新疆维族妇女感染的HPV 型别分布的研究

目的:探讨人乳头瘤状病毒(Human papillomavirus,HPV)在新疆南部维族妇女人群中的型别分布情况。方法:以年龄在30-59岁的新疆伽师县夏普吐勒乡维吾尔族妇女人群为基础进行筛查,签署知情同意书后,采集受试者宫颈脱落细胞,利用PCR和基因芯片技术检测HPV DNA并分型。结果:共2473名妇女入选。HPV总的感染率为9.1%,高危型中HPV-16的感染率最高为6.9%,其他高危型的感染率从高到低依次为:HPV-59、HPV-56、HPV-18、HPV-33、HPV-58、HPV-51、HPV-31、HPV-45、HPV-52、HPV-68、HPV-35、HPV-39。低危型中HPV11感染率最高,其他低危型的感染率从高到低依次为HPV-42、HPV-43、HPV-6、HPV-53、HPV-66、HPV-73。HPV-44、-83、-MM4没检测到。多重感染率为34.2%。结论:新疆维吾尔族妇女人群中以HPV16感染为主,其次为HPV59、56、18、33等。HPV59可能是新疆维吾尔族妇女较易感染的类型。体现了新疆维吾尔族妇女感染HPV的特殊性。     

人乳头瘤病毒疫苗的研究进展

人乳头瘤病毒（human papillomavirus,HPV）是宫颈癌的主要致病因子,宫颈癌已成为继乳腺癌之后引起女性死亡的第二大癌症,在一些发展中国家宫颈癌已经成为女性癌症死亡率的首位。另外,HPV还与很多其他癌症及皮肤、粘膜疣等常见疾病有关。因此,研究和开发预防和治疗宫颈癌及相关疾病的疫苗十分必要,本文综述了HPV预防性疫苗和治疗性疫苗的研究进展,以期为HPV疫苗的研究提供参考。     

Effectiveness of bivalent and quadrivalent human papillomavirus vaccination in Luxembourg

BackgroundIn Luxembourg, the human papillomavirus (HPV) vaccination program introduced in 2008, provided either bivalent (BV) or quadrivalent (QV) vaccines to girls aged 12–17 years. Here, we estimate the effectiveness of BV and QV vaccines combined and separately in reducing type-specific HPV prevalence eight years after the introduction of the vaccination program.MethodsA cross-sectional prevalence study was conducted among women aged 18–29 years in 2015-2017. Seven hundred sixteen participants were recruited at family planning centres or private gynaecology practices in Luxembourg. Vaccination records were verified in the social security database. Cervical samples were tested using the Anyplex II HPV28 assay. Vaccine effectiveness was estimated using logistic regression.ResultsIn total, 363/716 (50.7%) participants were HPV positive with any HPV and 209/716 (29.2%) with carcinogenic HPV genotypes. HPV vaccination offered high protection against HPV16/18 (adjusted odds ratio (AOR) = 0.13; 95% CI 0.03-0.63), HPV6/11 (AOR = 0.16; 95% CI 0.05-0.48) and cross-protection against HPV31/33/45 (AOR = 0.41; 95% CI 0.18-0.94). The AORs were generally enhanced when only considering vaccination before sexual debut corresponding to AORs: 0.05 (95% CI 0.00-0.88), 0.08 (95% CI 0.02-0.36) and 0.20 (0.06-0.65) against HPV16/18, HPV6/11 and HPV31/33/45, respectively. We observed significant protection against carcinogenic genotypes included in nonavalent vaccine for BV (AOR = 0.29; 95% CI 0.13-0.67), but not for QV (AOR = 0.81; 95% CI 0.47–1.40) (heterogeneity Chi² P = 0.04).ConclusionsOur study suggests high effectiveness of HPV vaccination against HPV6/11, HPV16/18 and a cross-protection against HPV31/33/45. Vaccination effectiveness was slightly higher for women vaccinated before sexual debut.     

Squamous anal cancer: Patient characteristics and HPV type distribution

BackgroundInfection with high risk human papillomavirus (HPV) is strongly associated with anal cancer. However, detailed studies on HPV type distribution by gender and age are limited.MethodsRetrospective study of 606 invasive anal cancers diagnosed between 1990 and 2005 in two large urban areas of the province of Québec, Canada. Cases were identified from hospitalization registry. Patient characteristics were collected from medical files. Archived anal squamous cancer specimens were available from 96 patients and were tested for HPV DNA and typing. Variant analysis was performed on 16 consecutive and 24 non-consecutive HPV16-positive samples to assess potential contamination during amplification.ResultsAmong the 606 patients with anal cancers, 366 (60%) were women. Median age at diagnosis was 63 years. HPV was detected in 88/96 (92%) of cases. HPV16 was the most frequent type detected in 90% of HPV-positive specimens. Other types including 6, 11, 18, 33, 52, 53, 56, 58, 62 and 82 were also found. HPV 97 was not detected. HPV prevalence was associated with female gender and younger age. No contamination occurred during amplification as shown by the subset of 41 HPV16-positive samples, as 37, 2 and 1 isolates were from the European, African and Asian lineages, respectively. The most frequent variants were G1 (n = 22) and the prototype (n = 12).ConclusionsWomen with anal cancer are at higher risk for anal HPV infection, and HPV infection, especially HPV16, is strongly associated with squamous anal cancer. Therefore, HPV vaccine could potentially prevent the occurrence of anal cancer in both men and women.     

553 例女性宫颈高危型HPV 感染与宫颈病变的相关性研究

目的:研究女性生殖道高危型人乳头瘤病毒(high risk human papilloma virus,hrHPV)感染现状及子宫颈上皮内瘤样病变(cervical intraepithelial neoplasia,CI N)的现患率,并分析该人群年龄与HPV感染及宫颈癌前病变的相关性以及hrHPV病毒载量与宫颈病变之间剂量-效应关系。方法:对553例30岁-78岁有性生活史的女性进行以人群为基础的横断面调查。对所有接受筛查的妇女均行宫颈HPV检测及电子阴道镜下病理活检,并以病理结果作为诊断子宫颈病变的金标准。资料采用EXCEL整理,利用x2检验和非条件Logistic回归分析危险因素和CI N的关系。结果:553例受检对象总体阳性检出率为18.0%。hrHPV感染情况在不同年龄段分布有显著差异,hrHPV病毒载量越高宫颈病变的程度越重。结论:不同年龄段hrHPV感染情况不同,hrHPV病毒载量与宫颈病变之间存在剂量-效应关系。     

Genetic diversity and phylogenetic analysis of HPV 16 & 18 variants isolated from cervical specimens of women in Saudi Arabia

Human papillomavirus (HPV) are well known to be associated with the development of cervical cancer. HPV16 and HPV 18 are known as high-risk types and reported to be predominantly associated with cervical cancer. The prevalence and genetic diversity of HPV have been well documented globally but, in the Kingdom of Saudi Arabia, data on HPV genetic diversity are lacking. In this study, we have analyzed the genetic diversity of both HPV16 and HPV18 based on their L1 gene sequence because L1 gene is a major capsid protein gene and has been utilized to develop a prophylactic vaccine. In January 2011–2012, a total of forty samples from cervical specimens of women in Saudi Arabia were collected. The association of HPV16, HPV18 was detected by polymerase chain reaction, sequenced and submitted to GenBank. The sequences identity matrix and the phylogenetic relationship were analyzed with selected HPVs. The highest sequence identity (99.5%) for HPV16 and (99.3%) for HPV was observed with selected HPVs. The phylogenetic analysis results showed that HPVs from Saudi Arabia formed a closed cluster with African, Asian, East Asian as well as American HPVs distributed into multiple linages from various geographical locations. The results provided the valuable information about genetic diversity, but there is an urgent need to generate full genome sequence information which will provide a clearer picture of the genetic diversity and evolution of HPVs in Saudi Arabia. In conclusion, the generated data will be highly beneficial for developing molecular diagnostic tools, analyzing and correlating the epidemiological data to determine the risk of cervical cancer and finally to develop a vaccine for Saudi Arabian population.     

A novel CD4 T-cell epitope described from one of the cervical cancer patients vaccinated with HPV 16 or 18 E7-pulsed dendritic cells

Previously, safety and immunogenicity of human papillomavirus type 16 (HPV16) or 18 E7-pulsed dendritic cells (DC) vaccinations were demonstrated in a dose-escalation Phase I clinical trial which enrolled ten patients diagnosed with stage IB or IIA cervical cancer (nine HPV 16-positive, one HPV 18-positive). The goal of the study was to define the T-cell epitopes of HPV 16 or 18 E7 protein in these patients in order to develop new strategies for treating HPV-associated malignancies. This was accomplished through establishing T-cell lines by stimulating peripheral blood mononuclear cells with autologous mature DC pulsed with the HPV 16 or 18 E7 protein, examining the T-cell responses using ELISPOT assays, and isolating E7-specific T-cell clones based on IFN-γ secretion. Then, the epitope was characterized in terms of its core sequence and the restriction element. Twelve T-cell lines from eight subjects (seven HPV 16-positive, one HPV 18-positive) were evaluated. Positive T-cell responses were demonstrated in four subjects (all HPV 16-positive). All four were positive for the HPV 16 E7 46-70 (EPDRAHYNIVTFCCKCDSTLRLCVQ) region. T-cell clones specific for the E7 47–70 region were isolated from one of the subjects. Further analyses revealed a novel, naturally processed, CD4 T-cell epitope, E7 58–68 (CCKCDSTLRLC), restricted by the HLA-DR17 molecule. This work was supported by the National Institutes of Health (R21CA094507). An erratum to this article can be found at     

553例女性宫颈高危型HPV感染与宫颈病变的相关性研究

目的：研究女性生殖道高危型人乳头瘤病毒（high risk human papilloma virus,hrHPV）感染现状及子宫颈上皮内瘤样病变（cervical intraepithelial neoplasia,CI N）的现患率,并分析该人群年龄与HPV感染及宫颈癌前病变的相关性以及hrHPV病毒载量与宫颈病变之间剂量-效应关系。方法：对553例30岁-78岁有性生活史的女性进行以人群为基础的横断面调查。对所有接受筛查的妇女均行宫颈HPV检测及电子阴道镜下病理活检,并以病理结果作为诊断子宫颈病变的金标准。资料采用EXCEL整理,利用x2检验和非条件Logistic回归分析危险因素和CI N的关系。结果：553例受检对象总体阳性检出率为18.0%。hrHPV感染情况在不同年龄段分布有显著差异,hrHPV病毒载量越高宫颈病变的程度越重。结论：不同年龄段hrHPV感染情况不同,hrHPV病毒载量与宫颈病变之间存在剂量-效应关系。     

A retrospective analysis of human papillomavirus (HPV) prevalence and genotype distribution among 25,238 women in Shanghai,China revealed the limitations of current HPV-based screening and HPV vaccine

BackgroundTo determine the human papillomavirus (HPV) type-specific prevalence and distribution among women with various age and cervical lesions in Shanghai, China. And to evaluate the carcinogenicity of different high-risk HPV (HR-HPV) and the efficacy of HR-HPV testing and HPV vaccine.MethodsThe clinical data from 25,238 participants who received HR-HPV testing (HPV GenoArray test kit, HybriBio Ltd) at the Affiliated Hospital of Tongji University from 2016 to 2019 were reviewed and analyzed using SPSS (version 20.0, Tongji University, China).ResultsThe overall prevalence of HPV was 45.57% in the study population, of which 93.51% were found HR-HPV infection. The three most prevalent HR-HPV genotypes were HPV 52 (22.47%), 16 (16.4%) and 58 (15.93%) among HPV-positive women, and HPV 16 (43.30%), 18 (9.28%) and 58 (7.22%) in women with histologically confirmed cervical cancer (CC). 8.25% of CC were found to be HPV negative. Only 83.51% of CC cases were related to the HPV genotypes covered by nine-valent HPV vaccine. HPV prevalence and genotype distribution varied with age and cervical histology. The odds ratios (OR) of HR-HPV for CC were also different, among which the top three types were HPV 45 [OR= 40.13, 95% confidence intervals (CI) 10.37–155.38], 16 (OR=33.98, 95%CI 15.90–72.60) and 18 (OR=21.11, 95%CI 8.09–55.09). The increase in the types of HPV infection did not increase the risk of CC correspondingly. As the primary cervical screening method, HR-HPV testing showed the high sensitivity (93.97%, 95%CI 92.00–95.49) but low specificity (42.82%, 95%CI 41.81–43.84).ConclusionsOur study provide the comprehensive epidemiological data on HPV prevalence and genotype distribution among Shanghai women with various cervical histology, which can not only serve as a significant reference for clinical practice, but also implicated the need of more effective CC screening methods and HPV vaccine covering more subtypes.     

人乳头状瘤病毒分型及病毒载量与宫颈病变的研究进展

宫颈癌作为目前妇女最常见的恶性肿瘤,新发病例在全球范围内仍处于上升趋势。已有实验证明,HPV感染是宫颈癌发生发展的必要条件。然而不同分型及其病毒载量高低仍会影响诊断率,从而影响病情,使之发展为持续性感染,最终演变为癌前病变或宫颈癌。目前,研究者对于高危型HPV病毒载量与宫颈病变之间是否存在依存关系尚存争议。研究表明,高病毒载量可能成为预测宫颈癌前病变进展的指标,然而对于病理诊断明确的宫颈癌患者而言,病毒载量并不能代表病理分期。本文综述了人乳头状瘤病毒,不同基因分型,病毒载量等因素对于宫颈病变及宫颈癌的相关性的近年研究进展,希望有助于指导临床工作。     

Lack of effects on fertility and developmental toxicity of a quadrivalent HPV vaccine in Sprague‐Dawley rats

Human papillomavirus (HPV) infection is one of the most common sexually transmitted diseases, with approximately half of the HPV‐infected people being adolescents and young adults. A recently developed quadrivalent HPV vaccine, GARDASIL^®, has been shown to be highly effective in the prevention of a number of HPV‐mediated diseases. The objective of the present study was to evaluate the potential effects of the vaccine on female fertility and F1 development, growth, behavior, and reproductive performance. In addition, anti‐HPV antibodies in the F0 females and F1 offspring were measured during the study. Two groups of 65 virgin Sprague‐Dawley rats were administered two or four intramuscular injections of the vaccine (full human dose of 0.5 mL at 5 and 2 weeks prior to mating, on Gestation Day [GD] 6, and Lactation Day [LD] 7; or GD 6 and LD 7 only). Additional groups of rats were administered phosphate‐buffered saline or Merck Aluminum Adjuvant (MAA) at the same four times. All females were mated to males of the same stock. Cesarean sections were performed on 22/group on GD 21, 22/group were allowed to deliver, and remaining females used for blood collections or replacements. F0 female fertility parameters were evaluated. An extensive number of prenatal, perinatal, and postnatal parameters were evaluated in the F1 generation. There were no unscheduled deaths during the study. There was no evidence of toxicity in the F0 females given either MAA or vaccine. There were no effects on the fertility or reproductive performance of the F0 females. There was no evidence of developmental toxicity to the F1 generation, including fetal body weight and morphology, postnatal growth and development, behavior, and reproductive performance. The quadrivalent vaccine induced a specific antibody response to the four HPV types in the F0 female rats following one or multiple injections. Antibodies against all four HPV types were transferred to the F1 generation during gestation and/or lactation, likely via the placenta and milk, respectively. The passively transferred antibodies persisted up to Postnatal Day 77 when they were last measured. These results demonstrate that this quadrivalent HPV vaccine had no detectable adverse effects in either the treated F0 female rats or the F1 generation. Birth Defects Res (Part B) 83:561–572, 2008. © 2008 Wiley‐Liss, Inc.     

用质量控制狭缝式杂交法（Coslot）检测妇女宫颈上皮中人乳头瘤病毒（HPV）DNA

本文报道一种能对ＨＰＶ进行较为准确分型的刚Ａ杂交程序和方法。它具有如下特点：１．由于简化了实验程序,标本消耗减少。因此该方法尤其适用于小组织活检标本（１－２ｍｇ）中ＤＮＡ同源序列检测;２．敏感性高。由于标本消耗减少,使ＨＰＶ检出率大为提高,与斑点杂交技术相比,敏感性提高１．６倍;３．型别特异性强。由于改变了实验条件及在实验过程中实行严格的质量控制,明显减少了不同ＨＰＶ型别的交叉反应,甚至在部分出现交叉反应的标本中,仍可通过强度对比来最后判定ＨＰＶ感染型别。应用该技术对１４３份宫颈炎标本、１７份宫颈不典型增生标本及２５份宫颈癌标本进行了ＨＰＶ１６型检测,阳性率分别为５０．０５％（７３／１４３）、５７％（２７／４７）及７６％（１９／２５）,并与Ｄｏｔｂｌｏｔ方法进行了比较。该方法在ＨＰＶ感染与宫颈癌关系的分子流行病学研究中具有重要意义和广阔的应用前景。     

HPV感染与宫颈癌预防的研究进展

宫颈癌在全球范围内依然是严重威胁妇女健康的常见恶性肿瘤之一。流行病学调查显示,高危型HPV持续感染是导致宫颈癌发病的主要原因,并且HPV感染具有显著的特点,因此,预防HPV感染是防治宫颈癌的主要途径。已明确性行为是促进HPV感染最重要的辅助因子,个体免疫力低下及年龄因素亦是促进HPV感染的重要因素。研究显示,在全球逐渐开展的预防措施包括对常见的高风险HPV类型进行预防性疫苗接种及包括HPV检测在内的宫颈癌筛查项目的实施已经在降低宫颈癌的发病率方面起到了很重要的作用。近些年来,人们越来越重视HPV检测在宫颈癌筛查程序中的应用。由于HPV的感染率具有显著的地域性差异,我们需要针对各地区的特点以完善相应的宫颈癌筛查程序,从而为宫颈癌防治工作的开展提供重要依据。     

5434例女性泌尿生殖道HPV感染情况及HPV亚型分析

目的了解泰顺地区女性易感人群人乳头瘤病毒(HPV)感染率和HPV亚型分型情况。方法收集2017年1月至2017年12月于泰顺县人民医院体检及就诊的女性患者泌尿生殖道标本,共5434例。采用PCR反向点杂交法进行HPV分型检测,并对结果进行分析。结果5434例标本中HPV阳性1895例,HPV阳性1309人次。高危型HPV检出1127例,占59.4%;低危型HPV检出356例,占18.8%。入选患者中单一感染935例,占71.4%;多重感染374例,占28.6%。高危型中检出最多的是HPV52型,共210例,占11.1%;其次是HPV58型137例,占7.2%;HPV16型124例,占6.5%;HPV18型62例,占3.3%。HPV52阳性患者集中于41~50岁,HPV53阳性患者集中于31~40岁,HPV16、HPV18阳性患者集中于20~40岁及>60岁。随着患者年龄的增长各HPV亚型的阳性率也在增长,且各年龄段患者均以高危型HPV感染为主(62.36%)。结论泰顺地区女性患者HPV亚型及患者年龄分布情况与我国南方女性HPV感染情况相符,HPV感染率随患者年龄增加而增高。高危型HPV检测对子宫颈癌等相关疾病的筛查及预防具有较高的应用价值。     

川西高原地区妇女人乳头瘤病毒感染现状及其与阴道微环境改变的关系

【背景】人乳头瘤病毒(human papillomavirus,HPV)感染与宫颈癌及癌前病变的发生密切相关,HPV疫苗是预防感染的重要手段,但HPV亚型分布具有明显的区域差异。【目的】了解川西高原地区妇女宫颈HPV感染现状,探讨其与阴道微环境改变之间的关系,为该人群HPV疫苗使用和宫颈癌预防工作提供指导。【方法】选取2017年11月至2019年5月在成都三六三医院体检和就诊的3 816例川西高原地区妇女,进行HPV基因分型、薄层液基细胞学检测(thinprep cytologic test,TCT)和阴道微环境检查,分析HPV感染率和基因型分布,并探讨高危型HPV (high risk HPV,hrHPV)感染与阴道微环境改变之间的关系。【结果】3 816例妇女的HPV总感染率为19.81%(756/3 816),hrHPV感染率为16.48%(629/3 816),HPV52在hrHPV感染中占比最高,达17.20%(135/785),后面依次为HPV58 (92/11.72%)、HPV16 (89/11.34%)、HPV53 (88/11.21%)、HPV18 (55/7.01%...     

CIN患者阴道微生态状况与HPV感染的关系分析

分析宫颈上皮内瘤变（CIN）患者阴道微生态与人乳头瘤病毒（HPV）感染的关系。

选取326例本院收治的CIN患者,采集阴道分泌物和宫颈脱落细胞,通过形态学和功能学检测阴道微生态状况,采用PCR-反向点杂交法进行HPV分型检测,采用二分类Logistic回归分析影响CIN患者HPV感染的因素。

CIN患者HPV阳性共256例,占78.5%,HPV阴性共70例,占21.5%。HPV阳性CIN患者白细胞计数＞10个/高倍视野、阴道清洁度Ⅲ-Ⅳ度、VVC阳性、AV阳性、BV阳性、pH值＞4.5、白细胞酯酶阳性、过氧化氢阳性占比高于HPV阴性患者,微生态正常者占比低于HPV阴性患者（P＜0.05）。二分类Logistic回归分析显示,VVC、BV、微生态正常和pH值为CIN患者HPV感染的影响因素。

CIN患者HPV感染率较高,且感染后存在阴道微生态失调,VVC、BV、微生态异常和pH值是CIN患者HPV感染的影响因素。

     

HPV分型及其高危亚型与宫颈病变的关系

目的研究HPV分型及其高危亚型与宫颈病变程度的关系。方法采用HPV基因分型技术对2014年7月至2015年8月在广州中医药大学第一附属医院妇科门诊就诊和体检的3 142例患者进行HPV分型检测,其中327例患者进行宫颈病理检查,分析高危型HPV感染与宫颈病变程度的关系。结果 3 142例受检对象中,21种HPV型别均被检测出,阳性检出者共969例,总检出率为30.80%,其中高危型741例,检出率为23.58%。随着宫颈病变级别的升高,高危型HPV阳性率也相应升高(P0.05),HPV16型感染阳性者宫颈癌发生率最高(P0.05)。高危型HPV阳性者中单一感染和多重感染与宫颈病变程度无关(P0.05)。结论高危型HPV亚型检测在宫颈病变预防和诊治中具有重要意义。     

兰州地区女性感染HPV及其基因类型分析

目的:了解兰州地区成年女性感染人乳头瘤病毒(humanpapillomavirus,HPV)及其基因类型分布状况,为本地区HPV分子流行病学研究提供理论依据。方法:利用PCR技术分别对100例妇科门诊就诊者进行HPV基因亚型检测。结果:100例样品中,HPV DNA检出率为19%(19/100),其中HPV16 DNA感染率15%(15/100),HPV58 DNA感染率3%(3/100),HPV18 DNA感染率2%(2/100),HPV 16与HPV18双重感染1例。结论:本地区成年女性HPV感染主要以HPV16多见,而HPV16与恶性肿瘤密切相关,因此对HPVDNA阳性者定期随访,有利于宫颈癌的防治。     

Low-dose radiation enhances therapeutic HPV DNA vaccination in tumor-bearing hosts

Current therapeutic approaches to treatment of patients with bulky cervical cancer are based on conventional in situ ablative modalities including cisplatin-based chemotherapy and radiation therapy. The 5-year survival of patients with nonresectable disease is dismal. Because over 99% of squamous cervical cancer is caused by persistent infection with an oncogenic strain of human papillomavirus (HPV), particularly type 16 and viral oncoproteins E6 and E7 are functionally required for disease initiation and persistence, HPV-targeted immune strategies present a compelling opportunity in which to demonstrate proof of principle. Sublethal doses of radiation and chemotherapeutic agents have been shown to have synergistic effect in combination with either vaccination against cancer-specific antigens, or with passive transfer of tumor-specific cytotoxic T lymphocytes (CTLs). Here, we explored the combination of low-dose radiation therapy with DNA vaccination with calreticulin (CRT) linked to the mutated form of HPV-16 E7 antigen (E7(detox)), CRT/E7(detox) in the treatment of E7-expressing TC-1 tumors. We observed that TC-1 tumor-bearing mice treated with radiotherapy combined with CRT/E7(detox) DNA vaccination generated significant therapeutic antitumor effects and the highest frequency of E7-specific CD8⁺ T cells in the tumors and spleens of treated mice. Furthermore, treatment with radiotherapy was shown to render the TC-1 tumor cells more susceptible to lysis by E7-specific CTLs. In addition, we observed that treatment with radiotherapy during the second DNA vaccination generated the highest frequency of E7-specific CD8⁺ T cells in the tumors and spleens of TC-1 tumor-bearing mice. Finally, TC-1 tumor-bearing mice treated with the chemotherapy in combination with radiation and CRT/E7(detox) DNA vaccination generate significantly enhanced therapeutic antitumor effects. The clinical implications of the study are discussed. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.