Hum Pathol

Destruction and loss of bronchial cartilage in cystic fibrosis.

Ogrinc G. Kampalath B. Tomashefski JF Jr.
Department of Pathology, Case Western Reserve University School of Medicine at MetroHealth Medical Center, Cleveland, OH 44109, USA.
We studied by means of serial sections of intact isolated bronchi, the distribution and morphology of bronchial cartilage in lobar and segmental airways of 6 patients with cystic fibrosis (CF). Findings were compared to those of 4 young adults without CF who served as controls. Compared to the controls, cartilage in CF airways extended for a shorter absolute distance along the bronchial tree and disappeared at a more proximal branching level. Loss of cartilage appeared to correlate with the severity of bronchiectasis. In proximal airways chronic inflammation, destruction and fibrous replacement of cartilage preceded its disappearance. Immunohistochemical staining indicated that cells of monocyte/macrophage lineage (CD68, MAC387 positive) were most closely associated with chondrolysis. Dystrophic calcification and ossification were more commonly seen in CF bronchi and dystrophic calcification was present even in the lobar branches. Destruction of bronchial cartilage is the result of sustained bronchial infection and chronic inflammation and is an additional contributory factor to bronchiectasis and airway instability in patients with CF.

Destruction and loss of bronchial cartilage in cystic fibrosis.

Ogrinc G. Kampalath B. Tomashefski JF Jr.
Department of Pathology, Case Western Reserve University School of Medicine at MetroHealth Medical Center, Cleveland, OH 44109, USA.
We studied by means of serial sections of intact isolated bronchi, the distribution and morphology of bronchial cartilage in lobar and segmental airways of 6 patients with cystic fibrosis (CF). Findings were compared to those of 4 young adults without CF who served as controls. Compared to the controls, cartilage in CF airways extended for a shorter absolute distance along the bronchial tree and disappeared at a more proximal branching level. Loss of cartilage appeared to correlate with the severity of bronchiectasis. In proximal airways chronic inflammation, destruction and fibrous replacement of cartilage preceded its disappearance. Immunohistochemical staining indicated that cells of monocyte/macrophage lineage (CD68, MAC387 positive) were most closely associated with chondrolysis. Dystrophic calcification and ossification were more commonly seen in CF bronchi and dystrophic calcification was present even in the lobar branches. Destruction of bronchial cartilage is the result of sustained bronchial infection and chronic inflammation and is an additional contributory factor to bronchiectasis and airway instability in patients with CF.

Iron-rich foci in chronic viral hepatitis.

Year 1998
Lefkowitch JH. Yee HT. Sweeting J. Green PH. Magun AM.
Department of Pathology, College of Physicians and Surgeons, Columbia University, New York, NY 10032, USA.
Stainable iron in the liver (hemosiderosis) is most commonly seen in individuals with homozygous genetic hemochromatosis, prior transfusion, hemolysis, porphyria cutanea tarda, and chronic alcohol-induced liver disease. In chronic viral hepatitis, however, significant hepatocellular hemosiderosis is uncommon. This report describes unusual foci of hepatocellular hemosiderosis ("iron-rich foci" or IRF) in liver biopsy specimens from three patients with chronic hepatitis with or without cirrhosis (two hepatitis C-related, one hepatitis B-related). IRF present within the lobular parenchyma or cirrhotic nodules contrasted sharply with the immediately adjacent hemosiderin-negative liver tissue. Serum iron indices were abnormal in all three patients, but homozygous hemochromatosis was ruled out based on the hepatic iron concentration and hepatic iron index for each case. These cases highlight the potential for irregular iron storage in chronic viral liver disease and possible confusion with genetic hemochromatosis. The possible pathogenesis of IRF and the relationship of iron storage to the outcome of interferon therapy in chronic viral hepatitis are discussed.

Keratin subsets and monoclonal antibody HBME-1 in chordoma: immunohistochemical differential diagnosis between tumors simulating chordoma.

Year 1998
O'Hara BJ. Paetau A. Miettinen M.
Department of Anatomy, Pathology and Cell Biology, Jefferson Medical College of Thomas Jefferson University, Philadelphia, PA, USA.
Thirty-five chordomas and more than 100 other tumors that have to be considered in the differential diagnosis, were immunohistochemically analyzed using a panel of antibodies including those to subsets of keratins (K), HBME-1, a monoclonal antibody recognizing an unknown antigen on mesothelial cells, and neuroendocrine markers. The patterns of immunoreactivities in chordoma were compared with those in renal cell carcinoma, colorectal mucinous adenocarcinoma, pituitary adenoma, skeletal chondrosarcoma, and extraskeletal myxoid chondrosarcoma (ESMC). Chordomas were consistently positive for keratin cocktail AE1/AE3, and for the individual keratins K8 and K19, and nearly always positive for K5, but they showed negative or only sporadic reactivity for K7 and K20. The keratin K8 and K19 reactivity was retained in those chordomas showing solid sheets of epithelioid, spindle cells, or cartilaginous metaplasia, and in one of two cases showing overtly sarcomatous transformation. In comparison, keratins were never present in skeletal chondrosarcoma, although K8 and to a lesser extent K19 were seen in occasional cases of ESMC with chordoid features. HBME-1 reacted strongly with chordoma and skeletal chondrosarcoma but was almost never positive in renal or colorectal carcinoma. These carcinomas lacked K5-reactivity, in contrast to chordoma. Chordomas were also consistently positive for neuron-specific enolase and occasionally focally for synaptophysin, but never for chromogranin. In contrast, pituitary adenomas regularly expressed the full spectrum of neuroendocrine markers and differed from chordoma by having a narrower repertoire of keratins, often showing negative or focal keratin 8- or AE1/AE3 reactivity and being almost always K19-negative. These findings indicate that chordoma can be immunohistochemically separated from tumors that can resemble it. Immunohistochemistry is especially useful in the diagnosis of small biopsy specimens that offer limited material for morphological observation.

Comparison of genetic alterations in colonic adenoma and ulcerative colitis-associated dysplasia and carcinoma.

Year 1998
Fogt F. Vortmeyer AO. Goldman H. Giordano TJ. Merino MJ. Zhuang Z.
Laboratory of Pathology, National Cancer Institute, Bethesda, MD, USA.
Carcinoma is an important complication of ulcerative colitis (UC) and develops from dysplastic precursor lesions. Genetic changes involved in the malignant transformation have not been fully characterized. We studied 19 cases of UC with high-grade dysplasia (HGD) and eight samples of associated carcinoma (CA). Microdissection of normal epithelium, epithelium at the site of chronic inflammation, HGD, and CA was performed. Polymerase chain reaction (PCR) amplification for loss of heterozygosity (LOH) of the following polymorphic microsatellites of putative tumor suppressor gene loci was done: APC (5q), DCC (18q), p16 (9p), p53 (17p), and 8p12. To compare genetic alterations, 22 typical adenomas of the colon were studied with the markers for APC and pl6 gene loci. The results indicated that LOH of p16 and p53 were present in nondysplastic epithelium, HGD, and CA. However, the LOH in nondysplastic epithelium was detected in some associated HGD, but not all. Whereas LOH of p16 was present in 7 of 14 cases of HGD (50%), it was noted in only 1 of 22 adenomas (5.0%). LOH in the APC and DCC gene loci in UC was noted in HGD with associated CA, but LOH of APC was not present either in cases of nondysplastic epithelium or in HGD alone. Conversely, LOH in APC was present in 4 of 19 colonic adenomas. We conclude that LOH of p53 and p16 in nondysplastic epithelium may be associated with chronic reparative processes. These changes may lead to susceptibility to further genetic damage involving the APC and DCC gene loci in the development of dysplasia and progression of CA in UC. The low frequency of LOH in the p16 gene (9p) in adenomas compared with dysplasia in UC combined with infrequent LOH in APC gene loci in cases of pure dysplasia in UC may support this combination of markers as a clinical test for the differentiation of polypoid dysplasia from adenomas in UC.

Value of the MOC-31 monoclonal antibody in differentiating epithelial pleural mesothelioma from lung adenocarcinoma.

Year 1998
Ordonez NG.
University of Texas M.D. Anderson Cancer Center, Houston 77030, USA.
MOC-31 is a monoclonal antibody that has recently become commercially available that recognizes an epithelial-associated, transmembrane glycoprotein often expressed in epithelial tumors. Although some authors have indicated that MOC-31 immunostaining can assist in distinguishing epithelial mesotheliomas from metastatic adenocarcinomas to the pleura, others have concluded that this marker has no value in separating these conditions. To determine whether MOC-31 immunostaining can assist in discriminating epithelial pleural mesothelioma from lung adenocarcinoma or from other carcinomas metastatic to the pleura, 38 epithelial pleural mesotheliomas, 40 pulmonary adenocarcinomas, 55 nonpulmonary adenocarcinomas, six squamous cell carcinomas of the lung (SCCLs), three small-cell lung carcinomas (SCLCs), 19 bronchial carcinoids (BCs), and 15 transitional cell carcinomas (TCCs) were studied. Reactivity was obtained in two (5%) of the mesotheliomas, in all 40 (100%) pulmonary adenocarcinomas, in 45 (82%) nonpulmonary adenocarcinomas, in six (100%) SCCLs, three (100%) SCLCs, 15 (83%) BCs, and 10 (67%) TCCs. The staining in the two positive mesotheliomas was restricted to a few cells, in contrast to the pulmonary adenocarcinomas and most of the other carcinomas where it was often strong and diffuse. It is concluded that MOC-31 can be useful in separating epithelial pleural mesothelioma from pulmonary adenocarcinoma or from other epithelial malignancies involving the pleura.

Immunohistochemical demonstration of MET overexpression in human intrahepatic cholangiocarcinoma and in hepatolithiasis.

Year 1998
Terada T. Nakanuma Y. Sirica AE.
Department of Pathology, Virginia Commonwealth University-Medical College of Virginia, Richmond 23298-0297, USA.
Expression of MET, the c-met-encoded receptor for hepatocyte growth factor, has not been investigated in proliferative biliary diseases of human liver, including hepatolithiasis and cholangiocarcinoma. Comparatively, we analyzed by immunohistochemistry the expression of MET in normal adult human livers (n = 20), normal postnatal preadult livers (n = 21), fetal livers (n = 36), hepatolithiatic livers (n = 32), and intrahepatic cholangiocarcinomas (n = 26). In normal adult livers, obvious MET immunoreactivity was not found in any cell types. In fetal liver, MET was weakly expressed in primitive biliary cells (ductal plate and immature bile ducts) and immature hepatocytes during 8 to 30 gestational weeks but was essentially negative thereafter. In hepatolithiasis, a condition of risk for cholangiocarcinoma development, MET was overexpressed in proliferated biliary cells in 26 of 32 cases (81%). In this nonneoplastic proliferative biliary condition, MET immunoreactivity was observed to be most prominent in the hyperplastic septal and large bile ducts of liver, and in the proliferated peribiliary glands associated with intrahepatic large bile ducts. In intrahepatic cholangiocarcinoma, MET overexpression in neoplastic biliary epithelium was observed in 15 of 26 cases (58%) and correlated with the degree of tumor differentiation, being highest in well-differentiated tumors and relatively low in poorly differentiated tumors. These data show for the first time that overexpression of MET is a common feature of hyperplastic and neoplastic biliary epithelial cells in human liver and suggest that MET/hepatocyte growth factor may be playing an important role in human biliary hyperplasia and in cholangiocarcinogenesis in vivo.

Clear cell papillary carcinoma of the liver: an unusual variant of peripheral cholangiocarcinoma.

Year 1998
Tihan T. Blumgart L. Klimstra DS.
Department of Pathology, Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
Cholangiocarcinomas may be extrahepatic or intrahepatic; the latter are further divided into hilar and peripheral types. Peripheral cholangiocarcinomas often resemble adenocarcinomas arising in other organs. Although clear cell changes may occur in hepatocellular carcinoma and extrahepatic cholangiocarcinoma, peripheral cholangiocarcinomas with clear cell change are rare. In such cases, an extrahepatic primary carcinoma must be excluded. We present a patient with a large, clear cell papillary carcinoma in the liver. Extensive workup of the patient for other possible primary sites including kidneys, adrenals, thyroid, prostate, or urinary bladder failed to indicate any other neoplasm. The patient is alive without evidence of disease 30 months after complete resection. The histological, immunohistochemical, and electron microscopic results were most consistent with a neoplasm in the cholangiocarcinoma family. To the best of our knowledge, a clear cell papillary peripheral cholangio carcinoma has not been described previously. This neoplasm may be related to the recently described clear cell carcinomas of the gallbladder and extrahepatic bile ducts.

Transforming growth factor alpha (TGFalpha) in hepatocellular carcinomas and adjacent hepatic parenchyma.

Year 1998
Nalesnik MA. Lee RG. Carr BI.
Division of Transplantation Pathology, University of Pittsburgh Medical Center, PA 15213, USA.
We examined stage T1 to T4 hepatocellular carcinomas (HCC) to determine whether transforming growth factor alpha (TGFalpha) presence differed between early- and late-stage HCC and between tumors with low and high proliferative rates. Paraffin sections from 36 HCC were evaluated for TGFalpha and the proliferation markers Kiel 67 antigen (Ki67) or proliferating cell nuclear antigen (PCNA) by immunoperoxidase staining. In 12 cases, double staining for TGFalpha and Ki67 was also performed. Eighty-one percent of tumors and 94% of adjacent liver sections contained TGFalpha. A trend toward inverse correlation was seen between the percentage of TGFalpha-positive tumor cells and the proliferative rate as determined by Ki67 staining. No clear correlation of TGFalpha to either tumor stage or percentage of PCNA-positive cells was seen. This study confirms the presence of TGFalpha in the majority of early- and late-stage HCC. Positivity within tumor tissue is consistent with autocrine or paracrine stimulation. A trend toward inverse correlation between TGFalpha-producing cells and the number of cycling cells suggests that rapidly proliferating tumors may consume this growth factor at an accelerated rate. Alternatively, other hepatic mitogens may have more functional significance in these latter tumors. Finally, the presence of TGFalpha in peritumoral hepatocytes suggests these cells as potential sources of paracrine stimulation for HCC.

Prognosis in malignant mesothelioma related to MIB 1 proliferation index and histological subtype.

Year 1998
Beer TW. Buchanan R. Matthews AW. Stradling R. Pullinger N. Pethybridge RJ.
Department of Histopathology, The Royal Hospital, Haslar, Hampshire, England.
The aims of this study were to evaluate the use of the proliferation marker MIB 1 and histological subtype as indicators of prognosis in malignant mesothelioma. Sections from 41 cases of malignant mesothelioma were histologically subtyped on hematoxylin and eosin sections and stained immunohistochemically for the proliferation marker MIB 1. A proliferation index was derived and the results compared with patient survival data. A statistically significant difference was found between the survival of patients having a low and high MIB 1 index (P < .001). Patients with tumors having a low MIB 1 index lived significantly longer than those with a high MIB 1 index. Patients with the spindle cell histological subtype of malignant mesothelioma had significantly shorter survival times than those with the epithelioid or mixed tumors (P < .01). The MIB 1 proliferation index and histological tumor subtype are useful markers of prognosis in malignant mesothelioma.

Human papillomavirus infection in esophageal carcinomas: a study of 121 lesions using multiple broad-spectrum polymerase chain reactions and literature review.

Year 1998
Poljak M. Cerar A. Seme K.
Institute of Microbiology and Immunology, Medical Faculty, Ljubljana, Slovenia.
To elucidate the putative role of human papillomavirus (HPV) infection in the etiology of esophageal cancer, 121 formalin-fixed, paraffin-embedded specimens originating from a non-high-incidence area for this carcinoma, from Slovenia, were screened for HPV infection using eight different polymerase chain reactions (PCR). Three different HPV consensus primer sets and four primer sets specific for HPV types 6, 16, and 18 failed to detect HPV DNA sequences in any of the tumor samples. Fragments of human beta-globin gene that served as internal controls were successfully amplified from 120 of 121 specimens. Our study confirms the opinion that most esophageal cancers originating from non-high-incidence geographic areas of this cancer are not associated with HPV infection. According to the studies reviewed, it is likely that HPV infection plays a much more significant role in esophageal carcinogenesis in those areas of the world with a high incidence of ESCC.

Adrenocortical tumor in a patient with familial adenomatous polyposis: a case associated with a complete inactivating mutation of the APC gene and unusual histological features.

Year 1998
Wakatsuki S. Sasano H. Matsui T. Nagashima K. Toyota T. Horii A.
Department of Molecular Pathology, Tohoku University School of Medicine, Sendai, Japan.
Familial adenomatous polyposis (FAP) is an autosomal dominant hereditary disorder caused by a germline inactivating mutation of the adenomatous polyposis coli (APC) gene. Patients with FAP sometimes develop various extracolonic manifestations including adrenocortical neoplasms. We present a 14-year-old boy with FAP who had an adrenocortical tumor with atypical histopathologic features, ie, sex-cord-like differentiation. Immunohistochemical studies of adrenal 4 binding protein (Ad4BP) and steroidogenic enzymes showed the capacity of these tumor cells to produce steroids. Genetic analysis of the tumor disclosed a two-hit mutation in APC: a germline 5-base pair deletion accompanied by a loss of the normal allele. Because there were no reports of genetic alterations in adrenocortical tumors developed in FAP patients, we examined 10 sporadic adrenal tumors (four carcinomas and six adenomas) for mutations in APC. However, no mutations were found in these 10 sporadic adrenal tumors. These results suggest that mutation of APC is also responsible for some fraction of the adrenocortical tumors: the tumor in this case is included.

Villin, cytokeratin 7, and cytokeratin 20 expression in pulmonary adenocarcinoma with ultrastructural evidence of microvilli with rootlets.

Year 1998
Tan J. Sidhu G. Greco MA. Ballard H. Wieczorek R.
Department of Pathology, New York Veterans Affairs Medical Center, New York 10010, USA.
Villin (V) is a glycoprotein of microvilli associated with rootlet formation. Most colonic adenocarcinomas have a V positive (+), cytokeratin (CK) 20 (+), CK7-negative (-) immunophenotype; most lung adenocarcinomas have a CK20(-), CK7(+) immunophenotype. The reports of villin immunoreactivity in lung adenocarcinoma range from 6% to 68% in studies using various fixations and varied anti-villin antibodies. Some lung adenocarcinomas have microvilli with rootlets leading to possible diagnostic confusion with metastatic colonic adenocarcinoma to lung. Nine primary lung adenocarcinomas with rootlets on ultrastructure (including four bronchioloalveolar carcinomas [BAC]), four metastatic lung adenocarcinomas with rootlets, nine metastatic colon adenocarcinomas to lung, and 10 randomly selected lung adenocarcinomas without rootlets (including five BAC), were immunostained with monoclonal antibodies to villin (1D2C3), CK7 (OV-TL12/30), and CK20 (Ks20.8) using a streptavidin peroxidase technique with heat-induced epitope retrieval. All primary lung adenocarcinomas with rootlets were CK7(+) CK20(-), and six of nine (67%) were V(+). Cytoplasmic villin positivity occurred in a diffuse--five of nine (56%), focal--two of nine (22%), or brush border pattern--two of nine (22%). Two of four metastatic lung adenocarcinomas with rootlets were V(+). One metastatic lung adenocarcinoma had a CK7(+), CK20(+), V(-) phenotype. All metastatic colonic adenocarcinomas were V(+), CK20(+), CK7(-), and 1 of 10 (10%) lung adenocarcinomas without rootlets was V(+), and all 10 were CK20(-), and CK7(+). In summary, villin positivity is more common in lung adenocarcinoma with rootlets (67%) than those without rootlets (10%). AU primary lung adenocarcinomas were CK7(+), CK20(-). The combination of villin, CK 7, and CK 20 is helpful in differentiating metastatic colon adenocarcinoma from lung adenocarcinoma with rootlets.

Multiple, unique, and common p53 mutations in a thorotrast recipient with four primary cancers.

Year 1998
Iwamoto KS. Mizuno T. Kurata A. Masuzawa M. Mori T. Seyama T.
Department of Radiobiology, Radiation Effects Research Foundation, Hiroshima, Japan.
Four primary cancers found at autopsy of a patient who received the thorium-based contrast agent Thorotrast 50 years ago and who was healthy up until a few months before his death from liver failure were analyzed for p53 mutations. The data suggest that the chronic alpha-irradiation may be a large causative factor. Multiple mutations were found in all the cancer tissues: two foci of a cholangiocellular carcinoma, a tubular adenocarcinoma of the stomach, a squamous cell carcinoma of the lung, and an adenocarcinoma of Vater's ampulla. The total number of point mutations detected were 13. Moreover, homozygous aberrations were detected in a large area of normal small intestine and noncancer liver tissues suggesting that nontumor cells which harbored p53 abnormalities gained a survival advantage and clonally expanded.

The interrelationship between tubular and papillary sectors of tubulo-villous colorectal adenomas: comparative morphometric analysis and evaluation of cell proliferation.

Year 1998
Reissenweber N. Gualco G. Ardao G. Velazquez S. Kliche I. Fosman E. Almeida E.
Department of Pathology, Clinical Hospital, University of Montevideo, Uruguay.
In a series of 100 colorectal adenomas, 23 tubulo-villous adenomas were individualized through the identification of papillae as structures persisting for more than 100 microm in serial sections with a connective axis lined with epithelial cells. In these adenomas, the tubular and villous areas with the highest dysplasia were selected, and a morphometric analysis was undertaken to assess the Index of Structural Atypia, the Nucleo-Glandular Index, and the Nuclear Stratification Index. The AgNor count and the proliferating cell nuclear antigen (PCNA) Label Index (LI) also were performed. The overall mean of each of these indexes was significantly higher in the villous sector than in the tubular one (P < .001). In 16 cases, the semi-objective method of dysplasia gradation showed a superior degree in the papillary sector, whereas it showed an equal degree in the remaining seven lesions. The AgNOR count was significantly different in all cases, with higher values in villous sectors (P < .05). With the exception of one case, this was confirmed by the PCNA LI. The Stratification Index showed significantly different values in 20 cases, whereas the other morphometric indexes showed a less discriminatory result. Our findings objectively show that the degree of dysplasia in tubulo-villous adenomas should be analyzed in the villous sector. The existence of heterogeneous cellular populations has been confirmed both in the structural organization of cells and in some basic parameters such as the cell proliferation rate in colorectal adenomas. Our findings suggest that the occurrence of villous architectural growth is a secondary event in a tubular adenoma. Enhanced cellular proliferation of the villous area allows the progressive substitution of tubular structures.

Aberrant crypt foci in human colons: distribution and histomorphologic characteristics.

Year 1998
Shpitz B. Bomstein Y. Mekori Y. Cohen R. Kaufman Z. Neufeld D. Galkin M. Bernheim J.
Department of Surgery A, Meir General Hospital and Tel Aviv University School of Medicine, Israel.
Aberrant crypt foci (ACF) are one of the earliest putative preneoplastic, and in some cases, neoplastic lesions in human colons. These microscopic lesions, identified on methylene blue-stained mucosa with a low-power-magnification microscope, are thought to be closely related to the earliest steps in multistage colonic tumorigenesis. We investigated the distribution pattern and histomorphological features of ACF in 74 patients with sporadic colorectal cancer. The distribution pattern shows a slightly higher prevalence with older age. The prevalence of the ACF in sigmoid colon was significantly higher in patients with colorectal cancer as compared with patients with benign colonic diseases. Also, significantly more ACF were detected in distal parts of the large bowel (descending, sigmoid colon, and rectum) than in proximal parts. Of 42 microdissected lesions, 12 were dysplastic and 30 were hyperplastic foci. The average size of dysplastic lesions was significantly larger than hyperplastic foci. More apoptotic bodies were found in dysplastic lesions. These lesions also showed an upward expansion of proliferative compartment and higher proliferation indices expressed as proliferating cell nuclear antigen-labeling index. Lymphoid follicles were frequently observed in the base of both hyperplastic and dysplastic foci (40% and 66.6%, respectively). The coincidence of lymphoid follicles was 2.5 to 8 times higher than expected. These features may be related to further progression of selected ACF during colorectal tumorigenesis.

Different patterns of DNA copy number changes in gastrointestinal stromal tumors, leiomyomas, and schwannomas.

Year 1998
Sarlomo-Rikala M. El-Rifai W. Lahtinen T. Andersson LC. Miettinen M. Knuutila S.
Haartman Institute, Department of Pathology, University of Helsinki, Finland.
It is not uniformly agreed whether gastrointestinal stromal tumors (GISTs) are phenotypical variants of leiomyomas (cellular leiomyomas) or whether they represent a separate, genotypically definable entity. In an attempt to solve this question, we examined immunohistochemically defined leiomyomas from the esophagus and uterus, gastric schwannomas, and benign gastrointestinal stromal tumors (GIST) by comparative genomic hybridization (CGH). All 14 leiomyomas (nine esophageal, five uterine) were actin- and desmin-positive but negative for CD34 and S100-protein. Changes in DNA copy numbers were seen only in three esophageal leiomyomas. Gains were observed in chromosomes 3, 4, 5, 8, and 17, whereas losses were seen in 16p. All schwannomas were positive for S100-protein and negative for actin, desmin, and CD34. In schwannomas, the only change by CGH was a gain in 11q in one case. The benign GISTs, all from the stomach, were positive for CD34 but negative for desmin and S100-protein; two cases were positive for actin. The CGH findings in the GISTs differed markedly from those in leiomyomas and schwannomas. Ten of the 13 cases (77%) showed DNA copy number losses in 14q, and additional or other losses were found in eight cases, most often in chromosome 22 (seven cases), 15 (three cases), and 1p (two cases). Furthermore, two of the GISTs showed gains in 5q. These results indicate that phenotypically undifferentiated GISTs are also genetically different from leiomyomas and schwannomas and support their classification apart from leiomyomas.

DNA index shift with disease progression in colorectal adenocarcinoma: a morphological and flow cytometric study.

Year 1998
Scott CA. Desinan L. Avellini C. Bardus P. Rimondi G. Rizzi V. Beltrami CA.
Institute of Pathology, University of Udine, Italy.
DNA index (DI) values seen in 86 sporadic colorectal adenocarcinomas were related to clinical, morphological, and disease progression features. DI, whose overall distribution was bimodal with peaks in the diploid and from hypotriploid to tetraploid ranges, was related to pathological lymph node staging (pN), staging, lymphoid reaction, and tubular configuration. With increasing severity in pathological features, an irregular shift in DI class prevalence was seen, with no steady increase from diploidy to higher degrees of aneuploidy. All UICC stage I tumors (13% of total) were aneuploid, 50% being hypertriploid; diploidy (35%) and hypertriploidy (22%) prevailed in stage II carcinomas (41% of total), diploidy (35%) and hypotriploidy (30%) in stage III (30% of total), and triploidy (33%) in stage IV (15% of total). Amongst features related to stage (lymphoid reaction, depth of neoplastic embolization, grading, tubular configuration, and polymorphism), few were associated with DI, and none influenced DI shift and class prevalence through the stages. The biological capabilities of colorectal adenocarcinoma in relation to stage are expressed by certain aneuploid DI classes (hypertriploidy: absence of extracolonic spread; hypotriploidy: lymph node metastases; triploidy: distant metastases). Diploidy is unrelated to criteria defining stage above I and predicts 50% of cases with development of metachronous metastases. Irregular DI class shift through the stages may be attributable to different pathways of cancerogenesis and disease progression in diploid versus aneuploid carcinomas. Alternatively, assuming that the diploid fraction in aneuploid tumors contains neoplastic cells, pure diploid carcinomas represent the selection of a vital clone that may give rise to a further mixed population whose aneuploid DI is different and best fitted to express the biological capabilities of that given stage.

Tracing the origin of adenocarcinomas with unknown primary using immunohistochemistry: differential diagnosis between colonic and ovarian carcinomas as primary sites.

Year 1998
Lagendijk JH. Mullink H. Van Diest PJ. Meijer GA. Meijer CJ.
Institute for Pathology, Free University Hospital, Amsterdam, The Netherlands.
To discriminate adenocarcinoma metastases originating from either colon or ovary, a panel of immunohistochemical markers was evaluated. For this purpose, paraffin sections from 157 primary and metastatic colonic and ovarian carcinomas were immunostained. These cases were divided into a learning group of 46 colonic and 54 ovarian carcinomas and a test group of 29 colonic and 28 ovarian carcinomas, including all metastatic tumors, among which were five with unknown primary site at the time of testing. The sections were immunostained with antibodies against carcinoembryonic antigen (CEA), cytokeratin 7 (CK7), cytokeratin 20 (CK20), CA125, vimentin, and CA19.9. Staining results were expressed as the product of staining intensity and percentage of positive tumor cells. Stepwise discriminant analysis was applied on the learning set to obtain a classification function for both tumors. The validity of the classification function was evaluated using the test set. There was considerable overlap in immunostaining for both tumor types, but colonic carcinomas were typically positive for CEA and CK20 and negative for CK7 and CA125. Ovarian carcinomas were typically positive for CK7 and CA125 and negative for CEA and CK20. In discriminant analysis, the best combination of markers appeared to be CK7 and CEA. Only one sample of the test group (2%) was misclassified. Taking learning and test groups together, 136 of the 157 samples (87%) were correctly classified with high posterior probability (PP > .8). However, from the 28 mucinous ovarian carcinomas, only 19 (68%) could correctly be classified with high PP. When excluding the nonmucinous ovarian carcinomas from the analysis, overall 87 of 103 (84.5%) of the samples were correctly classified (PP > .8) with a combination of CEA, CK7, and also vimentin. From the 28 mucinous ovarian carcinomas, only two (7%) were misclassified, and four could not be classified with sufficient certainty. In neither analysis did CK20, CA125, or CA19.9 emerge as discriminatory parameters. Based on the same data, an intuitive flow chart was constructed with which 129 of 157 cases could be classified (only one falsely) without further statistical analysis. The five metastases with an at first unknown primary could, according to the follow-up, all be classified correctly with high PP. Most ovarian carcinomas, including the mucinous ones, can be discriminated with high probability from colonic carcinomas using a panel of three antibodies directed against CEA, cytokeratin 7, and vimentin.

Diffuse lymphoplasmacytic acalculous cholecystitis: a distinctive form of chronic cholecystitis associated with primary sclerosing cholangitis.

Year 1998
Jessurun J. Bolio-Solis A. Manivel JC.
Department of Laboratory Medicine and Pathology, Fairview-University Medical Center and University of Minnesota, Minneapolis 55455, USA.
Inflammation of the gallbladder is known to occur in patients with primary sclerosing cholangitis (PSC). However, the histological features of this form of cholecystitis have not been adequately defined. The aim of this study was to compare the inflammatory lesions of PSC-associated cholecystitis with those present in other cholecystopathies. The cases consisted of 11 gallbladders from patients with PSC who underwent liver transplantation. As controls, gallbladders from liver transplant patients with primary biliary cirrhosis (n = 4) and other chronic nonbiliary hepatopathies (n = 8), and 13 cholecystectomies from patients with chronic cholecystitis with (n = 10) and without (n = 3) lithiasis, were studied. The following histological features were tabulated on coded slides: presence, depth of involvement, and distribution of the inflammatory infiltrate, predominant cell type, presence of lymphoid aggregates, epithelial damage, metaplastic changes (pyloric or intestinal), fibrosis, smooth muscle hypertrophy, and presence of Rokitansky-Aschoff sinuses. At variance with the wide range of histological abnormalities present in other forms of chronic cholecystitis, most PSC-related cholecystitis showed a diffuse infiltrate (6 of 11) rich in plasma cells (6 of 11) predominantly confined to the lamina propria (9 of 11). The combination of these three features was present exclusively in PSC (5 of 11 PSC cholecystitis compared with 0 of 25 controls; P = .001). In conclusion, this study suggests that a characteristic form of cholecystitis may develop in patients with PSC.

Alterations in colonic mucosal vessels in patients with cirrhosis and noncirrhotic portal hypertension.

Year 1998
Lamps LW. Hunt CM. Green A. Gray GF Jr. Washington K.
Department of Pathology, Vanderbilt University Medical Center, Nashville, TN 37232, USA.
Changes in intestinal mucosal microvasculature as a cause of lower gastrointestinal hemorrhage in patients with portal hypertension have been well documented clinically, but the analogous histomorphological changes have not been well characterized. The goal of this study was to evaluate qualitative and quantitative changes in colonic mucosal vessels in patients with cirrhosis or clinically evident portal hypertension and to correlate these changes with endoscopic and clinical findings. Colon biopsy or resection specimen slides from 46 patients with biopsy-proven cirrhosis (44 patients) or noncirrhotic portal hypertension (two patients) were reviewed. Immunoperoxidase stain for CD34 antigen was used to facilitate visualization of mucosal vessels, and vessel diameter was measured with a micrometer. Patients with inflammatory bowel disease were excluded. Twenty-four normal colon biopsy specimens served as controls. Mucosal vessels were divided into superficial, intermediate, and deep layers. As a group, the cirrhotic patients had a significantly higher mean diameter of vessels in all three layers. Qualitatively, increased numbers of small vessels and prominent branching were noted, especially in the superficial and intermediate layers. Tortuous, thick-walled vessels, suggesting arterialization of venules, were present in some cases. Eleven patients had endoscopic findings suggestive of vascular abnormalities, including erythematous mucosal patches, red macules, and telangiectasias. Eighteen had esophageal varices, and five had portal gastropathy. Nineteen patients had gastrointestinal (GI) bleeding, localized to the lower GI tract in 11. These qualitative and quantitative findings suggest that colonic mucosal vascular lesions are common in portal hypertension and may represent a potential source of clinically significant lower GI hemorrhage in these patients.

CD44V6 expression in human colorectal carcinoma.

Year 1998
Coppola D. Hyacinthe M. Fu L. Cantor AB. Karl R. Marcet J. Cooper DL. Nicosia SV. Cooper HS.
Department of Pathology, University of South Florida-Moffitt Cancer Center, Tampa 33612, USA.
CD44 is an adhesion molecule involved in cell-to-cell and cell-to-matrix interactions. This transmembrane glycoprotein exists in either standard or variant forms, originated by alternative splicing. One of the isoforms (CD44V6) has been shown, in some systems, to modify the metastatic potential of tumor cells. To investigate the role of this biomarker as possible prognostic antigen in colorectal cancer, we immunohistochemically analyzed the distribution of CD44V6 expression on formalin-fixed, paraffin-embedded tissues from resected colorectal cancers of 34 patients. The monoclonal antibody VFF7 against the amino acid sequence encoded by exon CD44V6 was applied using the avidin-biotin-peroxidase method. For each resected specimen, normal (N), adenomatous (AD), and carcinomatous (CA) colonic mucosa were tested. In 68% of the resected cases, these areas were present in the same slide, and in 76% of cases, nodal or liver metastases (MT) were available for evaluation. Adenomatous polyp biopsy specimens of 10 carcinoma-free patients were also tested. In selected cases, CD44V6 expression was also determined using the Western blot immunoprecipitation technique. CD44V6 immunoreactivity was detected in 100% of the ADs, and in 91% of CAs, but was mostly weak in only 38% of MTs (n=26). In 49% (n=35) of ADs, 11% (n=34) of CAs, and 4% of MTs (n=26), the stain was moderate to strong. CD44V6 immunoreactivity was predominantly membranous in ADs and cytoplasmic in MTs. In the CAs, both staining patterns were noted. Interestingly, the normal mucosa had a weak subnuclear localization of the stain. In the cases evaluated by Western blotting immunoprecipitation analysis, the level of CD44V6 protein expression was similar to that obtained by immunohistochemistry. No correlation was found with tumor type, stage, or patient survival. The predominant CD44V6 expression in ADs and CAs, but not in MTs, suggests that, in many cases, the expression of this adhesion molecule may be lost during the acquisition of migratory function by the tumor cells.

Massive macrophage lipid accumulation presenting as hepatosplenomegaly and lymphadenopathy associated with long-term total parenteral nutrition therapy for short bowel syndrome.

Year 1998
Perez-Jaffe LA. Furth EE. Minda JM. Unger LD. Lawton TJ.
Department of Pathology and Laboratory Medicine, University of Pennsylvania Medical Center, Philadelphia 19104-4283, USA.
We present a unique case of massive splenomegaly, hepatomegaly, and lymphadenopathy caused by lipid-laden macrophages in a 50 year old white female with short-bowel syndrome treated with long-term total parenteral nutrition. Using transmission electron microscopy and special stains we were able to show that the total parenteral nutrition lipid component was composed of lipid droplets and micelles morphologically identical to those found in lipid-laden macrophages which had accumulated in the patient's reticuloendothelial system leading to massive splenomegaly, hepatomegaly (without evidence of steatosis) and lymphadenopathy. While this phenomenon has been reported in animal models, no human cases have been previously reported.