Multidisciplinary approach in diagnosis of lung carcinoma.

AIM: To employ multidisciplinary approach in order to make the correct diagnosis of lung carcinoma clinically and morphologically mimicking lymphoma. METHODS: Immunostaining was performed by incubating tissue sections with appropriate antibodies, using the streptavidin-biotin technique. Antigen-antibody complexes were visualized with 3-amino-9-ethylcarbasole or diaminobenzidine hydrochloride substrate solution. We have investigated p53 gene mutations by polymerase chain reaction and DNA sequence analysis of exons 5, 6, 7, 8 and 9. RESULTS: Tumor cells expressed cytokeratin AE1/AE3, epithelial membrane antigen (EMA) and thyroid transcription factor-1 (TTF-1) without thyreoglobulin positivity. Further, tumor cells expressed neuroendocrine mar kers: synaptophysin, chromogranin A, neuron-specific enolase (NSE), CD56/NCAM, CD57/Leu-7 and protein gene product 9.5 (PGP9.5). P53 was also expressed. Diffuse large cell lymphomas of B and T cell origin were excluded. Direct sequencing analysis of exon 6 of the p53 gene revealed ATC to ACC mutation at codon 195. Final diagnosis of large cell lung neuroendocrine carcinoma (LCNEC) was established. CONCLUSIONS: Morphological pattern of tumor complied with large cell non-Hodgkin's lymphoma, but large cell lung carcinoma with neuroendocrine differentiation was proved immunohistochemically and confirmed by genetic analysis of p53 mutations in tumor tissue sample. Multidisciplinary approach in diagnosis of lung carcinoma is useful for its final diagnosis.

Prevalence of Y chromosome microdeletions in infertile men with severe oligozoospermia in Serbia.

AIM: The aim of this study was to determine the prevalence and type of microdeletions of the Y chromosome of men with severe oligozoospermia-ICSI candidates in the Serbian population and to compare our findings with those from other parts of the world. METHODS: In all patients spermiogram has been performed in order to determine the sperm concentration. Patients were subjected to detailed clinical, endocrinological and cytogenetic examinations. Microdeletion analysis was performed by polymerase chain reaction (PCR) on 203 patients with normal cytogenetic findings. The STS markers tested in each case were sY84, sY86 (AZFa); sY127, sY134 (AZFb); sY254, sY255 (AZFc). RESULTS: at least one of the STS markers was deleted in 11 of the 203 cases (5.4%). CONCLUSION: AZFc microdeletions were identified with a rather high prevalence in men with severe oligozoospermia ICSI candidates in Serbian population.

Molecular analysis of Y chromosome microdeletions in idiopathic cases of male infertility in Serbia.

The aim of this study was to detect frequency of microdeletions of Y chromosome in idiopathic cases of male infertility in Serbian population. Patients were subjected to detailed clinical, endocrinological and cytogenetic examinations. Ninety patients with normal cytogenetic findings with azoospermia and severe oligozoospermia were included in the study. In these patients microdeletion analysis was performed by multiplex polymerase chain reaction (PCR) method on DNA extracted from peripheral blood. In each case 6 markers in azoospermia factor (AZF) regions were tested: sY84, sY86 (AZFa); sY127, sY134 (AZFb); sY254, sY255 (AZFc). Deletions on Y chromosome were detected in 14 of 90 cases (15.6%), 9 with azoospermia and 5 with severe oligozoospermia. Of total number of 17 deletions, 11 (64.7%) were detected in AZFc region, 3 (17.6%) in AZFa region and 3 (17.6%) in AZFb region. Microdeletions in AZF region of Y chromosome, especially AZFc microdeletions, represent common genetic cause of idiopathic azoospermia and severe oligozoospremia in Serbian infertile men. Therefore, testing for Y chromosome microdeletions should be considered as an important element in diagnosis and genetic counseling of infertile men in Serbia and decisions regarding the assisted reproduction should be made based on the presence and type of AZF microdeletions.