nm23 is expressed in reactive mesothelium and is not useful for detection of malignant cells in serous effusions.

The cytological dilemma of distinguishing malignant cells from proliferating mesothelium has generated volumes of research but no clear consensus regarding an optimal staining panel. nm23 has been frequently described in malignant cells as a metastasis suppressor, but its mechanism of action and the relationship between nm23 expression, metastases, and prognosis is controversial. This is the first study to apply nm23 immunostaining in the setting of effusion cytology. One hundred samples of effusions (56 malignant and 44 benign) were immunostained with nm23 using the biotin-avidin technique and diaminobenzidine as a chromogen. Additionally, a mucicarmine stain was performed on most samples, all of which were evaluated for nm23 expression and mucin in a blinded fashion. After all the samples were reviewed, the diagnoses were disclosed and staining patterns evaluated. From the malignant cases, 51 of 56 cases were positive for nm23 in at least 5% of malignant cells, while 27 of 44 reactive mesothelium cases were also positive in at least 5% of cells. Of the malignant cases, all non-adenocarcinomas expressed nm23. We conclude that nm23 is a highly sensitive marker for detecting malignant cells. Its relatively high rate of expression in reactive mesothelium supports previous studies that suggest nm23 is expressed in proliferating cells.

Evaluation of the CD34 and CD10 immunostains using a two-color staining protocol in liver fine-needle aspiration biopsies.

Fine-needle aspiration (FNA) biopsies of liver masses may need immunohistochemistry to resolve difficult cases. We examined liver FNAs with CD10 and CD34 using a two-color staining protocol. Fifty-one cases of hepatocellular carcinoma (HCC) and 23 cases of liver metastases were stained first with CD10 using diaminobenzidine as a chromogen and then with CD34 using Fast Red. All cases were reviewed in a blinded fashion by two of the authors (R.I.Z. and Z.B.). Diagnoses were unblinded and staining patterns evaluated. Forty-eight of 51 cases of HCC stained for CD34 in a peripheral pattern and no cases of metastases stained for CD34 Seven of 23 cases of metastatic disease stained for CD10 in a diffuse cytoplasmic distribution and 29 of 51 HCCs stained in a canalicular pattern. Overall, the sensitivity of the CD10/CD34 combination was 98% and the specificity was 100% for HCC when appropriate staining patterns were observed. Two-color immunostaining with CD10 and CD34 may prove useful in distinguishing HCC from metastases.

Diagnostic value of CA 15-3 antibody in detecting metastatic adenocarcinoma.

OBJECTIVE: To determine the diagnostic value of CA 15-3 in detecting metastatic adenocarcinoma in body fluids using PreservCyt solution (Cytyc Corp., Boxborough, Massachusetts, U.S.A.) as collection fluid. STUDY DESIGN: Cytospin slides prepared from 72 cases with unequivocally benign or malignant diagnosis were studied. Of the cases studied, 34 were breast carcinomas, and 17 were benign pleural effusions. Slides were stained for CA 15-3 by using the avidin-biotin complex method. Cases were evaluated for the presence of membranous or cytoplasmic staining. The percentage of cells exhibiting strong staining was estimated for both breast carcinoma and all adenocarcinomas as a group. These results were compared with CA 15-3 staining exhibited by benign mesothelium. RESULTS: Ninety-one percent of the breast cancer cases studied showed a positive reaction with CA 15-3, while 6% of the benign mesothelium cases were positive (p < 0.01). The sensitivity of CA 15-3 was 91 % for breast carcinoma and 80% for all adenocarcinomas. Specificity was 94% for breast carcinoma and for all adenocarcinoma. CONCLUSION: CA 15-3 is a sensitive and specific marker for diagnosing adenocarcinoma in cytologic specimens using PreservCyt solution as collection fluid.

Low prevalence of loss of heterozygosity and SMAD4 mutations in sporadic and familial juvenile polyposis syndrome-associated juvenile polyps.

BACKGROUND: Juvenile polyps (JP) may develop sporadically or may be associated with the familial juvenile polyposis syndrome (FJPS). In FJPS, the epithelium is susceptible to dysplasia and, ultimately adenocarcinoma. However, the mechanisms involved in this transformation are unknown. Since the epithelium in colorectal carcinogenesis undergoes a stepwise genetic progression, the purpose of this study was to determine if loss of heterozygosity (LOH) abnormalities can aid in the differentiation between sporadic and FJPS-associated polyps. DESIGN: Ninety-one routinely-processed JP from three groups of patients were evaluated for this study. Group 1 included 39 polyps from 39 patients with a single JP and no personal or family history of FJPS; group 2 consisted of 24 polyps from 15 patients with 2-5 JP and no history of FJPS; and group 3 included 29 polyps from 22 patients with > or =5 polyps either with (7) or without (15) a family history of FJPS. Epithelium from typical, atypical, and overtly dysplastic polyps, when present (2 cases in group 3 only), were evaluated separately by microdissection and PCR analysis for LOH of APC, p53, 3p, 9p, and mutations in exon 9 of the SMAD4 gene. RESULTS: SMAD4 mutations were observed in 3 polyps from 2 patients in group 3 (10% of informative cases; p < 0.05 vs group 1), but not in any of the polyps from the other two groups. Overall, LOH of APC, p53, 3p, and 9p were detected in 1%, 15%, 10%, and 4% of JPs, but no differences were observed between the three clinical groups. Two polyps, both in group 3, with definite dysplasia did not show any genetic alterations. The morphologic appearance of the polyps was not a reliable feature in helping to differentiate sporadic from FJPS-associated polyps. CONCLUSIONS: LOH of APC, p53, 3p, and 9p may not be involved in the carcinogenic pathway of FJPS-associated polyps. SMAD4 gene mutations show a low sensitivity but a high specificity for FJPS.

Two-color immunostaining of liver fine needle aspiration biopsies with CD34 and carcinoembryonic antigen. Potential utilization in the diagnosis of primary hepatocellular carcinoma vs. metastatic tumor.

OBJECTIVE: To examine immunohistochemical staining of cell block material with antibodies against vascular marker CD34 and polyclonal carcinoembryonic antigen (pCEA) for their clinical utility as part of a 2-color staining protocol in fine needle aspiration (FNA) biopsy of liver masses to distinguish metastases from primary hepatocellular carcinoma (HCC). STUDY DESIGN: The authors obtained cell block material from 96 liver FNAs and performed simultaneous (i.e., "dual-color") immunohistochemical staining utilizing antibodies against vascular marker CD34 and pCEA. Cases were blinded and evaluated by the authors for staining pattern and intensity. A consensus was obtained, the results were unblinded, and the diagnoses were correlated. RESULTS: After staining, 89 cases had sufficient tissue for evaluation. Of the 19 HCC cases, 16 (84%) showed peripheral staining with CD34, and 13 (68%) showed a canalicular or mixed canalicular-cytoplasmic staining pattern for pCEA. Thirteen cases (68%) showed staining for both antigens. All HCC exhibited immunostaining for at least 1 antibody in an appropriate staining pattern. Of the 67 cases of metastatic malignancy, 5 (7%) showed a predominantly transgressing pattern of CD34 staining, 43 (64%) showed a predominantly cytoplasmic or mixed cytoplasmic-canalicular pattern of pCEA staining, and 2 cases (3%) showed staining for both antigens in a transgressing CD34 pattern and cytoplasmic pCEA pattern. None of the 3 normal liver tissue blocks showed staining with either antigen. CONCLUSION: Two-color immunohistochemical staining of liver cell block material obtained by FNA with antibodies to CD34 and pCEA can be helpful in differentiating metastatic tumors vs. primary HCC.

The beta subunit of human chorionic gonadotropin lacks specificity for malignant cells in serous effusions.

The cytologic diagnosis of malignancy is frequently straightforward. For difficult cases, multiple immunostains and immunostain panels have been investigated without consensus. beta-human chorionic gonadotropin (hCG) has been reportedly expressed in malignancies, but not in normal tissue. HCG also has been reported as a specific marker of metastases in serous fluids when detected with laboratory assays. We investigated the clinical utility of hCG in this cytologic setting. A total of 97 cases of benign and malignant effusions were studied. Each case was immunostained with monoclonal hCG using the avidin-biotin technique and diaminobenzidine as a chromogen. Additionally, a mucicarmine stain was performed on most cases. Cases were evaluated for hCG expression and mucin in a blinded fashion. After the cases were reviewed, the diagnoses were unblinded and staining patterns were evaluated. Of the 47 benign cases studied, 23 (49%) exhibited immunoreactivity to hCG in at least 5% of mesothelial cells present. In contrast, 28 of 44 (64%) adenocarcionomas exhibited a similar degree of immunostaining. In all, 21 (48%) of the adenocarcinomas were also positive for mucin; five of these mucin-positive cases were negative for hCG. The combination of mucin and hCG detected 33 of 44 (75%) adenocarcinomas. We conclude that hCG lacks the specificity for malignant cells to be of clinical use in effusion cytology.

Evaluation of host stem cell-derived cardiac myocytes in consecutive biopsies in long-term cardiac transplant patients.

BACKGROUND: Bone marrow-derived stem cells are involved in tissue formation in transplanted organs. In human renal cell transplants, recipient-specific cells have been shown to participate in regeneration of interstitial tissue. In hepatic transplants, hepatocytes with a recipient sex chromosome pattern have been observed. Recruitment of stem cells in repair and regeneration of cardiac myocytes has been demonstrated in experimental ischemia. Recently, cardiac antral myocytes of recipient origin have been shown to populate transplanted hearts in gender-mismatched transplants. METHODS: Using the fluorescence in situ hybridization (FISH) technique we examined serial post-transplant right ventricular biopsies and sections of ventricle (at time of autopsy) of 4 gender-mismatched female-to-male transplanted hearts (graft survival 0.5 to 10 years, mean 5.1 years) for Y chromosomes within cardiac myocytes. RESULTS: We detected recipient-specific sex chromosomal patterns in single rare cardiac myocytes in 1 case in a serial biopsy after 7 years of transplant survival. The other serial biopsies, as well as the final sections at autopsy, showed no recipient pattern chromosomes within the myocytes. In addition, 2 cases demonstrated Y chromosomes in the smooth muscle of intracardiac arteries. CONCLUSION: Development of recipient marrow-derived stem cells into functional myocytes in the ventricle of transplanted hearts is a rare feature. The higher percentage of stem cell population in the cardiac atria may be a feature of tissue repair and/or an early feature of transplant physiology. Similar to other transplant organs, recipient stem cells are involved in tissue neovascularization.

Histologic features of uterine leiomyomata treated with microsphere embolization.

BACKGROUND: Therapeutic embolization is a new treatment for symptomatic leiomyomata. Material available in the United States for uterine embolization includes precisely calibrated hydrophilic microporous microspheres made of a cross-linked acrylic copolymer. CASE: A 42-year-old woman with a longstanding history of symptomatic leiomyomatous uterus underwent uterine artery embolization with microspheres. Because of continuous intractable pelvic pain, an abdominal hysterectomy and bilateral salpingo-oophorectomy was performed. The gross and histologic findings included large amounts of microspheres and pus within the uterine cavity, infarcted leiomyomata with microspheres, and intravascular microspheres within hyalinized leiomyomas. CONCLUSION: Microsphere embolization leads to hyalinizing and ischemic necrosis with residual microsphere material in the myometrium, uterine cavity, fallopian tubes, and ovaries.

The clinical utility of the Das-1 monoclonal antibody in identifying adenocarcinoma of the colon metastatic to the liver in fine-needle aspiration tissue.

BACKGROUND: The cytologic diagnosis of adenocarcinoma in a liver mass usually is straightforward. Identifying where the adenocarcinoma arose from is much more problematic. The Das-1 immunostain is directed against a colon specific antigen and has shown excellent sensitivity and specificity for adenocarcinoma of the colon in surgical pathology studies. In the current study, the authors examined the clinical utility of the Das-1 immunostain in the setting of fine-needle aspiration cell block material from the liver. METHODS: The cell block material from 77 fine-needle aspiration biopsy specimens from the liver were studied. These included 17 hepatocellular carcinomas, 20 colon adenocarcinomas that were metastatic to the liver, and 40 other malignancies, predominantly adenocarcinomas, that were metastatic to the liver from a variety of primary tumor sites. Each case was stained with the Das-1 immunostain using the avidin-biotin complex method and evaluated in a blinded fashion for membranous and/or cytoplasmic staining. The diagnoses were unblinded and correlated with staining and clinical history. RESULTS: Thirteen of 20 metastatic colon carcinoma samples exhibited immunostaining whereas only 2 of the remaining 57 samples of malignancy exhibited immunostaining. CONCLUSIONS: The results of the current study suggest that the Das-1 immunostain may prove to be helpful in identifying adenocarcinomas in the liver as arising from the colon.

BCA-225 immunostain: is it of value in liver fine needle aspiration?

OBJECTIVE: To determine the utility of BCA-225 immunostain in differentiating hepatocellular carcinoma (HCC) from other malignant tumors in liverfine needle aspiration specimens. STUDY DESIGN: Cell block materialfrom 87fine needle aspirates of liver lesions, including 25 HCCs, 9 neuroendocrine carcinomas, 12 adenocarcinomas metastatic from the colon, 19 other metastatic adenocarcinomas, 13 other metastatic tumors and 2 cholangiocarcinomas, were immunostained with BCA-225 antibody using both the EnVision avidin-biotin method (Dako U.S.A., Carpinteria, California, U.S.A.), and the traditional avidin-biotin method, without antigen retrieval. Three independent observers evaluated the cases, and a consensus was reached. RESULTS: Strong immunostaining for BCA-225 in at least 10% of malignant cells was considered positive. Two of 25 HCCs, 9 of 19 adenocarcinoma metastases, 1 of 9 neuroendocrine carcinomas, 4 of 13 other metastases and 1 of 2 cholangiocarcinomas were positive. No colon metastases were considered positive. CONCLUSION: BCA-225 may help exclude the diagnoses of metastatic colon adenocarcinoma, neuroendocrine carcinoma and well- to moderately differentiated HCC in this cytologic setting.

The Das-1 immunostain is useful for discriminating metastatic colon adenocarcinoma from cholangiocarcinoma and hepatocellular carcinoma.

The distinction between cholangiocarcinoma, hepatocellular carcinoma and liver metastasis is important but at times difficult solely on microscopic appearances. The Das-1 immunostain exhibits specificity for colonic epithelium. However, its staining of cholangiocarcinomas and hepatocellular carcinomas has not been extensively studied. We evaluated the staining properties of the Das-1 immunostain in cholangiocarcinoma and hepatocellular carcinoma. Forty-four cholangiocarcinomas, 16 hepatocellular carcinomas, 17 colon adenocarcinomas metastatic to the liver and 3 benign biliary tumors were studied. The cases were stained with the Das-1 antibody following deparaffinization and rehydration. The slides were evaluated for membranous and/or cytoplasmic staining in a blinded fashion. The percentage of tumor cells exhibiting strong staining was estimated. Of the intra-hepatic cholangiocarcinomas, 4 of 36 (11%) and 2 of 16 (13%) hepatocellular carcinomas were positive for Das-1, though typically only in rare cells. In comparison, 7 of 8 (88%) extra-hepatic cholangiocarcinomas, 13 of 17 (77%) of metastatic colon carcinomas and 3 of 3 (100%) benign tumors of the biliary tract were strongly positive in a diffuse pattern. The Das-1 immunostain may play a useful role in evaluating liver malignancies.

Assessment of Glut-1 expression in cholangiocarcinoma, benign biliary lesions and hepatocellular carcinoma.

Hepatocellular carcinoma (HCC), cholangiocarcinoma (Chca) and benign bile ductule proliferations represent uncommon but important differential diagnoses in liver masses, especially if the patient has no known primary malignancy. The glucose transporter protein Glut-1 is commonly expressed in adenocarcinomas but its expression in HCC, Chca, and benign bile ductules has not been systematically investigated. Forty-two cases of Chca, 27 cases of benign bile ductule proliferations and 19 cases of HCC were stained with Glut-1. Cases were evaluated for a membranous staining pattern in tumor cells and the results compared. Twenty-one of 42 (50%) Chca stained with Glut-1 while no HCC or benign bile ductule proliferations did, neither did benign hepatocytes or portal triad structures. Glut-1 is a highly specific but insensitive stain for Chca. It may prove to be a helpful part of a diagnostic panel used to evaluate liver lesions.

Recipient-derived hepatocytes in liver transplants: a rare event in sex-mismatched transplants.

Bone marrow-derived stem cells have been shown to engraft and populate native tissues during repair and in transplanted animal tissues. Very few studies have been performed in humans to evaluate the possibility of stem cell engraftment in transplanted tissues. In human renal transplants, recipient cells have been demonstrated within vascular and interstitial structures. In a previous study of patients with hepatic transplants, hepatocytes with XY chromosome patterns have been detected in sex-mismatched female to male transplanted livers in a small number of cases. Because of the possibility of Y chromosome microchimerism of females with male offspring, we analyzed the presence of X and Y chromosomes in liver biopsies of 13 patients with sex-mismatched liver transplants (8 female to male, 5 male to female) and long transplant to biopsy intervals (1.2 to 12 years; mean, 4.5 years). We were able to detect recipient-specific sex chromosomal patterns in inflammatory cells by fluorescent in situ hybridization/immunohistochemistry combination within the liver parenchyma but not within hepatocytes. In conclusion, recipient engraftment of stem cells may be an early feature in liver transplant but may be an infrequent persistent feature in long-term grafts.

Immunohistochemical screening of mismatch repair genes hMLH1, hMSH2, and hMSH6 in dysplastic lesions of the colon.

Immunohistochemical decrease in staining for mismatch repair proteins may be seen in either microsatellite instability or inactivation (methylation) of mismatch repair proteins. Both are features of the malignant phenotype in a range of colorectal neoplasms. Expression of mismatch repair proteins in dysplastic lesions in ulcerative colitis (UC) has not been studied extensively. The authors analyzed protein expression of hMLH1, hMSH2, and hMSH6 in 18 cases of dysplasia-associated lesion or mass (DALM) in patients with UC and 10 sporadic adenomas. Immunohistochemical studies revealed adequate staining in almost all of the cases. A significant decrease in protein expression was seen in 2 DALM and 2 sporadic adenomas. The authors conclude that immunohistochemical studies of mismatch repair proteins can be applied to dysplastic lesions in UC with adequate staining results. Decreased wild type expression of hMLH1, hMSH2, and hMSH6 is not a feature of DALM in the setting of UC.

Genetic alterations in serrated adenomas: comparison to conventional adenomas and hyperplastic polyps.

Serrated adenoma is a recently described entity characterized by the presence of a hyperplastic (serrated) growth pattern combined with cytologic features of dysplasia. In contrast to conventional (nonserrated) adenomas, the molecular features of serrated adenomas have been poorly studied. Thus, it remains unclear if serrated adenomas are simply a morphologic variant of conventional adenomas or represent a different biologic entity. In this study, 46 serrated adenomas from 39 patients, 32 conventional (nonserrated) adenomas from 31 patients, and 18 hyperplastic polyps from 16 patients were evaluated for loss of heterozygosity (LOH) of APC, p53, p16, and 3p and for K-ras mutations of codons 12, 13, and 61 by polymerase chain reaction (PCR) analysis. Serrated adenomas demonstrated LOH of at least one genetic locus in 32.6% of cases. LOH of the APC gene, 3p, p53, and p16 was seen in 19.4%, 14.2%, 9.3%, and 13.8% of cases, respectively. K-ras mutations were observed in 18% of cases. Similar to serrated adenomas, conventional adenomas demonstrated at least one LOH event in 37.5% of cases and K-ras mutations in another 19% of cases. LOH of APC, 3p, p53, and p16 was observed in 22%, 33%, 5.8%, and 13.4% of cases, respectively. There were no significant differences in either the total number of genetic events or the presence of LOH of any of the individual markers between serrated adenomas and conventional adenomas. However, hyperplastic polyps showed LOH in 22% of cases and a single K-ras mutation (11%). The prevalence of LOH in hyperplastic polyps was lower than both serrated adenomas and conventional adenomas (P < .05). These results support the hypothesis that serrated adenomas represent a biologically similar morphologic variant of conventional adenomas.

Diagnostic utility of Glut-1 and CA 15-3 in discriminating adenocarcinoma from hepatocellular carcinoma in liver tumors biopsied by fine-needle aspiration.

BACKGROUND: Diagnosing liver tumors can be difficult in the setting of a poorly differentiated tumor or tumors with no known prior malignancy. Frequently, alpha-fetoprotein, carcinoembryonic antigen, factor VIII, and mucicarmine have been employed to distinguish hepatocellular carcinoma (HCC) from adenocarcinoma. However, these stains have their limitations. CA 15-3 and Glut-1 are expressed in a variety of carcinomas. To the authors' knowledge, their expression in HCC has not been studied extensively. The authors examined the clinical utility of CA 15-3 and Glut-1 in the setting of fine-needle aspiration biopsy samples from the liver. METHODS: Thirty-five cases of HCC and 59 cases of tumors metastatic to the liver were studied. These cases previously were studied with the hepatocyte paraffin-1 antibody. Each case was stained with CA 15-3 and Glut-1 using the avidin-biotin complex method. Each case was evaluated in a blinded fashion for membranous staining that was stronger than cytoplasmic or background staining. The diagnoses were unblinded and staining patterns were compared. RESULTS: CA 15-3 stained 43 of 59 metastatic carcinoma samples and 3 of 35 HCC samples. Glut-1 stained 34 of 59 metastases and 2 of 35 HCCs. Together, the 2 immunostains stained 51 of 59 metastases and 5 of 35 HCCs. Diagnostic accuracy was improved by adding hepatocyte paraffin-1 to the staining panel. CONCLUSIONS: CA 15-3 and Glut-1, especially in conjunction with hepatocyte paraffin-1, appear to be helpful in discriminating HCC from other carcinomas.

Genetic alterations of Carney complex are not present in sporadic cardiac myxomas.

Cardiac myxomas are the most frequent cardiac tumors and cause for significant morbidity and mortality. Recent evidence indicates that cardiac myxomas are, in fact, neoplasms rather than organized thrombi. Cardiac myxomas may present as solitary lesions or in association with the Carney complex. Carney complex has been linked to chromosome 2p16 and the PRKAR1A gene at 17q22-24. In this study, we analyzed sporadic cardiac myxomas to evaluate whether the genetic alterations seen in Carney complex are present in non Carney complex associated cardiac myxomas as well. We analyzed microdissected material from 13 patients with cardiac myxomas for the markers PRKAR1 9CA, D2S2153, D2S2251 and D2S123. None of the cases demonstrated loss of heterozygosity or definite band changes suggestive of microsatellite instability for any of the markers used. We conclude that sporadic cardiac myxomas are genetically not related to Carney complex and most likely do not represent an incomplete form of Carney complex.

Estudo do mecanismo de açäo do campo elétrico na incorporaçäo de cálcio iônico em células ósseas / Mechanism of action study of electric field in incorporation of ionic calcium in bone cells

Acredita-se que sinais elétricos endógenos afetem o netabolismo ósseo, sendo as respostas fisiológicas ao estímulo eletrico devidas a mecanismos celulares que envolvem variaçöes na concentraçäo citosólica de cálcio. No presente estudo esse efeito celular foi investigado através da estimulaçäo direta, por campo elétrico de intensidade fisiolgicamente significativa de 10 mV/cm e freqüência 1,t MHz, de células ósseas em cultura primária. Observou-se que o efeito e intermediado por canais de transporte de cálcio do tipo L