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The aim of this study was to determine whether detection of β-HPV gene products, as defined in epidermodysplasia verruciformis skin cancer, could also be observed in lesions from kidney transplant recipients alongside the viral DNA. A total of 111 samples, corresponding to 79 skin lesions abscised from 17 kidney transplant recipients, have been analyzed. The initial PCR analysis demonstrated that β-HPV-DNA was highly present in our tumor series (85%). Using a combination of antibodies raised against the E4 and L1 proteins of the β-genotypes, we were able to visualize productive infection in 4 out of 19 actinic keratoses, and in the pathological borders of 1 out of 14 squamous cell carcinomas and 1 out of 31 basal cell carcinomas. Increased expression of the cellular proliferation marker minichromosome maintenance protein 7 (MCM7), that extended into the upper epithelial layers, was a common feature of all the E4-positive areas, indicating that cells were driven into the cell cycle in areas of productive viral infections. Although the present study does not directly demonstrate a causal role of these viruses, the detection of E4 and L1 positivity in actinic keratosis and the adjacent pathological epithelium of skin cancer, clearly shows that β-HPV are actively replicating in the intraepidermal precursor lesions of kidney transplant recipients and can therefore cooperate with other carcinogenic agents, such as UVB, favoring skin cancer promotion.

Objectives (1) Hypothesis testing of the potency of rituximab (RTX) in preventing fibrotic complications and (2) assessing acceptability and feasibility of RTX in early systemic sclerosis (SSc). Methods A small, 24-month, randomised, double-blind, placebo-controlled, single-centre trial in patients with SSc diagnosed

For selecting therapy, it is important to distinguish different types of keratinocytic neoplasia. It is sometimes difficult to make histopathologic diagnosis, especially in organ transplant recipients (OTR) who develop numerous lesions.To investigate p16 immunostaining in different types of keratinocytic neoplasia in OTR, we studied 59 actinic keratoses (AK), 51 Bowen' s disease (BD), 63 squamous cell carcinomas (SCC), 16 benign keratotic lesions (BKL) from 31 OTR patients and 25 controls (eczema and psoriasis). Tissue sections were stained for HE and p16. We scored intensity, proportion and distribution of p16 positive lesional cells.In 19% of AK, 92% of BD, 35% of SCC and 12% of BKL more than 15% of lesional cells were p16-positive. In 16% of AK, 80% of BD, 18% of SCC and 13% of BKL strong p16 staining was observed. BKL, AK and SCC showed focal and patchy staining, BD showed diffuse pattern with strong staining of all atypical cells. Sparing of the basal layer was predominantly seen in BD. No control specimen showed p16-overexpression.p16 immunostaining shows a characteristic pattern in BD, but not in AK, SCC and BKL. It appears useful in recognizing BD, but not in differentiating between other keratinocytic neoplasia.

A 72-year-old woman was treated with a total postoperative radiotherapy dose of 55.86 Gy (to convert to rad, divide by 0.01) in 21 fractions after lumpectomy of a cT1bN0M0 mammary carcinoma. Two months after the last dose of radiation she developedpruriticpustules andpapuleson the skin of the ipsilateral right breast. She mentioned that shehad a cold sore once several years ago. Levoceterizinegavesomereliefof the itch.Physical examination findings showed pustules surrounding the areola on the right breast with an increasing number in the submammary fold (Figure 1). Quiz at jamaoncology.com Figure 1. Pustules and papules on the right breast

Background Anogenital warts are often presumed to represent nondysplastic or low-grade anal intraepithelial neoplasia (LGAIN). We previously demonstrated that up to 20% of intra-anal warts in HIV-positive men who have sex with men (MSM) contain regions of high-grade AIN (HGAIN). Objectives To determine the causative human papillomavirus (HPV) types of low- and high- grade dysplastic areas in warts from HIV-positive MSM. Methods A total of 42 intra-anal warts from 41 HIV-positive MSM were graded as nondysplastic, LGAIN or HGAIN. Whole-tissue sections (WTS) were analysed with the SPF10 polymerase chain reaction/LiPA25 HPV genotyping system. If the WTS contained multiple HPV types, dysplastic regions were isolated by laser capture microdissection (LCM) for HPV genotyping. Results Overall, 38 of 42 (91%) WTS tested positive for HPV DNA. Of these, 23 (61%) contained a single HPV type and 15 (39%) contained multiple HPV types. All LCM-selected regions contained no more than one HPV type. Ten of 42 (24%) WTS contained HGAIN disease, of which six (60%) were associated with a high-risk HPV (hrHPV) genotype. Twenty-three of 42 WTS contained LGAIN disease, of which two (9%) were associated with hrHPV. AIN lesions containing hrHPV types were identified using p16 staining. Conclusions LGAIN lesions can be caused by high-risk HPV genotypes and vice versa. We therefore recommend routine follow-up and treatment of all dysplastic intra-anal warts for HIV-positive MSM.

The classification of anal intraepithelial neoplasia (AIN) in mucosal biopsies is subject to considerable interobserver variability. Previous studies have shown that Ki-67 and p16/CDKN2A immunostains aid detection of dysplasia in biopsy samples from the uterine cervix. The aim of this study was to determine whether Ki-67 and p16/CDKN2A immunolabeling enhanced diagnostic accuracy in the assessment of anal mucosal biopsies from patients with suspected human papillomavirus (HPV) infection. The study consisted of 75 cases that were originally interpreted to represent normal anal transitional zone (n=15), fibroepithelial polyp (n=10), condyloma accuminatum (n=10), low-grade AIN (AIN1, n=20), and high-grade AIN (n=20), including 10 cases each of AIN2 and AIN3. The histologic features of all cases were re-reviewed and categorized based upon consensus agreement, which resulted in reclassification of 16 cases. Thus, the final study group consisted of 17 samples of normal anal transition zone, 14 fibroepithelial polyps, 6 condylomata accuminata, and 38 cases of AIN (11 AIN1, 16 AIN2, and 11 AIN3). Each case was tested for the presence of HPV DNA by a SPF10 polymerase chain reaction and LiPA25 genotyping assay and immunostained for Ki-67 and p16/CDKN2A. A positive Ki-67 result was defined as the presence of a cluster of at least 2 strongly stained epithelial nuclei in the upper two-thirds of the epithelial thickness. A positive result for p16/CDKN2A was defined as diffuse moderate-to-strong cytoplasmic and nuclear staining. None of the anal transition zone samples or fibroepithelial polyps showed Ki-67 or p16/CDKN2A staining. All condylomata and samples of AIN contained HPV DNA and showed positive Ki-67 labeling. All cases of high-grade AIN showed positive p16/CDKN2A staining. We conclude that Ki-67 labeling detects anal HPV-related changes with a high degree of sensitivity and specificity, whereas increased p16/CDKN2A staining is strongly associated with high-grade squamous neoplasia. These results indicate that a combination of these markers may aid interpretation of anal mucosal biopsy samples.

Contains fulltext : 87203.pdf (Publisher’s version ) (Closed access) Chlamydia trachomatis (Ct) comprises 3 serogroups and 19 serovars. Different genotyping methods are available to differentiate between the serovars. The aim of this study was to evaluate the sensitivity and discriminatory power of three genotyping methods, respectively Omp1 sequencing, the Ct Detection and genoTyping (DT) assay and the pmpH real-time PCR discriminating an LGV infection from a non-LGV infection. In total, 50 Aptima Combo 2 (AC2) Ct positive samples were selected and tested with the 3 genotyping methods. The Ct-DT assay detected 3 double Ct infections that caused a non interpretable result by Omp1 sequencing, while Omp1 sequencing has a higher discriminatory power that gave additional information about Ct genovariants. All three methods detected the 6 LGV samples. Although the pmpH real-time PCR detected all LGV infections, a substantial amount (24%) of non-LGV infections were missed. The sensitivity compared to AC2 Ct detection was 80% (95% CI 67-89%) for the Ct-DT assay, 72% (95% CI 58-83%) for Omp1 sequencing and 64% (95% CI 50-76%) for the pmpH real-time PCR. In conclusion, the Ct-DT assay is appropriate for serovar distribution studies, epidemiological studies and differentiation between an LGV and non-LGV Ct infection, while Omp1 sequencing is more appropriate for phylogenetic studies. The pmpH real-time PCR is suitable as second assay to differentiate between an LGV and non-LGV infection, but not as primary detection assay, due to its low sensitivity for non-LGV strains. 01 oktober 2010

Objective Chlamydia trachomatis (Ct) has been implicated as a co-factor in cervical carcinogenesis. The goal of the current study was to investigate if Ct may play a role in pathogenesis of cervical adenocarcinoma and, specifically, if there is a co-infection between Ct and Human Papillomavirus (HPV) in cervical adenocarcinomas. The second goal of the study was to determine the distribution of HPV genotypes in most recent cases of in-situ and invasive cervical adenocarcinomas. Methods Biopsies of 71 cervical adenocarcinomas (31 in-situ and 40 invasive) were tested for the presence of Ct using two novel PCR assays. In addition, all cases were tested for HPV using SPF10-PCR assay and genotyped using LIPA 25 test. Results None of the cases was found to be positive for Ct using two independent PCR assays. All lesions, however, were positive for HPV with the exception of a case of minimal deviation adenocarcinoma. Overall, 94.2% of cases were positive for either HPV16 ( n =44, 62.8%) or HPV18 ( n =20, 28.5%), or both ( n =2, 2.8%). Other single HPV types included HPV45 ( n =3, 4.2%) and HPV35 ( n =1, 1.4%). Conclusion The study demonstrated lack of co-infection between Human Papillomavirus and C. trachomatis in in-situ and invasive adenocarcinoma of the uterine cervix. The role of Ct as a carcinogenetic co-factor may be restricted to cervical squamous cell carcinomas. Accounting for type cross-protection, currently available HPV vaccines are likely to prevent close to 100% of HPV-positive cervical adenocarcinomas.

The current case report describes a 35-year-old man who presented with unilateral scrotal angiomas. The presence of unilateral scrotal angiomas was associated with an underlying varicocele on the ipsilateral side due to increased venous pressure. In case of unilateral scrotal angiomas further examination for underlying pathology is necessary.

Importance Cutaneous verruca vulgaris lesions (warts) and oral squamous cell papillomas are common lesions caused by human papillomavirus (HPV). Multiple reports have described cases of wart resolution following quadrivalent HPV vaccination. We report the case of a patient with chronic oral papillomas with resolution after quadrivalent HPV vaccination and perform a review of the literature. Observations An immunocompetent man in his 60s presented with chronic verrucous papules on the lips, tongue, and buccal mucosa refractory to multiple excisions. Biopsy showed squamous cell papilloma, and DNA sequencing revealed HPV-32. He received the quadrivalent HPV vaccine resulting in clearance of all lesions after 3 months. We found 8 reported cases of disseminated, recurrent warts with resolution after quadrivalent HPV vaccination. Improvement was seen within 4 weeks of vaccination, and resolution after 3 to 8 months. Conclusions and Relevance We report the case of recurrent oral papillomas caused by HPV-32 with complete resolution after quadrivalent HPV vaccination and reviewed reports of resolution of recalcitrant and disseminated warts after vaccination. Production of cross-protective immunoglobulins and cytotoxic T cells is a possible mechanism. There remains a critical need for randomized clinical trials to assess efficacy of quadrivalent HPV vaccination for treatment of oral squamous papillomas and cutaneous verruca vulgaris.

Organ transplant recipients (OTRs) have an increased risk of developing keratinocyte carcinomas (KCs). The aim of this study was to correlate infection with human papillomaviruses (HPVs) belonging to the beta genus (Beta-papillomavirus (Beta-PV)) at transplantation with later development of KCs. In a cohort study, sera collected between 1 year before and 1 year after transplantation of OTRs transplanted between 1990 and 2006 were tested for antibody responses against the L1 capsid antigen of Beta-PV and other HPV genera (Gamma-, Mu-, Nu-, and Alpha-PV) using multiplex serology. The OTRs were followed for a maximum of 22 years. Cox regression models with KC, squamous cell carcinoma (SCC), and basal cell carcinoma (BCC) as outcome variables were used. Out of 445 OTRs, 60 had developed KC: 14 developed only SCC, 24 only BCC, and 22 both types of KC. The time-dependent hazard ratio (HR) to develop either or both types of KC, adjusted for age, sex, and transplanted organ, in tested Beta-PV-seropositive OTR around the time of transplantation compared with Beta-PV-seronegative OTR was 2.9 (95% confidence interval (CI) 1.3–6.4). The HR for SCC was 2.9 (95% CI 0.99–8.5) and for BCC it was 3.1 (95% CI 1.2–8.0). There was also an association between Mu-PV seropositivity and KC, but there were no significant associations between other HPV genera tested and KC. A positive seroresponse for Beta-PV around transplantation significantly predicted the development of KC in OTRs up to 22 years later, providing additional evidence that infection with Beta-PV has a role in KC carcinogenesis.

Summary Background One-third of Dutch primary school children have cutaneous warts; each year around 20% of them seek medical treatment. However, little is known about the epidemiology of the types of human papillomavirus (HPV) causing these warts. Objectives To investigate the distribution of cutaneous wart-associated HPV types in three primary school classes by analysing skin swabs taken from warts, and the forehead, hand dorsum and sole of the foot of included children. Methods Using the hyperkeratotic skin lesion polymerase chain reaction/multiplex genotyping assay, each swab sample was used to genotype for 23 cutaneous wart-associated HPV types. Results Thirty-one (44%) of the 71 children had a total of 69 warts, with a maximum of six warts per child. In the wart swabs, HPV2, HPV27 and HPV57, members of Alphapapillomavirus species 4, were most frequently detected (27%, 32% and 14%, respectively), whereas HPV1 was only found in two plantar warts. The prevalence of HPV carriage, detected in swabs of clinically normal skin of the forehead, left hand and left sole was 80%, with the most prevalent types being HPV1 (59%), HPV2 (42%), HPV63 (25%) and HPV27 (21%). Conclusions Cutaneous wart-associated HPV types were highly prevalent in primary school children, but did not correlate with the HPV types in warts. In contrast to the existing literature, HPV1 was frequently detected on clinically normal skin but was much less frequent in warts.

Epidermodysplasia verruciformis (EV) is a rare, lifelong, autosomal recessive skin disease associated with an unusual susceptibility to infections with ubiquitous β-human papillomaviruses (β-HPVs), and in some cases also skin-tropic α genotypes. In this case report, HPV infection patterns were correlated with pathology and clinical manifestations of skin lesions from a patient with EV, without loss-of-function mutations in the EVER genes. HPV infection was investigated by both polymerase chain reaction (PCR) and laser capture microdissection (LCM) PCR, alongside immunofluorescence for the viral proteins E4 and L1. Analysis of eyebrow hair bulbs revealed multiple β-genus HPV infections, including HPV20 and HPV24, which were consistently found in all 11 skin lesions on the patient. Six lesions were also positive for the skin tropic α-genotype, HPV27. Clear-cut differences between two wart-like lesions, one caused by a skin-tropic α-genotype and the other by β-genotypes (as detected by LCM PCR) are shown, including the high cellular proliferation rate in β-HPV-induced lesions. Widespread expression of the early protein E4 was also evident in skin lesions positive for HPV20 by LCM PCR in both tumours and nearby intraepidermal proliferative areas. L1 expression was restricted to areas of intraepidermal proliferation showing productive infection. The patient's inability to control HPV infections is conclusive to the uncontrolled replication of few genotypes from both α and β genera, which cause proliferative lesions with clear-cut clinical and histological features.

Keratinocyte carcinomas are by far the most common malignancies seen in organ transplant recipients (OTR). Life-long immunosuppressive therapy is the most important risk factor for developing squamous cell carcinoma (SCC) in OTR. In the years after transplantation, OTR develop numerous warts and wart-like lesions followed by the development of SCC, which resembles the clinical picture of epidermodysplasia verruciformis patients in which human papillomavirus (HPV) infection was linked with skin cancer for the first time. HPV can be divided into genera and cause several distinct benign and (pre-) malignant diseases.There is evidence linking beta HPV infection with the development of SCC in OTR. However, the role of beta HPV in cutaneous SCC carcinogenesis is still enigmatic. Beta HPV is not integrated in the human cellular DNA and is not necessary for the maintenance of the malignant phenotype of cutaneous SCC. Whether different beta HPV types have different effects on cellular mechanisms and a combination of these HPV types may further increase the risk of cutaneous SCC is unknown. The carcinogenic effect, if present, is subtle and probably exerted early in carcinogenesis.

BACKGROUND High-grade anal intraepithelial neoplasia (AIN) is present in many human immunodeficiency virus (HIV)-positive men who have sex with men. The major etiologic factor is infection with an oncogenic human papillomavirus (HPV) genotype. We investigated whether individual components of high-grade AIN are caused by single HPV types. METHODS DNA was isolated from whole-tissue sections of 31 high-grade AIN that were recovered from 21 HIV-positive men who have sex with men. The SPF10 PCR/LiPA25 HPV genotyping system was used for DNA analysis. In whole-tissue sections with multiple HPV types, polymerase chain reaction was repeated in regions of AIN sampled by laser-capture microdissection. The results were compared with HPV types in anal swabs. RESULTS A single HPV type was observed in 15 (48%) of 31 whole-tissue sections. In an additional 14 whole-tissue sections, 1 HPV type was found in each lesion sample evaluated by laser-capture microdissection. Consequently, in 29 of 31 biopsy specimens (94%), a single HPV type was found in each lesional component studied. Two whole-tissue sections contained collision regions, each with 2 HPV types. HPV16 was presumed to be causative in 14 of 31 biopsy specimens (45%). More than half of the anal swabs did not contain all causative HPV types. CONCLUSIONS Individual components of high-grade AIN are caused by single HPV types (the so-called one lesion, one virus concept). HPV16 is causative in

Although the role of oncogenic human Alpha-papillomaviruses (HPVs) in the development of mucosal carcinomas at different body sites (eg cervix, anus, oropharynx) is fully recognized, a role for HPV in keratinocyte carcinomas (KCs; basal and squamous cell carcinomas) of the skin is not yet clear. KCs are the most common cancers in Caucasians, with the major risk factor being ultraviolet (UV) light exposure. A possible role for Beta-HPV types (BetaPV) in the development of KC was suggested several decades ago, supported by a number of epidemiological studies. Our current review summarizes the recent molecular and histopathological evidence in support of a causal association between BetaPV and the development of KC, and outlines the suspected synergistic effect of viral gene expression with UV radiation and immune suppression. Further insights into the molecular pathways and protein interactions used by BetaPV and the host cell is likely to extend our understanding of the role of BetaPV in KC.

Objective Cervical cancer is caused by carcinogenic human papillomavirus (HPV) infections. Prior to the introduction of HPV vaccination in Suriname, we performed a cross-sectional study to estimate the prevalence of and determinants for genital carcinogenic HPV infections. Methods Women were recruited at a family planning (FP) clinic and a sexually transmitted infections (STI) clinic. Vaginal swabs were used for HPV genotyping by the SPF 10 PCR-DEIA-LiPA 25 system. Logistic regression was used to identify determinants for carcinogenic HPV infection. Results The prevalence of any HPV was 54.2% and of carcinogenic HPV was 27.9% among 813 women attending the FP clinic. Among the 188 women attending the STI clinic, the prevalence of any HPV (76.1%) and of carcinogenic HPV (40.4%) was significantly higher. HPV52 was the most prevalent genotype in both clinics. The prevalence of HPV16 and/or 18 was 6.4% in the FP clinic and 12.2% in the STI clinic. The following determinants were independently associated with carcinogenic HPV infection among women visiting the FP clinic: ≥2 recent partners (OR 1.53; 95% CI 1.13 to 2.06), Chlamydia trachomatis co-infection (OR 1.89; 95% CI 1.32 to 2.70), disassortative ethnic sexual mixing (OR 1.50; 95% CI 1.13 to 1.99) and ethnic group (OR 1.90; 95% CI 1.27 to 2.85 for Creole and OR 1.67; 95% CI 1.06 to 2.62 for mixed ethnicity, both compared with Hindustani). No independent determinants were found among women visiting the STI clinic. Conclusions Carcinogenic HPV is highly prevalent among women in Suriname, and not equally distributed among ethnic groups. These data provide a baseline to assess possible shifts in the prevalence of HPV genotypes following vaccination.

Background Chlamydia trachomatis is the most prevalent bacterial sexually transmitted infection (STI) worldwide. A strong link between C. trachomatis serogroup/serovar and serological response has been suggested in a previous preliminary study. The aim of the current study was to confirm and strengthen those findings about serological IgG responses in relation to C. trachomatis serogroups and serovars. Methods The study population (n = 718) consisted of two patient groups with similar characteristics of Dutch STI clinic visitors. We performed genotyping of serovars and used titre based and quantitative commercially available ELISA kits (medac Diagnostika) to determine specific serum IgG levels. Optical density (OD) values generated by both tests were used to calculate the IgG titres (cut-off 1:50). Analyses were conducted stratified by gender. Results We observed very significant differences when comparing the median IgG titres of three serogroups, B, C and I: in women for B vs. C: p

Paraneoplastic dermatoses may be the first manifestation of a malignancy. Rapid recognition is therefore important.A 72-year-old woman who had been treated in the past for metastasised neuroendocrine carcinoma and had undergone curative treatment for stage-1c endometrial carcinoma developed three paraneoplastic dermatoses over a 2-year period: acanthosis nigricans, tripe palms and acquired hypertrichosis lanuginosa. Half a year later, cutaneous metastases derived from the endometrial carcinoma treated 12 years prior were discovered.The development of three paraneoplastic dermatoses in a short period of time is rare. Acanthosis nigricans, tripe palms and acquired hypertrichosis lanuginosa can develop in association with both neuroendocrine and endometrial carcinoma. The sudden development of paraneoplastic dermatoses is frequently a forewarning of progression of the malignancy.

SUMMARYThe prevalence of Chlamydia trachomatis varies between ethnic groups in The Netherlands. It is, however, unknown whether this is associated with specific serogroups. The objective of this study was to determine whether serogroup distribution is associated with ethnic origin in the region of The Hague, The Netherlands. Serogroups of 370 microbiologically confirmed C. trachomatis-positive samples were analysed. The samples were obtained from 247 women and 123 men between January and October 2008, of self-reported Dutch Caucasian, Dutch Antillean, Surinamese, N. African/Turkish or other descent. We observed a difference in serogroup distribution comparing Dutch Caucasian women to Dutch Antillean women (χ2 for distribution P = 0·035). Serogroup C was more common in Dutch Antillean women, whereas serogroup B was less common (P = 0·03). This difference was not observed for Dutch Antillean men. The observed difference in distribution of C. trachomatis serogroups between ethnic groups is relevant for further transmission studies.

Objective It has been reported that approximately 10% of low grade squamous intraepithelial lesions (LSIL) progress to high grade squamous intraepithelial lesions (HSIL) within a 2-year follow up. The factors related to lesion progression are currently unknown. The aim of the study was to identify prognostic markers of the course of LSIL. This retrospective study was designed to correlate regression, persistence and progression of biopsy-proven LSIL with patients’ age, HPV genotypes and immunohistochemical expression of the main cell cycle regulating proteins: p53, pRb, p16, and Ki-67. Study design A total of 584 consecutive patients with biopsy proven LSIL and 2-year follow-up were included in the age analysis. HPV genotyping was performed in 328 LSIL cases using the SPF10 PCR-LiPA25 (version 1), 238 LSIL cases were immunostained for Ki-67 and p16, and 101 cases were immunostained for pRb and p53. Results The odds of LSIL persistence and progression were significantly higher in women 30–39, 40–49 and 50+ years old, as compared to women 20–29 years old (OR 1.89, 2.52 and 2.39, respectively). The odds of persistence and progression were higher in women infected with HPV16, 18, 33 and 52 (OR 3.5, 3.1, 3.5 and 2.9, respectively). There were no significant differences in expression of immunomarkers (p16, p53, pRB and Ki-67) between the lesions that regressed versus the lesions that persisted or progressed. Conclusions Patients 30 years of age and older have a higher risk of LSIL progression or persistence as compared to 20–29 year olds. In addition, HPV genotyping, but not the cell cycle markers, may aid in prognosis of LSIL course.

A 95-year-old woman presented with a progressive purple blue asymptomatic tumorous plaque on her forehead, extending as a macula to the upper face, possibly arisen after a fall. Histology showed atypical epitheloid cells with vascular-lumen formation. The patient was diagnosed with cutaneous angiosarcoma. She was treated palliatively with radiotherapy.

A 86-year-old woman with a medical history of disseminated conjunctival melanoma presented with a progressive blue discolouration of the skin, which was initially attributed to a radiation dermatitis. However, histopathological examination of palpable firm nodules demonstrated lymphangitis melanomatosis.

Individuals with two red hair colour (RHC)-MC1R genetic variants have light skin and blond/reddish hair and, in comparison with those without such alleles, are at an increased risk of developing melanoma. Our study investigated the association of RHC variants and the Total Dermo-scopy Score (TDS), and the items that make up the TDS, in those with atypical naevi and melanomas from high risk melanoma patients. Eight hundred and seventy-six atypical naevi and 21 melanomas were scored according to the TDS system and MC1R polymorphisms were determined. Analyses revealed that several TDS items including pigment network, dark-brown colour and streaks were more frequently observed in atypical naevi from individuals without RHC variants, while structureless areas were more often observed in individuals with two RHC variants. Finally, no significant difference in TDS was detected in atypical naevi from individuals with two RHC variants compared to those without RHC. Clinicians should be aware of a different dermoscopic naevus pheno-type in patients with light blond or RHC MC1R variants.

A28-year-old woman presented with a 3-month history of asymptomatic bumps on both eyebrows (Figure 1A). She had had cosmetic tattooing of these areas approximately 1.5 year before the manifestation of the lesions and depilated her eyebrow hairs using tweezers. Her medical history was positive for diabetes mellitus type 2, for which she used metformin. She denied other symptoms such as shortness of breath. On physical examination, multiple hard yellowish–red papules were observed arranged in a linear fashion within the tattoo ink. When examined using diascopy an apple-jelly color was observed. The remaining skin showed no other abnormalities. The clinical differential diagnosis included a granulomatous foreign body reaction, sarcoidosis, or xanthomas. A 3-mm punch biopsy of the left eyebrow revealed a granulomatous dermatitis without necrosis (Figure 2). Serum analyses (angiotensin-converting enzymes [ACEs] and lysozyme) and a chest X-ray were normal, excluding sarcoidosis. A diagnosis of a delayed granulomatous reaction to tattoo ink was made. The lesions were injected using Dermojet with triamcinolone acetonide A10 1:1 diluted with lidocaine. When the patient visited the hospital 6 weeks after the triamcinolone acetonide injection for follow-up review, all lesions had resolved (Figure 1B). The lesions did not recur during 4 months of follow-up.

Background Lymphogranuloma venereum (LGV) proctitis is caused by Chlamydia trachomatis (Ct) genotype L and is endemic among men who have sex with men (MSM) in western society. Genotype L infections need to be distinguished from non-LGV (genotypes A-K) Ct infections since they require prolonged antibiotic treatment. For this purpose, an in-house developed pmpH based LGV polymerase chain reaction (PCR) test is used at the Amsterdam STI outpatient clinic. We investigated retrospectively the anal Ct genotype distribution, and the frequency of concomitant genotype infections in MSM infected with LGV and non-LGV Ct infections. To detect concomitant Ct genotype infections, the pmpH LGV PCR and genoTyping Reverse Hybridization Assay (Ct-DT RHA) were used. Methods A total of 201 Ct positive rectal swabs from MSM were selected, which were previously diagnosed as either LGV (n = 99) or non-LGV Ct infection (n = 102) according to the algorithm of Ct detection by the commercially available Aptima Combo 2 assay followed by an in-house pmpH LGV PCR. The samples were retested with the commercially available Ct-DT RHA, which differentiates between 14 major genotypes and is able to detect concomitant Ct genotypes. Results Excellent genotyping agreement was observed between the Ct-DT RHA and the pmpH LGV PCR (Kappa = 0.900, 95%CI = 0.845-0.955, McNemar's p = 1.000). A concomitant non-LGV genotype was detected in 6/99 (6.1%) LGV samples. No additional LGV infections were observed with the Ct-DT RHA among the non-LGV Ct group. In the non-LGV group genotype G/Ga (34.3%) was seen most frequent, followed by genotype D/Da (22.5%) and genotype J (13.7%). All LGV infections were caused by genotype L2. Conclusions Concomitant non-LGV genotypes do not lead to missed LGV proctitis diagnosis. The pmpH LGV PCR displayed excellent agreement with the commercially available Ct-DT genotyping RHA test. The genotypes G/Ga, D/Da and J were the most frequent non-LGV Ct strains in MSM.

Cofactors might affect the risk of the rare progression from infection with carcinogenic human papillomavirus (HPV) to cervical premalignancy to invasive cancer. Some studies have observed that Chlamydia trachomatis infection is associated with increased risk for cervical cancer. In a large prospective cohort, we assessed the role of C trachomatis in cervical premalignancy and addressed confounding by HPV.We identified 182 women with prevalent and 132 women with incident histological cervical intraepithelial neoplasia grade 2 (CIN2), grade 3 (CIN3), or cervical cancer (CIN2+) in the Costa Rica HPV Natural History Study. Control subjects were 995 (approximately 10% of the 10 049) subjects who were randomly selected from the same study. Cervical HPV status at enrollment was determined by MY09/MY11 polymerase chain reaction amplification and dot-blot hybridization. The presence of C trachomatis DNA in cervical exfoliated cells at enrollment was determined by a novel serovar-specific polymerase chain reaction-based C trachomatis detection and genotyping assay. Plasma drawn at enrollment from each subject was used to determine C trachomatis immunoglobulin G (IgG) status. Logistic regression was used to examine the association between C trachomatis and CIN2+, taking into account possible confounding by HPV.C trachomatis positivity at enrollment was associated with CIN2+ and concurrent and subsequent carcinogenic HPV infection. To account for confounding by HPV status, we restricted the analysis to women positive for carcinogenic HPV DNA at enrollment and found no association between C trachomatis status (as assessed by DNA or IgG) at enrollment and combined prevalent and/or incident CIN2+ (for C trachomatis DNA positivity, odds ratio = 0.77, 95% confidence interval = 0.42 to 1.41; for C trachomatis seropositivity, odds ratio = 1.09, 95% confidence interval = 0.85 to 1.41).We found no association between C trachomatis status, as assessed by DNA or IgG, and risk of cervical premalignancy, after controlling for carcinogenic HPV-positive status. Previous positive associations between C trachomatis and cervical premalignancy could have been caused, in part, by an increased susceptibility to HPV infection.

Reactive arthritis (previously known as Reiters syndrome) is an inflammatory arthritis that is a type of spondyloarthropathy. The disease consists of the classical triad of arthritis, urethritis and conjunctivitis, but mucocutaneous abnormalities also frequently appear: balanitis circinata, keratoderma blennorrhagicum, aphthous ulcers in the mouth and nail disorders. These skin lesions are mainly found in reactive arthritis induced by Chlamydia trachomatis (Ct). Reactive arthritis is often triggered by a sexually transmitted infection (Chlamydia trachomatis) or an enteric infection (such as Salmonella or Shigella). It is thought that human antibodies against the pathogen cross-react with the HLA antigen (mainly HLA-B27). To distinguish between reactive arthritis and psoriatic arthritis, screening of the urine or synovium for Ct infection should be carried out. Acute reactive arthritis is treated with NSAIDs as the first choice. In addition, patients may receive an intra-articular injection of glucocorticoids. The mucocutaneous abnormalities respond well to topical glucocorticoids. Although in the Netherlands a Ct induced reactive arthritis is not yet treated with antibiotics, a recent published clinical trial in patients with a chronic Ct induced reactive arthritis showed a significant reduction in complaints in the group treated with a combination of antibiotics for 6 months, compared to the placebo group. Active genitourinary Ct infection should be treated with antibiotics, the first choice being azithromycin 1000 mg as a single dose. It is important that the patient's partner is tested at the same time and if necessary treated simultaneously to prevent reinfection.

Objective The objective of the study was to compare the distribution of HPV genotypes and HPV16 variants in glandular and squamous cervical neoplasia. Methods Cases of endocervical adenocarcinoma in-situ (AIS, n =33) invasive adenocarcinoma (ADCA, n =55), cervical intraepithelial neoplasia-3 (CIN3, n =130) and squamous cell carcinoma (SCC, n =60) were collected at the New York Hospital and tested for HPV using SPF 10 PCR-LIPA 25 (version 1) assays and for HPV16 variants using a multiplex PCR and reverse hybridization assay. Results There was a difference between the spectrum of HPV genotypes detected in glandular and squamous neoplasia: 13 different HPV genotypes were detected in CIN3 as single infections and 11 in SCC, while only 4 single genotypes were detected in AIS and 3 in ADCA. The most common single HPV types in CIN3 were HPV16, 31, and 52 (56.9%, 10%, 8.4%, respectively). In SCC the most common were HPV16, 18 and 31 (70%, 6.5%, 4.9%). In AIS, HPV16, 18, 45 and 35 accounted for 69.7%, 27.2%, 3%, 3% of cases. The three single types in ADCA were HPV16 (43.6%), HPV18 (41.8%) and HPV45 (10.9%). European variants of HPV16 were the most common in CIN3 (83.8%), SCC (71.4%) and AIS (73.9%). In ADCA the Asian American (AA) variant was the most common (41.7%) followed by European variants (33.3%). AA variant was also detected in 17.4%, 4.1%, and 2.4% of HPV16 positive AIS, CIN3 and SCC, respectively. Conclusion Asian American variant of HPV16, HPV18 and HPV45 are preferentially associated with cervical adenocarcinoma as compared to squamous cell carcinoma .

The aims of this study were to compare a novel PCR-based Chlamydia trachomatis detection and genotyping (Ct-DT) assay with the FDA-approved, commercially available C. trachomatis detection Hybrid Capture 2 (HC2) assay and to investigate the C. trachomatis serovar distribution among young women in a rural Costa Rican study population. A total of 5,828 sexually active women participating in a community-based trial in Costa Rica were tested for C. trachomatis by HC2. A sample of 1,229 specimens consisting of 100% HC2 C. trachomatis -positive specimens ( n = 827) and a random sample of 8% HC2 C. trachomatis -negative specimens ( n = 402) were tested with the Ct-DT assay. Agreement between the two assays was determined by the unweighted kappa statistic. Discrepant specimens were tested with a second commercially available test (COBAS TaqMan). The Ct-DT-positive specimens were further analyzed with the Ct-DT genotyping step to investigate the distribution of 14 different C. trachomatis serovars (A, B/Ba, C, D/Da, E, F, G/Ga, H, I/Ia, J, K, L1, L2/L2a, and L3). After accounting for the sampling fraction selected for Ct-DT testing, crude agreement with the HC2 assay was 98% and the kappa was 0.92 (95% confidence interval [CI], 0.89 to 0.97). The 33 discordant samples that were further analyzed with the COBAS TaqMan test showed better agreement with the Ct-DT assay (31/33, P < 0.001). Among the 806 Ct-DT-positive samples, serovar E was the most common serovar (31%), followed by serovars F and D (both 21%) and serovar I (15%). In conclusion, the novel Ct-DT assay permits reliable detection and identification of C. trachomatis serovars.

Contains fulltext : 53601.pdf (Publisher’s version ) (Open Access) Chlamydia trachomatis (Ct) comprises distinct serogroups and serovars. The present study evaluates a novel Ct amplification, detection, and genotyping method (Ct-DT assay). The Ct-DT amplification step is a multiplex broad-spectrum PCR for the cryptic plasmid and the VD2-region of ompl. The Ct-DT detection step involves a DNA enzyme immunoassay (DEIA) using probes for serogroups (group B, C, and intermediate) and the cryptic plasmid, permitting sensitive detection of 19 Ct serovars (A, B/Ba, C, D/Da, E, F, G/Ga, H, I/Ia, J, K, L1, L2/L2a, and L3) without any cross-reactivity with other Chlamydia species and pathogenic bacteria or commensal organisms of the genital tract. Ct-positive samples are analyzed by a nitrocellulose-based reverse hybridization assay (RHA) containing probes for the 19 different serovars and for the cryptic plasmid. The sensitivity of the PCR-DEIA on clinical specimen is equivalent to that of the Cobas TaqMan assay [kappa = 0.95 (95% confidence interval = 0.92 to 0.99)]. Using the RHA, 98% of the Ct-DT detection step-positive samples could be typed. Analysis of cervical swabs from Uganda and The Netherlands revealed that the most common serovars in Uganda are G/Ga (45%), E (21%), K (13%), and F (8%), and in The Netherlands serovars E (38%), F (23%), G/Ga (11%), and D/Da (7%) were most common. Thus, multiplex broad-spectrum PCR in combination with DEIA and RHA permits highly sensitive and specific detection and identification of Ct serovars.

Background and Aim: Cervical cancer is caused by high-risk (HR) Human Papilloma Virus (HPV) infections. The mortality rate of cervical cancer in Suriname is 8/100.000 per year. Current HPV vaccinations protect against cancers induced by HR-HPV 16 and –18, and are not yet available in Suriname. We estimated prevalence of, and risk factors for urogenital HR-HPV infections among women from five major ethnic groups in Paramaribo, Suriname in a pre-vaccination era. Methods Between July 2009 and February 2010, women aged ≥ 18 years were recruited at a Family Planning clinic and STI clinic. Vaginal swabs were collected and general HPV detection was performed using the highly sensitive broad-spectrum SPF10 PCR and DNA enzyme immunoassay (DEIA). Subsequently, DEIA-positive samples were genotyped by the LiPA25 reverse hybridization assay (Labo Bio-medical Products, The Netherlands), targeting 25 different genotypes. Logistic regression analysis was used to identify determinants of HPV infection. Results 1001 women were included and tested for HPV of whom 584 (58%) were DEIA positive. Of the positive samples 472 (81%) could be typed by LiPA25. HR-HPV types were detected in 303 (30%) of all samples. HPV type 52 was the most prevalent genotype (n = 79; 8%). Of the HR-HPV, 46 (15%) had only HR-HPV types 16 and/or 18, covered by available vaccines. Adjusted for age the following independent associations with HR-HPV were found; ≥ 2 recent partners (OR = 1.5, 95% CI = 1.1–2.2), Chlamydia trachomatis co-infection (OR = 2.3, 95% CI = 1.5–3.5) and ethnic group (OR = 2.1, 95% CI = 1.4–3.1 for Creole; OR = 2.0, 95% CI = 1.2–3.3 for Maroon; OR = 2.8, 95% CI = 1.8–4.4 for Mixed race compared to Hindustani). Conclusions HR-HPV 52 is the most prevalent type in Suriname and HR-HPV is not equally distributed among ethnic groups. 85% of HR-HPV infections involve types that are not covered by the 2 currently available vaccines against cervical cancer. These data provide a basis for possible shifts in HPV genotype prevalence following vaccination.

Background Prevention and treatment of anal intraepithelial neoplasia (AIN) in HIV+ men who have sex with men (MSM) is subject of discussion. Knowledge on causative HPV types is crucial in understanding AIN and in vaccination studies. However, data on AIN-specific HPV are limited and whole tissue sections (WTS) often show multiple HPV infections. In this study, we analysed whether WTS and subsequent laser capture micro-dissection (LCM) with HPV PCR genotyping accurately detects type-specific HPV DNA in individual areas of high grade (HG)AIN. Methods 31 WTS with HGAIN of 21 HIV+ MSM were analysed by the SPF10 PCR/LiPA25 (version 1) HPV genotyping system. In case of multiple HPV types, PCR was repeated in selected areas of AIN, isolated by LCM. Results WTS PCR showed a single HPV type in 17 (55%). In the remaining 14 WTS sections with multiple HPV types, PCR was repeated in LCM-isolated dysplastic areas (median: 4 per WTS). In 12 of 14 these samples, the number of HPV types could be reduced to single HPV types within discrete areas of a lesion, resulting in a total of 29 (17+12), in which (components of) HGAIN show a singe HPV type. HPV 16 was found in 14/29 (48%), HPV 18 in 3 and HPV 58 in 3. The remaining HPV types that could be linked to a lesional area were HPV 26, 31, 35, 39, 52, 53, 54, 59, 67, 68/73, 74, 91 and one indeterminate HPV type. Conclusion WTS PCR and subsequent LCM PCR is accurate in detecting lesion specific HPV types in AIN and it seems that 94% of the AIN-lesions (on macroscopic or microscopic level) are caused by a single HPV type. Apart from HPV 16, the predominant type, a wide range of other HPV types are responsible for HGAIN, which has consequences for vaccination development.

Background Anal intraepithelial neoplasia (AIN) is present in the majority of HIV+ men who have sex with men (MSM). The main aetiological factor is human papilloma virus (HPV). We analysed whether HPV PCR genotyping of whole tissue sections (WTS) and subsequent laser capture microdissection (LCM) detects type-specific HPV DNA in individual areas of HGAIN, and compared these lesional HPV genotypes with HPV types in anal swabs. Methods: DNA was isolated from anal swabs and WTS of 31 HGAIN biopsies obtained from 21 HIV+ MSM and analysed by the SPF10 PCR/LiPA25 HPV genotyping system. In case of multiple HPV types per WTS, PCR was repeated in selected areas of AIN obtained by LCM. Results: A single HPV type was observed in 15 (48%) of 31 WTS. In the 16 WTS with multiple HPV types, LCM-isolated dysplastic areas were studied. In 14 of the 16 WTS, one single HPV type was demonstrated within each discrete area of the lesion, resulting in a total of 29 (15+14) biopsies in which a single HPV type was found in (all components of) the lesion. HPV 16 was found in 14 of 31 biopsies (45%). In almost half the biopsies, lesional HPV types were not detected in the anal swabs. Conclusions: WTS PCR and subsequent LCM PCR is accurate in detecting lesion-specific HPV types in HGAIN and in 94% of the biopsies a single HPV was found in (each component of) the HGAIN lesion. In the majority, HPV16 was not the causative type and HPV genotyping of anal swabs was not useful in detecting the lesion-specific type.

Chlamydia trachomatis serovars are divided into three serogroups, namely serogroup B, serogroup I (Intermediate) and serogroup C, and subsequently into 19 different serovars. Worldwide, serogroup B is the most prevalent followed by serogroup I. Clear differences have been observed in the duration of infection and growth kinetics between serovars from different serogroups in murine and cell culture models. Reasons for these observed differences are bacterial and host related, and are not well understood. The aim of this study was to determine the differences in immunoglobulin (Ig) G responses between the three serogroups in a group of patients infected with different serovars. Serovars were assessed from 235 C. trachomatispositive patients and quantitative IgG responses were determined. Analyses of variance were used to compare the IgG responses between the three serogroups. Of the serovars, 46% were B group (with serovar E the most prevalent: 35.3%), 39.6% were I group and 14.3% were C group. A highly significant difference in serologic response was shown when comparing the mean IgG concentrations (AU/mL) of patients having serovars in the most prevalent serogroup compared to the other serogroups: B = 135, C = 46 and I = 60 (B vs. C and B vs. I, P < 0.001). In conclusion, the most prevalent serovars generate the highest serologic responses.