We extend the data available on B19 detection in the thyroid to show a high correlation of virus in another cohort of PTC and HT at the protein level. We also show, for the first time, B19 infection of much more highly aggressive ATC/undifferentiated tumors. Nuclear to cytoplasmic shift in B19 protein in cancer tissue suggests a possible link between B19 and thyroid cancer pathogenesis/progression.

Chlamydia are Gram negative, obligate intracellular bacterial organisms with different species causing a multitude of infections in both humans and animals. Chlamydia trachomatis is the causative agent of the sexually transmitted infection (STI) Chlamydia, the most commonly acquired bacterial STI in the United States. Chlamydial infections have also been epidemiologically linked to cervical cancer in women co-infected with the human papillomavirus (HPV). We have previously shown chlamydial infection results in centrosome amplification and multipolar spindle formation leading to chromosomal instability. Many studies indicate that centrosome abnormalities, spindle defects, and chromosome segregation errors can lead to cell transformation. We hypothesize that the presence of these defects within infected dividing cells identifies a possible mechanism for Chlamydia as a cofactor in cervical cancer formation. Here we demonstrate that infection with Chlamydia trachomatis is able to transform 3T3 cells in soft agar resulting in anchorage independence and increased colony formation. Additionally, we show for the first time Chlamydia infects actively replicating cells in vivo. Infection of mice with Chlamydia results in significantly increased cell proliferation within the cervix, and in evidence of cervical dysplasia. Confocal examination of these infected tissues also revealed elements of chlamydial induced chromosome instability. These results contribute to a growing body of data implicating a role for Chlamydia in cervical cancer development and suggest a possible molecular mechanism for this effect.

Human erythrovirus B19 (EVB19) is a small, pathogenic DNA virus that has been associated with a wide range of illnesses. The primary site of replication is in bone marrow-derived erythroid progenitor cells, but EVB19 DNA has been detected in a wide range of organs. Recently, studies have linked EVB19 to thyroid cancers and other thyroid diseases. Previous studies from multiple laboratories have detected EVB19 capsid proteins in Graves' disease, Hashimoto's thyroiditis, and thyroid cancer tissues. Data on viral gene expression and mechanism of infection in the thyroid are lacking. To investigate EVB19 infection and persistence in the thyroid, previously archived adult and pediatric tissue sections were examined for EVB19 DNA, RNA, and capsid proteins, as well as EVB19 receptor P-antigen and co-receptor α5β1 integrin. EVB19 DNA and protein were detected in a majority of tissues examined (87% and 68%, respectively). Detection was similar in adult and pediatric samples. Quantification of viral genomes revealed no significant difference in the amount of viral DNA in benign, cancerous, or metastatic thyroid tissues. EVB19 capsid RNA was detected in 67% of the tissues examined, confirming at least low-level viral gene expression. Immunohistochemical staining for P-antigen and α5β1 detected the receptor and co-receptor most frequently on normal thyroid epithelial cells. EVB19 capsid staining could be detected in tumors lacking viral receptors. These results suggest that normal thyroid epithelial cells are the initial target for EVB19 infection in the thyroid and allow for continued persistence in both normal and cancerous thyroid tissues.

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Proteomic profiles of tumor protein expression by the surface enhanced laser desorption-ionization time of flight (SELDI-TOF) methodology have been shown to have a potential usefulness for protein discovery as well as screening, diagnosis, prognosis and therapeutic considerations of cancer from several organ systems. Fineneedle aspiration (FNA) specimens from tumor samples is an accepted method to diagnose the cells of interest but often can be a limited assessment due to quantity of the sample. The current use of fresh or rapidly frozen specimens for proteomic profiling can be burdensome for clinicians to collect and submit specimens. The current study tests the hypothesis that placement of FNA and other cytological material in PreservCyt s may be an acceptable method of sample handling for protein profiling evaluation by this method though it may not be suitable for true protein discovery or characterization. Excised fresh breast tissue for evaluation and/or treatment of a variety of breast lesions were sampled by FNA technique and placed into PreservCyt s . These samples were then homogenized under denaturing conditions and evaluated by the SELDI-TOF methodology. Most samples collected showed a satisfactory quantity of protein for analysis by the SELDI-TOF methodology. Protein patterns from a variety of benign and malignant lesions revealed reproducible patterns on triplicate testing. Benign lesions had similar protein patterns across age groups in this limited series that may have potential diagnostic significance. In conclusion, FNA of breast tissue placed in PreservCyt s is a potentially acceptable method of sample handling for evaluation by the SELDI-TOF methodology for establishment of reproducible protein patterns. Preliminary results from a spectrum of breast lesions suggest these patterns may have potential for ancillary testing for diagnostic consideration of breast lesions. This collection methodology could simplify sample gathering for further testing of all types of cytological specimens by the SELDI-TOF methodology. Larger studies will be needed to assess this methodology as a diagnostic aid.

Context.—The uses of monoclonal antibodies via immunochemistry have been reported frequently within the literature using various methodologies with applications to cytology specimens. The direct application of immunochemistry to cytology may have a variety of pitfalls that the general pathologist familiar with its application to histology may be unaware of when applying it prospectively to patient specimens. Objective.—To review common pitfalls when applying immunochemistry to cytology specimens and to suggest approaches to the more common differential dilemmas that apply to a variety of cytology specimens that could be seen in a general pathology practice. Data Sources.—The authors' own experiences of applying immunochemistry to cytopathology specimens within an academic setting along with supportive data from the literature. Conclusions.—Immunochemistry can be used to increase the predictability of a cytology diagnosis if care is taken with the cytology sample preparation methodology and there is judicious use of select monoclonal antibody panels to support a specific cytology diagnosis. Up-to-date evidence-based antibody databases should be used when selecting antibody panels.

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