Anemia mielotísica por osteopetrose em um cão: relato de caso / Mielotisic anemia caused by osteopetrosis in a dog: case report

A osteopetrose é uma doença rara, caracterizada pelo aumento generalizado da densidade óssea. Tem como característica principal, a reabsorção osteoclástica defeituosa, resultando no acúmulo de massa óssea. Além disso, pode ocorrer retardo do crescimento, desnutrição progressiva, anemia e caquexia. O presente relato descreve o caso de uma cadela, com aproximadamente nove meses de idade, sem raça definida, com histórico de apatia e disorexia. Hemogramas seriados demonstraram pancitopenia persistente. Vários exames laboratoriais foram realizados para excluir doenças como erliquiose, leishmaniose e cinomose, porém todos foram negativos. O mielograma constatou hipocelularidade relativa por provável aplasia/hipoplasia medular. Exames radiográficos evidenciaram o aumento da radiopacidade óssea e hipertrofia da região cortical, e a necropsia confirmou a redução do canal medular. O exame histopatológico confirmou a osteopetrose. Conclui-se que a osteopetrose pode causar comprometimento na produção de células sanguíneas, sendo uma causa rara de anemia mielotísica, além de alterações neurológicas secundárias à má-formação dos ossos do crânio.

Osteopetrosis is a rare disease characterized by generalized increase in bone density. The defective osteoclastic resorption results in the accumulation of bone mass. Furthermore, there may be growth delay, progressive malnutrition, anemia and cachexy. This report describes the case of a nine month old, mixed breed, female dog presented with apathy and disorexia. Laboratory tests were performed to rule out Ehrlichiosis, Leishmaniasis, and Canine distemper, but all were negative. Hemogram showed persistent pancytopenia and myelogram showed relative hypocellularity, probably due to spinal cord aplasia/hypoplasia. Radiographs showed increased bone opacity and hypertrophy of the cortical region, and the necropsy confirmed the reduction of the medullary canal. Histopathological examination confirmed osteopetrosis. In conclusion, osteopetrosis can compromise blood cell production, and is a rare cause of mielotisic anemia. It can also cause neurological deficits due to cranium bone deformation.

Relevance of the rates of PCNA, Ki-67 and p53 expression according to the epithelial compartment in cervical lesions.

In order to assess further biological evidence for similarities among the "diagnostic classes" of cervical lesions, which are now a matter of international discussion in the search for a uniform classification, the purpose of this study was to characterize the immunoexpression of cell proliferation markers (proliferating cell nuclear antigen, PCNA and Ki-67) and protein p53. Each marker was individually quantified in basal, intermediate, and superficial epithelial compartments presenting chronic cervicitis (CC) accompanied by the cytopathic effects of infection by human papillomavirus (CCHPV) or not (CC), as well as in cervical intraepithelial neoplasia (CIN) grades I, II, and III. A total of 100 patients were evaluated and the positive nuclei were counted separately, including all extensions of the available epithelium. The percentage of PCNA- and Ki-67-positive cells increased with increasing grade of the cervical lesions, although PCNA immunoreactivity was always greater than the immunoreactivity observed with Ki-67 antigen. The immunoexpression of p53 protein was found to be weak, with no remarkable behavior in any specific "diagnostic class". The differences in cell proliferation markers found herein further emphasize the progressive loss of epithelial layer organization in the course of the development of preneoplastic changes in cervical squamous epithelium. Furthermore, difficulties in morphologically distinguishing "borderline lesions" persist when cell cycle markers are studied, further supporting the suggestion to consider the lesions of CCHPV and CIN I together as only one diagnostic class. Conversely, the different immune profile found between CIN II and III further supports the validity of the subdivision of CIN into three groups.

Adenocarcinoma in females detected in serous effusions. Cytomorphologic aspects and immunocytochemical reactivity to cytokeratins 7 and 20.

OBJECTIVE: To investigate the usefulness of assessing the immunoreactivity of cytokeratins 7 (CK7) and 20 (CK20) as well as several cytomorphologic parameters in effusions with metastatic adenocarcinomas in the search for the primary site of the tumor. STUDY DESIGN: From the files of the Pathology Department, A. C. Camargo Hospital, we studied cytologic smears from 73 metastatic adenocarcinomas originally from the breast, 63 from the ovary, 40 from the lung and 32 from the stomach, looking for morphologic parameters that could have discriminant potential in suggesting the primary site in a routine situation, including intranuclear inclusions, prominent nucleoli, mitosis, signet-ring cells, psammoma bodies, nuclear crease, binucleation and multinucleation, papillary features, acinar profile (including ball cells) and single cells. Immunoreactions were performed with monoclonal antibodies to CK7 (OV-TL 12/30) and CK20 (Ks 20.8) and included morphologic analysis. Both analyses were studied in a blind fashion regarding the primary site of the tumors. RESULTS: Positivity ratios for breast, ovary, stomach and lung cases were 67.6%, 63.5%, 29.7% and 45.5%, respectively, for CK7 and 17.2%, 15.8%, 13.5% and 32.2%, respectively, for CK20. Discriminant analysis of morphologic and immunocytochemical parameters had an error rate of 42.9% in recognizing the primary site and a Wilk's lambda of .7290. CONCLUSION: The more efficient parameter with discriminant function was the papillary appearance showed by CK7, which should be used in further studies with a similar scope. The set of parameters used in this study were insufficient to discriminate the primary site of female adenocarcinomas in effusions with significant accuracy.

Cytomorphologic evidence of human papillomavirus infection in smears from the irradiated uterine cervix.

OBJECTIVE: To report the cytomorphologic evidence of human papillomavirus (HPV) infection in cervical smears from women treated with radiotherapy for carcinoma of the uterine cervix. STUDY DESIGN: From January 1986 to December 1993, 32 cervical cytologic smears (CCSs) from women who had received radiotherapy (Rtx) for squamous cell cervical carcinoma at A.C. Camargo Cancer Hospital were selected for study due to the presence of signs of HPV infection. Review of the files of these patients showed 22 additional samples, which we included in this study. The avidinbiotin-immunoperoxidase technique was performed on previously stained slides using polyclonal rabbit antibovine papillomavirus type 1 to confirm the HPV infection. RESULTS: Positive reactions were found in 18 cases (27 samples), mainly in cells with classic koilocytotic features, but also in epithelial cells that lacked cytopathic effects. Only one case, a biopsy from a verrucous lesion observed after Rtx, was found to be positive for HPV 6/11 DNA by in situ hybridization. CONCLUSION: Cytologic and immunohistochemical positivity for HPV, as well as the reaction with a HPV 6/11 DNA probe, strongly suggested that HPV infection was acquired or expressed after radiotherapy. Further studies are needed to assess the mechanisms responsible for such an association.

Cytological diagnosis of serous effusions in a cancer hospital in Brazil.

The performance of cytological diagnosis in serous effusions was evaluated through a historical large series study which was carried out at Department of Surgical Pathology of Hospital A.C. Camargo. Files from the period of 1966 to 1990 were reviewed. Out of 4297 serous effusions samples (from 3379 patients), 2520 were pleural, 1763 ascitic and 14 pericardiac, with cytological report of malignancy in 917, 688 and 4 cases, respectively. Cytological diagnoses were confirmed true or false after correlation with the final diagnosis of the patients (clinical follow-up and/or histological evaluation). The results observed were 1982 false-negative, 21 false-positive, 1588 true-positive and 468 true-negative cases. Suspicious (161 or 3.75%) and inconclusive (77 or 1.79%) cases were not considered for statistical evaluation. Sensitivity, specificity, efficiency, as well as positive and negative predictive values were 44.5%, 95.7%, 50.1%, 98.7% and 20%, respectively. These values in pleural and ascitic effusions, separately showed similar performance, which was not observed in pericardiac samples due to small casistics. These data show low sensitivity and negative predictive values which contrast with the high specificity and positive predictive values that can be partially explained by the methodological limitations and invasive features of neoplasias.

Cytologic diagnosis of melanoma in serous effusions. A morphologic and immunocytochemical study.

The diversity of melanoma patterns greatly impairs the interpretation of malignant cells in effusion samples. The presence of melanin pigments greatly helps determine the histogenetic origin of the tumor, but unfortunately many cases do not exhibit this feature. We reviewed cases with a definitive diagnosis of melanoma in order to identify some useful characteristics of the morphologic examination of effusions. We also subjected the effusions to the HMB45 immunoreaction to determine the diagnostic usefulness of this monoclonal antibody. The study was performed on 21 effusion samples containing malignant cells, and the main cytologic findings were similar to those on other neoplasms except for the presence of melanin pigment. The HMB45 immunoreaction was very sensitive, confirming the diagnosis in 14 of 18 cases (77.8%). Melanin pigments seem to be useful markers for melanoma in effusions, and HMB45 can be used as an ancillary method in the differential diagnosis.

Washed-biopsy cytology of abdominal lesions.

The accuracy of an alternative method of preparing cytologic smears from material collected under laparoscopic examination was studied. "Washed-biopsy" cytology was performed by saline solution lavage of the fragments using a cytocentrifuge. Rich and well-preserved cellularity was obtained. Among 51 cases, cytology revealed 12 benign, 3 suspicious and 36 malignant cases, including 9 true negatives, 3 false negatives and 36 true positives. Suspicious cases were excluded from the analysis. The sensitivity, specificity, positive and negative predictive values, and efficiency, when comparing the cytologic to the definitive diagnoses, were, respectively, 92.3%, 100.0%, 100.0%, 75.0% and 93.7%. We recommend the routine use of the washed-biopsy method in laparoscopic examinations to improve the accuracy of cytologic investigations.

The role of fine needle aspiration cytology in the diagnosis of lesions of the head and neck excluding the thyroid and salivary glands.

Fine-needle aspiration cytology (FNAC) was performed in 273 cases with suspicious masses of the head and neck, excluding the thyroid and salivary gland, at the A.C. Camargo Hospital from 1983 to 1989. The most frequently punctured site was the lymph nodes (n = 178), and the most common histologic diagnosis of malignancy was squamous cell carcinoma (n = 43). FNAC results were: 105 positive, 6 suspicious, 135 negative, and 27 unsatisfactory cases. There was a correlation between cytology and histology in 217 cases, with 102 true-positive, 76 true-negative, 1 false-positive, and 7 false-negative cases. Suspicious and unsatisfactory cases were not considered for this index. Analysis of the results showed an efficiency of 95.6%, sensitivity of 93.5%, and specificity of 98.7%. Positive and negative predictive values were respectively 99.0% and 91.5%.

[Comparison of the methods of cytobrush and Ayre spatula in the concentration of endocervical cells]. / Comparação dos métodos de "cytobrush" e espátula de Ayre na concentração de células endocervicais.

A comparison between two sampling methods of cervicovaginal material--cytobrush and Ayre's spatula--was performed to verify the endocervical cell concentration in a group of 163 asymptomatic women. Age ranged from 18 to 57 years, with an average of 30. Smears of both methods showed satisfactory cellularity for study. Cytobrush showed endocervical cells (columnar and/or metaplastic) in 100 cases (61.3%) and Ayre's spatula in 39 (20.2%). Cytobrush showed more efficient results in terms of endocervical cell concentration; according to literature, this can be an important factor in the sensitivity of the cytologic method to detect intraepithelial neoplasia.

Cytology of pulmonary samples after cancer radiation therapy.

Radiation-induced cytomorphologic atypia is presented. The cellular changes produced by radiation therapy were studied in cytologic samples from eight patients with bronchogenic carcinoma. Pulmonary specimens were obtained from sputum, and washing and brushing examination in patients before and after radiation. In all 48 samples with 213 slides were reviewed by light microscopy. An increased number of metaplastic cells and macrophages, as well as a high leukocyte concentration in post-radiation samples were observed in the majority of the patients. Nuclear and cytoplasmic vacuolization were the most frequent damage observed in the post-radiation group. The alterations of epithelial cells, which are often found in the cervix, were rarely observed in the pulmonary specimens.

Fine needle aspiration cytology in the diagnosis of non-neoplastic diseases of head and neck masses.

In this study, 3 patients with head and neck masses, which were suspicious of neoplastic compromise, were submitted to FNAC investigation. Two cases showed specific infection by Paracoccidioidomycosis and one by Actinomycosis. The cytologic findings were in agreement with histopathologic diagnosis and clinical follow-up; that results showes that FNAC importance in the diagnosis of non-neoplastic masses could be explored in ambulatorial routine.

Detection of Chlamydia trachomatis cervical infection: a comparison of Papanicolaou and immunofluorescent staining in smears obtained by Ayre's spatula and cytobrush.

Two methods of staining (Papanicolaou versus direct immunofluorescence) and two methods of collection of the samples (Ayre's wooden spatula versus cervical Cytobrush) were compared in order to verify the efficiency in detecting Chlamydia trachomatis (CT) infections in the female genital tract. Out of 166 asymptomatic patients, 59 were positive for CT by means of direct immunofluorescence: 36 were detected in Cytobrush samples, 16 in Ayre's spatula samples, and 7 in the samples collected by both methods. Papanicolaou smears showed "moth-eaten" features suggestive of CT infection in a great number of metaplastic cells present in 35 cases: 24 collected by Cytobrush, 4 by Ayre's spatula and 7 by both methods. Our data show that Cytobrush is more efficient that Ayre's spatula in concentrating cellular material. It is thus possible to detect CT infection with more accuracy by means of direct immunofluorescence, and to suspect CT infection in smears collected by means of Cytobrush and stained by Papanicolaou's method.

Herpes simplex virus in postradiation cervical smears. A morphologic and immunocytochemical study.

From January 1987 to August 1988, cytomorphologic criteria of both herpes simplex virus (HSV) and radiation effects were observed in Papanicolaou smears from 3 of 1,340 patients who had received radiotherapy for squamous cell carcinoma of the cervix. Avidin-biotin immunoperoxidase staining, using a rabbit IgG polyclonal HSV antibody, confirmed the presence of HSV antigen in those three postradiation smears. Both multinucleated molded cells and epithelial cells that lacked cytopathic effects were positive for HSV. Three other postradiation smears from these cases were similarly positive for HSV antigen; the one preradiation smear was negative. In situ hybridization and immunoperoxidase studies on sections from the preradiation biopsies were negative: severely altered neoplastic cells showed no reactivity. The absence of HSV markers in the preradiation specimens suggests that the HSV infections were secondary to the radiotherapy; further studies are needed to prove this association and to assess the possible mechanisms. These cases clearly indicate that the overlapping features of radiation and viral effects (such as multinucleation) may be present simultaneously.

[Fine needle aspiration cytology in the diagnosis of head and neck tumors]. / Citologia de punção aspirativa por agulha fina no diagnóstico de tumores de cabeça e pescoço.

The authors studied 166 head and neck tumors of patients admitted to the A.C. Camargo Hospital using the method of fine needle aspiration cytology (FNAC) in order to correlate these findings with pathohistological diagnoses. The correlation was possible in 81 cases: 3 cases were false negative and 1 was false positive. FNAC sensibility was 90.6%, specificity was 97.9%, and efficiency was 95%.

Chlamydia trachomatis in cervical uterine irradiated cancer patients.

Twenty-two women with uterine carcinoma with morphological alterations postradiation treatment suggestive of Chlamydia trachomatis infection were analyzed by direct immunofluorescence. The diagnosis was confirmed in 12 (54.4%) cases. Discussions of the presence of Chlamydia in irradiated cells were not found in the literature. This finding may signify a posttherapeutic morbidity or a preradiation infection.

[Chlamydia trachomatis in smears of irradiated uterine cervix: cytologic and immunocytochemical aspects]. / "Chlamydia trachomatis" em esfregaços de colo uterino irradiado: aspectos citológicos e imunocitoquímicos.

Some authors recently suggested the role of Chlamydia trachomatis as a potentiating agent of female genital tract neoplasias. Cervicovaginal smears of women with uterine carcinoma treated with radiotherapy are analyzed in the present work. A series of such smears showed morphological changes suggestive of Chlamydia infection with the Papanicolaou stain. Confirmation of the presence of such agent was done using an avidin-biotin-peroxidase technique. A higher prevalence of Chlamydia in the group of patients with carcinoma of the uterus treated with irradiation than in the population without ionizing treatment was verified. The presence of cells with radiotherapic changes infected by Chlamydia is a new finding in cytology.

[Urinary cytodiagnosis in urothelial neoplasms. A study of 678 cases]. / Citodiagnóstico urinário nas neoplasias uroteliais: estudo de 678 casos.

Reliability of urinary cytology was studied in 678 cases in the pathology department of the A.C. Camargo Hospital from 1966 to 1988. Cytologic diagnosis was compared to the final diagnosis of the patients (histopathologic diagnoses, cystoscopy, and clinical data). The rate of sensitivity, specificity, predictive value (positive and negative), and efficiency was obtained. Patients ranged from 2 months to 88 years, mean age being 59.9, among 446 men and 180 women. Cytologic diagnoses produced 139 positive cases (20.5%), 459 negative (67.7%), 21 "suspected" (3.0%), and 59 unsatisfactory cases (8.8%). There were 306 true negative cases (58.4%), 88 false-negative (16.8%), 126 true-positive (24.0%), and only 4 false-positive (.76%). The authors did not prepare a statistical correlation for the "suspected" and unsatisfactory cases; in 74 cases there was not information enough to perform the necessary comparison. The rate of sensitivity was 59.0%, specificity 98.7%, predictive positive value 96.9%, predictive negative value 77.6%, and efficiency 82.4%.