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We report the case of a newborn with acute myeloid leukaemia, who developed perineal necrotizing fasciitis due to Pseudomonas Aeruginosa, after twenty days of life. Following surgical debridement, she was effectively treated with topical negative pressure therapy (V.A.C.(R) device) with silver foam dressings, this achieved complete closure in thirteen days. Negative pressure therapy should be considered when conventional wound care fails to achieve complete wound closure, even in neonates.

Background: Pemphigus vulgaris is an autoimmune disease characterized by blisters and widespread erosions, involving skin and mucous membranes, caused by autoantibodies to desmoglein 1 and 3. This pathology is associated with increased morbidity and mortality if untreated. The treatment of pemphigus vulgaris requires multiple immunosuppressive agents, but often it is particularly resistant., Objective: To evaluate the efficacy and safety of rituximab therapy in refractory pemphigus vulgaris., Methods: Five patients diagnosed as having pemphigus vulgaris were treated with anti-CD20 monoclonal antibody (rituximab). Each patient was treated with rituximab intravenously at a dosage of 375 mg per square metre of body surface area once weekly for 4 weeks., Results: All the patients presented clinical resolution. No adverse effects were observed. It is important to observe the clinical evolution in the future, but our experience is still limited to a short lifetime and follow-up., Conclusion: In our experience rituximab has been an effective and safe treatment for refractory pemphigus vulgaris.

Five immunodominant Treponema pallidum recombinant polypeptides (rTpN47, rTmpA, rTpN37, rTpN17, and rTpN15) were blotted onto strips, and 450 sera (200 from blood donors, 200 from syphilis patients, and 50 potentially cross-reactive) were tested to evaluate the diagnostic performance of recombinant Western blotting (recWB) in comparison with in-house whole-cell lysate antigen-based immunoblotting (wclWB) and T. pallidum hemagglutination (MHA-TP) for the laboratory diagnosis of syphilis. None of the serum specimens from blood donors or from potential cross-reactors gave a positive result when evaluated by recWB, wclWB, or MHA-TP. The evaluation of the immunoglobulin G immune response by recWB in sera from patients with different stages of syphilis showed that rTmpA was the most frequently identified antigen (95%), whereas only 41% of the specimens were reactive to rTpN37. The remaining recombinant polypeptides were recognized as follows: rTpN47, 92.5%; rTpN17, 89.5%; and rTpN15, 67.5%. The agreement between recWB and MHA-TP was 95.0% (100% with sera from patients with latent and late disease), and the concordance between wclWB and MHA-TP was 92.0%. The overall concordance between recWB and wclWB was 97.5% (100% with sera from patients with secondary and late syphilis and 94.6 and 98.6% with sera from patients with primary and latent syphilis, respectively). The overall sensitivity of recWB was 98.8% and the specificity was 97.1% with MHA-TP as the reference method. These values for sensitivity and specificity were slightly superior to those calculated for wclWB (sensitivity, 97.1%, and specificity, 96.1%). With wclWB as the standard test, the sensitivity and specificity of recWB were 98.9 and 99.3%, respectively. These findings suggest that the five recombinant polypeptides used in this study could be used as substitutes for the whole-cell lysate T. pallidum antigens and that this newly developed recWB test is a good, easy-to-use confirmatory method for the detection of syphilis antibodies in serum.

A surface immunofluorescence assay (SIFA) using live spirochetes was analyzed and compared with Western blot (WB), fluorescent treponemal antibody absorption (FTA-ABS), microhemagglutination (MHA-TP), and Treponema pallidum immobilization (TPI) assays for detecting serum antibodies to T. pallidum in patients with syphilis, in disease controls, and in healthy subjects. SIFA and WB were 99% sensitive (99 of 100 positive specimens) and specific (140 of 140 negative specimens); FTA-ABS showed a sensitivity and a specificity of 90 and 89% (90 of 100 positive and 125 of 140 negative specimens), respectively. MHA-TP showed a sensitivity of 84% (84 of 100 positive specimens) and a specificity of 98.5% (138 of 140 negative specimens). Finally, TPI had a sensitivity of 52% (52 of 100 positive specimens) and a specificity of 100% (140 of 140 negative specimens). The T. pallidum SIFA was therefore highly specific, showing no equivocal reactivities with control sera, and sensitive. The results suggest the possible use of SIFA as a confirmatory test in the serologic diagnosis of syphilis.

Sensitivity and specificity of IgG detection by Western blotting performed with a lysate of Treponema pallidum whole cells were compared with those of the most common assays used in the laboratory diagnosis of syphilis, i.e. fluorescent treponemal antibody absorption test (FTA-ABS) and treponemal haemagglutination assay (TPHA). Thirty-five serum samples obtained from twenty-one patients with a clinical diagnosis of early syphilis, based on the presence of typical chancre or skin or mucous membrane lesions, were studied. In addition, thirty blood samples from donors, ten sera positive for Borrelia burgdorferi and five positive for Leptospira interrogans were tested as controls. The clinical diagnosis was the reference method used to compare the performance of the serological tests. Western blotting performed with a sensitivity and specificity of 100%, whereas the corresponding sensitivity and specificity for FTA-ABS were 88.5% and 98%, respectively. The performance of TPHA showed a sensitivity of 86% and a specificity of 100%.

Background and Objectives: The ligase chain reaction is an in vitro DNA amplification technique that exponentially amplifies selected DNA sequences., Goal: To evaluate a ligase chain reaction assay for the detection of Chlamydia trachomatis cryptic plasmid DNA (LCx Chlamydia) in patients routinely attending a sexually transmitted disease center in Italy., Study Design: Urethral or cervical swabs were obtained from 501 consecutive patients (334 men and 167 women). The samples were assayed in parallel with LCx Chlamydia and conventional tissue culture; discordant results were further assayed by direct immunofluorescence and a ligase chain reaction with alternate primers., Results: After resolution of discordant results, the LCx method showed a sensitivity, specificity, positive predictive value, and negative predictive value of 100%, 99.3%, 96.7%, and 100% in men; 100%, 100%, 100%, and 100% in women; and 100%, 99.5%, 97.1%, and 100% overall, respectively. By comparison, the sensitivity of tissue culture was 81.4% in men, 50% in women, and 77.6% overall., Conclusions: The automated LCx method is sensitive, fast, and accurate and represents a useful diagnostic tool for C. trachomatis infection, even in low and medium prevalence populations.

OBJECTIVE: The purpose of the present study was to evaluate an in vitro DNA amplification assay named the ligase chain reaction (LCR) for the detection of Chlamydia trachomatis cryptic plasmid DNA in urine from men and women, in comparison with urethral swab culture in men and cervical swab culture in women. METHODS: 591 patients (394 men with urethritis and 197 female sex partners) attending a center for sexually transmitted diseases in northern Italy between January 1994 and January 1995 were enrolled in this study. A cervical swab was collected from women and a urethral swab from men for standard tissue cell culture. From each patient 20 mL of the first stream of the urine (FVU), taken at least 2 h after the last urination, were collected for LCR analysis. Discrepant results were further analyzed by direct fluorescence and a LCR with alternative primers. RESULTS: In men the prevalence of C. trachomatis infection by urethral culture was 13.45% and, after resolution of discordant results, the LCR method performed on FVU showed a sensitivity, specificity, positive predictive value and negative predictive value of 89.4%, 100%, 100% and 98.2%, respectively; the sensitivity of tissue cell culture was 92.8%. In female sex partners, the prevalence of C. trachomatis infection by cervical culture was 3.04%; LCR detected eight true positive samples, two more than tissue cell culture, and no false-negative results. CONCLUSION: LCR analysis of FVU is a rapid, non-invasive technique and represents a good alternative to tissue cell culture. Further study is needed to investigate possible LCR inhibitors present in urine samples.

We compared a commercially available PCR assay (Amplicor, Roche, Switzerland) and tissue culture isolation for the detection of C. trachomatis in urethral and/or endocervical swabs. Of the 200 patients studied (130 men and 70 women) PCR and tissue culture gave concordant results in 199 cases; in one case PCR was positive and culture negative. The Amplicor PCR assay proved fast and sensitive and suitable for routine use in most clinical microbiology laboratories.

A surface immunofluorescence assay (SIFA) was analyzed and compared with a conventional indirect immunofluorescence assay (IFA) and whole-cell enzyme-linked immunosorbent assay (ELISA) for detecting immunoglobulin G (IgG) antibodies to Borrelia burgdorferi in sera from patients with Lyme disease. Fifty-five patients with syphilis and 33 patients with rheumatoid arthritis were used as disease controls. The sensitivity of the SIFA was low during the acute phase of Lyme disease (sera from seven of nine patients presenting with erythema chronicum migrans were negative during the first 2 months of illness); later, seroconversion was observed in all patients at various times during convalescence. Sera from five patients with complicated Lyme disease were strongly positive. SIFA was found to be highly specific, since sera from all patients with secondary or latent syphilis and patients with rheumatoid arthritis did not react in the test. Strong cross-reactivity occurred when these sera were tested in conventional IFA and ELISA; sera from 38 (69%) patients with syphilis were positive by IFA and sera from 51 (93%) patients were positive by ELISA, whereas 7 (21%) and 12 (36%) of the serum samples from patients with rheumatoid arthritis were positive by IFA and ELISA, respectively. Immunoblot analysis of SIFA-positive sera showed that the 31- and 34-kDa outer surface proteins (proteins A and B, respectively) of B. burgdorferi were the major reactive antigens involved in the test. The results support a role for SIFA in the investigation of complicated Lyme disease as well as in the differentiation of Lyme disease from other diseases associated with B. burgdorferi cross-reactive antibodies.

An ultrastructural investigation of two patients suffering from acrodermatitis chronica atrophicans revealed in the small dermal vessels swelling of the endothelial cells, material of probably plasmatic origin accumulated in the subendothelial area, and sleeves of basement membrane-like material on a concentric perivascular layout.

The authors report the results of treatment with ketoconazole in 8 patients with chronic mucocutaneous candidiasis (CMC). The drug, administered in the dose of 200 mg once a day orally for a period of time varying from 2 to 12 months, led to improvement in or elimination of clinical symptoms in all patients. One patient had a relapse on suspension of treatment, but this regressed rapidly on resumption of ketoconazole. In 7 cases there were no side-effects. In one case there was an increase in serum liver enzymes which disappeared spontaneously 7 days after suspension of treatment. These results appear encouraging in view of the difficulty of treating this disease, which is often resistant to conventional antifungal therapy.

The virological course of herpes zoster infection in 42 otherwise normal hosts was studied by virus isolation and antibody titration. Varicella-zoster virus (VZV) was isolated from vesicle fluid from all three patients examined on the first day of the vesicular eruption and from five out of six examined on the second day. The isolation rate fell to one out of six patients on the seventh day of illness and VZV was not isolated from patients at a later stage of the illness. IgG antibodies were detected by IFAMA and ELISA, in sera from all the patients by the end of the first week of illness; IgG antibody titres were highest during the second and the third weeks. IgM antibodies to VZV were detected in sera from six of the 42 patients with herpes zoster after fractionation by ion-exchange chromatography.

The Authors report the results of a study on the etiology of non-gonococcal urethritis (NGU) carried out on 180 male patients with NGU and on 50 healthy volunteers matched for age and sex. Chlamydia trachomatis infection was proven by culture in 54 (30%) of the NGU-patients and in none of the healthy controls. Ureaplasma urealyticum was recovered from men both with and without urethritis. Bacteria other than C. trachomatis were not found in NGU-patients in a significant percentage. All C. trachomatis-positive NGU-patients presented antibodies against C. trachomatis, by indirect fluorescence, while only 54% of C. trachomatis-negative NGU-patients and 22% of healthy volunteers, were positive. A further serological screening of 78 women with no history of urethritis or cervicitis and 44 children less than 10 years of age was carried out in order to investigate the prevalence of serum antibodies against C. trachomatis in groups of healthy population in Italy.

The inhibitory activity of neutralizing sera on Herpes simplex virus type 1 (HSV-1) plaque enlargement (PE) is easily detected using the S variant of low passage clinical isolates (Mannini-Palenzona et al., 1985b; Costanzo et al., 1986). The same sera show little or no activity on PE of the product of the S variant rapid in vitro conversion, the L variant, and laboratory strains HSV-1 (F) and (MP). No significant difference was found in the inhibitory activity of sera from healthy individuals with no history of recurrence and patients with recurrences.