Maria Cannone

University of Milan, Milano, Lombardy, Italy

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Publications (23)40.9 Total impact

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    ABSTRACT: The rare occurrence of histology-proven cervical intraepithelial neoplasia grade 3 (CIN 3) or invasive cancer with a negative HC2 result is known. Tissue blocks of 37 cases of histology-diagnosed CIN 3+ with a concomitant negative HC2 test were genotyped to investigate the human papillomavirus (HPV) status within the lesion. We considered 1,976 cervical excision specimens performed with concomitant HC2 test. Of these, 37 histology-confirmed CIN 3+ resulted HC2 negative. Thirty-three paraffin blocks, derived by the cervical excision, could be genotyped for high- (HR) and low-risk (LR) HPV genotypes. Detailed histology showed 30 CIN 3, 2 squamous cell invasive carcinomas, and 5 invasive adenocarcinomas. One specimen resulted not amplifiable at the genotyping. Twenty-two cases (68.7%) were positive for HR-HPV types, either in single (n = 17) or multiple HR-HPV infection (n = 5). Most of the HR-HPVs found were 16 or 18. Ten cases (31.3%) were negative for HR-HPV types; 5 of these were positive for probable HR-HPV types, not detectable with HC2 HR-probes, 1 was positive to LR-HPV types, while 1 had HPV-69/71. Three cases were negative for HPV DNA, either high or low risk. Of the rare cases of CIN 3+ lesions with concomitant negative HC2 test, 69% are true failures in HR-HPV detection. One third of HC2-negative CIN 3+ is related to the presence of other HPV genotypes not covered by the HC2 panel or to undetectable HPV in the lesion; both these rare occurrences were already described in large cancer series and partially explain the occurrence of HPV-negative CIN 3+.
    Journal of Lower Genital Tract Disease 08/2013; 18(1). DOI:10.1097/LGT.0b013e3182909f86 · 1.11 Impact Factor
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    ABSTRACT: Genetic and environmental factors are thought to contribute to the etiology of the autoimmune disease myasthenia gravis (MG). Viral involvement has long been suspected, but direct evidence of involvement has not been found. We recently reported that Toll-like receptor 4 (TLR4)-a key activator of innate immunity-was overexpressed in the thymus of some patients with MG, suggesting that thymic infection by pathogens might be involved in MG pathogenesis. We searched for evidence of intrathymic infection in patients with MG. Twenty-seven MG thymuses (6 involuted, 7 hyperplastic, 5 thymitis, and 9 thymoma) previously tested for TLR4 expression, 18 nonpathologic control thymuses, and 10 pathologic control thymuses from patients without MG (8 thymoma and 2 hyperplastic) were analyzed for cytomegalovirus, varicella-zoster virus, herpes simplex virus types 1 and 2, eubacteria, respiratory syncytial virus, and enteroviruses using PCR techniques. Immunohistochemistry and double immunofluorescence were used to detect enterovirus capsid protein VP1 in thymic specimens and analyze TLR4 expression in VP1-positive cells. Poliovirus was detected in 4 MG thymuses (14.8%; 2 thymitis and 2 thymoma). No virus was detected in any control thymus. A linear correlation between plus and minus strand poliovirus RNA levels was observed in all 4 thymuses, suggesting persistent thymic infection. VP1 protein was detected in the cytoplasm of CD68-positive macrophages scattered through thymic medulla in all PV-positive thymuses. VP1 and TLR4 colocalized in infected cells. Poliovirus-infected macrophages are present in thymus of some patients with myasthenia gravis, suggesting a viral contribution to the intrathymic alterations leading to the disease.
    Neurology 04/2010; 74(14):1118-26. DOI:10.1212/WNL.0b013e3181d7d884 · 8.30 Impact Factor
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    ABSTRACT: With the support of the independent humanitarian organization "Amici Fundation Terra Nueva" in Quito, Ecuador, we evaluated the feasibility of a cytologic screening program sustained by volunteers on the field and in Italy. 250 women underwent a cervical Pap-test. The women with a positive Pap-smear were re-called for visual inspection with acetic acid (VIA), whereas those with a negative smear were invited for a new Pap-test after 3 years. To obtain samples for molecular assays, cytologic material was removed from slides, submitted to DNA extraction and amplified by nested PCR of the L1 region of HPV DNA. PCR-positive samples were sequenced. RESULTS. Six (2.6%) samples showed squamous intra-epithelial lesions (SILs): 4 low grade and 2 high grade SILs were present in women more than 40 years old. The overall rate of successful DNA recovery on a per-slide basis was 96.5%. High grade SILs were characterized by HPV 16 and 18 co-infection. HPV 16 was detected in one low grade SIL. HPV-DNA was detected in 11 smears (4.95%): in all 6 SILS and in 5 of the 216 negative smears. Independent humanitarian organizations could play a role in supporting national screening programs offering skilled field professionals and technical support by scientists operating in their countries. Our molecular technique has the potential to provide important epidemiological information in many resource-poor areas of developing countries.
    Pathologica 04/2009; 101(2):76-9.
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    ABSTRACT: We performed a technical and cost-effectiveness analysis of quantitative reverse transcription-polymerase chain reaction (Q-RT-PCR) for the assessment of HER2 in breast cancer. We evaluated 44 frozen and 55 formalin-fixed paraffin-embedded (FFPE) breast carcinoma specimens by Q-RT-PCR, immunohistochemical analysis, and fluorescent in situ hybridization (FISH). Immunohistochemical and FISH analyses were performed on individual slides and on tissue microarray. Costs of techniques were calculated to study 1 case and 10 or 40 cases. Q-RT-PCR provided reliable data in frozen and FFPE specimens, which were significantly correlated. HER2 messenger RNA levels were significantly stratified in agreement with immunohistochemical data (P < .05). There was complete concordance between Q-RT-PCR and immunohistochemical results for negative and strongly positive (3+) cases. The intermediate immunohistochemical group (2+), including FISH+ and FISH- cancers, could also be stratified by Q-RT-PCR. Cost analysis documented the advantage of Q-RT-PCR in all US Food and Drug Administration-approved assays. Our data support the use of Q-RT-PCR for testing breast cancer specimens to select patients for HER2 inhibitory therapy.
    American Journal of Clinical Pathology 04/2008; 129(4):563-70. DOI:10.1309/1AKQDQ057PQT9AKX · 3.01 Impact Factor
  • Clinical Immunology 01/2006; 119. DOI:10.1016/j.clim.2006.04.253 · 3.99 Impact Factor
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    ABSTRACT: To analyze a procedure for breast cancer that requires performance of a molecular test before extensive intraoperative examination, fine needle aspiration cytology (FNAC) of surgically removed sentinel lymph nodes (SLNs). The diagnostic accuracy of extensive histologic examination and immunohistochemistry (IHC) of 101 SLNs from 98 breast carcinoma patients were compared with that of the evaluation of 2 specific mRNA markers by reverse transcriptase polymerase change reaction (mammaglobin and MUC-1). Cell specimens were obtained by FNAC of the SLNs immediately before freezing. Metastases were detected on frozen sections in 19 cases (18.81%). IHC on serial sections confirmed the metastases and showed micrometastases or isolated tumor cells in 24 SLNs (23.76%). Mammaglobin was expressed in 20 FNAC specimens (19.80%). MUC-1 assay was positive in 11 cases only (10.89%). This technique allows a complete histologic examination without sacrifice of part of the SLN and at the same time is a valuable diagnostic adjunct to the detection of occult tumor cells. Moreover, it is less expensive and time consuming than extensive IHC.
    Acta cytologica 01/2006; 50(3):271-6. DOI:10.1159/000325953 · 1.56 Impact Factor
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    ABSTRACT: Thymic abnormalities are present in approximately 80% of myasthenia gravis (MG) patients, and the thymus seems to be the main site of autosensitization to the acetylcholine receptor. In view of findings that the innate immune system can generate an autoimmune response, we studied the expression of Toll-like receptors (TLRs) 2 to 5, key components of innate immunity signaling pathways, in 37 thymuses from patients with autoimmune MG. TLR4 mRNA levels were significantly greater in thymitis (hyperplasia with diffuse B-cell infiltration) and involuted thymus than in germinal center hyperplasia and thymoma. By immunohistochemistry and confocal microscopy, cells positive for TLR4 protein were rarely detected in thymoma. However, in thymitis TLR4 protein was mostly found on epitheliomorphic (cytokeratin-positive) cells located in close association with clusters of acetylcholine receptor-positive myoid cells in thymic medulla and also at the borders between cortical and medullary areas. B cells were never TLR4-positive. TLR4 protein was also present in remnant tissue of involuted thymus. This is the first finding of a possible link between innate immunity and MG. We speculate that in a subgroup of MG patients, an exogenous or endogenous danger signal may activate the innate immune system and give rise to TLR4-mediated mechanisms contributing to autoimmunity.
    American Journal Of Pathology 08/2005; 167(1):129-39. DOI:10.1016/S0002-9440(10)62960-4 · 4.60 Impact Factor
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    ABSTRACT: The possibility of a pressure monitoring system by differential pressure sensors to detect contaminant effects on cellular cultures metabolic activity is discussed using Saccharomyces cerevisiae, lymphocyte, and AHH1 cell cultures. Metabolic (aerobic and anaerobic) processes in cells are accompanied by CO(2) production that induces changes in pressure values when cells are cultured in sealed vessels. These values are subsequently converted in voltage units and plotted pressure dynamics versus time. This procedure leads to a standard curve, typical of the cellular line, which characterizes cellular metabolism when all parameters are controlled, such as temperature and nutrients. Different phases appear in the S. cerevisiae differential pressure curve: an initial growth up to a maximum, followed by a decrement that leads to a typical "depression" (pressure values inside the test-tubes are lower than the initial one) after about 35 h from the beginning. The S. cerevisiae differential pressure curve is successfully used to test the effects of chemical (Amuchina, trieline) and physical (UV radiation, blue light, magnetic fields) contaminants. The same technique is applied to lymphocytes and AHH1 cultures to investigate the effects generated by a 72-h exposure to a 50-Hz, 60-microT electromagnetic field. Lymphocyte samples, cultured in a PHA medium, grow less than control ones, but exhibit a greater metabolic activity: changes in the exposure system configuration influence neither sample growth differences nor metabolic response variations between control and irradiated samples, while all the other irradiation parameters remain constant. Control and irradiated lymphocyte samples, without PHA in culture medium, show the same behavior both during irradiation and metabolic test. AHH1 control and irradiated samples show no difference both in growth percentage during irradiation and in metabolic activity. Different cell cultures respond to the same stimulus in different manners.
    Journal of Biomedical Optics 01/2004; 9(5):1074-88. DOI:10.1117/1.1782591 · 2.75 Impact Factor
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    ABSTRACT: A wealth of data point to Delayed Luminescence (DL) as a good candidate for early and reliable detection technique in neoplastic cells and tissues sorting. Aiming at a DL experimental set up for such a kind of information, a testing technique for morphological analysis should be provided. This could certify the early identification of pathologies and abnormalities in cells and tissues by DL. DL technique may be coupled with FIB (Focused Ion Beam) imaging analysis to give a correlated, both spectroscopic and morphological investigation, at the submicron scale. A strong link among others has been reported to exist between DL signal characteristics and cytoskeleton structure and dynamics: FIB (Focused Ion Beam) imaging is for the moment being the best non invasive check at all and it can detect morphological alterations as early as possible since its resolution can go down to 2-5 nm. The cells, that can be highlighted by the fast DL and slow and efficient FIB, can be in parallel analysed by a metabolic manometric technique that uses differential pressure sensors: the different cellular activity of normal and abnormal cells can be recorded and this allows fast and non-invasive investigations, although requiring a minimal number of cells. In addition it"s possible to study, by the confocal microscopy spectroscopic analysis, DNA fragments, exploiting the optical characteristics of a dye, like ethidium bromide, to detect dynamic and conformational changes in DNA chains. These changes can be artificially induced in cells (e.g. by irradiation) or found in neoplastic cells. The acquired experience allows an independent check of spectroscopic, morphologic and metabolic testing by a control on nucleic acid defects. These four techniques may be used together creating a "protocol" in order to permit an early and reliable alterations diagnosis of cells and tissues, guaranteeing an high accuracy standard.
    Proceedings of SPIE - The International Society for Optical Engineering 10/2003; DOI:10.1117/12.500201 · 0.20 Impact Factor
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    ABSTRACT: The possibility of using a pressure monitoring system based on differential pressure sensors to detect contaminant effects on cellular cultures metabolic activity is discussed using Saccharomyces cerevisiae cell cultures: differential pressure curves' shape, starting slope and maximum are affected both by physical and chemical contamination. Aim of the present study is the investigation of the effects generated by a 72h exposition of Saccharomyces cerevisiae, human lymphocytes and AHH1 cellular line cultures to 50Hz, 60(mu) T electromagnetic field. No significant differences have been recorded between irradiated and control yeast samples. On other hand irradiated lymphocytes samples, cultures in a PHA medium, grow less than control ones, but exhibit a greater metabolic activity: changes in the exposure system configuration influence neither sample growth differences nor metabolic response variations between control and irradiated samples. Control and irradiated lymphocyte samples, without PHA in culture medium, show the same behavior both during irradiation and metabolic test. AHH1 control and irradiated samples show no difference both in growth percentage during irradiation and in metabolic test. Different cell cultures respond to the same stimulus in different manners.© (2002) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.
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    ABSTRACT: The possibility of using a pressure monitoring system based on differential pressure sensors to detect contaminant effects on cellular cultures metabolic activity is discussed using Saccharomyces cerevisiae cell cultures: differential pressure curves' shape, starting slope and maximum are affected both by physical and chemical contamination. Aim of the present study is the investigation of the effects generated by a 72h exposition of Saccharomyces cerevisiae, human lymphocytes and AHH1 cellular line cultures to 50Hz, 60(mu) T electromagnetic field. No significant differences have been recorded between irradiated and control yeast samples. On other hand irradiated lymphocytes samples, cultures in a PHA medium, grow less than control ones, but exhibit a greater metabolic activity: changes in the exposure system configuration influence neither sample growth differences nor metabolic response variations between control and irradiated samples. Control and irradiated lymphocyte samples, without PHA in culture medium, show the same behavior both during irradiation and metabolic test. AHH1 control and irradiated samples show no difference both in growth percentage during irradiation and in metabolic test. Different cell cultures respond to the same stimulus in different manners.
    Proceedings of SPIE - The International Society for Optical Engineering 06/2002; · 0.20 Impact Factor
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    ABSTRACT: The results are discussed of a systematic investigation into the electromagnetic field (EMP) exposure consequences on human lymphocytes. These artificial fields have intensities comparable to the Earth's magnetic field, and are used for exposures up to 4 days. Different and complementary techniques are used to safely assess the consequences of EMFs on the cells; in particular, morphology, metabolism, and population dynamics are investigated. The recourse to ultramicroscopy, pressure monitoring in sealed bottles, atomic mass spectroscopy, and cytofluorimetry techniques give good insight into the EMF-induced changes. A statistically significant deviation of irradiated samples with respect to control samples is reported. A critical analysis and a survey of similar experiments reported in the literature led us to examine the experimental setup with attention to the geometry of the irradiation system. Yeast cells were used as a model system to statistically test the different steps in the overall procedure, thanks to information gathered during a radiobiology experiment performed at the Rutherford Appleton Laboratory. Finally, the role of different magnetic field detectors in the reproducibility of the experiments is carefully discussed.
    Electromagnetic Biology and Medicine 03/2001; 20(1):81-106. DOI:10.1081/JBC-100103162 · 0.77 Impact Factor
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    ABSTRACT: The histopathologic diagnosis of cutaneous tuberculosis (CTB) is often troublesome, because there are several other entities (tuberculids, demodicidosis, granulomatous rosacea, and acne agminata) that may display granulomatous inflammation with caseation necrosis. The current study describes four cases of granulomatous disease of the face. The final diagnosis (assessed on the basis of the clinical response to therapy) was CTB in three cases and granulomatous rosacea in one case. Histologically, epithelioid granulomas were a constant feature; in one case of CTB, they displayed a palisading (granuloma annulare-like) arrangement. Caseation necrosis was a prominent feature only in the case of granulomatous rosacea. Routinely processed biopsy specimens were evaluated with nested polymerase chain reaction (nPCR) for Mycobacterium tuberculosis (MBT) DNA. The correlation between nPCR results and clinical outcome was less than optimal; in fact, one case showed an excellent clinical response to the antituberculous drug therapy despite the absence of MBT DNA amplification. In granulomatous diseases of the face, the importance of evaluating not only nPCR but the overall clinicopathologic picture so as to avoid diagnostic misinterpretations is emphasized.
    American Journal of Dermatopathology 03/2001; 23(1):8-15. DOI:10.1097/00000372-200102000-00002 · 1.43 Impact Factor
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    ABSTRACT: Human lymphoblastoid cells of normal origin and from genetic instability syndromes, i.e. Fanconi anemia (FA) group C and ataxia telangectasia, were continuously exposed to extremely low frequency magnetic field (ELF-MF). We report that ELF-MF, though not perturbing cell cycle progression, increases the rate of cell death in normal cell lines. In contrast, cell death is not affected in cells from genetic instability syndromes; this reflects a specific failure of the apoptotic response. Reintroduction of complementation group C in FA cells re-established the apoptotic response to ELF-MF. Thus, genes implicated in genetic instability syndromes are relevant in modulating the response of cells to ELF-MF.
    FEBS Letters 02/2001; 487(3):397-403. DOI:10.1016/S0014-5793(00)02376-0 · 3.34 Impact Factor
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    ABSTRACT: The results are discussed of a systematic investigation on the electromagnetic field exposure consequences on human lymphocytes. These artificial fields have intensities comparable with the Earth magnetic field one, and are used for exposures up to 4 days. Different and complementary techniques are used to safely assess the consequences of ElectroMagnetic Fields (EMF) on the cells; in particular morphology, metabolism and population dynamics are investigated. The recourse to ultra microscopy, pressure monitoring in sealed bottles, atomic mass spectroscopy. Far IR Fourier Transform and cytofluorimetry techniques give a good insight in the EMF induced changes. A statistically significant deviation of irradiated samples with respect to the control ones are reported. A critical analysis and a survey of similar experiments reported in literature lead us to the exam of the experimental set up with attention to the geometry of the irradiation system. Finally the role of different magnetic field detectors in the reproducibility of the experiments will be carefully discussed.
    Proceedings of SPIE - The International Society for Optical Engineering 04/2000; DOI:10.1117/12.382041 · 0.20 Impact Factor
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    ABSTRACT: To evaluate the usefulness of a nested polymerase chain reaction (PCR) for Mycobacterium tuberculosis complex on routinely stained cytologic samples from patients with extrapulmonary tuberculosis. Nested PCR for the detection of a fragment of the IS6110 insertion sequence of M tuberculosis complex was applied to Ziehl-Neelsen-negative archival cytologic slides of serous effusions (pleural [n = 7], peritoneal [n = 1] and pericardial [n = 1]) and a lymph node fine needle aspirate (n = 1) from nine human immunodeficiency virus (HIV)-positive patients with autopsy-proven active extrapulmonary tuberculosis. Malignant effusions and aspirates from nine HIV-positive patients with non-Hodgkin's lymphoma and pleural effusions from seven HIV-negative patients with heart failure were used as controls. DNA was extracted after removing the coverslip and gently scraping the cytologic sample from the slides. In all cases, enough DNA was obtained for PCR without any significant loss of integrity, as demonstrated by PCR positive for HLA-Dq. PCR for M tuberculosis was positive in 8 of the 10 samples (80%) from patients with tuberculosis but also in three samples (30%) from HIV-positive patients in the control group. None of the samples from the HIV-negative patients was positive. PCR for M tuberculosis can be reliably performed on archival cytologic slides from extrapulmonary samples, but although it is highly sensitive, it may lead to positive results in immunocompromised patients without any sign of active tubercular disease.
    Acta cytologica 01/2000; 44(6):1023-8. · 1.56 Impact Factor
  • 01/2000; 44:1023-1028. DOI:10.1159/000328591
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    ABSTRACT: Abstract Objectives. To evaluate a possible mechanism of human immunodeficiency virus (HIV) and human papillomavirus (HPV) interaction, we have identified the cervical tissue compartments that harbor HIV.Materials and Methods. We studied 39 paraffin-embedded, cervical conization specimens with high-grade cervical intraepithelial neoplasia (CIN3) occurring in HIV-infected women. From selected intraepithelial HPV-positive nonulcerated specimens (confirmed by in situ hybridization), we obtained serial 4- to 5-μm-thick sections that were stained with hematoxylin and eosin, anti-S100 protein, and anti-CD4. The presence of intramucosal Langerhans' cells or dendritic cells or CD4-positive cells was recorded. Three consecutive, nonmicrodissected, full-thickness sections of the same specimens were used for polymerase chain reaction (PCR) analysis (group A). Three other uncovered, consecutive sections from the same blocks were examined with an inverted microscope, and full-thickness specimens of mucosa were dissected from the underlying cervical stroma, were gently removed, and were used for PCR (group B). The quality of DNA was checked by HLA-DQα amplification; then a nested PCR for HIV proviral DNA was performed.Results. Of group A, 5 of 39 cases (12.8%) were positive, whereas HIV was not detected in the microdissected sections of group B, with or without intraepithelial Langerhans' or CD4 cells.Conclusions. HIV does not affect cervical epithelium. The absence of infected Langerhans' or dendritic cells (or both) indicates a migration to the proximal lymph nodes of the infected cells. The absence of HIV proviral DNA in the CIN infiltrated by CD4 cells could be due to the low number or absence of infected CD4 cells.
    Journal of Lower Genital Tract Disease 09/1999; 3(4):254 - 259. DOI:10.1046/j.1526-0976.1999.34007.x · 1.11 Impact Factor
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    ABSTRACT: Objectives: To evaluate a possible mechanism of human immunodeficiency virus (HIV) and human papillomavirus (HPV) interaction, we have identified the cervical tissue compartments that harbor HIV. Materials and Methods: We studied 39 paraffin-embedded, cervical conization specimens with high-grade cervical intraepithelial neoplasia (CIN3) occurring in HIV-infected women. From selected intraepithelial HPV-positive nonulcerated specimens (confirmed by in situ hybridization), we obtained serial 4- to 5-[mu]m-thick sections that were stained with hematoxylin and eosin, anti-S100 protein, and anti-CD4. The presence of intramucosal Langerhans' cells or dendritic cells or CD4-positive cells was recorded. Three consecutive, nonmicrodissected, full-thickness sections of the same specimens were used for polymerase chain reaction (PCR) analysis (group A). Three other uncovered, consecutive sections from the same blocks were examined with an inverted microscope, and full-thickness specimens of mucosa were dissected from the underlying cervical stroma, were gently removed, and were used for PCR (group B). The quality of DNA was checked by HLA-DQa amplification; then a nested PCR for HIV proviral DNA was performed. Results: Of group A, 5 of 39 cases (12.8%) were positive, whereas HIV was not detected in the microdissected sections of group B, with or without intraepithelial Langerhans' or CD4 cells. Conclusions: HIV does not affect cervical epithelium. The absence of infected Langerhans' or dendritic cells (or both) indicates a migration to the proximal lymph nodes of the in [horizontal ellipsis] (C)1999The American Society for Colposcopy and Cervical Pathology
    Journal of Lower Genital Tract Disease 01/1999; 3(4):254-259. DOI:10.1097/00128360-199910000-00008 · 1.11 Impact Factor
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    ABSTRACT: Tuberculous cervicitis (TC) is a rare disease the diagnosis of which is based on the microscopic and/or cultural recognition of mycobacteria. In recent years, the polymerase chain reaction (PCR), especially with double-round amplification ("nested" PCR [nPCR]), has been increasingly used for rapid detection of mycobacteria in clinical samples. The present case is the first example of tuberculosis diagnosed with the aid of nPCR amplification of mycobacterial DNA fragments on smeared and Papanicolaou-stained cytologic material. First detected on vaginal smears, the amplicon IS6110 was subsequently identified also on paraffin-embedded tissue sections. The technique described here could also be applied to aspiration cytology smears to give rapid and accurate information on mycobacterial infections.
    Acta cytologica 01/1999; 43(2):308-12. DOI:10.1159/000331000 · 1.56 Impact Factor