96 research outputs found
Cancer of the uterine cervix and human papillomavirus infection
Human papillomaviruses (HPVs) have emerged as the principal sexually transmitted causal agents in the development of cancer of the uterine cervix in women. They also cause a variety of benign lesions, warts, intraepithelial neoplasia and anogenital, oral and pharyngeal papillomas. Presently, more than 100 HPV genotypes have been identified in humans, and about one-third of them have been sequenced. Of these, while HPV types 16 and 18 are considered to be the high-risk types, HPV 6 and 11 are the low-risk types in the development of cervical cancer. Evidence for causal role of HPV in the development of cervical neoplasia comes from the etiological and epidemiological observations together with the experimental findings of the molecular pathways elicited by HPV-transforming genes. Further evidence in favour of papillomavirus as the carcinoma virus comes from the findings of presence of HPV infections in cancers of oral, esophageal, larynx and nonmelanoma skin cancers. The oncogenic potentials of the virus have been attributed to its E6 and E7 genes. The products of these two genes stimulate cell proliferation by activating the cell-cycle-specific proteins and interfere with the functions of cellular growth-regulatory proteins, p53 and Rb. Identification and characterization of several human pathogenic HPV types warrant prevention of viral infection through vaccination or therapeutic intervention which could eventually control infection and expression of human pathogenic papillomaviruses
Socio-demographic profile of copper-T beneficiaries in the family planning out-patient department of a teaching hospital: a record-based descriptive study
Background: Intra uterine contraceptive devices (IUCD) used as a spacing method is one of the main strategies of the family welfare programme, as they are among the safest and most effective, affordable and convenient reversible contraceptives available. The objectives of the study were to study the socio-demographic details of beneficiaries accepting Cu-T in the family planning OPD of the medical college, relation of IUCD insertion time with respect to menses or delivery and its outcome.Methods: After IEC approval, a descriptive, complete enumeration study of recorded data from IUD register from 2006 to 2015 (n=1141) was carried out from the IUCD registers of the family planning out-patient department (OPD) of a medical college.Results: Beneficiaries had a significantly lower literacy rate (p<0.05) and a lower employment rate (p<0.01) than their husbands. 447 (39.4%) women accepted IUCD before 1 year from their last delivery. In 20 women, IUCDs were expelled, while in 32 (2.8%), they were wasted. The difference between the couples having no male children and those having at least 1 male child opting for IUCD was statistically highly significant (p<0.01).Conclusions: Cu-T is being well utilised as a spacing method in the Family Welfare Component of the Reproductive and Child Health Programme. Evidence of Preference for a male child can be seen in this study
A simple 'paper smear' method for dry collection, transport and storage of cervical cytological specimens for rapid screening of HPV infection by PCR
Human papillomaviruses (HPVs) are major pathogens associated with the development of cancer of the uterine cervix, the most common malignant tumour of women world-wide. Reliable diagnosis of HPV infection, particularly the 'high-risk' types (16/18), may facilitate early identification of 'high-risk' populations for developing cervical cancer and may augment the sensitivity and specificity of primary cervical cancer screening programmes by complementing the conventional Pap test. A simple paper smear method has been developed for dry collection, transport and storage of cervical smears/scrapes at room temperature for subsequent detection of HPV DNA by PCR assay. Imprint biopsies, blood and fine-needle aspirates were also collected by this method. The cervical scrapes or other body fluids were smeared (within 0.5-1 cm diameter) and dried on to sterile small slides made of Whatman 3MM filter paper, and stored individually at room temperature or at 4°C. A small piece (2-3 mm) of the paper smear was punched or cut out with a sterile surgical blade, boiled in an eppendorf tube containing 50 µl of distilled water for 5 min and used directly for PCR amplification. The quality and quantity of DNA derived from paper smears and the results of PCR amplifications for HPV type 16, BRCA1 and p53 genes were identical to those obtained from the same samples following collection in PBS, storage (-70°C) and phenol-chloroform-based DNA extraction. DNA was stable in the paper smears for up to a year, whether stored at room temperature or at 4°C. This method is simple, rapid and cost-effective, and can be effectively employed for large-scale population screening, especially for regions where the specimens are to be transported from distant places to the laboratory
Detection of human papillomavirus DNA sequences in cancer of the urinary bladder by in situ hybridisation and polymerase chain reaction
Objective: To evaluate the prevalence of "high risk" human papillomavirus type 16 (HPV 16) in transitional cell carcinoma of the urinary bladder. Materials and Methods: The study included 10 biopsy specimens from male patients of transitional cell carcinoma of the urinary bladder for the detection of HPV DNA sequences. Specimens were collected from the Urology Clinic of the K.G. Medical College Hospital, Lucknow, India. Detection of HPV DNA was carried out by tissue in situ hybridisation (a single copy gene localisation method) using 3H-labelled HPV DNA probe and also by polymerase chain reaction (PCR) techniques using primers to HPV 16 upstream regulatory region (URR). RESULTS--Out of 10 cases of transitional cell carcinoma of the urinary bladder, "high risk" HPV 16 DNA was detected only in one (10%) by using in situ hybridisation whereas two cases (20%) were found to be positive by polymerase chain reaction. Conclusion: Our results suggest that the rare occurrence of HPV in bladder carcinoma may not have a causal relation with the viral infection
Human papillomavirus DNA sequences in adenocarcinoma of the uterine cervix in Indian women
Background: Infection with human papillomavirus (HPV) is considered to be the principal causal agent in the development of squamous cell carcinoma of the uterine cervix. Although adenocarcinoma of the cervix originates adjacent to the squamous epithelial neoplastic lesions, the etiopathogenesis of adenocarcinoma is not yet clearly understood. Recent studies have raised more controversy, rather than answering the question of whether specific HPV infection also plays a role in the development of adenocarcinoma of the cervix. Molecular DNA hybridization techniques were used to detect HPV types prevalent in both adenocarcinoma and squamous cell carcinoma of the uterine cervix, which is the most common cancer in Indian women. Methods: Histologically confirmed, formaldehydefixed, paraffin-embedded tissue sections from 12 cases of adenocarcinoma and 30 cases of squamous cell carcinoma of the uterine cervix were analyzed retrospectively or the presence of HPV DNA types 6b, 11, 16, and 18 by Southern blot hybridization and in situ hybridization. Results: Of 12 adenocarcinomas, 5 (41.67%) tumors were positive for HPV DNA. All five cases were positive for HPV 16, and two (16.6%) of these were hybridized again to the HPV 18-specific DNA probe. All tumors were negative for HPV 6b and 11. In addition, no biopsy specialnens were positive after hybridization with a mixed probe of HPV 31, 33, 35, 39, and 45. These results were compared to those obtained for 30 squamous cell carcinomas of the cervix. Although 20 (66%) were exclusively positive for HPV 16 and 6 (20%), more tumors were of HPV 16 related types as detected under nonstringent conditions of hybridization, only one (3%) was positive for HPV 18. The results of in situ hybridization were found to be in good agreement with those of Southern blotting. Conclusions: HPV 16 is the type present almost exclusively in squamous cell carcinoma of Indian women. A higher frequency of HPV 16 in adenocarcinoma of Indian women, in contrast to HPV 18, as reported from other regions, may be attributed to geographic variation rather than to histologic differences only, and both HPV 16 and 18 may be present in adenocarcinoma of the uterine cervix
Molecular Characterization and Phylogenetic Study of Coxsackievirus A24v Causing Outbreaks of Acute Hemorrhagic Conjunctivitis (AHC) in Brazil
Coxsackievirus A24 variant (CA24v) is the most prevalent viral pathogen associated with acute hemorrhagic conjunctivitis (AHC) outbreaks. Sixteen years after its first outbreak in Brazil, this agent reemerged in 2003 in Brazil, spread to nearly all states and caused outbreaks until 2005. In 2009, a new outbreak occurred in the northeast region of the country. In this study, we performed a viral isolation in cell culture and characterized clinical samples collected from patients presenting symptoms during the outbreak of 2005 in Vitória, EspÃrito Santo State (ES) and the outbreak of 2009 in Recife, Pernambuco State (PE). We also performed a phylogenetic analysis of worldwide strains and all meaningful Brazilian isolates since 2003.Sterile cotton swabs were used to collect eye discharges, and all 210 clinical samples were used to inoculate cell cultures. Cytopathic effects in HEp-2 cells were seen in 58 of 180 (32%) samples from Vitória and 3 of 30 (10%) samples from Recife. Phylogenetic analysis based on a fragment of the VP1 and 3C gene revealed that the CA24v causing outbreaks in Brazil during the years 2003, 2004 and 2005 evolved from Asian isolates that had caused the South Korean outbreak of AHC during the summer of 2002. However, the 2009 outbreak of AHC in Pernambuco was originated from the reintroduction of a new CA24v strain that was circulating during 2007 in Asia, where CA24v outbreaks has been continuously reported since 1970.This study is the first phylogenetic analysis of AHC outbreaks caused by CA24v in Brazil. The results showed that Asian strains of CA24v were responsible for the outbreaks since 1987 and were independently introduced to Brazil in 2003 and 2009. Phylogenetic analysis of complete VP1 gene is a useful tool for studying the epidemiology of enteroviruses associated with outbreaks
Touchdown General Primer (GP5+/GP6+) PCR and optimized sample DNA concentration support the sensitive detection of human papillomavirus
BACKGROUND: The GP5+/GP6+ PCR assay is a well-established HPV detection technique. This study has examined the effects of incorporating 'hot start' and 'touchdown' steps into the protocol. In addition, dTTP was substituted with dUTP to permit contamination control measures against carry-over PCR product. METHODS: Firstly, HPV-16 was amplified from SiHa cell DNA (0.1 ng–100 ng) diluted in a background of C-33A DNA (100 ng-2 μg). Secondly, the detection of small quantities (15ag-1.5pg) of HPV recombinant plasmids (types 16, 31, 33, 45, 51, 52, and 56) diluted in C-33A DNA was investigated. Thirdly, clinical sample DNA extracts (cervical smears, formalin-fixed vaginal lesions and breast tumors) were tested for HPV. Six different PCR protocols were assessed. HPV was detected by gel electrophoresis, and by Southern and dot blot hybridization. RESULTS: HPV detection sensitivity was dependent on the total amount of DNA in a PCR. Touchdown protocols supported HPV-16 detection from 1 ng or 0.5 ng SiHa cell DNA in a background of 2 μg or 1 μg C-33A DNA respectively, and from 0.1 ng of SiHa cell DNA (~28 copies HPV-16) in 500 ng or 100 ng background DNA. Under standard GP5+/GP6+ annealing conditions, HPV-16 went undetected when the DNA content of a PCR was 2 μg or 1 μg, and with 500 ng C-33A DNA the sensitivity limit was 1 ng SiHa cell DNA. HPV recombinant plasmids were each detected with high (albeit varying) sensitivity by a touchdown protocol. HPV-31 was better amplified under standard annealing conditions (1.5fg in 100 ng background DNA) than by a touchdown approach (15fg detection limit). HPV-52 was not amplified by the standard protocol at the dilutions tested. Seventeen different HPV types were demonstrated in 47/65 (72%) abnormal cytology samples recorded as HPV negative by standard GP5+/GP6+ conditions. Twenty-one different HPV types were recorded in 111/114 (97%) vaginal lesions. Multiple infections were also detectable using a touchdown approach. Of 26 breast tumors, 5 (19%) tested HPV positive by the standard assay and 15/26 (58%) using a touchdown protocol. CONCLUSION: Touchdown modification of the GP5+/GP6+ PCR assay enables the detection of HPV undetected under regular assay conditions. The use of standardized DNA quantities in a PCR rather than standard sample volumes containing arbitrary amounts of DNA is supported. A touchdown approach may be beneficial as an analytical test for the re-evaluation of (apparently) HPV negative abnormal cervical cytological or histological samples, and for investigating the association of HPV with disease conditions at diverse organ sites. The clinical utility of a touchdown approach for HPV detection requires further investigation as increased assay analytical sensitivity may not necessarily equate with improved clinical sensitivity or specificity
Presence of papillomavirus sequences in condylomatous lesions of the mamillae and in invasive carcinoma of the breast
BACKGROUND: Viruses including Epstein–Barr virus (EBV), a human equivalent of murine mammary tumour virus (MMTV) and human papillomavirus (HPV) have been implicated in the aetiology of human breast cancer. We report the presence of HPV DNA sequences in areolar tissue and tumour tissue samples from female patients with breast carcinoma. The presence of virus in the areolar–nipple complex suggests to us a potential pathogenic mechanism. METHODS: Polymerase chain reaction (PCR) was undertaken to amplify HPV types in areolar and tumour tissue from breast cancer cases. In situ hybridisation supported the PCR findings and localised the virus in nipple, areolar and tumour tissue. RESULTS: Papillomavirus DNA was present in 25 of 29 samples of breast carcinoma and in 20 of 29 samples from the corresponding mamilla. The most prevalent type in both carcinomas and nipples was HPV 11, followed by HPV 6. Other types detected were HPV 16, 23, 27 and 57 (nipples and carcinomas), HPV 20, 21, 32, 37, 38, 66 and GA3-1 (nipples only) and HPV 3, 15, 24, 87 and DL473 (carcinomas only). Multiple types were demonstrated in seven carcinomas and ten nipple samples. CONCLUSIONS: The data demonstrate the occurrence of HPV in nipple and areolar tissues in patients with breast carcinoma. The authors postulate a retrograde ductular pattern of viral spread that may have pathogenic significance
Prevalence of enteroviruses in children with and without hand, foot, and mouth disease in China
High-risk human papillomavirus infections in breast cancer in Syrian women and their association with Id-1 expression: a tissue microarray study
High-risk human papillomaviruses (HPVs) could be important risk factors for breast carcinogenesis and metastasis. Based on this hypothesis, we recently studied the effect of E6/E7 onco-proteins of high-risk HPV type 16 in two non-invasive human breast cancer cell lines, BT20 and MCF7; we reported that E6/E7 converts these cell lines to invasive cells. This is accompanied by an overexpression of Id-1, which is an important regulator of breast metastasis. In this investigation, we examined the presence of high-risk HPVs (16, 18, 31, 33 and 35) and the expression of their E6 onco-protein as well as their correlation with Id-1 gene expression, using polymerase chain reaction (PCR) and tissue microarray (TMA) analysis, respectively, in a cohort of 113 Syrian breast cancer patients. We found that high-risk HPV types 16, 18, 31, 33 and 35 are present in 8.84, 9.73, 7.07, 55.75 and 37.16% of our samples, respectively, which represent invasive breast cancers. Overall, 69 (61.06%) of the 113 samples are HPV positive; among these specimens 24 tissues (34.78%) are coinfected with more than one HPV type. Furthermore, we report that the expression of the E6 onco-protein of these high-risk HPVs is correlated with Id-1 overexpression in the majority of invasive breast cancer tissue samples. Our data suggest that high-risk HPV infections are associated with human breast cancer progression in Syrian women
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