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  • 1
    Keywords: CANCER ; tumor ; carcinoma ; human ; CLASSIFICATION ; EXPOSURE ; RISK ; SITE ; PROTEIN ; PROTEINS ; TUMORS ; PATIENT ; DNA ; INFECTION ; FAMILY ; RISK-FACTORS ; SKIN ; MR ; papillomavirus ; ASSOCIATION ; antibodies ; IN-SITU ; risk factors ; PATHOGENESIS ; human papillomavirus ; VIRUS-LIKE PARTICLES ; HUMAN-PAPILLOMAVIRUS ; CARCINOMAS ; case-control studies ; squamous cell carcinoma ; INDIVIDUALS ; sensitivity ; RENAL-TRANSPLANT RECIPIENTS ; basal cell carcinoma ; glutathione-S-transferase ; CELL CARCINOMA ; case-control study ; population-based case-control study ; ASSOCIATIONS ; case control studies ; INTERVAL ; TECHNOLOGY ; RISK-FACTOR ; CANCERS ; population-based ; IMMUNOCOMPETENT INDIVIDUALS ; E6 PROTEIN ; multiplex serology ; PLUCKED EYEBROW HAIRS
    Abstract: Background. Although infection with human papillomaviruses (HPVs) is a major risk factor for several epithelial cancers, an etiologic relationship between HPV and keratinocyte cancers, such as squamous cell carcinomas (SCCs) and basal cell carcinomas (BCCs), remains unclear. Methods: In a population-based case-control study of 252 SCC case patients, 525 BCC case patients, and 461 control subjects, we used multiplex serology to detect antibodies in plasma samples against 16 HPV types from phylogenetic genera alpha, beta, and mu. Multiplex serology is a new method that is based on fluorescent bead technology and allows simultaneous detection of antibodies against up to 100 different in situ affinity-purified recombinant HPV proteins. Data on sun sensitivity, outdoor exposure, and other risk factors for keratinocyte cancers were collected through personal interviews. Odds ratios (ORs) with 95% confidence intervals (CIs) were calculated via unconditional logistic regression models. Results: Overall, we detected HPV antibodies more frequently in SCC patients than in control subjects (OR = 1.6, 95% CI = 1.2 to 2.3), but we found no difference in HPV seropositivity between BCC case patients and control subjects (OR = 0.8, 95% CI = 0.6 to 1.1). Among HPV types, seropositivity to HPV types in genus beta (OR = 1.5,95% CI = 1.0 to 2.1), particularly HPV 5 (OR = 1.8,95% CI = 1.0 to 3.1), was associated with SCC risk. Individuals with tumors on chronically sun exposed sites were more likely to be seropositive for beta HPV types than individuals with SCC at other anatomic sites. The highest SCC risk was associated with positivity for multiple HPV types and, among individuals seropositive for HPV beta, a tendency to sunburn; however, the associations had limited statistical precision. Conclusions: Our findings support a role for HPV types from the genus beta in the pathogenesis of SCC
    Type of Publication: Journal article published
    PubMed ID: 16537831
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  • 2
    Keywords: CANCER ; Germany ; human ; CLASSIFICATION ; DIAGNOSIS ; SYSTEM ; SYSTEMS ; DISEASE ; DISEASES ; PATIENT ; DNA ; INFECTION ; papillomavirus ; ALPHA ; IDENTIFICATION ; CERVICAL-CANCER ; PCR ; REGION ; human papillomavirus ; genotyping ; HIGH-RISK ; HPV ; HUMAN-PAPILLOMAVIRUS ; sensitivity ; MANAGEMENT ; RE ; METAANALYSIS ; methods ; USA ; microbiology ; AGREEMENT ; UPSTREAM ; HPV types ; KAPPA ; COINFECTION
    Abstract: Human papillomavirus (HPV) DNA detection and typing are important for diagnosis and management of HPV-associated diseases. One of the most commonly used PCR methods, GP5+/6+, shows weaknesses in amplifying certain types. To circumvent this limitation, we developed and validated broad-spectrum primers targeting the GP5+/6+ region. The addition of eight upstream and two downstream BSGP5+/6+ (BS) primers improved amplification of plasmids of 14 genital HPV types 10- to 1,000-fold versus GP5+/6+ PCR without altering sensitivity for the 10 others. For these 24 types, an analytic sensitivity of 〈= 1,000 plasmid copies in the presence of 100 ng cellular DNA was obtained. Additionally, we integrated an internal beta-globin PCR into both HPV PCR systems, allowing simultaneous DNA quality control without affecting the sensitivity of HPV detection. Furthermore, we describe five additional low-risk HPV probes used in multiplex HPV genotyping (MPG) for simultaneous identification of all 15 high-risk, 3 putative high-risk, and 9 low-risk HPV genotypes. The performance of BSGP5+/6+ multiplexed with beta-globin primers was compared to that of standard GP5+/6+ with DNA from 1,112 cervical scrapings. There was 79% overall agreement (kappa = 0.816). BSGP5+/6+ was significantly more sensitive than GP5+/6+ for detection of HPV 30, 39, 42, 44, 51, 52, 53, 68, 73, and 82, detecting 212 additional HPV infections and increasing the proportion of multiple infections from 17.2 to 26.9% in cancer patients. In conclusion, BSGP5+/6+ multiplexed with beta-globin PCR provides an improvement in type-specific amplification sensitivity and homogeneity compared to, GP5+/6+ and offers simultaneous internal control of DNA quality. BSGP5+/6+-MPG, therefore, is suitable for epidemiologic and also diagnostic applications
    Type of Publication: Journal article published
    PubMed ID: 18199790
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  • 3
    Keywords: CELLS ; tumor ; CELL ; human ; COMMON ; DISEASE ; SITES ; PROTEINS ; SAMPLE ; SAMPLES ; TUMORS ; TIME ; PATIENT ; DNA ; SKIN ; papillomavirus ; antibody ; IN-SITU ; LESIONS ; COPY NUMBER ; human papillomavirus ; GENOTYPES ; HPV ; REPLICATION ; glutathione-S-transferase ; PSORIASIS ; EPIDERMODYSPLASIA-VERRUCIFORMIS ; hair ; GENOTYPE ; NONMELANOMA SKIN-CANCER ; USA ; PLUCKED EYEBROW HAIRS ; CLINICAL-ASPECTS ; HAIRS ; HUMAN-PAPILLOMAVIRUS-DNA
    Abstract: Epidermodysplasia verruciformis (EV) is a rare disease, characterized by cutaneous warts and associated with a strong predisposition to beta-genus human papillomavirus (HPV). Earlier studies reported high copy numbers of HPV-DNA in nearly all skin tumors from EV patients, but neither HPV replication status in non-lesional skin nor anti-HPV seroreactivity in these patients have been reported yet. We therefore performed a comprehensive viral load analysis for the more common beta-HPV types on skin samples and plucked eyebrow hairs from four EV patients treated at our dermatology department. The results clearly demonstrate that they carry a multiplicity (up to eighteen types) of beta-HPV genotypes in both skin sites. Worthy of note, a high intrapatient concordance for specific types between hair bulbs and skin biopsies was observed and the same beta-PV profile was maintained over time. Viral load analysis revealed a load range between less than one HPV-DNA copy per 100 cells to more than 400 HPV-DNA copies per cell in both eyebrow hairs and skin proliferative lesions. Evaluation of seroreactivity to beta-HPV types in the four EV patients revealed that antibodies against the 16 beta-HPV were significantly more prevalent and showed higher titers than in the controls
    Type of Publication: Journal article published
    PubMed ID: 18923444
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  • 4
    Keywords: SPECTRA ; CANCER ; carcinoma ; CELL ; Germany ; human ; EXPOSURE ; HISTORY ; POPULATION ; RISK ; GENOME ; radiation ; RESPONSES ; DNA ; INFECTION ; CARCINOGENESIS ; SKIN ; papillomavirus ; antibodies ; antibody ; LESIONS ; WOMEN ; MEN ; RISK FACTOR ; human papillomavirus ; HPV ; HUMAN-PAPILLOMAVIRUS ; SQUAMOUS-CELL CARCINOMA ; NETHERLANDS ; squamous cell carcinoma ; INDIVIDUALS ; sensitivity ; NATURAL-HISTORY ; RENAL-TRANSPLANT RECIPIENTS ; glutathione-S-transferase ; SERUM ; CELL CARCINOMA ; EPIDERMODYSPLASIA-VERRUCIFORMIS ; development ; RISK-FACTOR ; SQUAMOUS-CELL ; SUN EXPOSURE ; virology ; SEROPREVALENCE ; biotechnology ; CUTANEOUS HUMAN PAPILLOMAVIRUSES ; CONFIDENCE ; SCC ; PAPILLOMAVIRUS TYPES
    Abstract: Solar UV radiation is the main risk factor for cutaneous squamous cell carcinoma (SCC), but infections with skin human papillomavirus (HPV) types have also been linked to the development of SCC. Little is known about the natural history of these infections and whether the seroprevalence of skin HPV types is affected by ambient or individual levels of sun exposure. This study investigated this by analysing sera for antibodies to 26 skin HPV types from five phylogenetic genera obtained from 807 healthy individuals from the Netherlands, Italy and Australia, countries with strong differences in sunlight intensity. Overall HPV seroprevalence, was similar across the three countries (50-57% for beta-HPV types, 40-48% for gamma-HPV types), and the most frequent beta-HPV and gamma-HPV types were the same in all countries. The highest seroprevalences; for 24 of the 26 skin HPV types were observed in Italy (114 types) and Australia (ten types). Seroprevalence among men was generally higher than among women, and the male sex was significantly associated with both beta-HPV [odds ratio (OR) 2.81, 95 % confidence interval (CI) 1.64-4.821 and gamma-HPV (OR 2.42, 95% CI 1.40-4.18) antibodies in Australia. The only measure of sun sensitivity or UV exposure significantly associated with skin HPV seroprevalence was found for weekend sun exposure in Australia and beta-HPV antibodies. It was concluded that type spectra and HPV seroprevalence are similar in countries with different sunlight intensity, and that levels of UV exposure do not play a strong role in the development of skin HPV antibodies in this study population
    Type of Publication: Journal article published
    PubMed ID: 19386782
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  • 5
    Keywords: CANCER ; EXPRESSION ; COMBINATION ; Germany ; DIAGNOSIS ; screening ; RNA ; transcription ; DNA ; INFECTION ; MARKER ; ACID ; LESIONS ; PROGRESSION ; MALIGNANCIES ; PATTERNS ; ASSAY ; WOMEN ; cervical cancer ; CERVICAL-CANCER ; TYPE-16 ; HIGH-RISK ; HPV ; TRANSFORMATION ; HUMAN KERATINOCYTES ; HUMAN-PAPILLOMAVIRUS TYPE-16 ; L1 ; specificity ; INFECTIONS ; PRECURSORS ; ONCOLOGY ; PATTERN ; papillomaviruses ; MESSENGER-RNAS ; development ; E2 PROTEIN ; transcriptome ; Lead ; Type ; OVERTREATMENT ; SEQUENCE-BASED AMPLIFICATION
    Abstract: Infections with high-risk human papillomaviruses (HPV), mainly HPV type 16, can cause malignant transformation of the human cervical epithelium and the development of cervical cancer (CxCa). A rapid and precise diagnosis of the precancerous lesions by conventional cytology or HPV DNA tests remains difficult and often leads to overtreatment. We quantitatively analyzed the HPV16 transcriptome of 80 HPV16 DNA-positive cervical scrapes classified as mild cytologic grade, including no intraepithelial lesion or malignancy (NIL/M; normal, n = 25) and low-grade squamous intraepithelial lesion (LSIL; n = 24), and severe cytologic grade, including high-grade squamous intraepithelial lesion (HSIL; n = 24) and CxCa ( n = 7), with novel nucleic acid sequence-based amplification-Luminex assays. In severe lesions, HPV16 E6*II and E1C encoding transcripts were strongly upregulated, whereas spliced E1(boolean AND)E4 and L1 encoding transcripts were markedly downregulated. Using a combination of the four marker transcripts, 100% of CxCa and 67% of HSIL cases were correctly identified as severe, and 74% of LSIL and 92% of NIL/M samples as mild cytologic grade. Compared with a commercially available HPV E6/E7 mRNA assay, the specificity of the marker combination for discriminating severe and mild cytologic lesions increased from 23% to 83%. In conclusion, we identified a novel HPV16 RNA pattern for grading of cervical lesions with a potentially high diagnostic value for the primary screening of CxCa precursors and the triage of cervical lesions. Cancer Res; 70(1); 249-56. (C) 2010 AACR
    Type of Publication: Journal article published
    PubMed ID: 20028865
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  • 6
    Keywords: CELLS ; RISK ; TISSUE ; DNA ; ASSOCIATION ; antibodies ; antibody ; PCR ; human papillomavirus ; HPV ; SQUAMOUS-CELL CARCINOMA ; HEAD ; L1 ; PREVALENCE ; POLYMERASE CHAIN-REACTION ; glutathione-S-transferase ; NECK-CANCER ; HUMAN-PAPILLOMAVIRUS HPV
    Abstract: According to PCR, the prevalences of human papillomavirus (HPV) DNA were 6.3% (13 of 206) in tonsillitis or hypertrophic tonsillar tissues and 0.6% (1 of 174) in exfoliated cells from normal tonsils. HPV-16 was the only type detected in tonsillar tissues, but it did not appear to lead to L1 antibody production
    Type of Publication: Journal article published
    PubMed ID: 15750119
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  • 7
    Keywords: Germany ; human ; HYBRIDIZATION ; DNA ; papillomavirus ; IDENTIFICATION ; AMPLIFICATION ; ASSAY ; WOMEN ; REPRODUCIBILITY ; CERVICAL-CANCER ; PCR ; human papillomavirus ; HIGH-RISK ; HUMAN-PAPILLOMAVIRUS ; sensitivity ; RE ; ASSAYS ; HIGH-THROUGHPUT ; TECHNOLOGY ; DNA HYBRIDIZATION ; PAP
    Abstract: Typing of human papillomaviruses (HPV) by DNA hybridization procedures, such as reverse line blot (RLB) assay, is sensitive and well validated. However, the application of these assays to high-throughput analyses is limited. Here, we describe the development of multiplex human papillomavirus genotyping (MPG), a quantitative and sensitive high-throughput procedure for the identification of multiple high- and low-risk genital HPV genotypes in a single reaction. MPG is based on the amplification of HPV DNA by a general primer PCR (GP5+/6+) and the subsequent detection of the products with type-specific oligonucleotide probes coupled to fluorescence-labeled polystyrene beads (Luminex suspension array technology). Up to 100 different HPV types can be detected simultaneously with MPG, and the method is fast and labor saving. We detected all 22 HPV types examined with high specificity and reproducibility (the median interplate coefficient of variation was below 10%). Detection limits for the different HPV types varied between 100 and 800 pg of PCR products. We compared the performance of MPG to an established RLB assay on GP5+/6+-PCR products derived from 94 clinical samples. The evaluation showed an excellent agreement (kappa = 0.922) but also indicated a higher sensitivity of MPG. In conclusion, MPG appears to be highly suitable for large-scale epidemiological studies and vaccination trials as well as for routine diagnostic purposes
    Type of Publication: Journal article published
    PubMed ID: 16455905
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  • 8
    Keywords: CANCER ; CELL ; human ; screening ; POPULATION ; PROTEIN ; PROTEINS ; DNA ; INFECTION ; MARKER ; papillomavirus ; NEOPLASIA ; prevention ; HEALTH ; ASSAY ; NUMBER ; AGE ; WOMEN ; cervical cancer ; cervical intraepithelial neoplasia ; CERVICAL-CANCER ; MARKERS ; PCR ; human papillomavirus ; TYPE-16 ; FRANCE ; HIGH-RISK ; HPV ; HUMAN-PAPILLOMAVIRUS ; POPULATIONS ; UNITED-STATES ; intraepithelial neoplasia ; vaccination ; L1 ; ABNORMALITIES ; PREVALENCE ; ONCOLOGY ; RE ; GRADE ; PROTOCOL ; biomarker ; pooled analysis ; USA ; cancer research ; CANCERS ; population-based ; serology ; PUBLIC-HEALTH ; SEROPREVALENCE ; GENERAL-POPULATION ; HPV types ; HPV vaccination ; PARTNER ; HPV-16 ; HPV PREVALENCE SURVEYS ; SOUTH-KOREA
    Abstract: Data on human papillomavirus (HPV) and cervical cancer burden in Central Asia are scarce. To investigate HPV infection in Ulaanbaatar, the capital of Mongolia, we obtained cervical cell specimens from a population of 969 women ages 15 to 59 years. DNA of 44 HPV types was detected using a GP5+/6+ PCR-based assay. Seropositivity for L1 proteins of HPV 16, 18, 31, 33, 45, 52, and 58 was assessed using multiplex HPV serology. Cytologic abnormalities were detected in 127 women (13.1%), among whom 6 cervical intraepithelial neoplasia grade 3 and 2 invasive cervical cancers were diagnosed. Overall HPV DNA prevalence was 35.0%, being highest (48.5%) in women ages 〈25 years. High-risk types were detected in 24.5% of women. HPV DNA prevalence declined with age but remained 〉25% in all age groups. HPV seroprevalence was also very high (38.0%) and increased steadily from 33.2% to 48.9% in women ages 〈25 and 50 to 59 years, respectively. However, the proportion of women positive for both HPV markers of any individual HPV type was low. HPV16 was the most frequently detected type by PCR (6.1%), serology (23.0%), or both (2.1%). Lifetime number of sexual partners and induced abortions were shown to be directly associated with HPV DNA and/or seroprevalence. HPV prevalence in Ulaanbaatar was higher than that detected by similar HPV testing protocols in other populations in Asia or elsewhere and would suggest an important, yet unquantified, cervical cancer burden. Improving cervical cancer prevention, through screening and HPV vaccination, is an important public health issue for Mongolia
    Type of Publication: Journal article published
    PubMed ID: 18628425
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  • 9
    Keywords: CANCER ; tumor ; carcinoma ; CELL ; COMBINATION ; human ; EPIDEMIOLOGY ; RISK ; TISSUE ; TUMORS ; validation ; PATIENT ; DNA ; INFECTION ; MARKER ; BIOMARKERS ; TISSUES ; SKIN ; papillomavirus ; antibodies ; antibody ; skin cancer ; MARKERS ; PCR ; human papillomavirus ; HPV ; HUMAN-PAPILLOMAVIRUS ; SQUAMOUS-CELL CARCINOMA ; squamous cell carcinoma ; NORMAL TISSUE ; SERIES ; sensitivity ; specificity ; NONMELANOMA SKIN CANCERS ; RENAL-TRANSPLANT RECIPIENTS ; BIOPSY ; SKIN-CANCER ; basal cell carcinoma ; glutathione-S-transferase ; SERUM ; CELL CARCINOMA ; ELISA ; ONCOLOGY ; EXTENSION ; ARRAY ; EPIDERMODYSPLASIA-VERRUCIFORMIS ; HIGH PREVALENCE ; hair ; TUMOR TISSUE ; biomarker ; TECHNOLOGY ; ACTINIC KERATOSES ; NORMAL SKIN ; technique ; USA ; RISK-FACTOR ; CANCERS ; SQUAMOUS-CELL ; nonmelanoma skin cancer ; IMMUNOCOMPETENT INDIVIDUALS ; NOV ; validation studies ; BASAL-CELL ; MICROARRAY PRIMER EXTENSION ; MULTIPLEX PCR
    Abstract: Cutaneous human papillomavirus (HPV) may be associated with the development of nonmelanoma skin cancer (NMSC), as suggested by reports of HPV DNA in NMSC tumors. HPV has also been investigated as an NMSC risk factor in epidemiologic studies, although findings vary across studies that used different biomarkers of HPV infection in normal tissues. To identify appropriate biomarkers for use in future epidemiologic studies, we conducted a sampling validation study. NMSC tumor tissue was obtained from 20 patients with pathology-confirmed basal or squamous cell carcinoma of the skin, in addition to several normal tissues, including eyebrow hairs, normal skin swabs obtained using multiple techniques, normal skin punch and shave biopsies, and serum for antibody measurement. Presence of cutaneous HPV DNA in tissues was measured with multiplex PCR using HPV type-specific primers and array primer extension (APEX) for HPV typing. Antibody detection was based on glutathione-S-transferase capture ELISA in combination with fluorescent bead technology. Using HPV DNA in tumor tissues as a gold standard, sensitivity and specificity were calculated for each measure of HPV infection in normal tissues. beta-Papillomavirus DNA was observed in tumor tissues in 60% of patients. The normal skin punch biopsy demonstrated optimal sensitivity (75%) and specificity (75%). Biomarkers obtained using less-invasive techniques demonstrated poor specificity when considered individually, although specificity improved when biomarkers were combined. Based on the current case series, the combinations of antibodies + eyebrow hairs or antibodies + eyebrow hairs + Dacron swabs are the optimal, minimally invasive markers of cutaneous HPV infection for use in epidemiologic studies. (C) 2008 Wiley-Liss, Inc
    Type of Publication: Journal article published
    PubMed ID: 18729188
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  • 10
    Keywords: CANCER ; carcinoma ; CELL ; human ; COMMON ; COHORT ; EPIDEMIOLOGY ; HISTORY ; POPULATION ; RISK ; PROTEINS ; TIME ; DNA ; INFECTION ; RISK-FACTORS ; ANTIGEN ; SKIN ; papillomavirus ; ASSOCIATION ; SUSCEPTIBILITY ; antibodies ; antibody ; AGE ; WOMEN ; MEN ; RISK FACTOR ; human papillomavirus ; VIRUS-LIKE PARTICLES ; HPV ; HUMAN-PAPILLOMAVIRUS ; POPULATIONS ; case-control studies ; squamous cell carcinoma ; L1 ; PREVALENCE ; glutathione-S-transferase ; SERUM ; CELL CARCINOMA ; case control study ; case-control study ; RECIPIENTS ; HPV 16 ; TECHNOLOGY ; ACTINIC KERATOSES ; NONMELANOMA SKIN-CANCER ; HISTOLOGY ; USA ; UNIT ; RISK-FACTOR ; SQUAMOUS-CELL ; IMMUNOCOMPETENT INDIVIDUALS ; serology ; SEROPREVALENCE ; cutaneous squamous cell carcinoma (SCC) ; HPV types ; human papillomavirus (HPV) ; Genital warts ; CONFIDENCE ; organ transplant recipients ; SCC
    Abstract: A case-control study was conducted in 140 people with histology proven cutaneous squamous cell carcinoma (SCC) and 454 controls, nested within 2 cohorts of organ transplant recipients (OTR) recruited in London and Oxford between 2002 and 2006. All participants had a skin examination, completed a questionnaire and had serum tested for antibodies against the L1 antigen of 34 HPV types using Luminex technology. SCC was more common in men than women (odds ratio [OR] = 1.7, 95% confidence interval [CI]: 1.1-2.8, p = 0.02) and in people with susceptibility to burn easily (OR = 3.0, 95%CI: 1.9-4.8; p 〈 0.001). The risk increased with increasing age (p-trend 〈 0.001), increasing time since transplant (p-trend 〈 0.001), increasing self-reported number of sunburns as a child (p(trend) 〈 0.001) and with the presence of viral warts (p 〈 0.001). As expected, antibodies against HPV 16 were associated with a self-reported history of an abnormal cervical smear among women (OR 5.1, 95%CI: 2.6-10.2) and antibodies against HPV 6 were associated with a self-reported history of genital warts (OR 4.0, 95%CI: 2.2-7.2). However, no clear associations between any of the HPV types examined (including cutaneous betaHPVs) and SCC were identified. For example, the seroprevalence of HPV 5 was 15% among cases and 9% among controls (p = 0.09) and the seroprevalence of HPV 8 was 23% among cases and 21% among controls (p = 0.6). Nor was seropositivity to multiple types associated with SCC. These serological data do not provide evidence for a role for HPV in the aetiology of cutaneous SCC among OTR in two UK-based populations. (C) 2009 UICC
    Type of Publication: Journal article published
    PubMed ID: 19588489
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