on the colonic mucosa surrounding an adenocarcinoma in the biopsies collected during eight years of follow-up. Spirochaetes were found attached to normal mucosa, to hyperplastic and to adenomatous polyps, but not to the epithelium of the carcinoma. The rectal biopsy collected during the last follow-up colonoscopy was subjected to histopathology and to a specific examination for brachyspires, demonstrating the presence of B. pilosicoli DNA. This report could stimulate microbiological investigations during the follow-up of colonic polyps in order to explain whether the persistence of abdominal symptoms in such patients could be caused by a colonic spirochaetosis susceptible to eradication by a targeted therapy.

Case report: detection of rotavirus RNA in the cerebrospinal fluid of a child with rotavirus gastroenteritis and meningism.

Although case reports have described detection of rotavirus (RV) in extraintestinal sites such as the liver, kidney, and central nervous system (CNS) of children with RV gastroenteritis, CNS localization in RV infection seems to be rare. RT-PCR and nucleotide sequencing detected a G1P[8] strain in the stool and cerebrospinal fluid (CSF) samples of a patient with concurrent RV-associated enteritis and CNS signs. Upon sequence analysis, the viruses detected in the CSF was identical to the virus detected in the stools. In the VP7- and VP4-based phylogenetic dendograms the strain clustered within the G1-Ic sub-lineage and the P[8]-III lineage. This study supports the hypothesis that RV infection was able to spread from the intestinal tract to the CNS, and likely played a role in the onset of neurological disease.

Hepatitis C virus core antigen: analytical performances, correlation with viremia and potential applications of a quantitative, automated immunoassay.

BACKGROUND: Testing for hepatitis C virus core antigen (HCV Ag) may represent a complementary tool to anti-HCV and HCV-RNA in the diagnosis and monitoring of HCV infection. OBJECTIVE: To evaluate the performance characteristics of the automated Abbott ARCHITECT HCV Ag assay. STUDY DESIGN: Five sites analyzed over 3000 routine serum samples from populations at different risk, comparing HCV Ag results with anti-HCV screening and supplemental assay results and with HCV-RNA. RESULTS: The HCV Ag assay showed a specificity of 100%, a good precision (CV<10%) and excellent dilution linearity (r>0.999). The sensitivity (3 fmol/L) corresponds to 700-1100 IU/mL of HCV-RNA. A non-linear correlation with HCV-RNA was found: r=0.713 vs. Siemens bDNA (523 specimens), r=0.736 vs. Roche Cobas TaqMan (356 specimens) and r=0.870 vs. Abbott Real-Time PCR (273 specimens). HCV Ag quantitation was equally effective on different HCV genoypes (239 for genotype 1/1a/1b/1c, 108 for genotype 2/2a/2c, 86 for genotype 3/3a, 50 for genotype 4/4a/4c/4d). Testing of subjects at high risk for HCV and with potential or actual impairment of the immune system identified 2 cases negative for anti-HCV and positive for HCV Ag on 361 hemodialyzed (0.6%) and 7 cases on 97 (7.2%) among transplant recipients. HCV Ag positivity anticipated anti-HCV seroconversion in all three cases of acute hepatitis C. CONCLUSIONS: HCV Ag may be used as reflex testing on anti-HCV positive individuals to confirm or exclude an active infection, and on subjects with acute hepatitis or belonging to high risk groups.

Similar diagnostic performances of antigen detection and nucleic acid detection of Cryptosporidium spp. in a low-prevalence setting.

The diagnostic value of a real-time PCR assay for the detection of Cryptosporidium spp. in fecal samples was assessed as compared to the combination of the immunocromatographic assay (IC) and immunofluorescence assay (IF) currently performed in our laboratory for the diagnosis of cryptosporidiosis. On a total of 1040 samples collected from 2006 to 2010 and belonging to 533 patients suspected of having an intestinal parasitosis, Cryptosporidium spp. was detected in 31 samples (belonging to 12 patients) by IC and IF; the real-time PCR assay revealed Cryptosporidium spp. DNA in 5 additional samples for a total of 36 samples (13 patients). The real-time PCR assay exhibited higher sensitivity than IC and IF; however, its application to the diagnosis of cryptosporidiosis should be evaluated by every single laboratory, depending on the availability of trained personnel, financial resources, and the cost/effectiveness related to the prevalence of cryptosporidiosis.

Clinical and molecular observations of two fatal cases of rotavirus-associated enteritis in children in Italy.

Two fatal cases of infantile rotavirus enteritis occurred in northern Italy in 2005. Both children were severely dehydrated, and death was related to severe cerebral edema. Histological examination demonstrated extensive damage of the intestinal epithelium, villous atrophy or blunting, and macrophage infiltration. The two rotavirus strains were of the G1P[8] type and the long electropherotype. The 2005 G1P[8] rotaviruses differed in the NSP4, VP3, VP4, and VP7 genes from G1P[8] rotaviruses circulating in 2004, suggesting the onset of a new G1P[8] strain in the local population.

Presence of anti-Borrelia burgdorferi antibodies and Borrelia burgdorferi sensu lato DNA in samples of subjects in an area of the Northern Italy in the period 2002-2008.

Lyme borreliosis (LB) is a multisystemic disease caused by Borrelia burgdorferi sensu lato complex transmitted to humans by Ixodes ticks and whose epidemiology is poorly investigated in Europe. In this study an epidemiologic survey on the prevalence of the causative agent of such infectious disease was performed in the area of Parma (Northern Italy) during 2002-2008. Serum samples belonging to 2336 patients and cerebrospinal fluid samples (belonging to 42 of the same patients) were analyzed for serologic diagnosis of LB. Direct laboratory assays [polymerase chain reaction (PCR) and cultivation] were performed on samples belonging to patients with the clinical suspicion of LB. The seroprevalence was 0.55% considering the subjects with both anti-B. burgdorferi IgG and IgM. The samples tested by culture and PCR were all negative except for a tick removed from the skin of a healthy man. The results suggest that infection by B. burgdorferi in this area quite rarely occurs.

Differential infectious entry of human influenza A/NWS/33 virus (H1N1) in mammalian kidney cells.

In this report we focused our interest on the early events of the replication cycle of NWS/33 human influenza A (NWS) virus in MDCK (canine), LLC-MK2 (simian), and NSK (swine) kidney cells, with different susceptibility upon infection. We have previously demonstrated that actin organization induces restriction to viral replication during the early stages of NWS virus infection in simian kidney cells. To explore how cell endocytic mechanisms are hijacked by NWS virus and may modulate the outcome of viral infection, the effect of drugs affecting selectively the entry via clathrin-coated pits, caveolar/raft-dependent endocytosis and macropinocytosis was analyzed. Results point to critical differences in terms of internalization pathways exploited by NWS virus to enter the examined cell models. Moreover, we show that some ways of entry do not allow an effective virus internalization, depending on the cell type. Understanding how specific cell functions/components may regulate early phases of viral replication allows us to deepen our knowledge on influenza virus infection and provides new insights for anti-viral researches.

Cell-cycle-dependent localization of human cytomegalovirus UL83 phosphoprotein in the nucleolus and modulation of viral gene expression in human embryo fibroblasts in vitro.

The nucleolus is a multifunctional nuclear compartment widely known to be involved in several cellular processes, including mRNA maturation and shuttling to cytoplasmic sites, control of the cell cycle, cell proliferation, and apoptosis; thus, it is logical that many viruses, including herpesvirus, target the nucleolus in order to exploit at least one of the above-mentioned functions. Recent studies from our group demonstrated the early accumulation of the incoming ppUL83 (pp65), the major tegument protein of human cytomegalovirus (HCMV), in the nucleolus. The obtained results also suggested that a functional relationship might exist between the nucleolar localization of pp65, rRNA synthesis, and the development of the lytic program of viral gene expression. Here we present new data which support the hypothesis of a potentially relevant role of HCMV pp65 and its nucleolar localization for the control of the cell cycle by HCMV (arrest of cell proliferation in G1-G1/S), and for the promotion of viral infection. We demonstrated that, although the incoming pp65 amount in the infected cells appears to be constant irrespective of the cell-cycle phase, its nucleolar accumulation is prominent in G1 and G1/S, but very poor in S or G2/M. This correlates with the observation that only cells in G1 and G1/S support an efficient development of the HCMV lytic cycle. We propose that HCMV pp65 might be involved in regulatory/signaling pathways related to nucleolar functions, such as the cell-cycle control. Co-immunoprecipitation experiments have permitted to identify nucleolin as one of the nucleolar partners of pp65.

Pancreatic pseudocyst-inferior vena cava fistula causing caval stenosis, left renal vein thrombosis, subcutaneous fat necrosis, arthritis and dysfibrinogenemia.

AIM: We describe the case of a 38 year old man, with a story of alcohol abuse, who developed a very painful nodular subcutaneous fat necrosis, fever and polyarthritis, denying any abdominal symptoms due to a pancreatic pseudocyst-inferior vena cava fistula. MATERIAL OF STUDY: The authors discuss the unusual and protracted course with intermittent hyperamylasemia and hyperlipasemia related to clinical manifestations such as subcutaneous fat necrosis, polyarthritis, pleural effusion and dysfibrinogenemia, and vascular complications as inferior vena cava stenosis and left renal vein thrombosis without abdominal symptomatology. RESULTS: After ultrasonograms and CT Scans showing a 3-4 cm cyst at the pancreatic head with a solid bud protruding into the pseudocystic cavity, and an ERCP showing a communication between the pancreatic duct and the pseudocyst but failing in demonstrating the vascular fistula, the patient underwent a Roux-en-y pseudocyst-jejunostomy and suture of the caval communication leading to complete recovery with normalization of laboratory findings. DISCUSSION: In our case, the locally sclerosing activity of the enzymes in the endothelium led to a communication between the inferior vena cava and the pseudocyst and to a complete thrombosis of the left renal vein and to a stenosis of the inferior vena cava itself The fluctuance of the symptomatology severity was probably due to an intermittent opening of the passage between pseudocyst and vena cava. Such a clinical case, to the author knowledge, has never been reported. CONCLUSION: When in presence of very high levels of amylasemia and lipasemia in spite of the paucity of abdominal symptomatology, and the onset of unusual complications such as panniculitis, pleural effusion, arthritis and coagulative disorders, a pancreatic pseudocyst-inferior vena cava fistula should be kept in consideration during diagnosis.

Evaluation of a real-time polymerase chain reaction assay for the detection of Dientamoeba fragilis.

The diagnostic value of a real-time polymerase chain reaction (PCR) assay targeting the 5.8S rDNA of Dientamoeba fragilis was investigated as compared with conventional parasitologic methods including cultivation testing 959 fecal samples from 491 patients attending a tertiary-care hospital and suspected of having an intestinal parasitosis. The real-time PCR assay revealed 117 additional D. fragilis-positive samples as compared with conventional methods, showing 100% sensitivity and specificity in our experience. On the whole, D. fragilis infection was detected in 186 samples from 105 patients (21.4%, third in frequency among the diagnosed intestinal parasitoses). The evaluated real-time PCR assay represents an effective tool to obtain both an accurate diagnosis and a reliable epidemiologic picture of dientamoebiasis.

Norovirus RNA in the blood of a child with gastroenteritis and convulsions--A case report.

Potential extra-intestinal spread is an important issue in understanding the pathogenesis of NoV disease. A previously healthy 14-month-old boy was admitted to the Pediatric Emergency Department of the University-Hospital of Parma, Italy, for afebrile convulsions in a gastroenteritis episode. Bacterial culture and microscopic examination on cerebrospinal fluid (CSF) yielded negative results as well as PCRs and reverse-transcription PCRs (RT-PCRs) for neurotropic viruses performed either on CSF or plasma. Stools were subjected to electron microscopy and conventional cell culture, yielding negative results. NoV was found in stools and plasma by nested RT-PCR targeting the NoV polymerase gene. The nucleotide sequences obtained from the two specimens showed 100% identity, demonstrating that the strain invading the blood stream was from the intestine, and, in comparison with GenBank sequences, they belonged to NoV genotype GII.4, "2006b" variant. The child had no abnormal electrolyte balance and no fever that could justify seizures, encouraging the hypothesis that NoV could be the cause of the neurologic disorder. These findings further induce to review the current concept of human NoV focused on intestinal infection.

Evaluation of a real-time polymerase chain reaction assay for the laboratory diagnosis of giardiasis.

A real-time polymerase chain reaction (PCR) assay was evaluated in comparison with the combination of conventional methods (microscopic examination and antigen detection assay) during the period 2006 to 2008 on 771 fecal samples belonging to 386 patients to assess its usefulness for an accurate laboratory diagnosis of giardiasis. The real-time PCR assay detected Giardia intestinalis DNA in 195 samples (106 patients), including 26 samples (21 patients) negative by the conventional assays. Among the 21 patients, in 8 cases, giardiasis was previously diagnosed also by conventional methods in additional samples of the same patients, whereas in 13, it would have been undiagnosed if real-time PCR assay was not used. The real-time PCR assay demonstrated a detection limit of 2 cysts per reaction and 100% specificity and sensitivity compared to conventional methods. A genotype analysis targeting the beta-giardin gene allowed to identify 53 samples (23 patients) containing genotype A and 59 samples (45 patients) containing genotype B.

Modulatory effect of rRNA synthesis and ppUL83 nucleolar compartmentalization on human cytomegalovirus gene expression in vitro.

The nucleolus is a nuclear domain involved in the biogenesis of ribosomes, as well as in many other important cellular regulatory activities, such as cell cycle control and mRNA processing. Many viruses, including herpesviruses, are known to exploit the nucleolar compartment during their replication cycle. In a previous study, we demonstrated the preferential targeting and accumulation of the human cytomegalovirus (HCMV) UL83 phosphoprotein (pp65) to the nucleolar compartment and, in particular, to the nucleolar matrix of lytically infected fibroblasts; such targeting was already evident at very early times after infection. Here we have investigated the possible effects of rRNA synthesis inhibition upon the development of HCMV lytic infection, by using either actinomycin D or cisplatin at low concentrations, that are known to selectively inhibit RNA polymerase I activity, whilst leaving RNA polymerase II function unaffected. Following the inhibition of rRNA synthesis by either of the agents used, we observed a significant redistribution of nucleolar proteins within the nucleoplasm and a simultaneous depletion of viral pp65 from the nucleolus; this effect was highly evident in both unextracted cells and in nuclear matrices in situ. Of particular interest, even a brief suppression of rRNA synthesis resulted in a very strong inhibition of the progression of HCMV infection, as was concluded from the absence of accumulation of HCMV major immediate-early proteins within the nucleus of infected cells. These data suggest that a functional relationship might exist between rRNA synthesis, pp65 localization to the nucleolar matrix and the normal development of HCMV lytic infection.

Primary appendiceal tumors: report on 10 cases.

We report our experience on 10 patients with primary tumors of the appendix treated at our institution from 1998 to 2005. There were 5 women and 5 men, with a mean age of 59.1 years. Laparotomy was performed in 4 cases; whereas, the other 6 patients underwent laparoscopic exploration: Three operations were completed laparoscopically, and 3 were converted to laparotomy. Six tumors were malignant, and the remaining were benign. Proportions of perioperative and late mortality were both 10%. Two of the four patients with benign tumors died from causes unrelated to the appendiceal neoplasm. The 6 patients with malignant tumors and the other 2 with benign disease were alive and disease free after a mean follow-up of 43 months. Despite the rarity of appendiceal primary tumors, surgeons should be aware of these neoplasms for making correct treatment decisions. We stress the importance of laparoscopic exploration in the management of appendiceal masses.

Molecular characterization of group C rotaviruses detected in children in Italy.

BACKGROUND: Group C rotavirus (GCRV) infection has been described worldwide in children and adults either as sporadic cases or large outbreaks of gastroenteritis but GCRV epidemiology is still unclear. OBJECTIVE: To acquire molecular information on GCRV infection in children hospitalized with gastroenteritis in Italy. STUDY DESIGN: Stools positive for rotavirus-like-particles by electron microscopy during the 2004-2005 surveillance for rotavirus infection in Parma, Italy, were screened by polyacrylamide gel electrophoresis. GCRV strains detected were characterized by sequence and phylogenetic analysis of the genes encoding the capsid proteins VP4, VP6 and VP7. RESULTS: Two of 856 samples (0.23%; 0.7% of 273 samples containing rotavirus-like particles) contained GCRV. These Italian strains were virtually identical in the 3 genes and were closely related to human strains identified in Asia, rather than to strains of European origin. CONCLUSIONS: These results confirm the circulation of GCRVs in Europe and demonstrate genetic heterogeneity among European GCRVs. Inclusion of GCRVs in the diagnostic algorithms for childhood diarrhoea could be helpful in monitoring temporal changes in the GCRV epidemiology, likely under the influence of the changing demographic dynamics.

Molecular characterization of VP4, VP6 and VP7 genes of a rare G8P[14] rotavirus strain detected in an infant with gastroenteritis in Italy.

In this study, the molecular characterization of a rare G8P[14] group A rotavirus (GARV) strain detected in Northern Italy during the 2004-2005 epidemiological rotavirus season is described. Two hundred and seventy three rotavirus-like particle positive stools out of 856 stools from children (31.9%) hospitalized with gastroenteritis were analyzed using polyacrilamide gel electrophoresis and 271 GARVs were genotyped by VP7 and VP4 specific RT-PCRs. One strain (PR/1300/04) with a long electropherotype (e-type) displayed the G8 specificity and was VP4 un-typeable. The P and the subgroup (SG) specificities were determined by sequencing the VP4 and the VP6 gene, respectively. The PR/1300/04 strain exhibited P[14] and SGI specificities. By sequence and phylogenetic analyses of the VP4, VP6 and VP7 amplicons, the PR/1300/04 VP4 and VP6 genes were demonstrated to be of human rotavirus origin, with the VP4 gene closely related to the human Italian PA169 strain (G6P[14]), while the VP7 gene was of animal origin (bovine). These data suggest that the Italian PR/1300/04 strain could be a reassortant between a PA169-like Italian strain with P[14] specificity, long e-type and SGI, and a G8 animal strain. The increasing number of reports of atypical GARVs in humans suggests that interspecies transmission of genes greatly contributes to the GARV genetic evolution.

Evaluation of Toxoplasma gondii immunoglobulin G (IgG) and IgM assays incorporating the newVidia analyzer system.

The new Vidia system is a fully automated system based on chemiluminescence and antigen bound to magnetic microparticles, which allows a fast measurement of Toxoplasma gondii-specific immunoglobulin G (IgG) and IgM levels. The analytical performances of the Vidia Toxo IgG and IgM assays were compared with those of the automated Vidas, AxSYM, and Liaison Toxo IgG and IgM assays. The comparative evaluation was performed utilizing 204 frozen sera belonging to 166 subjects and 201 fresh sera collected from 198 subjects. For the Vidia Toxo IgG system, the sensitivities were 100% in both the retrospective and prospective studies, and specificities were 98.39% in the retrospective study and 100% in the prospective study, respectively. The sensitivities of the other three Toxo IgG assays were 100%, and the specificities ranged from 96.77% to 100%. For the Vidia Toxo IgM assay, the sensitivity and specificity were 100% in both the retrospective and prospective studies. The overall sensitivities and specificities of the other three Toxo IgM assays ranged from 80% to 100% and from 99.44% to 100%, respectively. In our study, the Vidia system revealed excellent sensitivity (100% for both IgG and IgM assays) and good specificity (99.25% for IgG and 100% for IgM assays).

An 8-year survey on the occurrence of imported malaria in a nonendemic area by microscopy and molecular assays.

Our study aimed to describe the occurrence of imported malaria in a nonendemic area (Parma, Italy) during the period 2000 to 2007, comparing the data obtained by microscopy and molecular assays targeting plasmodial 18S subunit rRNA gene. The prevalence of imported malaria in Parma was 21.8% by microscopy and 22.7% by polymerase chain reaction (PCR). Plasmodium falciparum accounted for 81.1% of the cases, followed by Plasmodium ovale (8.8%), Plasmodium vivax (3.8%), and Plasmodium malariae (1.9%). Mixed infections accounted for 4.4% of the cases. In this study, PCRs proved to be more sensitive and specific than microscopy and changed the picture of malaria epidemiology in Parma, detecting additional cases of malaria undiagnosed by microscopy and allowing speciation of plasmodia in cases misidentified by microscopy. Generally, imported malaria cases reflect the number of immigrants who visit their native countries, in particular, West Africa, explaining the increased prevalence of P. ovale cases among non-P. falciparum infections in Parma.

Evaluation of rubella virus immunoglobulin G (IgG) and IgM assays with the new Vidia instrument.

For rubella virus immunoglobulin G (IgG) and IgM detection, Vidia assays were compared to Vidas, AxSYM, and Liaison assays with 419 serum samples. Only Vidia produced a sensitivity of 100% for IgG and IgM. Vidia specificities were 98.4% for IgG and 99.8% for IgM, versus Vidas specificities of 100 and 99.3%. Vidia IgG and IgM assays performed equally well.

Myiasis of the scalp due to Dermatobia hominis in a traveler returning from Brazil.

This article describes a case of myiasis by Dermatobia hominis diagnosed in a young Italian man returning from a vacation through Brazil. Considering the increasing number of travels to tropical and subtropical areas, clinicians in nonendemic areas must think about the possibility of imported unusual infestations during their daily practice.