Incidence and risk factors for respiratory tract bacterial colonization and infection in lung transplant recipients.

To evaluate incidence of and risk factors for respiratory bacterial colonization and infections within 30 days from lung transplantation (LT). We retrospectively analyzed microbiological and clinical data from 94 patients transplanted for indications other than cystic fibrosis, focusing on the occurrence of bacterial respiratory colonization or infection during 1 month of follow-up after LT. Thirty-three percent of patients developed lower respiratory bacterial colonization. Bilateral LT and chronic heart diseases were independently associated to a higher risk of overall bacterial colonization. Peptic diseases conferred a higher risk of multi-drug resistant (MDR) colonization, while longer duration of aerosol prophylaxis was associated with a lower risk. Overall, 35% of lung recipients developed bacterial pneumonia. COPD (when compared to idiopathic pulmonary fibrosis, IPF) and higher BMI were associated to a lower risk of bacterial infection. A higher risk of MDR infection was observed in IPF and in patients with pre-transplant colonization and infections. The risk of post-LT respiratory infections could be stratified by considering several factors (indication for LT, type of LT, presence of certain comorbidities, and microbiologic assessment before LT). A wider use of early nebulized therapies could be useful to prevent MDR colonization, thus potentially lowering infectious risk.

Telemedicine in pediatric wound care.

BACKGROUND: Telemedical wound care is one of the possible applications of teledermatology. The treatment of pediatric wounds needs frequent and periodic assessments of their local status and adjustment of dressings choice. MATERIALS AND METHODS: We present our experience using telemedicine in the successful assessment and treatment of 19 pediatric patients at the OPBG, Rome . Photographs with a digital camera were taken and sent weekly by mail to a wound care specialist in Rome. This allowed the expert to diagnose and evaluate the wounds periodically. RESULTS: In the shown cases, telemedicine allowed us to have an immediate evaluation and therapy adjustment. The quality of the images was good enough that the physician could regularly evaluate the status of the wound and immediately give his feedback to the parents. Of these 19, 13 patients (68%) experienced a wound resolution during the remote monitoring period. The satisfaction of the parents detected at 3, 6 and 12 months was found to be respectively 57%, 71%, 84%. CONCLUSION: Reducing transportation to the hospital to obtain a specialist advice, wound teleconsultation lowers health care costs and improve the quality of life for pediatric patients and their family, while still maintaining a high quality of pediatric wound care.

Increased phospholipase activity in Helicobacter pylori strains isolated from patients with gastric carcinoma.

PURPOSE: Phospholipase activity, one of Helicobacter pylori pathogenicity factors, has not been investigated enough, so far, although it may induce a remarkable damage to the gastric mucosa. In the present work, we have compared the whole phospholipase activity of H. pylori strains isolated from patients with gastric carcinoma with that of strains isolated from dyspeptic patients without gastric carcinoma. METHODS: We measured the phospholipase activity of one distinct H. pylori colony isolated from each of 10 patients with gastric carcinoma and 10 controls, dyspeptic patients without endoscopic and histological signs of gastric carcinoma. We also determined the phospholipase activity of 20 additional strains isolated from different areas of neoplastic and non-neoplastic tissue of two patients with gastric carcinoma, the cagA and vacA positive G27 and 328 wild strains and their respective vacA and cagA negative isogenic mutants. The whole phospholipase activity of strains was determined by measuring the release of (14)C-labeled palmitic acid from the radioactive l-3-phosphatidylcholine, 1,2-di[1-(14)C]palmiloyl substrate; results were expressed in pmol of palmitic acid per mg of protein. RESULTS: H. pylori strains isolated from patients with gastric carcinoma had levels of phospholipase activity significantly higher than those of strains isolated from controls (99.37 [S.D. 40.45] versus 34.46 [S.D. 16.46], P<0.001). In patients with gastric carcinoma, the mean phospholipase activity of strains isolated from neoplastic tissue was similar to that of strains isolated from non-neoplastic tissues (123.02 [S.D. 44.36] and 115.77 [S.D. 81.48], respectively. Interruption of cagA gene caused a ca. 20% reduction of phospholipase activity (36.38 versus 45.22 of the wild strain); that of vacA caused no reduction of phospholipase activity (26.53 and 25.37 of the wild strain). CONCLUSIONS: The infection by H. pylori strains that produce high levels of phospholipase may increase the risk of developing gastric carcinoma. We hypothesise that indirect products of phospholipase activity, such as prostaglandins, leukotrienes and lysophospholipids, may mediate carcinogenesis.

Cloning and sequencing of the F gene of live attenuated Urabe Am9 mumps virus.

A cDNA clone encoding the entire F gene of the live attenuated mumps virus (MpsV) strain, Urabe Am9, has been isolated and sequenced. The F gene sequence shows significant homology with the one reported for the wild-type MpsV Miyahara strain.

Evaluation of rubella virus E2 and C proteins in protection against rubella virus in a mouse model.

An animal model is described that can provide further information for evaluating novel vaccines against rubella virus (RV). A group of mice was immunized with the lysate of insect cells infected by a recombinant baculovirus expressing E2 and C proteins of RV. Another group of mice was immunized with the RA27/3 rubella vaccine. After 2 weeks, both groups of mice were challenged intramuscularly with live RV and the blood was drawn after 8, 24, 48 and 72 h. The presence of rubella challenge virus in an unnatural host, such as the mouse, was monitored by RT-PCR. The mice immunized with the RA27/3 rubella vaccine were the only ones able to inhibit the challenge virus replication, E2 and C proteins, which alone are not sufficient to protect animals against RV, served as a negative control for a protective vaccine against RV that expresses E1 protein of RV.

Localization of a new neutralizing epitope on the mumps virus hemagglutinin-neuraminidase protein.

Four protein fragments which span the entire hemagglutinin-neuraminidase protein (HN) of mumps virus were expressed in HeLa cells and cell extracts were tested for their capability to induce neutralizing antibodies in mice. Fragment HN3 (aa 213-372) was able to induce the production of hemagglutination-inhibiting and neutralizing antibodies. When a subfragment of HN3, the synthetic peptide NSTLGVKSAREF (aa 329-340 of HN) was used for immunization, hemagglutination-inhibiting and neutralizing antibodies against mumps wild type virus but not against the Urabe Am9 vaccine virus were raised. The peptide could, therefore, contain a new epitope, which may be critical for protective host humoral immune response.

Multiplex PCR for rapid and differential diagnosis of Mycoplasma pneumoniae and Chlamydia pneumoniae in respiratory infections.

A duplex polymerase chain reaction (PCR) was developed for the simultaneous detection of Chlamydia pneumoniae and Mycoplasma pneumoniae. A study of 163 respiratory specimens from in-patients of the "Centre Hospitalier et Universitaire de Nancy" showed the good sensitivity of this duplex PCR allowing the detection of C. pneumoniae and M. pneumoniae from 8 and 13 patients, respectively, whereas the culture was negative for C. pneumoniae for all the samples and positive for M. pneumoniae only in 9 cases. The value of these results has been confirmed by running on the same samples specific nested PCRs for these two microorganisms that gave the same results. Thus, the proposed duplex amplification technique may facilitate the diagnosis of infection by these two agents that are difficult to isolate.

Detection by multiplex polymerase chain reaction and typing of Chlamydia trachomatis isolates.

The multiplex polymerase chain reaction (PCR) was applied for the detection of the Chlamydia trachomatis chromosome and plasmid. The multiplex PCR demonstrated a sensitivity of 0.8 fg of chlamydial DNA, corresponding to the detection of about 5 copies of the plasmid. Analysis of 195 genital specimens collected randomly from a female population, showed that the multiplex PCR is more sensitive and rapid than culturing for detecting Chlamydia trachomatis. Moreover, sequencing of the II variable domain of the omp1 gene, directly from DNA of the clinical specimens, appears to be a simple and rapid method for determining serovar isolates.

Helicobacter pylori determinants of pathogenicity.

The bacterium Helicobacter pylori colonises the stomach of man and induces a strong inflammatory response. Differences in the possession of pathogenicity determinants by H. pylori isolates could account in part for the different clinical outcomes of infection. The main H. pylori pathogenic factors, i.e. urease, the cytotoxin VacA, and the genes involved in virulence contained in the pathogenicity island (PAI) cag, may promote tissue damage and ulceration, and could contribute to gastric cancer development. Strains with the mosaic vacA allelic type s1a/m1 and possessing the cag insertion are considered endowed with increased inflammatory potential, and are more likely to be isolated from patients with peptic ulcer and gastric cancer. The presence in H. pylori cag PAI of operons involved in the stimulation of gastric epithelial cells to secrete high levels of inflammatory cytokines, in mobilisation of DNA, and formation of secretory mechanisms and conjugation apparati, could contribute to increase the risk of gastric cancer development in patients infected by this microorganism.

A syndrome of bilateral hemorrhage of the thalamus and myocarditis with fatal course.

We report the case of a 41-year-old patient with bilateral hemorrhage of the thalamus, leading to death. Post-mortem examination showed acute myocarditis. Neuropathological study showed perivascular infiltrates in affected thalamic regions. Laboratory investigation failed to find any causal agent. We hypothesize an infective agent, affecting the heart and thalamus, as the cause of this syndrome. Diaschisis due to the strategic anatomical position of the thalamus may have been responsible for coma state and death.

Papillary carcinoma on a thyroglossal duct cyst: diagnostic problems and therapeutic dilemma. A case report.

Thyroglossal duct cysts are one of the most common congenital abnormalities of the cervical region. Complications of these swellings are rare, and among these, appearance of a carcinoma has also been noted. We present a case of papillary carcinoma arising in a thyroglossal duct cyst in 20-year-old woman with a swelling of about 4 cm, located at the middle region of the neck over the hyoid bone. Our patient was treated using a modified Sistrunk operation, in which thyroidectomy proved crucial for the correct diagnosis and continuation of appropriate treatment. Our case confirms the difficulty in distinguishing a primitive thyroglossal duct carcinoma from a synchronous metastatic papillary carcinoma of the thyroid. This dilemma often remains unresolved.

Rapid identification of Toscana virus by nested PCR during an outbreak in the Siena area of Italy.

The sand fly-transmitted Toscana virus is recognized as an etiologic agent of an aseptic meningitis with a long convalescence. This infection has been reported overall in many tourists or in a seronegative population circulating in endemic Mediterranean areas (Italy, Portugal, Egypt, and Cyprus). We report a cluster of acute Toscana virus infections in the local population during the summer of 1995. Twenty-one clinical cases of meningitis were investigated for the presence of Toscana virus by nested PCR performed on the S segment of the virus RNA extracted from cerebrospinal fluid samples.

Intracellular life.

Intracellular parasites and endosymbionts are present in almost all forms of life, including bacteria. Some eukaryotic organelles are believed to be derived from ancestral endosymbionts. Parasites and symbionts show several adaptations to intracellular life. A comparative analysis of their biology suggests some general considerations involved in adapting to intracellular life and reveals a number of independently achieved strategies for the exploitation of an intracellular habitat. Symbioses mainly based on a form of syntrophy may have led to the establishment of unique physiological systems. Generally, a symbiont can be considered to be an attenuated pathogen. The combination of morphological studies, molecular phylogenetic analyses, and palaeobiological data has led to considerable improvement in the understanding of intracellular life evolution. Comparing host and symbiont phylogenies could lead to an explanation of the evolutionary history of symbiosis. These studies also provide strong evidences for the endosymbiogenesis of the eukaryotic cell. Indeed, an eubacterial origin for mitochondria and plastids is well accepted and is suggested for other organelles. The expansion of intracellular living associations is presented, with a particular emphasis on peculiar aspects and/or recent data, providing a global evaluation.

Immunopotentiating of mucosal and systemic antibody responses in mice by intranasal immunization with HLT-combined influenza virosomal vaccine.

The mucosal vaccination strategy against influenza has been investigated by using influenza virosomal vaccine (IRIV) combined with two different adjuvants: the procholeragenoid (PCG) and the Escherichia coli heat labile toxin (HLT). A comparative study has been carried out on mice administered intranasally with these different formulations of influenza vaccine. PCG appears less effective than HLT in inducing an IgG response, but both the adjuvants elicit mucosal adjuvant activity inducing s-IgA in the upper respiratory tract. On the contrary, only HLT when administered intranasally to mice with influenza virosomes stimulates the production of s-IgA in the lower respiratory tract thereby providing a better protection against primary infection of the respiratory system. Both HLT and PCG enhance the production of IFN-gamma in the respiratory tract, nevertheless HLT appears more efficacious as a mucosal adjuvant.

Evidence of Toscana virus variants circulating in Tuscany, Italy, during the summers of 1995 to 1997.

The Toscana virus can cause neurological infection in adults. This study of 112 cases of acute meningitis which occurred during the summers of 1995, 1996, and 1997 demonstrated the presence of viral RNA in the cerebrospinal fluid of 56 patients. Their sequence analysis shows four variants of the Toscana virus.

Rapid detection and typing of circulating mumps virus by reverse transcription/polymerase chain reaction.

The reverse transcription/polymerase chain reaction (RT-PCR) was used to amplify a 129-bp fragment of the mumps virus F gene from strains circulating in the Siena area from 1993-1995. The nucleic acid was amplified directly from the samples; no growth in cell culture was required. Nucleotide sequence analysis and the comparison with other virus strains enabled the typing of the detected viruses. There appears to be more than one lineage of mumps virus circulating at any given time in the same location. A PCR assay coupled with the sequencing of the 5' end of the F gene seems to be a convenient method for characterizing mumps virus strains. This method, useful in diagnosis, also appears to be suitable for epidemiological studies.

Rapid detection of different RNA respiratory virus species by multiplex RT-PCR: application to clinical specimens.

BACKGROUND: The polymerase chain reaction (PCR) applied in diagnostic and epidemiologic investigations is very useful for sensitivity, specificity and time saving. OBJECTIVE: We have developed a method for the detection of genomic RNA of two different species of virus, the influenza A virus (IA) and the respiratory syncytial virus (RS), which are responsible for clinical similarities. We applied this multiplex RT-PCR protocol on clinical specimens. STUDY DESIGN: We describe a method which allows rapid diagnosis by performing a single retro-transcriptase (RT) reaction associated with the PCR (multiplex RT-PCR) on different genomes in a single sample. We have evaluated the sensitivity and the specificity of the multiplex test on positive controls, then, on RNA extracted from clinical specimens harvested from 15 children with respiratory symptoms during the spring-winter season 1997. RESULTS AND CONCLUSIONS: The multiplex RT-PCR protocol, applied to respiratory specimens, allows the investigation of RNA IA virus and RS virus in a single sample at the same time. The detection of the etiologic viral agent is rapid and it is possible to evaluate incidental simultaneous infections.

Mycoplasma pneumoniae non-pulmonary infection presenting with pharyngitis, polyarthritis and localized exanthem.

We report a case of pharyngitis, polyarthritis and localized exanthem in acute Mycoplasma pneumoniae infection not involving the lower respiratory tract. Diagnosis was made by means of a particle agglutination test and IgM/IgG indirect immunofluorescence assay. This case describes a clinical complex never reported before and suggests the need for a high index of suspicion in cases of atypical presentation of M. pneumoniae infection.