Cervical cancer screening programs in low-income communities. Experiences from Ecuador. Low cost detection of HPV infection in a developing country.

OBJECTIVES: 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. METHODS: 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. CONCLUSION: 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.

Pathological, clinical and prognostic characteristics of breast cancer in Central Sudan versus Northern Italy: implications for breast cancer in Africa.

AIMS: In patients of Black African ethnicity, breast cancer is reportedly characterized by aggressive, poorly differentiated phenotype(s). To highlight possible differences between breast cancer in indigenous sub-Saharan African and European patients, two breast cancer case series, from Central Sudan (Khartoum) and Northern Italy (Milan), were compared for clinicopathological characteristics, expression of oestrogen receptor (ER), progesterone receptor (PR), Her-2/neu, basal cytokeratin (CK) 5/6 and CK17, and breast cancer subtypes. METHODS AND RESULTS: After careful antigen retrieval, 114 and 138 consecutive formalin-fixed paraffin-embedded (FFPE) breast cancer cases from the Radiation and Isotope Centre (Khartoum) and from MultiMedica (Milan), respectively, were screened by immunohistochemistry for ER, PR, Her-2/neu, CK5/6 and CK17. Compared with the Italian patients, the Sudanese patients were younger (P < 0.0001) and their tumours were larger (P < 0.0001), more advanced in stage (P < 0.00001), higher grade (P < 0.00001) and more frequently positive for nodal metastases (P < 0.00001). ER expression varied between the two series (P < 0.0008), but no significant differences were found for PR (P < 0.32), combined hormone receptors (P < 0.12), Her-2/neu (P < 0.09), CK5/6 (P < 0.1), CK17 (P = 0.4), combined basal CK status (P = 1) or breast cancer subtypes (P = 0.12). CONCLUSION: The differences between the Sudanese and Italian breast cancer series reflect stage at diagnosis rather than intrinsic biological characteristics. This may have relevant implications for breast cancer prevention and treatment in Africa.

Facial granulomatous diseases: a study of four cases tested for the presence of Mycobacterium tuberculosis DNA using nested polymerase chain reaction.

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.

Normal and cancer-prone human cells respond differently to extremely low frequency magnetic fields.

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.

Nested polymerase chain reaction on vaginal smears of tuberculous cervicitis. A case report.

BACKGROUND: 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. CASE: 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. CONCLUSION: The technique described here could also be applied to aspiration cytology smears to give rapid and accurate information on mycobacterial infections.

Human Immunodeficiency Virus Localization in Human Papillomavirus-Related, High-Grade Squamous Intraepithelial Lesions of the Cervix in Women with HIV Infection: Microdissection and Molecular Analysis on Formalin-Fixed and Paraffin-Embedded Specimens.

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-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 ….

Local impairment of immunoreactivity in HIV-infected women with HPV-related squamous intraepithelial lesions of the cervix.

AIMS AND BACKGROUND: The aim of this study was to compare the local immune response in two groups of patients with high-grade cervical intraepithelial squamous lesions (SIL): one with HIV infection and the other with HPV infection alone. MATERIALS AND METHODS: 16 conization specimens (8 from HIV-infected and 8 from non-HIV-infected patients) of HPV-related, high-grade SIL were selected. The specimens from non-HIV patients were considered as controls. The total number of Langerhans cells, CD4 and CD8 cells present in 10 field areas (3.120 mm2) was recorded in each case. In HIV patients CD4 and CD8 peripheral counts were performed immediately before surgery. RESULTS: The CD4/CD8 ratio never exceeded 0.71, whereas the lowest ratio in controls was 0.81: this difference was statistically significant (P = 0.0009). The mean number of Langerhans cells was markedly reduced in the high-grade SILs in the HIV patients in comparison with controls (P = 0.001). The number of CD4 cells and the CD4/CD8 ratio correlated with the peripheral CD4 count (P = 0.001 and 0.02). CONCLUSIONS: In our study a marked local impairment of cervical immunoreactivity was observed, which may play a major role in the progression of these lesions in HIV-infected women.

Calretinin. A selective marker of normal and neoplastic mesothelial cells in serous effusions.

OBJECTIVE: To document that a polyclonal antiserum to calretinin, a 29-kd calcium-binding protein, consistently decorates normal and tumor mesothelial cells in cytologic preparations. STUDY DESIGN: Thirty-three archival cytologic specimens from eight patients with histologically confirmed malignant mesothelioma and 13 from patients with metastatic serous effusions were destained and then immunostained with anticalretinin antiserum. For investigation of cell suspensions, four pleural fluids were incubated with anticalretinin antiserum. After cytocentrifugation the specimens were stained in accordance with the alkaline phosphatase anti-alkaline phosphatase (APAAP) method. For electron microscopic examination the cell suspensions were then incubated with gold-labeled antirabbit antibody. RESULTS: The diagnostic sensitivity of this new immunocytochemical approach reached 100% for the eight malignant mesotheliomas investigated. Only 3 of the 13 adenocarcinomas metastatic to the serous membranes included in this study were weakly reactive, accounting for 81% specificity. Binding of anticalretinin antiserum to living mesothelial cells was consistently documented in all four cases investigated. CONCLUSION: Calretinin is a very useful marker for positive identification of normal and tumor mesothelial cells in serous effusions.

[Detection of mycobacterium tuberculosis DNA using nested polymerase chain reaction in lymph nodes with sarcoidosis, fixed in formalin and embedded in paraffin]. / Ricerca del DNA del Mycobacterium tuberculosis mediante nested Polymerase Chain Reaction in linfonodi con sarcoidosi fissati in formalina e inclusi in paraffina.

An involvement of mycobacteria in the pathogenesis of sarcoidosis has often been hypothesized, but not confirmed reproducibly. In this study we applied a nested Polymerase Chain Reaction (PCR) to the insertion sequence IS6110 for detection of Mycobacterium tuberculosis (MT) DNA in formalin-fixed and paraffin-embedded tissues from patients with sarcoidosis, with tuberculosis and atypical mycobacteriosis confirmed by culture, and from negative control samples. MT-DNA could be detected in 2/30 samples of sarcoidosis, in 10/10 tuberculoses, in 0/5 atypical mycobacterioses and in 0/10 negative controls. Nested PCR confirmed its high sensitivity and specificity in detecting MT-DNA on archival histopathological specimens. From our results we conclude that in the granulomatous lesions of sarcoidosis MT-DNA is only sporadically demonstrable and probably it doesn't play a role in the pathogenesis of this disease.

Immunocytochemical detection of progesterone receptors. A study in a patient with primary pulmonary hypertension.

Primary pulmonary plexogenic arteriopathy (PPPA) is one of the principal conditions in which pulmonary hypertension may be clinically unexpected. It occurs in the lung vessels in the absence of any demonstrable cause. Its high incidence in women of childbearing age combined with reports of disease following delivery of a child or assumption of oral contraceptives suggest that hormonal factors may play a role in the pathogenesis of PPPA. The suspicion that the pulmonary vascular lesions occurring in PPPA could represent the effect of a hormonal mediated vascular hyperreactivity prompted the evaluation of the steroid hormone receptor status on lung tissue obtained from a women suffering from this disease who had a double-lung transplantation. By the immunocytochemical method performed on formalin fixed, paraffin-embedded lung tissue, we showed the presence of progesterone receptors (PR) in the nuclei of the myofibroblasts forming the arterial obstructive intimal proliferations and of the spindle cells present in the walls of the plexiform lesions. To enhance the staining and to facilitate the observation, we used a microwave-based antigen unmasking technique. The lack of estrogen receptors and the presence of PR could have increased, in the case, the sensitivity of the pulmonary muscular arteries to vasoconstrictory compounds. We hypothesize that on this substrate of a presumptive steroid-mediated vasoconstriction the sequence of the histologic lesions characteristic of pulmonary vascular hypertensive disease could have developed.