In-depth characterization of a new patient-derived xenograft model for metaplastic breast carcinoma to identify viable biologic targets and patterns of matrix evolution within rare tumor types.

Metaplastic breast carcinoma (MBC) is a rare breast cancer subtype with rapid growth, high rates of metastasis, recurrence and drug resistance, and diverse molecular and histological heterogeneity. Patient-derived xenografts (PDXs) provide a translational tool and physiologically relevant system to evaluate tumor biology of rare subtypes. Here, we provide an in-depth comprehensive characterization of a new PDX model for MBC, TU-BcX-4IC. TU-BcX-4IC is a clinically aggressive tumor exhibiting rapid growth in vivo, spontaneous metastases, and elevated levels of cell-free DNA and circulating tumor cell DNA. Relative chemosensitivity of primary cells derived from TU-BcX-4IC was performed using the National Cancer Institute (NCI) oncology drug set, crystal violet staining, and cytotoxic live/dead immunofluorescence stains in adherent and organoid culture conditions. We employed novel spheroid/organoid incubation methods (Pu·MA system) to demonstrate that TU-BcX-4IC is resistant to paclitaxel. An innovative physiologically relevant system using human adipose tissue was used to evaluate presence of cancer stem cell-like populations ex vivo. Tissue decellularization, cryogenic-scanning electron microscopy imaging and rheometry revealed consistent matrix architecture and stiffness were consistent despite serial transplantation. Matrix-associated gene pathways were essentially unchanged with serial passages, as determined by qPCR and RNA sequencing, suggesting utility of decellularized PDXs for in vitro screens. We determined type V collagen to be present throughout all serial passage of TU-BcX-4IC tumor, suggesting it is required for tumor maintenance and is a potential viable target for MBC. In this study we introduce an innovative and translational model system to study cell-matrix interactions in rare cancer types using higher passage PDX tissue.

Human steroidogenic factor-1 (hSF-1) regulates progesterone biosynthesis and growth of ovarian surface epithelial cancer cells.

The majority of cancers derived from ovarian surface epithelial (OSE) cells are lethal. Estrogens promote proliferation of OSE cells, whereas progesterone inhibits proliferation and promotes apoptosis of OSE cells. Human steroidogenic factor-1 (hSF-1) induction of the steroidogenic acute regulatory protein (StAR) gene, and the steroidogenic enzymes CYP11A1 and HSD3B2 is central to progesterone biosynthesis. Whereas hSF-1 and StAR are expressed in human ovarian surface epithelial (HOSE) cells, hSF-1 and StAR protein were not expressed in a panel of malignant ovarian cancer cell lines (SKOV-3, BG-1, and Caov-3), and in human OSE cells immortalized by SV40 large T antigen (IOSE-121). Transient expression of hSF-1 in SKOV-3 cells activated the expression of StAR, p450scc and 3betaHSD-II mRNAs, and induced progesterone biosynthesis. Additionally, hSF-1 suppressed proliferation and promoted apoptosis of SKOV-3 cells and suppressed SKOV-3 cell growth induced by ERalpha and estradiol. These findings suggest that hSF-1 is central to progesterone biosynthesis in OSE cells. Human SF-1 may decrease OSE cancer cell numbers directly by apoptosis, and indirectly by opposing estradiol-induced proliferation. These findings are consistent with the hypothesis, that down-regulation of hSF-1 contributes to progression of ovarian epithelial cancers.

Chemoprevention of NMU-induced rat mammary carcinoma with the combination of melatonin and 9-cis-retinoic acid.

In experimental trials using the N-nitroso-N-methylurea (NMU)-induced rat mammary tumor model, a significant decrease in tumor incidence (to 5%) was observed in rats treated with melatonin and 9-cis-retinoic acid (9 cRA) compared to controls (55%). Although 9cRA alone decreased tumor incidence to 26%, this response did not reach statistical significance. Tumor incidence was significantly inhibited to 20% in the animals that received melatonin and 9cRA on alternating days. Latency to tumor onset was prolonged in animals receiving either of the combination treatments compared with controls, and tumor multiplicity was also significantly decreased.

Fine-needle aspiration of cytomegalovirus sialadenitis in a patient with acquired immunodeficiency syndrome: pitfalls of diff-quik staining.

A case of cytomegalovirus (CMV) sialadenitis in a patient with acquired immunodeficiency syndrome (AIDS) diagnosed by fine-needle aspiration biopsy (FNAB) is reported. Although a diagnosis of CMV sialadenitis can be established on Diff-Quik-stained slides, the characteristic viral changes are the best appreciated on slides stained by the Papanicolaou (Pap) technique. Differential pitfalls are discussed. It needs to be stressed that clinically unsuspected diagnosis of CMV infection may uncover congenital or acquired defects of cellular immunity, and particularly AIDS. Diagn. Cytopathol. 2000;22:101-103.

Disseminated coccidioidomycosis. The role of cytology in multidisciplinary clinical approach and diagnosis.

A case of clinically unsuspected disseminated coccidioidomycosis diagnosed by different cytologic approaches and confirmed by mycological culture is reported. An African-American man presented with a clinical picture of pneumonia not responding to antibiotics. He subsequently developed a large neck mass and was found to have mediastinal and hilar adenopathy highly suspicious of a neoplastic process. Fine needle aspiration biopsy of the neck mass, followed by flexible bronchoscopy, was performed. Various cytologic approaches and techniques in rapid diagnosis of suspicious masses are discussed.

Alveolar macrophage apoptosis and TNF-alpha, but not p53, expression correlate with murine response to bleomycin.

Apoptosis is considered to be a protective mechanism that limits lung injury. However, apoptosis might contribute to the inflammatory burden present in the injured lung. The exposure of mice to bleomycin (BLM) is a well-established model for the study of lung injury. BLM exposure induces DNA damage and enhances tumor necrosis factor (TNF)-alpha expression in the lung. To evaluate the importance of alveolar macrophage (AM) apoptosis in the pathogenesis of lung injury, we exposed BLM-sensitive (C57BL/6) and BLM-resistant (BALB/c) mice to BLM (120 mg/kg) and studied the induction of apoptosis [by light-microscopy changes (2, 8, 12, 24, 48, and 72 h) and annexin V uptake by flow cytometry (24 h)], the secretion of TNF-alpha (measured by ELISA), and the expression of p53 (by immunoblotting) in AM retrieved from these mice. BLM, but not vehicle, induced apoptosis in AM from both murine strains. The numbers of apoptotic AM were significantly greater (P < 0.001) in C57BL/6 mice (52.9%) compared with BALB/c mice (40.8%) as demonstrated by annexin V uptake. BLM induction of apoptosis in AM was preceded by an increased secretion of TNF-alpha in C57BL/6 but not in BALB/c mice. Furthermore, double TNF-alpha receptor-deficient mice, developed on a C57BL/6 background, demonstrated significantly (P < 0.001) lower numbers of apoptotic AM compared with C57BL/6 and BALB/c mice. BLM also enhanced p53 expression in AM from both murine strains. However, p53-deficient mice developed BLM-induced lung injury, exhibited similar lung cell proliferation (measured as proliferating cell nuclear antigen immunostaining), and accumulated similar amounts of lung hydroxyproline (65 +/- 6.9 microgram/lung) as did C57BL/6 (62 +/- 6.5 microgram/lung) mice. Therefore, AM apoptosis is occurring during BLM-induced lung injury in a manner that correlates with murine strain sensitivity to BLM. Furthermore, TNF-alpha secretion rather than p53 expression contributes to the difference in murine strain response to BLM.tumor necrosis factor; strain susceptibility

Cytologic nuclear grade of malignant breast aspirates as a predictor of histologic grade. Light microscopy and image analysis characteristics.

OBJECTIVE: To determine if cytologic nuclear grade characteristics combined with image analysis assessment of morphometric nuclear parameters (1) correlate with the modified Scarff-Bloom-Richardson grading system and (2) discriminate between low and high nuclear grades of invasive ductal carcinoma. STUDY DESIGN: Fifty-four fine needle aspiration biopsies (FNABs) of breast carcinoma were evaluated for five morphologic nuclear grade characteristics. In addition, four morphometric, standardized object measurements were analyzed by an image analysis system. Corresponding biopsies of invasive ductal carcinoma (46 cases) were independently evaluated with the Scarff-Bloom-Richardson grading system, modified into low (scores 3-6) and high (scores 7-9) grades. RESULTS: An overall agreement of 82% was reached by three of four cytopathologists for each of five morphologic characteristics. There was a strong correlation (r = .8059, P < .0001) between cytologic nuclear grade and modified histologic grade. Only pleomorphism, nucleoli and sum optical density retained their statistical significance in distinguishing low from high grade ductal carcinomas. These three characteristics also had the strongest correlation with cytologic nuclear grade. CONCLUSION: Cytologic nuclear grade from aspirates of ductal carcinoma can be a predictor of the modified histologic grades of Scarff, Bloom and Richardson. Nuclear morphology reinforced by image morphometry may separate these tumors into low and high nuclear grade categories.

Adrenal cortical carcinoma with a liver metastasis in a child. Report of a case with fine needle aspiration findings and image analysis of nuclear DNA.

BACKGROUND: Adrenal cortical carcinoma is a rare and highly malignant neoplasm. Liver metastases from it may require special differential techniques in addition to cytologic findings. CASE: A 14-month-old child had adrenal cortical carcinoma and a liver metastasis confirmed by fine needle aspiration biopsy (FNAB). Histologic and clinical criteria of malignancy only partially applied to this tumor. Immunohistochemical analysis of both the primary tumor and metastasis supported an adrenal cortical origin. Other ancillary tests-DNA ploidy, proliferation index (proliferating cell nuclear antigen, Ki-67) and p53 protein immunolocalization-were utilized in a diagnostic sequence. Flow cytometric and image analysis of DNA ploidy of the primary tumor gave similar results. Assessment of DNA content of the adrenal cortical carcinoma and liver metastasis by image analysis showed identical patterns. CONCLUSION: The applicability of new diagnostic techniques to FNAB material may provide new objective measures of the biologic potential of adrenal cortical carcinoma. The validity of these tests is enhanced with the use of image-based quantitative approaches that provide greater reproducibility and objectivity of their results.

Fine needle aspiration of basal cell adenocarcinoma of the parotid gland. Report of a case with assessment of DNA ploidy in aspirates and tissue sections by image analysis.

BACKGROUND: Basal cell adenocarcinoma of the parotid gland is a low grade malignant neoplasm. It has cytologic features of basal cell adenoma and a histologically infiltrative growth pattern of malignant tumors with perineural and vascular invasion. CASE: Fine needle aspiration biopsy findings of basal cell adenocarcinoma of the parotid gland in a 77-year-old male were supplemented by DNA ploidy analysis. CONCLUSION: No single cytologic feature was found to unequivocally distinguish this lesion from basal cell adenoma and/or solid variant of adenoid cystic carcinoma. Therefore, for diagnostic purposes, we grouped all three lesions under the term basal cell tumor. Evaluation of DNA content of tumor cells revealed diploid histograms in both cytologic material and paraffin-embedded tissue. Infiltrative tumor nests, the histologic basis for differentiating basal cell adenocarcinoma from adenoma, showed the same diploid pattern. Though DNA quantitation may not discriminate basal cell adenoma from basal cell adenocarcinoma, it may prove useful in separating them from adenoid cystic carcinoma, which is considered to be a tumor with high malignant potential.

Kids speak out on adoption: a multiage bookwriting group for adopted children with special needs.

This article describes the conceptualization and development of the Saturday Club for Adopted Kids (a four-session, multiage, bookwriting group for adopted children and young people) and the benefits of offering such a project-oriented group in which adoptees can tell their stories and share their experiences in an environment that supports their recovery from early trauma and offers them a way to transform the meaning of their personal losses into social action to help others. Club participants produced a book, Kids Speak Out on Adoption, based on their experiences, perceptions, and feelings.

Cytologic diagnosis of ductal versus lobular carcinoma of the breast.

OBJECTIVE: To determine the value of cytology in the differential diagnosis of ductal versus lobular carcinoma of the breast. STUDY DESIGN: In this study we examined 11 cytologic parameters in fine needle aspiration (FNA) cytology specimens from 52 patients who underwent surgery and had subsequent histologic diagnoses. Eighty-eight percent (46 cases) were infiltrating ductal carcinoma, and 12% (6 cases) were invasive lobular carcinoma. RESULTS: Of the 11 cytologic parameters only chromatin pattern (P < .0001), nuclear size (P < .004) and overall cell size (P < .004) showed statistically significant differences between the two groups. Nuclear chromatin was coarsely granular only in cases of ductal carcinoma, while fine granularity could be seen in both ductal and lobular tumors. An automated morphometric system was used to determine the nuclear and overall cell size. The granularity of nuclear chromatin in ductal carcinoma cells did not correlate significantly with nuclear and overall cell size; therefore, they should be considered independent cytologic parameters. CONCLUSION: The cytologic differential diagnosis of ductal versus lobular carcinoma is difficult; based on this study, the presence of coarsely granular chromatin, nuclear size > 44 microns2 and cell size > 82 microns2 are the only features diagnostic of ductal versus lobular carcinoma.

Fine needle aspiration of prostatic rhabdomyosarcoma. A case report demonstrating the value of DNA ploidy.

A case of primary rhabdomyosarcoma of the prostate diagnosed by fine needle aspiration biopsy and confirmed with immunocytochemistry is described. Evaluation of DNA content of tumor cell nuclei by image analysis was performed to complement the diagnosis and clinical stage information and to further predict the tumor response to chemotherapy. Diploid tumors should prompt clinicians to use a more aggressive chemotherapeutic protocol to achieve as favorable a response to therapy as that seen in aneuploid tumors. The DNA content obtained with differently fixed and stained specimens yielded similar results, further confirming the wide applicability of image analysis as a diagnostic/prognostic tool in prospective and retrospective studies in pathology.

Unusual presentation of a large tension bronchogenic cyst in an adult.

A routine chest radiograph in a 20 year old man revealed a giant air filled cavity of the left hemithorax under tension. At thoracotomy a large left lower lobe intraparenchymal cyst required lobectomy and the pathological findings were consistent with a bronchogenic cyst. Although tension bronchogenic cysts are common in infants, this case demonstrates their unusual presentation in an adult.

Proctitis cystica profunda in paraplegics. Report of three cases.

Proctitis cystica profunda is a benign disease of the rectal mucosa that can be mistaken for rectal carcinoma both grossly and microscopically. Symptoms may consist of blood or mucus in the stool, diarrhea, tenesmus, or rectal pain. The disease has never been reported in a paraplegic population before, but the proposed etiology makes this group seem to be at high risk. We report three cases in our paraplegic population and discuss the nature of the disease as well as its treatment.

Anti-estradiol immunoperoxidase labeling of nuclei, not cytoplasm, in paraffin sections, determines estrogen receptor status of breast cancer.

We evaluated a commercially available polyclonal antibody to 17 beta-estradiol as the basis for an estrogen receptor (ER) assay of breast carcinoma in formalin-fixed paraffin tissues and then compared it with both the ER-ICA antibody in serial paraffin sections and the biochemical assay of corresponding fresh tissue. Using the estradiol antibody, 49 of 50 cases showed some cytoplasmic staining; 38 cases had nuclear staining. Sensitivity and specificity for different proportions of positive nuclear and cytoplasmic staining were calculated using receiver-operator characteristic curves. The optimum correlation with the biochemical assay was obtained with nuclear staining alone. Greater than 30% nuclear positivity as a cut-off point yielded a sensitivity of 76% and a specificity of 82%. The corresponding ER-ICA values in 38 cases yielded a sensitivity of 93% and a specificity of 56%. The methodology for the ER-ICA assay was more technically demanding in paraffin sections than that of the estradiol antibody and considerably more expensive. This study is the first to show that with nuclear staining only, and not cytoplasmic staining, as the parameter of positivity, the immunocytochemical assay of ER with anti-17 beta-estradiol antibody in routinely processed, formalin-fixed, archival material is an accurate and specific method for the determination of the ER status of breast carcinoma.