HPV Genotype Prevalence and Success of Vaccination to Prevent Cervical Cancer.

BACKGROUND: Nearly 500,000 new cases of cervical cancer are estimated annually worldwide. Three vaccines are currently licensed to prevent cervical cancer. The success of vaccination depends mainly on the prevalence of HPV genotypes, and many cases of HPV infection have been diagnosed after vaccination. Our aim was to search for HPV genotyping in cervical samples to verify the proportion of women that remain susceptible to infection even after vaccination. METHODS: 21,017 liquid-based cervical (LBC) specimens were received for cytology and HPV detection from 2015 to 2018. Before slide preparations for cytology, a 1,000-µL aliquot was taken from the LBC fixative and subjected to automated DNA extraction and multiplex PCR followed by capillary electrophoresis to detect and classify HPV. RESULTS: HPV was detected in 895 (4.3%) specimens. The most prevalent genotype was HPV-16, followed by HPV-58 and HPV-66. A total of 258 (28.8%) cases were positive for high-risk (HR)-HPV types (66, 59, 39, 56, 30, 35, 53, 51, 68, 82, and 70) that are not covered by the HPV vaccines. CONCLUSION: A significant proportion of HPV types detected in cytological specimens are representative of HR-HPV not covered by the available vaccines. The health system should be aware of the considerable percentage of women who are not being immunized and will continue to need cervical cancer screening.

Involvement of the central nervous system in neuroblastomas: A potential direct pathway.

Although frequently disseminated to other anatomical sites, neuroblastoma (NB) is rarely reported as involving the central nervous system (CNS), which may reflect insufficient research in poorly controlled systemic disease. Here we demonstrate the involvement of the CNS in patients with NB over 18 months of age at diagnosis of extensive systemic disease. Meningeal metastases were observed even in the presence of complete systemic control. Although no improvement in patient's survival was observed, radiotherapy was effective in preventing CNS recurrence after observation of actual or previous dural disease. In conclusion, this study uncovered the uncommon pathologic involvement of the CNS in children with advanced NB and underscores the meningeal surface as a potential pathway for this to occur.

Retinoic acid inhibits dendritic cell differentiation driven by interleukin-4.

All-trans-retinoic acid (atRA) appears to affect Th1-Th2 differentiation and its effects on immune responses might also be mediated by dendritic cell (DC). Nonetheless, studies have been showing contradictory results since was observed either induction or inhibition of DC differentiation. Our aim was to investigate atRA action on human monocyte derived DC differentiation. For this purpose we tested pharmacological and physiological doses of atRA with or without cytokines. Cell phenotypes were analyzed by flow cytometry and function was investigated by phagocytosis and respiratory burst. DC, positive control group, was differentiated with GM-CSF and IL-4 and maturated with TNF-alpha. We demonstrated that atRA effects depend on the dose used as pharmacological doses inhibited expression of all phenotypic markers tested while a physiological dose caused cell differentiation. However, atRA combined or not with cytokines did not promote DC differentiation. In fact, atRA was detrimental on IL-4 property as a DC inductor.

Cdx2, cytokeratin 20, thyroid transcription factor 1, and prostate-specific antigen expression in unusual subtypes of prostate cancer.

There are some unusual histologic variants of prostate carcinoma, including mucinous, signet-ring cells, and ductal carcinomas that can metastasize in a problematic way and simulate lung, colorectal, or bladder primaries. Currently, antibodies that are organ-specific have been used in the routine surgical pathology practice. Our aim is to study the profile of expression of Cdx2, thyroid transcription factor 1 (TTF1), and cytokeratin 20 (CK20) in prostate cancer with unusual histologic finding. Twenty-nine prostate adenocarcinomas with unusual histologic findings were submitted to immunohistochemistry with prostate-specific antigen (PSA), CK20, Cdx2, and TTF1 antibodies. There were 7 mucinous, 5 ductal, 2 signet-ring cells, and 15 usual acinar adenocarcinomas with focal mucinous differentiation. To compare the results with usual acinar adenocarcinomas, we studied 10 primary and their respective lymph node metastases in a tissue microarray, 2 unusual metastatic adenocarcinomas, and 6 usual acinar high-grade carcinomas. For tumors with special histologic finding, Cdx2 was expressed by 9 (31.0%) mucinous, signet-cell, or with focal mucinous differentiation. Thyroid transcription factor 1 was moderately positive in mucinous differentiation areas of 2 (6.9%) adenocarcinomas. Cytokeratin 20 was expressed by 9 (31.0%) tumors, among them, 3 ductal adenocarcinomas. Prostate-specific antigen was positive in 28 (96.6%) cases and negative in 1 ductal adenocarcinoma. There was only 1 worrisome ductal adenocarcinoma that was strongly CK20 positive and PSA negative. Almost one third of mucinous prostate carcinomas express Cdx2. Cytokeratin 20 can be positive also in one third of prostate carcinomas, especially the ductal type. Pathologist should be alert when evaluating immunohistochemical profiles of unusual histologic findings of prostate cancer, mostly in distant sites.

Prostate cancer detection at rebiopsy after an initial benign diagnosis: results using sextant extended prostate biopsy.

INTRODUCTION: Sextant prostate biopsy remains the standard technique for the detection of prostate cancer. It is well known that after a diagnosis of small acinar proliferation (ASAP) or high grade prostate intraepithelial neoplasia (HGPIN), the possibility of finding cancer is approximately 40% and 30%, respectively. OBJECTIVE: We aim to analyze follow-up biopsies on patients who initially received a benign diagnosis after exclusion of HGPIN and ASAP. METHODS: From July 2000 to December 2003, 1177 patients were submitted to sextant extended prostate biopsy in our hospital. The mean patient age was 65.5 years old, and the median number of fragments collected at biopsy was 13. HGPIN and ASAP were excluded from our study. We only considered patients who had a diagnosis of benign at the first biopsy and were subjected to rebiopsies up until May 2005 because of a maintained suspicion of cancer. RESULTS: Cancer was initially detected in 524 patients (44.5%), and the diagnosis was benign in 415 (35.3%). Rebiopsy was indicated for 76 of the latter patients (18.3%) because of a persistent suspicion of cancer. Eight cases of adenocarcinoma (10.5%) were detected, six (75%) at the first rebiopsy. Six patients were submitted to radical prostatectomy, and all tumors were considered clinically significant. CONCLUSION: Our data indicate that in extended prostate biopsy, the first biopsy detects more cancer, and the first, second, and third rebiopsies after an initial benign diagnosis succeed in finding cancer in 7.9% (6/55), 5.9% (1/15) and 20% (1/4) of patients, respectively.

Validation of a New Low-Cost, Methanol-Based Fixative for Cervical Cytology and Human Papillomavirus Detection.

OBJECTIVE: To test the performance of a new fixative for pap smear collection for liquid-based cervical cytology, CellPreserv® and compare it with the commercially available, PreservCyt® used in the diagnosis and detection of human papillomavirus (HPV). METHODS: Seven hundred twenty five women participated in this study after signing an informed consent. The specimens were collected using a traditional device, agitated in PBS, and equally divided in both fixatives. The slides were prepared routinely, stained by Papanicolaou, examined blindly by 2 cytologists, and reviewed by one cytopathologist. To search for HPV, 1,000 µL from each fixative was taken and processed by polymerase chain reaction. RESULTS: Considering the adequacy of samples, both fixatives had similar results - 0.33 and 0.32% of the cases unsatisfactory for PreservCyt® and CellPreserv®, respectively. Considering the 701 satisfactory cases and comparing the new fixative to the traditional fixative, there was 99.3% concordance between both. The results regarding the HPV detection was 100% concordant between the 2 fixatives. CONCLUSION: The new methanol-based fixative, CellPreserv®, is cheaper and equally efficient for treating cervical cancer screening and for HPV detection, and can be safely used by the health system prevailing in low-income countries.

Helicobacter pylori and cagA gene detected by polymerase chain reaction in gastric biopsies: correlation with histological findings, proliferation and apoptosis.

CONTEXT AND OBJECTIVE: The virulence of Helicobacter pylori (HP) in gastroduodenal disease is related to pathogenicity islands (cagPAI) present in some strains. Infection with cagPAI induces IL-8 secretion, increases epithelial cell proliferation and may be important in carcinogenesis. Our objective was to detect HP and the cagA gene (cagPAI marker) by polymerase chain reaction (PCR) and to correlate these results to histological findings, epithelial cell proliferation and apoptosis. DESIGN AND SETTING: Retrospective, at the Surgical and Molecular Pathology Laboratory, Hospital Sírio-Libanês. METHODS: DNA samples isolated from 164 gastric biopsies were used for HP detection by PCR. cagPAI+ was identified in HP+ cases by cagA gene amplification. All cases were submitted to immunohistochemistry to evaluate cell proliferation, and TUNEL to detect apoptosis. Statistical analysis was performed to compare results. RESULTS: HP was detected in 67.7% of the patients, with good correlation between HP infection and moderate to severe gastritis, gastric ulcer and MALT lymphoma. There was a correlation between cagPAI+ strains and severe gastric diseases including cancer. The risk of gastric ulcer, adenocarcinoma and MALT lymphoma was 8.8 times higher for cagPAI+ patients. cagPAI+ infection was related to higher proliferation rates. The proliferation/apoptosis index was significantly higher for cagPAI+ patients. CONCLUSION: Cell growth deregulation in cagPAI+ patients could be demonstrated by the difference in the proliferation index. We believe that this explains the carcinogenic role of Helicobacter pylori.

[High resolution chromoendoscopy in the differential diagnosis of neoplastic and non-neoplastic polyps]. / Colonoscopia de alta resolução com cromoscopia no diagnóstico diferencial dos pólipos neoplásicos e não-neoplásicos.

BACKGROUND: Magnifying colonoscopy brought the possibility of precise histologic diagnosis of colorectal lesions through their surface appearance. Despite the high accuracy of magnifying colonoscopy it is a specialized and expensive equipment not available in most medical centers. Due to these reasons the use of conventional colonoscopy with chromoscopy has been raised because this produce can reproduce most of the information previously obtained by magnifying colonoscopy. AIM: To determine the role of high resolution colonoscopy and indigo carmine chromoscopy for differential diagnosis between neoplastic and non-neoplastic colorectal lesions through measurements of accuracy, sensitivity, specificity, positive and negative predictive values. PATIENTS / METHODS: It was performed a prospective study. Seventy-four colorectal polyps were evaluated in 54 patients. A high resolution Olympus Exera CFQ 160L colonoscope was used. After the identification of the lesions, they were dyed with indigo carmine 0,2% and classified according to Kudo's classification by a single observer. After resection, the polyps were submitted to histopathological examination. RESULTS: The endoscopic findings were compared to histopathologic results. The accuracy of the method was 79,7%, sensibility of 88,8%, specificity of 55%, positive predictive value of 84,2% and a negative predictive value of 64,7%. CONCLUSION: We can conclude that we must be careful to apply high resolution colonoscopy and chromoscopy because adenomatous lesions can be misdiagnosed as non-neoplastic.

miRNA analysis of prostate cancer by quantitative real time PCR: comparison between formalin-fixed paraffin embedded and fresh-frozen tissue.

OBJECTIVE: Micro RNA (miRNA) is a class of small noncoding RNA that plays a major role in the regulation of gene expression, which has been related to cancer behavior. The possibility of analyzing miRNA from the archives of pathology laboratories is exciting, as it allows for large retrospective studies. Formalin is the most common fixative used in the surgical pathology routine, and its promotion of nucleic acid degradation is well known. Our aim is to compare miRNA profiles from formalin-fixed paraffin embedded (FFPE) tissues with fresh-frozen prostate cancer tissues. METHODS: The expression of 14 miRNAs was determined by quantitative real time polymerase chain reaction (qRT-PCR) in 5 paired fresh-frozen and FFPE tissues, which were representative of prostate carcinoma. RESULTS: There was a very good correlation of the miRNA expression of miR-let7c and miR-32 between the fresh-frozen and FFPE tissues, with Pearson's correlation coefficients of 0.927 (P = 0.023) and 0.960 (P = 0.010), respectively. For the remaining miRNAs, the correlation was good with Spearman correlation coefficient of 0.638 (P < 0.001). CONCLUSION: Analysis of miRNAs from routinely processed and stored FFPE prostate tissue is feasible for some miRNAs using qRT-PCR. Further studies should be conducted to confirm the reliability of using stock tissues for miRNA expression determination.

Establishment and characterization of androgen-independent human prostate cancer cell lines, PcBra1, PcBra2, and PcBra3.

One of the main obstacles for understanding biological events involved in cancer is the lack of experimental models for in vitro studies especially for prostate cancer (PC). There are a limited number of PC cell lines being the majority originated from metastatic tumors mostly acquired from American Tissue Cell Culture which demands importation an expensive and bureaucratic process. Also it is well known that there are ethnic differences between populations concerning the behavior of tumors and the research based on cell lines derived from Brazilians should be interesting. Our aim was to develop tumor cell lines from primary PC.

Establishment and characterization of human bladder cancer cell lines BexBra1, BexBra2, and BexBra4.

Bladder cancer (BC) is the fourth most common cancer in the USA. In Brazil, BC represents 3% of the total existing carcinomas in the population and represents the second highest incidence among urological tumors. The majority of bladder cancer cell lines available were derived from Caucasians and established in the seventies or eighties. Thus, neoplasia development in these cells likely occurred in environment conditions vastly different than today. In the present study, we report the establishment and characterization of three Brazilian bladder cancer cell lines (BexBra1, BexBra2, and BexBra4). These cell lines may be helpful for dissecting the genetic and epigenetic aspects that trigger the progression of BC. Moreover, the development of a Brazilian representative of the disease will allow us to investigate the potential inter-racial differences of malignancy-associated phenotypes in bladder cancer.

Benign glandular inclusion in obturator lymph node of a man treated for prostate carcinoma.

Benign glandular inclusions in lymph nodes are extremely rare in men. Their identification is essential because it changes dramatically the prognosis and therapy of neoplasms. Described herein is the first case of benign glandular inclusion in an obturator lymph node dissected during a radical prostatectomy for treatment of prostate adenocarcinoma. A 60-year-old man underwent radical prostatectomy and obturator-hypogastric lymph node dissection for treatment of prostate adenocarcinoma. Benign glandular inclusion was found in microscopic examination. The lesion was characterized by two glandular spaces lined by a single, cuboid, benign epithelium localized in the sinus of one of four dissected lymph nodes. Immunohistochemistry showed mesothelial differentiation. Pathologists should be aware of benign glandular inclusion in obturator lymph nodes dissected during a radical prostatectomy for treatment of prostate cancer in order to avoid the incorrect diagnosis of metastatic disease.

Paratesticular rhabdomyoma.

The fifth case of paratesticular rhabdomyoma is described in a 55-year-old man. The histology revealed a tumor consisting of round or spindle cells, with deeply acidophilic cytoplasm and cross striations. An inguinal resection was performed, and the patient is well with no local recurrence or metastasis 13 months after surgery.

Prostate cancer detection at rebiopsy after an initial benign diagnosis: results using sextant extended prostate biopsy

INTRODUCTION: Sextant prostate biopsy remains the standard technique for the detection of prostate cancer. It is well known that after a diagnosis of small acinar proliferation (ASAP) or high grade prostate intraepithelial neoplasia (HGPIN), the possibility of finding cancer is approximately 40 percent and 30 percent, respectively. OBJECTIVE: We aim to analyze follow-up biopsies on patients who initially received a benign diagnosis after exclusion of HGPIN and ASAP. METHODS: From July 2000 to December 2003, 1177 patients were submitted to sextant extended prostate biopsy in our hospital. The mean patient age was 65.5 years old, and the median number of fragments collected at biopsy was 13. HGPIN and ASAP were excluded from our study. We only considered patients who had a diagnosis of benign at the first biopsy and were subjected to rebiopsies up until May 2005 because of a maintained suspicion of cancer. RESULTS: Cancer was initially detected in 524 patients (44.5 percent), and the diagnosis was benign in 415 (35.3 percent). Rebiopsy was indicated for 76 of the latter patients (18.3 percent) because of a persistent suspicion of cancer. Eight cases of adenocarcinoma (10.5 percent) were detected, six (75 percent) at the first rebiopsy. Six patients were submitted to radical prostatectomy, and all tumors were considered clinically significant. CONCLUSION: Our data indicate that in extended prostate biopsy, the first biopsy detects more cancer, and the first, second, and third rebiopsies after an initial benign diagnosis succeed in finding cancer in 7.9 percent (6/55), 5.9 percent (1/15) and 20 percent (1/4) of patients, respectively.

Helicobacter pylori and cagA gene detected by polymerase chain reaction in gastric biopsies: correlation with histological findings, proliferation and apoptosis

CONTEXTO E OBJETIVO: A virulência de Helicobacter pylori em doenças gastroduodenais está relacionada à presença de ilha de patogenicidade (cagPAI) que ocorre em algumas cepas. A infecção pelo cagPAI induz a secreção de IL-8, aumenta a proliferação epitelial, podendo ter um papel importante na carcinogênese. Nosso objetivo foi detectar HP e o gene cagA (marcador de cagPAI) pela técnica de PCR (polymerase chain reaction), correlacionando com os achados histológicos, de proliferação e apoptose. TIPO DE ESTUDO E LOCAL: Estudo retrospectivo, no Laboratório de Patologia Cirúrgica e Molecular do Hospital Sírio Libanês. MÉTODOS: DNA isolado de 164 biópsias gástricas foi submetido a PCR para detecção de HP. Os casos positivos foram submetidos a nova reação para identificação do gene cagA. Pela técnica de imunohistoquímica foi analisada a proliferação celular e, pela TUNEL, a apoptose. RESULTADOS: HP foi detectado em 67,7% dos pacientes. Houve correlação entre a presença do HP e o diagnóstico de gastrite moderada ou grave, úlcera e linfoma do tipo MALT. Houve correlação entre cagPAI+ e a doença gástrica grave, incluindo o câncer. O risco de úlcera, adenocarcinoma ou linfoma MALT para os portadores de cagA+ foi de 8,8. Infecção pelo cagPAI correlacionou-se com aumento na taxa de proliferação. O índice proliferação/apoptose foi significantemente maior para os pacientes cagPAI+. CONCLUSÕES: Uma desregulação do crescimento celular nos pacientes cagPAI+ foi demonstrada pela diferença do índice de proliferação, que acreditamos pode explicar o papel carcinogênico da bactéria.

Colonoscopia de alta resoluçäo com cromoscopia no diagnóstico diferencial dos pólipos neoplásicos e näo-neoplásicos / High resolution chromoendoscopy in the differential diagnosis of neoplastic and non-neoplastic polyps

RACIONAL: Com o advento da colonoscopia com magnificaçäo de imagem, tem se mostrado possível a identificaçäo da natureza histopatológica das lesöes colorretais através de suas características da forma que as glândulas se abrem na superfície da mucosa (PITS). Apesar de ser método com alto índice de acurácia para o diagnóstico diferencial entre lesöes neoplásicas e näo-neoplásicas, trata-se de equipamento especializado, de custos elevados, näo disponível na grande maioria dos centros médicos. Diante dessa realidade, a colonoscopia convencional com cromoscopia vem adquirindo espaço, pois acredita-se que pode reproduzir grande parte das informações obtidas até entäo pela colonoscopia com magnificaçäo de imagem. OBJETIVO: Determinar o papel da colonoscopia de alta resoluçäo com cromoscopia no diagnóstico diferencial entre pólipos neoplásicos e näo-neoplásicos através dos seguintes índices estatísticos: acurácia, sensibilidade, especificidade, valores preditivos positivo e negativo. PACIENTES E MÉTODO: Realizou-se estudo prospectivo, onde foram avaliadas 74 lesöes polipóides colorretais em 54 pacientes. Foi utilizado o colonoscópio Olympus Exera CFQ 160L de alta resoluçäo. Após identificadas, as lesöes foram coradas com índigo carmim 0,2 por cento e analisadas conforme classificaçäo descrita por Kudo, por um único examinador. Após ressecados, os pólipos foram encaminhados para estudo anatomopatológico. RESULTADOS: Os achados endoscópicos foram comparados com os resultados histopatológicos. A acurácia do método foi de 79,7 por cento, sensibilidade de 88,8 por cento, especificidade de 55 por cento, valor preditivo positivo de 84,2 por cento e valor preditivo negativo de 64,7 por cento. CONCLUSÄO: Pode-se concluir que se deve ter cautela na aplicaçäo clínica da colonoscopia de alta resoluçäo com cromoscopia, pois lesöes adenomatosas podem ser interpretadas como näo-neoplásicas