Agreement and internal quality assurance of the Neubauer hemocytometer and Makler chamber for human sperm concentration determination.

OBJECTIVE: The Neubauer hemocytometer, as well as the Makler chamber, are devices commonly used in andrology laboratories. The present study aimed to verify if both methods yield comparable results, and whether they can be used interchangeably to determine sperm concentration. METHODS: Sperm and latex beads concentration measurements were performed with the Neubauer hemocytometer and the Makler chamber. Fixed and proportional biases were estimated, and the method agreement was determined by assessing sperm concentration results with the Bland and Altman plot. The Coefficient of Variation (CV) and relative bias were calculated as an index of precision and accuracy, respectively, by measuring latex beads target concentrations in both chambers. RESULTS: The Makler chamber systematically overestimated the Neubauer hemocytometer concentration measurements by a mean of -7.99%, with limits of agreement (LOA) between -41% to 25.61% (p<0.001). The fixed bias was found for concentration values inferior to 40 x 106/ml range (p<0.001), but not higher concentration results (p>0.05). Measurements with the Neubauer hemocytometer showed the greatest consistency in the study with the CV ranging from 3.01% to 6.67%; while the CV with the Makler chamber ranged from 8.46% to 25.64%. The relative bias for the Neubauer hemocytometer determinations varied from 0.12% to 8.40%, while for the Makler chamber varied from 7.6% to an overestimation of 38.0%. CONCLUSIONS: Measurements made with the Makler chamber demonstrated more variability and a higher degree of overestimation. The Makler chamber is a poor substitute to the Neubauer hemocytometer for evaluation of oligozoospermic samples, although both chambers render similar results for highly concentrated samples.

DNA-ICM as an adjuvant method applied on oral cytological specimens.

OBJECTIVE: This study aimed to evaluate cytology diagnosis accuracy using adjuvant methods in clinical routine for oral cancer. STUDY DESIGN: This prospective study was conducted on 98 patients with clinically potentially malignant or malignant oral cavity lesions. One oral lesion smear was taken from each patient using a cytobrush before biopsy and stored at PreservCyt Thinprep. Samples were cytologically analyzed, and DNA ploidy measurement was performed on the same slide. The diagnostic methods' accuracy was then calculated. RESULTS: In clinical inspection, 61 patients had suspicious lesions for malignancy, whereas 37 had potentially malignant disorders. Cytology associated with DNA image cytometry presented a sensitivity of 81.2% and specificity of 90.9%. When analyzing lesions located in high-risk sites to oral malignancies individually, cytology associated with DNA image cytometry presented a sensitivity of 88.2%, specificity of 100.0%, accuracy of 90.0%, and Kappa value of 0.77 (CI 95%: 0.48-1.00). CONCLUSIONS: Association between cytology and DNA image cytometry is an objective and non-invasive diagnostic method that demonstrated high sensitivity and specificity in diagnosing malignant epithelial squamous cell transformation in the oral cavity.

DNA image cytometry parameters to identify high-grade cervical lesions.

OBJECTIVE: Evaluate the performance of different DNA image cytometry (DNA-ICM) ploidy parameters in the categorisation of DNA-ICM results and identification of high-grade cervical intraepithelial neoplasia or worse (≥ CIN2). METHODS: Cervical samples from 232 women were collected for DNA-ICM analysis and biopsy confirmation. Five DNA parameters were used to define DNA aneuploidy: number of cells with exceeding events (EE) over 2.5cEE, 4cEE, 5cEE and 9cEE, and aneuploid stemlines. DNA-ICM results were categorised as normal, suspicious, and abnormal. RESULTS: For individual DNA ploidy parameters, sensitivity values for 50 cells with 2.5cEE, 45 cells with 4cEE, 1 cell with 9cEE and aneuploid stemline were 72.95%. 54.1%, 69.67% and 54.1%, while specificity values were 80.0%, 90.0%, 89.09% and 95.45%, respectively. For the 5cEE parameter, the sensitivity values for 1, 2, 3, 4 and 5 cells were 93.44%, 85.25%, 81.97%, 77.87% and 75.41%, while specificity values were 46.36%, 63.64%, 74.55%, 76.36% and 80.91%, respectively. For categorised DNA-ICM results, a suspicious result showed superior sensitivity than an abnormal result (87.70% vs 82.79%, P = 0.031), but lower specificity (54.55% vs 75.45%, P < 0.001). Both types of DNA-ICM result were statistically significantly different from a normal result (P < 0.05). CONCLUSION: For prognostic purposes, 1 cell with 9cEE, 45 cells with 4cEE and aneuploid stemline are the best parameters with which to categorise an abnormal DNA-ICM result, followed by 50 cells with 2.5cEE and 4 cells with 5cEE. For screening purposes, 10 cells with 2.5cEE, 10 cells with 4cEE, and 2 cells with 5cEE are suitable parameters with which to categorise a suspicious DNA-ICM result.

DNA ploidy measurement and human papillomavirus in abnormal cervical cytology.

OBJECTIVE: To compare the efficacy of high-risk human papillomavirus (HR-HPV) and DNA image cytometry (DNA-ICM) status for identifying high-grade cervical intraepithelial neoplasia or worse (≥CIN2). METHODS: This cross-sectional study was performed in women undergoing follow-up procedure after a previous abnormal cervical cytology. Cervical cells were collected for HPV detection and DNA ploidy measurement. Biopsy samples were taken for histological confirmation. Sensitivity and specificity values for ≥CIN2 detection with HR-HPV and DNA-ICM were determined. RESULTS: HR-HPV was present in 74.5% of the women. The most frequent HPV infection was HPV 16, followed by HPV 31, 33 and 58. Aneuploidy was observed in 60.6% of all cases. Referral cytology revealed 78.0% sensitivity and 68.6% specificity for detecting a ≥CIN2 lesion. The HR-HPV test alone showed 92.7% sensitivity, albeit it was not statistically different from DNA-ICM (88.1%, P > .05). Positivity for HPV or DNA-ICM resulted in 100% sensitivity. Higher specificity was observed for the combination of HR-HPV and DNA-ICM (88.6%), with no difference from DNA-ICM alone (85.7%, P > .05). CONCLUSION: DNA-ICM or HR-HPV positivity identified all cases of ≥CIN2 in women undergoing follow-up procedure after a previous abnormal cervical cytology. Routine cervical cancer screening could be improved by the incorporation of DNA-ICM as a complementary method to primary screening to identify which women need closer follow-up.

Detecção de HPV utilizando iniciadores MY09/MY11 e GP5+/GP6+ em pacientes com alterações citológicas e/ou colposcópicas / HPV detection using primers MY09/MY11 and GP5+/GP6+ in patients with cytologic and/or colposcopic changes

Introduction: Cervical cancer is one of the most common diseases among women, and cause considerable morbidity and mortality. Considering that cervical cancer is an important neoplasia in northeastern Brazil, and the prevalence of high-risk human papillomavirus (HPV) is directly associated with it, this work had aimed to correlate the cytological and/or colposcopic findings with HPV infection status, and verify the performance of MY09/MY11 and GP5+/6+ primers for HPV detection. Material and method: Patients in this study were from Penedo-AL, a city with high level of poverty (poverty rate of 60.62%). Out of 70 patients with cytological and/or colposcopic changes, 32 agreed to participate in the study. Results: Regarding cytology, 21 (30%) patients presented atypical squamous cells of undetermined significance (ASC-US); 20 (29%), atypical glandular cells of undetermined significance (AGUS); 12 (17%), low-grade intraepithelial lesion (LSIL); five (7%), high-grade intraepithelial lesion (HSIL); and 12 (17%), positive colposcopy. From these, 27 (84%) presented the band gene encoding for human ß-globin. From the 27 patients, eight (30%) were positive for HPV. The results showed that the deoxyribonucleic acid (DNA) of HPV was detected in 15% and 30% by using MY-PCR and GP +-PCR, respectively. Conclusion: This study suggests that more than one type of oligonucleotide primer should be used in clinical samples to increase sensitivity for the detection of HPV...

Introdução: O câncer cervical é uma das doenças mais frequentes entre mulheres, e causa considerável morbidade e mortalidade. Com base nos fatos de que o câncer cervical é uma neoplasia importante no nordeste brasileiro, e que a prevalência do papilomavírus humano (HPV) de alto risco está diretamente associado a ele, este trabalho teve como objetivos correlacionar os achados citológicos e/ou colposcópicos com status de infecção de HPV e verificar o desempenho dos iniciadores MY09/MY11 e GP5+/GP6+ para detecção do HPV. Material e métodos: Os pacientes deste estudo foram de Penedo-AL, uma cidade com elevado nível de pobreza (índice de pobreza de 60,62%). Do total de 70 pacientes com alterações citológicas e/ou colposcópicas, 32 aceitaram participar do estudo. Resultados: Com relação a citologia, 21 (30%) pacientes apresentaram células escamosas atípicas de significado indeterminado (ASC-US); 20 (29%), células glandulares atípicas de significado indeterminado (AGUS); 12 (17%), lesão intraepitelial de baixo grau (LSIL); cinco (7%), lesão intraepitelial de alto grau (HSIL); e 12 (17%), colposcopia positiva. Destas, 27 (84%) apresentaram banda do gene que codifica para ß-globina humana. Das 27 pacientes, oito (30%) apresentaram positividade para o HPV. Os resultados mostraram que o ácido desoxirribonucleico (DNA) do HPV foi detectado em 15% e 30%, usando MY-PCR e GP +-PCR, respectivamente. Conclusão: Este estudo sugere que mais de um tipo de iniciador de oligonucleotídeo deve ser utilizado em amostras clínicas para aumentar a sensibilidade na detecção do HPV...

Evaluation of adherence to treatment by patients seen in a psychosocial care center in northeastern Brazil

In the treatment of mental disorders, nonadherence to medication, the main cause of psychiatric morbidity, is observed in about 50 percent of the cases and is responsible for numerous losses. This study evaluated adherence to drug treatment by patients seen in a Psychosocial Care Center (CAPS) in northeastern Brazil. Adherence to treatment was evaluated using the Haynes-Sackett and Morisky-Green-Levine tests. All patients registered in the CAPS were included in the study (n= 101). Only 11.88 percent of the patients adhered to drug treatment. The main reasons not to use medication were: oblivion (68.83 percent), feeling unwell after taking the medication (54.22 percent), not having money to buy the medication (43.83 percent), not finding the medication in the public health service (39.94 percent) and fear of harm that might be caused by the drug (28.90 percent). Furthermore, 85.1 percent of the patients did not know their diseases, 88.1 percent did not know their treatment, 86.4 percent did not feel good when they took their medication, and 88.1 percent took their medication incorrectly. The results revealed that the lack of information about diseases and drugs used, the nuisance posed by drug therapy and the low access to medications reduce adherence to treatment and, consequently, treatment effectiveness.

No tratamento de desordens mentais, a não-adesão ao tratamento ocorre em cerca de 50 por cento dos casos e é responsável por inúmeros prejuízos, além de ser a principal causa de morbidade psiquiátrica. O presente estudo objetivou avaliar a adesão ao tratamento medicamentoso de pacientes atendidos em um Centro de Atenção Psicossocial (CAPS) no Nordeste do Brasil. A avaliação da adesão ao tratamento foi feita através dos testes de Haynes-Sackett e Morisky-Green-Levine. Todos os pacientes cadastrados no CAPS foram incluídos no estudo (n = 101). Observou-se que apenas 11,88 por cento dos pacientes aderiram ao tratamento medicamentoso. As principais razões para o não uso dos medicamentos foram: esquecimento (68,83 por cento), sentir-se mal após a ingestão de medicamentos (54,22 por cento), não ter dinheiro para comprar medicamentos (43,83 por cento), não encontrar os medicamentos no serviço público de saúde (39,94 por cento) e medo dos danos causados pelos medicamentos (28,90 por cento). Além disso, observou-se que 85,1 por cento dos pacientes não conheciam suas doenças, 88,1 por cento não conheciam seus tratamentos, 86,4 por cento não se sentiam bem quando usavam medicamentos e 88,1 por cento usavam os medicamentos incorretamente. Os resultados demonstram que a falta de informação sobre doenças e medicamentos, os danos decorrentes da terapia medicamentosa e o baixo acesso aos medicamentos comprometem a adesão ao tratamento e, consequentemente, a eficácia do tratamento.

9p21 Deletion in the diagnosis of malignant mesothelioma in serous effusions additional to immunocytochemistry, DNA-ICM, and AgNOR analysis.

BACKGROUND: The diagnosis of malignant mesothelioma (MM) in serous effusions is difficult but may be achieved by the application of adjuvant methods. METHODS: The authors cytologically diagnosed 33 effusions as suspicious or positive for MM cells by using DNA-image cytometry (DNA-ICM), immunocytochemistry and AgNOR analysis. The authors further detected 9p21 deletions by chromosomal fluorescence in situ hybridization (FISH). In addition, 31 cases of metastatic carcinomas and 39 of tumor cell-negative effusions were investigated. All diagnoses were confirmed by histologic and/or clinical follow-up. RESULTS: DNA aneuploidy was found in 71% of MMs, 100% of metastatic carcinomas, and in none of the negative effusions. Calretinin was positive in 100% of MMs, in none of the metastatic carcinomas, and in 94.9% of negative effusions. BerEP4 showed positivity in 15.6% of MMs, 87.1% of metastatic carcinomas, and in none of the negative effusions. With AgNOR analysis, 89.3% of MMs and 96.7% of metastatic carcinomas showed >or=2.5 AgNOR dots as satellites and >or=4.5 as total AgNOR counts. 9p21 deletions were demonstrated in 90.9% of MM cases, 45.2% of metastatic carcinomas, and in none of the negative effusions. By cytology alone, 81.8% of MMs were identified unequivocally. Addition of DNA-ICM improved the prevalence of tumor cell detection to 87.9% and of AgNOR analysis to 97%. The introduction of 9p21 deletions by FISH improved this prevalence to 100%. CONCLUSIONS: Because of these results, the authors propose the sequential application of immunocytochemistry, DNA-ICM, and AgNOR analysis to establish a cytological diagnosis of malignant mesothelioma in serous effusions. In persistent doubtful diagnoses, the authors recommend fluorescence in situ hybridization to analyze the 9p21 deletion.