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Biomedical subjects

F Chiarini

Publications and source records attributed to F Chiarini.

At least 19 recordsLinked to original sources

Gut-associated bacterial microbiota in paediatric patients with inflammatory bowel disease.

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.

Adolescent↗

Karyotype studies in South American species of Solanum subgen. Leptostemonum (Solanaceae).

Mitotic chromosome numbers and karyotypes of 13 South American species (12 native and one naturalized) from four sections of SOLANUM subgen. LEPTOSTEMONUM were studied. Chromosome numbers of S. ACERIFOLIUM, S. AENICTUM, S. CONDITUM, S. CONSIMILE, S. INCARCERATUM, and S. PLATENSE are reported for the first time. The number 2n = 24 was found in most species, while 2n = 22 was found in S. MAMMOSUM and S. PLATENSE. The latter is the second SOLANUM with this unusual number. Satellites are always present and were visible in more than 50 % of the cells studied. Karyotypes are symmetrical: M and SM chromosomes are common, whereas ST chromosomes are rare. The karyotypes of S. AENICTUM, S. MAMMOSUM, and S. PANICULATUM are comparatively asymmetrical. Species can be distinguished by a combination of chromosome number, karyotype formulae, karyotype length, the position of satellites in a particular chromosome pair, and asymmetry indices. The phenogram obtained does not reflect the sectional arrangements or the systematic affinities of the species studied. In sect. ACANTHOPHORA, increased asymmetry is associated with derived characters (strong andromonoecy, winged seeds, mammiform fruits). Diversification in the subgenus is suggested to be related to visible chromosome rearrangements and cumulative, cryptic structural changes may have also played a relevant evolutionary role.

Biological Evolution↗

Extended Spectrum Beta-Lactamase-producing Klebsiella pneumoniae outbreaks during a third generation cephalosporin restriction policy.

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.

Cephalosporins↗

Infectious agents in tissues from spontaneous abortions in the first trimester of pregnancy.

Some evidence suggests that intrauterine infection plays a major role in the pathogenesis of early pregnancy loss, but the implication and prevalence of microrganisms in the aetiology of spontaneous abortion during the first trimester of pregnancy has not yet been well established. In this study, we analysed the tissues relative to the product of conception from abortions during the first trimester (51 spontaneous abortions and 56 voluntary pregnancy interruptions) in women attending the Gynecological Sciences Perinatology and Puericulture Department of "Policlinico Umberto I". Specimens were investigated by cultural methods for the presence of yeasts, gram positive, gram negative bacteria, and genital mycoplasma. By molecular diagnostic procedures, DNA sequences of Chlamydia trachomatis, herpes simplex viruses, adenovirus, human papillomaviruses and human polyomaviruses BK and JC were searched. None of these agents could be found in voluntary pregnancy interruption samples, with the exception of 3.6% of specimens positive for adenovirus, whereas spontaneous abortion tissues were positive for at least one microrganism by 31.5%. Data analysis showed the occurrence of both monomicrobial and polymicrobial infections.

Abortion, Spontaneous↗

The role of intravenous immunoglobulin therapy in autoimmune and inflammatory disorders.

The first use of immunoglobulin therapy, historically, was in 1890 when Emil von Behring developed effective antiserum against diphtheria toxin, but only in the early 1970s technological advancements in the fractionation of plasma lead to the production of Ig preparations which could be administered intravenously. Intravenous Ig products are a mainstay for disorders such as: primary immunodeficiency, serious infections, autoimmune and inflammatory disorders. During autoimmune and systemic inflammatory disease IVIg exhibits a number of immune modulatory activities such as: Fc Receptor-mediated effects, modulation of complement, modulation of cytokine production, superantigens neutralization, antibodies neutralization by idiotype network, increased catabolism of IgG, but also biologic effects of other molecules present in IVIg preparations. Recent understanding about IVIg composition and mechanism of action can explain its therapeutic effect in autoimmune and inflammatory disorders. Nevertheless it is important to underline that IVIg is a heterogeneous product and it is difficult to determine the exact mechanism of its activities in every disease. The increased use of IVIg in the treatment of autoimmune disorders outlined the issue of tolerability. Undesiderable effects to IVIg occurs in less than 5% of patients.

Autoimmune Diseases↗

Abnormal "low grade" transformation zone: current diagnostic gold standard.

The aim of this work was to examine different methods of investigation in the diagnosis of the abnormal "low grade" transformation zone of the portio. Over a period of one year 41 patients subjected to colposcopic examination underwent exo-endocervical sampling for oncologic evaluation and for detection of viral and bacterial infections (HPV, HSV, adenovirus, mycoplasmas and chlamydia trachomatis), as well as portio biopsy. A 65.8% correlation was found between cytology and the HPV-DNA test results, while histology and the presence of the HPV virus agreed in 51.4% of cases. In those cases in which minimal histological alterations were found (koilocytosis) a high percentage of HPV negativity was found. In discordant negative cytologic tests that were however positive for HPV by PCR, the genotypes identified were always 6 and 11.

Base Sequence↗

Herpes simplex virus type 2 modulates the susceptibility of human bladder cells to uropathogenic bacteria.

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.

Bacterial Adhesion↗

Detection of viral and bacterial infections in women with normal and abnormal colposcopy.

Signs and symptoms of sexually-transmitted diseases (STD) do not allow any etiological diagnosis in women. Colposcopic findings are seldom pathognomic. Consequently, the microbiology laboratory with the recent availability of molecular diagnostic tools is required to detect the infectious bacterial and/or viral agents involved in STD. In cervical samples of women submitted to gynaecological screening for past or present signs and symptoms of inflammation and with different colposcopic findings, we searched by molecular approaches Chlamydia trachomatis, Mycoplasma genitalium, herpes simplex virus type 1 and 2, adenovirus and 45 genotypes of papillomaviruses and, by cultural methods Mycoplasma hominis and Ureaplasma urealyticum. Colposcopy permitted us to divide the studied population into three groups: 48 women had negative colposcopic findings, 50 presented signs of flogosis and 100 resulted positive for an abnormal transformation zone (ANTZ) and/or for HPV colposcopic findings. Results obtained by microbiological assays indicated that the prevalence of infectious agents did not always correlate with colposcopy. Double and triple infections were found in groups 2 and 3, with mycoplasmas being the most common microrganisms present in association and quite almost copresent with papillomaviruses.

Adult↗

Simultaneous detection of HPV and other sexually transmitted agents in chronic urethritis.

BACKGROUND: Many pathogens may be responsible of Non Gonococcal Urethritis (NGU) with the possible occurrence of symptomatic and asymptomatic mixed viral and bacterial infections. In particular, genital papillomaviruses (HPVs) have been searched since they are linked to both benign and malignant lesions of the penis and urethra and the presence of a potential male carried state has received limited scrutiny while the screening of sexually active females has received substantial attention. METHODS: In male patients affected by chronic NGU, the presence of DNA of Chlamydia trachomatis, herpes simplex virus (HSV) type 1 and 2 and human papillomaviruses by PCR and the occurrence of Gram positive and Gram negative micro-organisms, of Mycoplasma hominis and Ureaplasma urealyticum, by conventional cultural methods have been investigated. RESULTS: Results obtained indicated a high percentage of mixed infections, up to 36%. Genital HPV DNA was detected in 31% of specimens positive for two or more agents, and HSV DNA was detected in 10% of studied population. CONCLUSIONS: The concomitant presence of different infectious agents could determine latent, sub-clinical or chronic infections with periodic reactivation. In particular results suggest that HPV and HSV may stimulate cytokine production which can up regulate the expression of other infectious agents and may be responsible for latent chlamydial infections characterised by the persistence of this micro-organism in an altered form, viable but in a culture negative state. Therefore an increased awareness of mixed infections is relevant to define the management and treatment of chronic urethritis.

Adult↗

In vitro study of a double infection by herpes simplex virus type 2 and Chlamydia trachomatis.

Sexually transmitted diseases (STDs) may recognize multiple etiological agents. Among them, Chlamydia trachomatis (Ct) and herpes simplex virus type 2 (HSV-2) cause symptomatic, subclinical and asymptomatic infections of the urogenital tract which can lead to serious sequelae. In the present study the coinfection and superinfection by Ct and HSV-2 in epithelial cultured cells from human cervix (HeLa 229) are described. A double infection, followed by the intracellular synthesis of chlamydial and viral antigens, was established. Both synergistic and interfering phenomena were recorded: viral antigen synthesis resulted increased whereas Ct inclusion bodies were produced to a lower extent.

Antigens, Bacterial↗

Lack of association between clinical and environmental isolates of Pseudomonas aeruginosa in hospital wards.

Seventy-three environmental and clinical isolates of Pseudomonas aeruginosa recovered from a single hospital over a 6-month period were compared for epidemiological type characteristics. Environmental isolates were obtained from sinks, taps and water, in rooms where patients were treated. The strains represented only six O-antigenic types and 8.2% of them were not typable. Serotype 011 was most frequent in the environment, whereas serotypes 06, 012 and 02,5 predominated among clinical isolates. More than 60% of all isolates belonged to four pyocin types (1, 10, 33 and 45), and approximately 80% were phage typable. Environmental isolates were more sensitive to antibiotics than clinical isolates. There was little correspondence between the types of strains of P. aeruginosa isolated from patients and those isolated from the environment. However, isolates of identical type were frequently recovered from different patients within the same clinic and were found to be related in time and location. We conclude that the environment was not an important source of P. aeruginosa infection and that transfer of organisms was mainly from patient-to-patient.

Bacterial Typing Techniques↗

Chlamydia trachomatis genitourinary infections: laboratory diagnosis and therapeutic aspects. Evaluation of in vitro and in vivo effectiveness of azithromycin.

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.

Azithromycin↗

[Infection of the vascular prosthesis. The diagnostic, preventive and treatment protocols: preliminary results].

A number of strategies for the management of vascular graft infections are described in literature. Nevertheless, this serious compliance is still burdened with high rates of morbidity and mortality. At the present the principal efforts are therefore directed towards prevention and early diagnosis. In this view, the authors propose their protocol of study, on the basis of the encouraging results till now obtained.

Bacteria↗

Virulence determinants in Pseudomonas aeruginosa strains from urinary tract infections.

A total of 121 uropathogenic Pseudomonas aeruginosa strains were examined for production of several virulence-related factors. These strains were distributed in five predominant O-serotypes, i.e. O 4, O 12, O 11, O 6 and O 5, which accounted respectively for 23.9, 23.1, 12.3, 8.2 and 5.7% of isolates. Pyochelin and pyoverdin siderophores were produced by most of the isolates, defective variants occurring at very low frequency (2.4% for pyochelin and 7.4% for pyoverdin). Adherence to uroepithelial cells and production of cytotoxins was demonstrated in 52.8 and 67.7% of the strains, respectively, with higher frequencies for epidemiologically related strains belonging to serotypes O 4 and O 12. Titration of total proteases, elastase and phospholipase C revealed a high degree of heterogeneity among isolates. However, examination of individual O-serotypes by exoenzyme production showed that elevated levels of total proteases and elastase were characteristics of serotypes of minor numerical importance, i.e. O 1, O 10, O 11 and O 17, whilst low levels of elastase were produced by strains belonging to the predominant serotypes, namely O 4 and O 12. Moreover, epidemiologically related strains belonging to serotypes O 4 and O 12 appeared more homogeneous than the whole serogroup, when compared with other groups on the basis of exoenzyme levels.

Bacterial Adhesion↗

A family of bombinin-related peptides from the skin of Bombina variegata.

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.

Amino Acid Sequence↗

Epidemiological typing of uropathogenic Pseudomonas aeruginosa strains from hospitalized patients.

One hundred and twenty-one clinical isolates of Pseudomonas aeruginosa from patients with hospital-acquired urinary tract infections (UTIs) were studied to determine their major epidemiological markers, including API 20NE profile, O-serotype, pyocin type, phage type, lysogenic state and antibiotic susceptibility. Serotypes O4, O12, O11, O6 and O5 were found with a high frequency, accounting respectively for 23.9%, 23.1%, 12.3%, 8.2% and 5.7% of isolates. Pyocin type 10 was most common (32.2%) followed by types 1 (10.7%), 33 (7.5%) and 105 (4.1%); subtype h was predominant being characteristic of 34.7% of isolates. Most of the strains (69.4%) were either not phage typable or sensitive to phages 68 and 119x. Resistance to gentamicin, tobramycin, amikacin, imipenem and ciprofloxacin was more frequent among strains belonging to serotype O12. The O-serotypes were combined with API 20NE profiles, pyocin and phage types, lysogenic states and antibiotic resistance in order to identify epidemiologically related clones. Within predominant serotypes--O4 and O12--most strains displayed similar but not identical type characteristics, whereas other serotypes were less homogeneous. Our results support the concept that a combination of current typing techniques allows the identification of epidemiologically related P. aeruginosa isolates.

Bacterial Typing Techniques↗

Role of membrane glycosphingolipids as Pseudomonas aeruginosa adhesin receptor in rabbit bladder mucosa.

The adhesiveness of a mucous strain of Pseudomonas aeruginosa to rabbit bladder mucosa was studied after preincubation of the microorganism with several glycolipids with different carbohydrate moieties to investigate their importance in the interaction with bacterial adhesins. Vesical cells were also treated with lectins (limulin and soybean) to confirm the role of saccharides as membrane receptor. The results obtained showed that galactose-containing glycolipids were able, in varying degrees, to reduce bacterial binding. The most active compounds were glycosphingolipids with negatively charged terminal groups. Lectin treatment of bladder mucosal cells confirmed the importance of galactose and sialic acid as mucosal cell membrane receptors for P. aeruginosa.

Adhesins, Bacterial↗