Quantifying synovitis in rheumatoid arthritis using computer-assisted manual segmentation with 3 Tesla MRI scanning.

PURPOSE: To investigate the reliability, validity and feasibility of a computer-assisted manual segmentation method for determining the synovial membrane volume as a surrogate measure for synovitis in patients with rheumatoid arthritis (RA). MATERIALS AND METHODS: The 3 Tesla (T) MRI scans were acquired in 22 early RA and 16 established RA patients. Synovial membrane volumes in postcontrast T1w axial images at three wrist joint regions were determined by two nonradiologist observers using a computer-assisted manual segmentation method. RESULTS: Intraobserver reliability, measured by intraclass correlation coefficients (ICCs), was excellent in the early (ICC = 0.99) and established (ICC = 0.99) RA cohorts. Interobserver reliability (mean ICC [95% Confidence Interval]) was moderate to excellent in the early and established RA groups (ICCs = 0.87 [0.68,0.94] and 0.88 [0.66, 0.96], respectively). There was a strong correlation between the synovial membrane volumes derived by segmentation and the RA MRI scoring system (RAMRIS) scores for synovitis at all joints in the early (Spearman rho = 0.86-0.96) and established (Spearman rho = 0.85-0.93) RA cohorts. The entire segmentation technique took 19 to 21 min per patient. CONCLUSION: Measurement of MRI synovitis using a computer-assisted manual segmentation method demonstrated excellent intraobserver and very good interobserver reliability, content validity (represented by its strong correlation with RAMRIS synovitis), and moderate feasibility.

Rare AU-1-like G3P[9] human rotaviruses with a Kun-like NSP4 gene detected in children with diarrhea in Italy.

Three G3P[9] rotaviruses, detected in children hospitalized with gastroenteritis in Palermo, Italy, were found to be genetically related to strains of either human or feline origin in the VP7, VP4, and VP6 genes. In contrast, in the NSP4 gene the viruses resembled G2P[4] human strains, suggesting a reassortment between AU-1-like and Kun-like strains.

[Choroidal metastasis as a presenting feature of breast carcinoma]. / Métastase choroïdienne révélant un cancer du sein.

Choroidal metastasis is a rare presenting feature of breast carcinoma. A 48-year-old woman presented with blurred vision of the right eye related to choroidal metastasis. Diagnostic work-up disclosed breast carcinoma with multiple metastases of the liver and lungs. Initial cerebral computed tomography scan was normal. During the follow-up, generalized seizure leaded to the diagnosis of multiple calcified cerebral metastasis. In 15 to 30 percent of cases, choroidal metastasis reveals a solid tumor, usually of the lung or the breast. Cerebral metastasis are common in breast cancer, but rarely calcified.

Monocyte and lymphocyte apoptosis resistance in acute and chronic brucellosis and its possible implications in clinical management.

This study evaluated the level of susceptibility of monocytes and lymphocytes to spontaneously induced and CH11-induced apoptosis in 16 patients with Brucella infection. The expression of some immunological and apoptotic markers was evaluated. Before therapy, monocytes showed a high level of resistance to spontaneously induced or CH11-induced apoptosis in all patients. In patients with acute infection, this resistance persisted for 10-20 days after treatment was initiated, then decreased; in chronically infected patients, it persisted after 45 days of treatment. Lymphocytes were also more resistant to CH11-induced apoptosis. The level of activated CD8(+) T lymphocytes was high in patients with acute infection. The data indicate that the CD95-mediated apoptotic pathway is not involved in CH11 resistance. Lymphocytes are not infected by Brucella, so their resistance to apoptosis may be due to a soluble factor released by infected monocytes. The evaluation of levels of susceptibility to CH11-induced apoptosis in monocytes may be used to test the effectiveness of the therapy.

Enteropathogens associated with childhood diarrhea in Italy. The Italian Study Group on Gastrointestinal Infections.

BACKGROUND: Infectious diarrheal diseases remain an important cause of childhood morbidity in industrialized countries. The knowledge of the etiology and epidemiology of childhood diarrhea in a given area is needed to plan any measure designed to prevent or ameliorate diarrheal illness and to develop practice guidelines for the most appropriate stool examination procedures. METHODS: We evaluated 618 children with diarrhea and 135 controls prospectively for viral, bacterial and parasitic enteric pathogens. Diarrheagenic Escherichia coli was identified by gene probes specific to different virulence factors. Stool filtrates were examined for the presence of free bacterial toxins by a cell culture cytotoxicity assay. Clinical and epidemiologic data were recorded and analyzed in relation to microbiologic findings. RESULTS: Enteropathogens were identified in 59% of children with diarrhea and in 10.4% of asymptomatic controls. The agents mainly associated with disease were rotavirus (23.6%), Salmonella (19.2%) and Campylobacter (7.9%). Rotavirus was significantly more frequent among children observed as inpatients whereas Campylobacter was significantly more common in outpatients. Infections with diarrheagenic E. coli, Shigella flexneri, yersinia enterocolitica, Cryptosporidium and Giardia were observed in a limited number of patients. The clinical presentation of children was not sufficiently characteristic to permit presumptive diagnosis of a specific pathogen. conversely the presence of blood and/or leukocytes in stools had a high positive predictive value for Salmonella or Campylobacter infection. CONCLUSION: The results of this study will be useful for planning strategies to prevent and control diarrheal diseases in our country.

Mitochondrial disruption and apoptosis in lymphocytes of an HIV infected patient affected by lactic acidosis after treatment with highly active antiretroviral therapy.

AIMS: Highly active antiretroviral therapy (HAART) can induce an increase in lactic acid concentrations that seems to be caused by mitochondrial dysfunction induced by the interaction of nucleoside reverse transcriptase inhibitors (NRTIs) with DNA polymerase gamma in the mitochondria. Mitochondrial alterations have been described in liver and muscle cells of NRTI treated human immunodeficiency virus (HIV) infected patients. Because lymphocytes are the main target for HIV and because mitochondria are involved in apoptosis, we studied mitochondrial morphology and apoptosis in the lymphocytes of an HIV infected patient with severe lactic acidosis after treatment with stavudine, didanosine, and indinavir. METHODS: The patient was a 39 year old woman. After two years of treatment she developed rapid weight loss with severe fat wasting, peripheral neuropathy, and hyperlacticaemia, which persisted after treatment withdrawal. The numbers and the morphology of the mitochondria were evaluated by electronic microscopy; the percentage of apoptotic cells was calculated by flow cytometry after staining with annexine V and by fluorescent microscopy after staining with ethidium bromide and acridine orange. RESULTS: The numbers of mitochondria in the lymphocytes were greatly decreased when compared with the lymphocytes of healthy individuals. The most important mitochondrial morphological alterations were swelling and the disruption of cristae and internal mitochondrial structure. These alterations were more evident during the period in which lactic acid values were very high. Moreover, a high percentage of apoptotic lymphocytes was seen. Morphological examination conducted one week after the normalisation of lacticaemia showed a pronounced increase in the number of mitochondria. The morphological alterations were no longer evident, although the size of each mitochondrion was smaller than normal. Moreover, the percentage of apoptotic cells was lower than 5%. CONCLUSIONS: This report describes important morphological alterations in lymphocyte mitochondria in an HIV infected patient during a severe phase of HAART induced hyperlacticaemia. These alterations persisted for several weeks after treatment withdrawal and were associated with an increase in lymphocyte apoptosis. Considering the important role of mitochondria in the apoptotic pathway, the increase in lymphocyte apoptosis may be a consequence of proapoptotic factors released from altered mitochondria.

Molecular epidemiology of astrovirus infection in Italian children with gastroenteritis.

Abstract A 1-year study involving 157 gastroenteritis samples was conducted to investigate the role of human astrovirus (HAstV) as a cause of gastroenteritis in Italian children aged < 2 years. The overall incidence of HAstV was 3.1%. Most cases occurred between March and May, and four of the five isolates were of the HAstV-1 type, the other being HAstV-3. Analysis of genetic variability showed that the three HAstV-1 isolates collected in 2000 clustered together, but separately from the 1999 isolate. The results indicated that HAstV should be considered as a potential diarrhoeal pathogen in Italian children.

Streptococcus milleri group infection associated with digestive fistula in patients with vascular graft: report of seven cases and review.

We described seven patients with Streptococcus milleri group aortic (six patients) or vena cava (one patient) graft infection secondary to a vasculo-digestive fistula. Time between vascular graft setting and first clinical signs varied from eight months to more than thirteen years. Six patients had fever. Three patients presented with recurrent fever for more than nine months and in two of these cases, delay before diagnosis was long because repeated blood cultures were sterile. Three patients had abdominal pain and/or digestive haemorrhage. Abdominal CT-scan S. milleri was not contributive for the diagnosis in four patients. Streptococcus anginosus was isolated in four patients, Streptococcus constellatus in three patients. One patient died before surgical management. The other six patients were cured by a surgical management associated with a prolonged antibiotic (lactams) treatment. S. milleri group graft infections are rare (or misdiagnosed) while we found only 4 similar cases in the English medical literature. We conclude that a peri-prosthetic infection secondary to a digestive fistula must be insistently searched (and blood cultures must be repeated many times) in any patient with an aortic (or any other vascular) graft presenting prolonged or recurrent fever or acute digestive symptoms.

Diversity of human rotaviruses detected in Sicily, Italy, over a 5-year period (2001-2005).

Rotavirus infection was detected in 39.9% of 1030 children hospitalized with gastroenteritis in Palermo, Italy, in the period 2001-2005. Rotavirus strains belonging to G1, G4 and G9 types were continually detected, with G1 being the most common type in 2001, 2002 and 2004. A G4 epidemic occurred in 2003, while G9 was predominant in 2005. G2 strains displayed a low prevalence, except in 2003. G3 rotaviruses accounted for 2.7-17% of the gastroenteritis episodes in 2002-2005. The P-type of a subset of 166 strains confirmed the circulation of the usual G/P combinations, but single G1P[6], G9P[9] and G6P[9] strains were also found.

Identification of group A porcine rotavirus strains bearing a novel VP4 (P) Genotype in Italian swine herds.

The VP4 gene of a G5 Italian porcine rotavirus strain, 344/04-1, was nontypeable by PCR genotyping. The amino acid sequence of the full-length VP4 protein had low identity (

Viral gastroenteritis in children hospitalised in Sicily, Italy.

The aim of the present study was to describe the epidemiologic and clinical characteristics of acute viral gastroenteritis in hospitalised Italian children. A total of 215 stool specimens were collected from January to December 2003 from patients hospitalised in Palermo for acute diarrhoea. Samples were tested for group A rotavirus, astrovirus, adenovirus, norovirus, enteropathogenic bacteria, and parasites. Rotaviruses, mostly belonging to types G1-G4, were detected in 25.1% of samples, astrovirus in 7%, adenovirus in 6%, norovirus in 18.6%, and bacterial agents in 17.2%. No parasitic infections were diagnosed. Mixed infections represented 9.8% of all cases. The mean and median ages of children with rotavirus gastroenteritis were lower than those of children with other viruses (p = 0.029), with the highest median ages being found in astrovirus-infected patients. Vomiting and dehydration were more frequent among patients with viral infection (p < 0.01), and the severity score was significantly higher for children infected with astrovirus or group A rotavirus (p = 0.008). Rotavirus was the leading cause of prolonged hospitalisation (p = 0.005). In conclusion, viruses were confirmed in Italy as the most common cause of severe enteric illness in childhood, with rotavirus types G1-G4, which correspond to those included in the rotavirus vaccines being developed, playing the main role. Routine testing should be introduced for noroviruses, since they seem to represent an important cause of sporadic paediatric gastroenteritis.

Identification of a novel VP4 genotype carried by a serotype G5 porcine rotavirus strain.

Rotavirus genome segment 4, encoding the spike outer capsid VP4 protein, of a porcine rotavirus (PoRV) strain, 134/04-15, identified in Italy was sequenced, and the predicted amino acid (aa) sequence was compared to those of all known VP4 (P) genotypes. The aa sequence of the full-length VP4 protein of the PoRV strain 134/04-15 showed aa identity values ranging from 59.7% (bovine strain KK3, P8[11]) to 86.09% (porcine strain A46, P[13]) with those of the remaining 25 P genotypes. Moreover, aa sequence analysis of the corresponding VP8* trypsin cleavage fragment revealed that the PoRV strain 134/04-15 shared low identity, ranging from 37.52% (bovine strain 993/83, P[17]) to 73.6% (porcine strain MDR-13, P[13]), with those of the remaining 25 P genotypes. Phylogenetic relationships showed that the VP4 of the PoRV strain 134/04-15 shares a common evolutionary origin with porcine P[13] and lapine P[22] rotavirus strains. Additional sequence analyses of the VP7, VP6, and NSP4 genes of the PoRV strain 134/04-15 revealed the highest VP7 aa identity (95.9%) to G5 porcine strains, a porcine-like VP6 within VP6 genogroup I, and a Wa-like (genotype B) NSP4, respectively. Altogether, these results indicate that the PoRV strain 134/04-15 should be considered as prototype of a new VP4 genotype, P[26], and provide further evidence for the vast genetic and antigenic diversity of group A rotaviruses.

Genetic variability among serotype G4 Italian human rotaviruses.

A total of 254 serotype GH rotavirus strains were detected in Palermo, Italy, from 1985 to 2003. Out of 38 serotype G4 strains selected for genetic analysis, 14 were recognized by genotyping as type G9. Strains confirmed to belong to the G4 type showed temporal patterns of genetic evolution in their VP7 and VP4 gene sequences, and the latest Italian G4 strains were distantly related to the reference vaccinal ST3 strain.

Sequence analysis of the VP7 and VP4 genes identifies a novel VP7 gene allele of porcine rotaviruses, sharing a common evolutionary origin with human G2 rotaviruses.

During an epidemiological survey encompassing several porcine herds in Saragoza, Spain, the VP7 and VP4 of a rotavirus-positive sample, 34461-4, could not be predicted by using multiple sets of G- and P-type-specific primers. Sequence analysis of the VP7 gene revealed a low amino acid (aa) identity with those of well-established G serotypes, ranging between 58.33% and 88.88%, with the highest identity being to human G2 rotaviruses. Analysis of the VP4 gene revealed a P[23] VP4 specificity, as its VP8* aa sequence was 95.9% identical to that of the P14[23],G5 porcine strain A34, while analysis of the VP6 indicated a genogroup I, that is predictive of subgroup I specificity. Analysis of the 10th and 11th RNA segments revealed close identity to strains of porcine and human origin, respectively. The relatively low overall aa sequence conservation (<89% aa) to G2 human rotaviruses, the lack of N-glycosylation sites that are usually highly conserved in G2 rotaviruses, and the presence of several amino acid substitutions in the major antigenic hypervariable regions hampered an unambiguous classification of the porcine strain 34461-4 as G2 serotype on the basis of sequence analysis alone. The identification of a borderline, G2-like, VP7 gene allele in pigs, while reinforcing the hypotheses of a tight relationship in the evolution of human and animal rotaviruses, provides additional evidence for the wide genetic/antigenic diversity of group A rotaviruses.

Rotavirus gastroenteritis in Italian children: can severity of symptoms be related to the infecting virus?

The aim of our study was to determine whether the severity of rotavirus gastroenteritis may be related to the different characteristics of infecting viral strains. The severity of clinical symptoms in 401 children with acute rotavirus gastroenteritis was assessed using a scoring system for frequency and duration of vomiting, diarrhea, and fever, as well as the patients' requirements for intravenous rehydration. Rotavirus strains were characterized by determining the electropherotype of their double-stranded RNA, the G type and subgroup by a panel of monoclonal antibodies, and the P type by reverse transcription-polymerase chain reaction. Strains with a short electropherotype, G2P[4] type, and subgroup I were associated with more-severe gastroenteritis and affected children older than those infected with strains with a long electropherotype, G1P[8] or G4P[8] type, and subgroup II. Minor differences in clinical symptoms were also detected in children infected with different long electropherotypes and with G1P[8] and G4P[8] specificities.

Comparison of serological methods for the detection of IgG and IgM antibodies to rubella virus.

Four commercial kits- the hemagglutination inhibition test Behring (HI-B), the HI Rubenosticon, the latex test Rubalex, the enzyme-linked immunosorbent assay (ELISA) ETI-RUBEK G--and a non-commercial hemolysis test in gel (SRH) were compared for the measurement of rubella IgG antibodies by testing 126 serum samples, submitted for rubella immune status determination. Two commercial ELISAs (ETI-RUBEK M and Rubazyme M) were also evaluated for the detection of rubella IgM antibodies on 64 sera from patients with suspected acute rubella infection. In the assay for rubella specific IgG antibodies, HI-B and ETI-RUBEK G tests exhibited the most sensitivity, whereas Rubenosticon and Rubalex tests showed more specificity. In the IgM evaluations, six false positive results were shown by Rubazyme M, whereas the ETI-RUBEK M kit was sensitive and specific. On the basis of all the determinations, a combination of two tests (HI-B and SRH) appears to be a good simple system to obtain valid indications in serological rubella diagnosis.

[Serological determination of anti-mumps immunity in a population sample from Palermo]. / Determinazione sierologica di immunita antiparotitica in un campione di popolazione palermitana.

A seroepidemiological survey was performed by an immunoglobulin G antibody-capture immunoassay on 1061 subjects from Palermo (Sicily), throughout the age range from birth up to 50 years. A proportional increase of seropositivity rate to mumps virus was shown in relation to increasing age, from 17.5% in the earliest years up to 60.3% in children aged 10 years. After the first decade of life, seropositivity rate reached a plateau staying constant up to the upper age classes explored. Given the relatively high frequency of complications upon mumps infection, it is suggested that an extended vaccination policy could cover a proportion of the non-immune adult subjects.

[Virological and serological study in the herpetic gengivostomatitis (author's transl)]. / Criteri virologici e sierologici per l'accertamento diagnostico di casi di stomatite erpetica.

Virological and serological investigations to verify herpetic etiology have been performed on 24 children and 1 adult with acute gengivostomatitis. Viral isolation and detection of antibodies against HSV, carried out by neutralization, complement fixation, and indirect immunofluorescence techniques, have confirmed this etiology in the 88% of cases.

Rotavirus persistence in cell cultures: selection of resistant cells in the presence of foetal calf serum.

The Lincoln strain of bovine rotavirus, cytocidal for bovine AU-BEK cells, can establish in the same cell cultures in the presence of foetal calf serum (FCS) a persistent infection that depends on selection of highly resistant cells. In fact, after the induction of the carrier state only a small fraction of the cell population was infected. The parental and the carried viruses are not demonstrably different, the cultures are resistant to superinfection by the homologous virus, the cultures can be cured by antiviral serum in the medium and uninfected resistant cell clones can be selected. The presence of FCS was essential during induction and maintenance of persistence. Its effects appear to be exerted not on the control of virus replication in fully sensitive cells but on the proliferation and selection of the resistant cells.

[Immunofluorescence reaction for rotavirus analysis in fecal specimens]. / La reazione di immunofluorescenza per la ricerca di rotavirus in campioni di feci.

Testing for rotavirus detection in faeces from infants with acute diarrhea can be carried out, with high sensitivity, by immunofluorescence on rhesus monkey kidney cell lines (MA-104). Preincubation of viral inoculum with trypsin enhances further the sensitivity. In the present study we tried to assess the optimal conditions for detection of viral antigens, by analyzing the expression of virus-specific proteins at different times of infection and the effects of trypsin presence during virus adsorption and replication. Maximal production of viral antigens was obtained by infecting the cells 4 hours after cell-seeding and incubating them for 18 hours afterwards. In experiments with simian rotavirus SA-11, pretreated with trypsin and then with a trypsin inhibitor, a marked reduction in the percentage of fluorescent cells was observed thus indicating a role of the enzyme both on the virus and on the cell layer.