Marco Cusini

Distribution of human herpesvirus-8 sequences throughout the spectrum of AIDS-related neoplasia

Objective:AIDS frequently associates with certain malignancies, including Kaposis sarcoma, non-Hodgkins lymphoma (NHL), and anogenital neoplasia. In this study we aimed to define the frequency of infection by human herpesvirus (HHV)-8 throughout the spectrum of AIDS-related neoplasia in Europe. Design:A tumour panel representative of the distinct types of AIDS-related neoplasms was tested for the presence of HHV-8 DNA sequences. Autologous uninvolved tissues were also tested in selected cases. Methods:The presence of HHV-8 DNA sequences was assayed by a combination of polymerase chain reaction followed by oligohybridization and Southern blot hybridization of genomic DNA with an HHV-8-specific probe. Results:HHV-8 sequences were detected in 100% of AIDS-related Kaposis sarcoma (all 35 cases). Among AIDS-related NHL, HHV-8 sequences selectively clustered with body-cavity-based lymphomas (BCBL; all three cases), although they were consistently negative in small non-cleaved cell lymphomas (none in 18 cases), diffuse large cell lymphomas (none in seven), or anaplastic large cell lymphomas (none in three). No HHV-8 sequences were found in cases of anogenital neoplasia (out of 14) or Hodgkins disease (out of three). HHV-8 DNA sequences were also positive in the uninvolved skin of all six AIDS-related Kaposis sarcoma patients, but not in the circulating lymphocytes of a BCBL patient. Positivity for HHV-8 sequences occurred in patients belonging to all major AIDS risk categories. Conclusions:These data confirm that HHV-8 sequences associate at high frequency with selected types of AIDS-related neoplasia, namely Kaposis sarcoma and BCBL, although they are consistently absent in other types of AIDS-NHL and AIDS-related anogenital neoplasia.

Mycoplasma genitalium in males with nongonococcal urethritis: Prevalence and clinical efficacy of eradication

Background: Mycoplasma genitalium is regarded as a potential pathogen of the human urogenital tract based on prevalence findings of several European studies. Goal: To determine the prevalence of M genitalium in urethral specimens of symptomatic patients with nongonococcal urethritis and from asymptomatic patients attending a sexually transmitted disease clinic in Milan, and to verify the clinical efficacy of M genitalium eradication by antibiotic treatment. Study Design: From May 1998 to late April 1999, a routine analysis for M genitalium by DNA amplification (polymerase chain reaction) was performed in patients attending the Institute of Dermatological Science in Milan. The authors examined urethral swabs from 178 symptomatic and 23 asymptomatic males. M genitalium‐positive patients were clinically and microbiologically tested after treatment with either doxycycline or azithromycin. Results: Among males with nongonococcal urethritis, M genitalium was detected in 14.0% of patients as the only agent; in 15.1% of patients in association with Chlamydia trachomatis and/or Ureaplasma urealyticum; and in 1 asymptomatic patient. In all symptomatic M genitalium‐positive patients, antibiotic treatment eradicated the infection and cured clinical symptoms. Conclusion: These data reveal the high prevalence of M genitalium in symptomatic patients, the rarity of asymptomatic carriers, the high susceptibility to antibiotic treatment, and the clinical efficacy of M genitalium eradication. Moreover, data confirm the etiologic role of M genitalium in inflammatory processes of the human urogenital tract in the Mediterranean area.

2016 European guideline on Mycoplasma genitalium infections

Mycoplasma genitalium infection contributes to 10–35% of non‐chlamydial non‐gonococcal urethritis in men. In women, M. genitalium is associated with cervicitis and pelvic inflammatory disease (PID). Transmission of M. genitalium occurs through direct mucosal contact. Asymptomatic infections are frequent. In women, symptoms include vaginal discharge, dysuria or symptoms of PID – abdominal pain and dyspareunia. In men, urethritis, dysuria and discharge predominates. Besides symptoms, indication for laboratory test is a high‐risk sexual behaviour. Diagnosis is achievable only through nucleic acid amplification testing (NAAT). If available, NAAT diagnosis should be followed with an assay for macrolide resistance. Therapy for M. genitalium is indicated if M. genitalium is detected or on an epidemiological basis. Doxycycline has a low cure rate of 30–40%, but does not increase resistance. Azithromycin has a cure rate of 85–95% in macrolide susceptible infections. An extended course appears to have a higher cure rate. An increasing prevalence of macrolide resistance, most likely due to widespread use of azithromycin 1 g single dose without test of cure, is drastically decreasing the cure rate. Moxifloxacin can be used as second‐line therapy, but resistance is increasing. Uncomplicated M. genitalium infection should be treated with azithromycin 500 mg on day one, then 250 mg on days 2–5 (oral), or josamycin 500 mg three times daily for 10 days (oral). Second line treatment and treatment for uncomplicated macrolide resistant M. genitalium infection is moxifloxacin 400 mg od for 7–10 days (oral). For third line treatment of persistent M. genitalium infection after azithromycin and moxifloxacin doxycycline 100 mg two times daily for 14 days can be tried and may cure 30%. Pristinamycin 1 g four times daily for 10 days (oral) has a cure rate of app. 90%. Complicated M. genitalium infection (PID, epididymitis) is treated with moxifloxacin 400 mg od for 14 days.

Human herpesvirus type 8 variants circulating in Europe, Africa and North America in classic, endemic and epidemic Kaposi's sarcoma lesions during pre-AIDS and AIDS era

Human herpesvirus-8 (HHV-8) variants have been found heterogeneously distributed among human populations living in diverse geographic regions, but their differential pathogenicity in Kaposis sarcoma development remains controversial. In the present study, HHV-8 variant distribution has been analyzed in classic, iatrogenic, endemic as well as epidemic Kaposis sarcoma (KS) during pre-AIDS and AIDS period (1971-2008) in countries with different KS incidence rate. DNA samples from cutaneous KS lesions of 68 patients living in Africa (n=23, Cameroon, Kenya and Uganda), Europe (n=34, Greece and Italy) and North America (n=11) have been subjected to PCR amplification of HHV-8 ORF 26, T0.7, K1 and K14.1/15, followed by direct nucleotide sequencing and phylogenetic analysis. Among the 23 African samples, the majority of HHV-8 ORF 26 variants clustered with the subtype R (n=12) and B (n=5). Conversely, the viral sequences obtained from 45 European and North European tumors belonged mainly to subtype A/C (n=36). In general, HHV-8 and K1 variant clustering paralleled that of ORF 26 and T0.7. Genotyping of the K14.1/15 loci revealed a large predominance of P subtype in all tumors. In conclusion, comparison of the HHV-8 sequences from classic or endemic versus AIDS-associated KS showed a strong linkage of the HHV-8 variants with specific populations, which has not changed during AIDS epidemic.

Seroprevalence of herpes simplex virus type 2 infection among attendees of a sexually transmitted disease clinic in Italy

Background and Objectives: An increased prevalence of herpes simplex virus type 2 (HSV‐2) infection has been recently observed in industrialized countries. Goal: To determine HSV‐2 seroprevalence in a high‐risk population in Italy. Study Design: A cross‐sectional study was performed to ascertain the HSV‐2 prevalence among 919 persons attending an STD clinic in northern Italy. A HSV‐2‐specific glycoprotein G‐2‐based immunoglobulin G enzyme‐linked immunoabsorbent assay (Gull/Meridian ELISA; Meridian Diagnostics, Cincinnati, OH) was used and validated against Western blot analysis. Results: A prevalence of 24.6% was found without differences between males and females. Seroprevalence increased with age and number of partners during the previous year. Compared with Western blot analysis, the Gull/Meridian ELISA showed a sensitivity of 91.9% and a specificity of 98%, and positive and negative predictive values of 93.9% and 97.4%, respectively. Conclusion: This is the first Italian survey of HSV‐2 infection conducted with a properly validated, Food and Drug Administration‐approved, type‐specific serologic method in a high‐risk population. It is likely that between one to three million adults are infected with HSV‐2.

Mucocutaneous manifestations in patients infected with human immunodeficiency virus

The mucocutaneous manifestations observed in 516 patients infected with human immunodeficiency virus attending an outpatient clinic for acquired immunodeficiency syndrome in Milan, Italy, from July 1985 to March 1987 are listed, and the clinicopathologic aspects of these disorders are reported. The prognostic significance of some human immunodeficiency virus-associated dermatoses is also discussed.

MDM2 and CDKN1A gene polymorphisms and risk of Kaposi’s sarcoma in African and Caucasian patients

A single-nucleotide polymorphism in the MDM2 promoter (SNP309; rs2279744) causes elevated transcription of this major negative regulator of p53 in several cancer types. We investigated MDM2 SNP309 and CDKN1A (p21/Waf1/Cip1) codon 31 (rs1801270) polymorphisms in 86 cases of cutaneous Kaposi’s sarcoma (KS) from African and Caucasian patients, and 210 healthy controls. A significant increase of the MDM2 SNP309 T/G genotype was observed among classic KS cases (odds ratio 2.38, 95% confidence interval 1.0–5.5). Frequencies of CDKN1A codon 31 genotypes were not significantly different between cases and controls. The results suggest that the MDM2 SNP309 G allele may act as a susceptibility gene for the development of classic KS in Caucasian patients.

Background review for the 2016 European guideline on Mycoplasma genitalium infections.

Mycoplasma genitalium is a cause of 10–35% of non‐chlamydial non‐gonococcal urethritis in men and in women, and is associated with cervicitis and pelvic inflammatory disease (PID). Transmission of M. genitalium occurs through direct mucosal contact. In women, symptoms include vaginal discharge, dysuria or symptoms of PID – abdominal pain and dyspareunia. In men, urethritis, dysuria and discharge predominates. Asymptomatic infections are frequent. In this review, we present the evidence base for the recommendations in the 2016 European guideline on M. genitalium infections and describe indications for testing, recommended diagnostic methods, treatment and patient management. The guideline was prepared on behalf of the European branch of The International Union against Sexually Transmitted Infections; the European Academy of Dermatology and Venereology; the European Dermatology Forum; the European Society of Clinical Microbiology and Infectious Diseases; the Union of European Medical Specialists. The European Centre for Disease Prevention and Control and the European Office of the World Health Organisation also contributed to their development.

Trend of ciprofloxacin resistance in Neisseria gonorrhoeae strains isolated in Italy and analysis of the molecular determinants

A total of 599 Neisseria gonorrhoeae strains collected in Italy in 2 periods, 2003 to 2005 and 2007 to 2008, were screened for ciprofloxacin susceptibility by Etest. Ciprofloxacin-resistant strains (49.7%) were characterized by i) serovar determination, patterns of mutation in gyrA, and parC genes (38%, randomly selected) and ii) N. gonorrhoeae multiantigen sequence typing (56% of the strains isolated from patients who declared their sexual orientation). The percentage of ciprofloxacin-resistant strains increased from 42 (2003-2005) to 58 (2007-2008); in the second period, strains with MIC value >32 microg/mL have been observed. Mutations in gyrA and parC genes were identified in the majority of strains (88%). Ciprofloxacin-resistant isolates among men who have sex with men (MSM) increased from 24% in 2003 to 2005 to 47% in 2007 to 2008. However, sequence types exclusively found among MSM were mostly due to a single strain. This is the first study in Italy combining N. gonorrhoeae ciprofloxacin susceptibility testing with molecular analyses and comparing the results over time.

Syphilis outbreak in Milan, Italy

Infectious syphilis has been considered a sensitive marker of risky sexual behaviour.1 Following a decline of syphilis in Western Europe,2,3 there has been a resurgence of infectious syphilis in many countries, with a number of outbreaks in men who have sex with men (MSM).4,5 The STD …