Body Cavity Fluids Research Articles

Cytological features of melanoma in exfoliative fluid specimens

Aims:To describe the cytological appearances of melanoma in fluid specimens and potential diagnostic pitfalls in interpreting such specimens, which have been infrequently reported in the literature.Methods:Cases of melanoma diagnosed at...

Rapamycin reduces primary effusion lymphoma progression by targeting VEGF production and VEGF responses

Kaposi's sarcoma herpesvirus (KSHV)-infected primary effusion lymphoma (PEL) typically presents as a malignant effusion in body cavities that disseminates to distant sites. There is a need for new therapies for PEL as most patients rapidly succumb in spite of high-dose chemotherapy. We examined the potential utility of Rapamycin for PEL treatment. We found that the downstream effectors of the mammalian target of Rapamycin (mTOR) p70S6k and S6 are constitutively phosphorylated in PEL cells, and that Rapamycin inhibits this constitutive p70S6k and S6 phosphorylation. Rapamycin reduces PEL proliferation but is not cytotoxic for PEL cells. Intraperitoneal injection of PEL cells in NOD/SCID mice causes experimental effusion lymphoma. Rapamycin delayed PEL development in this model, markedly reduced accumulation of ascites (p = 0.009), prevented formation of solid tumor masses, and a significantly extended mouse survival (P < 0.001). However, Rapamycin did not eradicate PEL in mice. We examined the mechanisms by which Rapamycin reduces PEL progression in this mouse model. Levels of VEGF, which promotes vascular permeability and is critical to the accumulation of body cavity fluids, were significantly reduced in ascites of Rapamycin-treated mice compared to controls (p = 0.009). Rapamycin inhibited VEGF-induced phosphorylation of VEGF receptor (R) 2 in endothelial cells and activation of the downstream effectors of VEGFR2 phosphorylation src and enos (endothelial nitric oxide synthase). Rapamycin did not alter KSHV genes transcription in PEL cells, and only insignificantly reduced levels of IL-10, the principal growth factor for PEL, in ascites of PEL-bearing mice. Reduction of VEGF secretion by PEL and impairment of endothelial cell responses to residual VEGF likely explain reduced accumulation of ascites in Rapamycin-treated mice. The failure of Rapamycin to significantly reduce IL-10 levels in PEL-bearing mice and to promote PEL cell death likely explain PEL persistence in mice treated with Rapamycin. The successful use of Rapamycin to reduce PEL effusion and disease progression by reducing VEGF secretion and endothelial cell responses to VEGF illustrates a novel application of mTOR inhibition that targets the tumor microenvironment rather than the tumor cells, and is applicable to the treatment of PEL and other malignancies characterized by ascites accumulation and increased vascular permeability.

Alkaline phosphatase, cytokeratin 7, cytokeratin 8 in the diagnosis of primary lung adenocarcinoma from 148 pleura fluids specimens.

Adenocarcinomas are the most common cause of malignancy in pleura fluids. Usual primary sites include the lung, breast, gastrointestinal tract, and genitourinary tracts. Predicting the site of origin of an adenocarcinoma can be difficult due to overlapping morphologic characteristics. We investigated the use of alkaline phosphatase (AP), Cytokeratin7 (CK7) Cytokeratin8 (CK8) to distinguish adenocarcinomas of lung in 148 body cavity fluid samples. Overall results for primary lung adenocarcinomas, demonstrated CK8 reactivity in 106 (72%) of 148 cases. 95 primary lung carcinoma samples (65%) were positive for CK7. AP was expressed in 81% of primary lung adenocarcinomas. Positive immunoreactivity for AP was characterized by a red, diffusely apical cytoplasmic staining in tumor cells that ocurred singly or in groups. There was a significant difference between AP, CK 7 and CK 8 expressions in primary lung adenocarcinomas (P=0.02; Chi-squared test). The sensitivity of AP, CK8, CK7 as a marker for primary lung adenocarcinomas were 82%, 72%, 64%, respectively. Thus the AP positive staining largely confirmed the cytologic diagnosis of lung adenocarcinoma.

Usefulness of EMA, GLUT-1, and XIAP for the Cytologic Diagnosis of Malignant Mesothelioma in Body Cavity Fluids

We compared the effectiveness of epithelial membrane antigen (EMA) with 2 newly described markers, X-linked inhibitor of apoptosis protein (XIAP) and an isoform of glucose transporter (GLUT-1), in the distinction between malignant mesothelioma (MM) and benign effusion (BE) in body cavity fluids. Immunohistochemical studies were performed on cell block sections from 35 cases of histologically confirmed MM and 38 BEs, using antibodies to EMA, XIAP, GLUT-1 (GLUT-1m, monoclonal; GLUT-1p, polyclonal). Results were graded based on the percentage of cells staining: negative (0%), 1+ (<25%), 2+ (25%-49%), 3+ (50%-74%), and 4+ (75%-100%). The performance of each marker was compared using receiver operating characteristic curve analysis. EMA demonstrated the best accuracy, with an area under the curve of 0.91 as compared with XIAP (0.67), GLUT-1m (0.74), and GLUT-1p (0.80). Based on these findings, EMA is a better marker than XIAP or GLUT-1 for the diagnosis of MM.

Evaluation of flow cytometric immunophenotyping and DNA analysis for detection of malignant cells in serosal cavity fluids

The serosal cavities are frequent sites of tumor metastasis. The distinction between carcinoma cells, inflammatory cells, and reactive or malignant mesothelial cells can be difficult in cytology. Multicolor flow cytometry (FCM) provides the opportunity to evaluate multiple antigens simultaneously, making it possible to characterize various cell populations. In this study, we aimed to assess the diagnostic accuracy of FCM immunophenotyping and DNA in comparison with serum tumor markers and classic cytology for detection of malignant cells in pleural and ascitic fluids. One hundred and nineteen samples of body cavity fluids were analyzed. Immunophenotyping was performed by four-color immunofluorescent staining using monoclonal antibodies against Ber-EP4, cytokeratin, CD3, and CD45. The DNA analysis by FCM was also performed. In addition, serum CA19-9, CEA, AFP, and CA125 were analyzed. Ber-EP4 marker had the highest sensitivity (73%) and specificity (95.5%) in the detection of carcinoma cells in serous fluid and correlated with cytology in most of cases (73%). The mean of DI differed statistically in patients with malignant effusions than in benign one. DI showed no difference in fluids due to infiltration of malignant epithelial cells or hematopoietic malignancy or due to hepatocellular carcinoma developing in cirrhotic liver. Thus, flow cytometry appears to aid not only in the detection of malignant cells but also in the characterization of cell type. On the other hand, although DNA ploidy examination had better sensitivity; it had no advantage over conventional cytopathological examination in identification of malignant cells.

腹膜悪性中皮腫の１例―細胞転写法およびセルブロック法の有用性―

背景 : 中皮腫の早期診断には体腔液細胞診が有用であるが, 診断に難渋する症例も多い. 体腔液細胞診のみで中皮腫と確定診断した腹膜悪性中皮腫の 1 例を経験したので報告する.症例 : 59 歳, 男性. 原因不明の腹水貯留にて来院した. PET で異常所見なし. 細胞所見は, 組織球などの炎症細胞を背景に核小体が肥大した 2 核∼多核の中皮細胞が多数みられ, 相互封入像も認めたことから中皮腫と推定診断した. 時間の経過とともに乳頭状集塊もみられた. 細胞転写法を用いた免疫細胞化学染色を行い, calretinin, CK5/6, mesothelin, D2-40, E-cadherin, EMA, p53 (陽性率 : 51%) 陽性, CEA, MOC31, Ber-EP4 陰性であった. さらに, 腹水から作製したセルブロックによる電顕検索で細長い微絨毛を豊富に認め, 中皮腫と確定診断した.結論 : 体腔液細胞診では診断に難渋する中皮腫でも, 細胞転写法を用いた複数の抗体による免疫細胞化学染色およびセルブロック法による電顕検索を行うことにより, 腹水材料のみでも悪性中皮腫と確定診断することは可能である.

A Liquid Concept—Do Classic Preparations of Body Cavity Fluid Perform Differently Than ThinPrep Cases?: Observations From the College of American Pathologists Interlaboratory Comparison Program in Nongynecologic Cytology

Newer liquid-based preparations differ morphologically from classic preparations (smears, filters, and cytocentrifuged preparations). Is adenocarcinoma more readily detected in liquid-based preparations? We reviewed responses from 16,750 fluid challenges of adenocarcinoma distributed in 2005 in the College of American Pathologists Interlaboratory Comparison Program in Nongynecologic Cytology (CAP NGC). To compare the performance of body cavity fluid liquid-based preparations with adenocarcinoma to that in classic preparations in the CAP NGC. Responses for ThinPrep challenges were compared with classic preparations for exact match diagnoses of adenocarcinoma from pelvic washes, pleural fluid, pericardial fluid, and peritoneal fluids in the 2005 CAP NGC. A total of 13,690 pathologists, 8345 cytotechnologists, and 5958 laboratories submitted responses to fluid challenges in 2005. Adenocarcinoma comprised 16,750 of the fluid challenges; 88% were classic preparations, and 12% were ThinPrep challenges. The exact match to the reference diagnosis of adenocarcinoma was seen in 77% of conventional preparations and 81% of ThinPrep challenges when a general category of "positive for malignancy" was assigned. When "suspicious for malignancy," an exact match diagnosis of adenocarcinoma was made in 5% and 4% of classic and ThinPrep challenges, respectively. ThinPrep challenges performed slightly better overall, but only pelvic washings and peritoneal fluids demonstrated statistically significant improved performance with ThinPrep challenges. Use of liquid-based preparation is widespread for nongynecologic preparations and performs as well, and sometimes better than, classic preparations in an interlaboratory comparison program.

A liquid concept--do classic preparations of body cavity fluid perform differently than ThinPrep cases? Observations from the College of American Pathologists Interlaboratory Comparison Program in Nongynecologic Cytology.

Abstract Context.—Newer liquid-based preparations differ morphologically from classic preparations (smears, filters, and cytocentrifuged preparations). Is adenocarcinoma more readily detected in liquid-based preparations? We reviewed responses from 16 750 fluid challenges of adenocarcinoma distributed in 2005 in the College of American Pathologists Interlaboratory Comparison Program in Nongynecologic Cytology (CAP NGC). Objective.—To compare the performance of body cavity fluid liquid-based preparations with adenocarcinoma to that in classic preparations in the CAP NGC. Design.—Responses for ThinPrep challenges were compared with classic preparations for exact match diagnoses of adenocarcinoma from pelvic washes, pleural fluid, pericardial fluid, and peritoneal fluids in the 2005 CAP NGC. Results.—A total of 13 690 pathologists, 8345 cytotechnologists, and 5958 laboratories submitted responses to fluid challenges in 2005. Adenocarcinoma comprised 16 750 of the fluid challenges; 88% were classic preparatio...

Development of germ cells and reproductive biology in the sipunculidPhascolosoma esculenta

Sipuncula are of increasing interest for fisheries and aquaculture in China. Sustainable harvests will rely on a better knowledge of reproductive characteristics and stock enhancement. Here, we investigated the structural characteristics of and seasonal changes in germ cell development of the sipunculid Phascolosoma esculenta from the south-eastern coast of Zhejiang, China. An annual survey of egg numbers in the coelom (body cavity) fluid by light and electron microscopy of the females indicates that P. esculenta is dioecious. No defined gonad but dissociated germ cells were found in the coelomic cavity during the 1-year observation. The germ cells showed multiplication and development in the coelomic cavity. Reproduction took place from May to September, with a peak in July and August. The oogenesis can be divided into four phases: cell proliferation, pre-vitellogenesis, vitellogenesis and egg envelope formation and maturation. The process of spermatogenesis can also be divided into four phases: cell multiplication, cell growth, cell maturation and metamorphosis. Monthly changes in the relative number of eggs in each stage indicate that P. esculenta lays eggs in batches. The sperm thrives in the coelomic fluid in the form of cell groups with patterns of genesis and release similar to those of the eggs. Eggs of P. esculenta were fertilized only when reaching the nephridium. The sex ratio was about 1:1 throughout the year.

AFM-based analysis of human metastatic cancer cells

Recently biomechanics of cancer cells, in particular stiffness or elasticity, has been identifiedas an important factor relating to cancer cell function, adherence, motility, transformationand invasion. We report on the nanomechanical responses of metastatic cancer cells andbenign mesothelial cells taken from human body cavity fluids using atomic forcemicroscopy. Following our initial study (Cross et al 2007 Nat. Nanotechnol. 2780–3), we report on the biophysical properties of patient-derived effusion cellsand address the influence of cell morphology on measured cell stiffness. Using acytocentrifugation method, which yields morphologically indistinguishable cells that can beprepared in 1 min and avoids any possible artifacts due to 12 h ex vivo culture, we findthat metastatic tumor cells are more than 80% softer than benign cells with adistribution over six times narrower than that of normal cells. Consistent with ourprevious study, which yielded distinguishable cell populations based on ex vivogrowth and morphological characteristics, our results show it is unlikely thatmorphology alone is sufficient to explain the difference in elastic moduli for thesetwo cell types. Moreover, analysis of non-specific cell adhesion inherent to tumorand normal cells collected from patients show surface adhesion of tumor cells is∼33% less adhesive compared to that of normal cells. Our findings indicate that biomechanical-basedfunctional analysis may provide an additional platform for cytological evaluation anddiagnosis of cancer in the future.

Retraction halos: Nature's dotting pen in effusion cytology processed by Ultrafast Papanicolaou stain

Effusion cytology is challenging when solitary adenocarcinoma cells resemble histiocytes or mesothelial cells, and when adenocarcinomas look like histiocytic hyperplasia. It is also challenging to locate rare tumor cells in markedly inflammatory background. Over a period of 9 years, 9,474 cases of body cavity fluids including 3,642 cases of pleural, 1,900 cases of peritoneal and 345 cases of pericardial effusions and 3,587 cases of pelvic washes were processed with Ultrafast Papanicolaou stain. Retraction halos around spherical solitary cells and tissue fragments were observed whenever the background was inflammatory, whereas absent around histiocytes or histiocytic aggregates, and inconspicuous around solitary mesothelial cells. Retraction halos were helpful when the tumor fragments are obscured by marked inflammation, the quantity of tumor fragments is sparse, or the tumor cells resemble histiocytes or mesothelial cells. In short, retraction halos are nature's dotting pen for effusion cytology and are particularly useful in challenging cases.

Significance of intercellular spaces (windows) in effusion fluid cytology: A study of 46 samples

The presence and nature of intercellular windows were studied on 46 body cavity fluid samples chosen on the basis of an unequivocal diagnosis on May-Grünwald-Giemsa (MGG)/Papanicolaou-stained smears and cell blocks. Of these, 24 cases had adenocarcinoma (AC) and seven had reactive mesothelium (RM) with 15 having distinct populations of both. Mucicarmine and PAS stains were used wherever indicated. The specificity of windows for predicting reactive mesothelium was evaluated. Intercellular windows were found in all cases of reactive mesothelium coinciding with the presence of the fuzzy peripheral microvillous borders. Surprisingly, as many as 17/39 (44%) of the adenocarcinomas also exhibited this feature, of which 13 had a distinctly visible evenly distributed ciliated cell membrane. In addition, 30/39 (77%) cases of AC exhibited a "window-like" appearance caused by cytoplasmic vacuolation. None of the adenocarcinoma clusters with true window formation showed positivity for the mucin stains, whereas all the clusters with pseudowindows caused by vacuolation were stained. Thus the specificity of intercellular windows for RM was merely 56%, though the sensitivity was 100%. On the other hand, the absence of windows was 100% specific for adenocarcinoma. Intercellular windows, though a feature of reactive mesothelial cell populations, can also be found in cases of ciliated adenocarcinomas and may not have a significant predictive value.

Pleomorphic lobular carcinoma in pleural fluid: Diagnostic pitfall for atypical mesothelial cells

Pleomorphic lobular carcinoma (PLC) is a subtype of infiltrating lobular carcinoma because of its dyscohesiveness, linear infiltration pattern, and lack of membranous E-cadherin staining. However, it differs from classic lobular carcinoma because of its high-grade cytology and more aggressive clinical behavior. In breast fine-needle aspiration biopsies, PLC can be confused with invasive ductal carcinoma, particularly the apocrine variant. In this report, we illustrate how metastatic PLC in body fluid specimens shows many of the same cytomorphologic changes that occur in reactive/atypical mesothelial cells. Fortunately, the immunohistochemical staining pattern of PLC can help to distinguish it from other possible diagnoses in the differential, such as reactive/atypical mesothelial cells and other metastatic neoplasms. However, the frequent apocrine features seen in this variant of breast carcinoma can cause nonspecific immunohistochemical positivity that may make the interpretation difficult. This is the first report illustrating the cytopathology and immunohistochemical findings of pleomorphic lobular carcinoma in body cavity fluid cytology. Our case highlights the important issues and pitfalls to be aware of when making this diagnosis.

Cytologic malignancy versus benignancy: how useful are the "newer" markers in body fluid cytology?

Abstract Context.—Differentiating reactive effusion, malignant mesothelioma, and metastatic adenocarcinoma in body cavity fluids can be challenging. Interpreting immunohistochemical markers in cell block preparations can be difficult because of nonspecific staining, focal staining, or poor staining quality. We selected a panel of conventional and newer markers to assess their utility in evaluating effusions. Objective.—To evaluate the efficacy of 5 immunohistochemical markers in the differential diagnosis of reactive mesothelial proliferation, malignant mesothelioma, and metastatic adenocarcinoma in body cavity fluids. Design.—A total of 72 formalin-fixed, paraffin-embedded cell block specimens from pleural and peritoneal effusions, including 5 mesotheliomas, 48 adenocarcinomas, and 19 benign effusions were stained with antibodies against calretinin, D2-40, XIAP, MOC-31, and WT1. Results.—All benign effusions and mesotheliomas demonstrated diffuse membranous staining with D2-40. All mesotheliomas displaye...

Cytologic Malignancy Versus Benignancy: How Useful Are the “Newer” Markers in Body Fluid Cytology?

Differentiating reactive effusion, malignant mesothelioma, and metastatic adenocarcinoma in body cavity fluids can be challenging. Interpreting immunohistochemical markers in cell block preparations can be difficult because of nonspecific staining, focal staining, or poor staining quality. We selected a panel of conventional and newer markers to assess their utility in evaluating effusions. To evaluate the efficacy of 5 immunohistochemical markers in the differential diagnosis of reactive mesothelial proliferation, malignant mesothelioma, and metastatic adenocarcinoma in body cavity fluids. A total of 72 formalin-fixed, paraffin-embedded cell block specimens from pleural and peritoneal effusions, including 5 mesotheliomas, 48 adenocarcinomas, and 19 benign effusions were stained with antibodies against calretinin, D2-40, XIAP, MOC-31, and WT1. All benign effusions and mesotheliomas demonstrated diffuse membranous staining with D2-40. All mesotheliomas displayed calretinin positivity, whereas only 58% of benign effusions stained focally with calretinin. MOC-31 was positive in all cases of adenocarcinoma, whereas all benign effusions and mesotheliomas were negative. All cases of the metastatic adenocarcinoma were negative for calretinin and D2-40. However, background reactive mesothelial cells were positive for calretinin and D2-40. Overall, D2-40 highlighted more mesothelial cells than calretinin. WT1 was positive in 50% of benign effusions, 60% of mesotheliomas, and 27% of adenocarcinomas. XIAP stained most mesotheliomas (80%), some adenocarcinomas (51%), and rare benign effusions (11%). MOC-31 and D2-40 were very sensitive and specific markers of epithelial and mesothelial cells, respectively. Compared with calretinin, D2-40 was a more sensitive marker of mesothelial cells. WT1 proved to be nonspecific. XIAP was not a sensitive marker for malignancy and had a limited value in cytology. We recommend using a panel to include MOC-31 and D2-40 to improve diagnostic accuracy in body cavity effusions.

The Cytopathology of Body Cavity Fluid

Cytologic examination of the body cavity fluid is very important because the specimens represent a significant percentage of nongynecologic samples and this cytologic examination may be the first, best or only chance for making the diagnosis of an underlying malignancy. The purposes of body cavity fluid examination are to correctly identify cancer cells and if possible, to identify the tumor types and primary sites when presented with unknown primary tumor sites. The most important basic differential diagnosis is that of benign and reactive disease vs malignant disease. Reactive mesothelial cells are a consistent population in body cavity fluid, and these are the most versatile cells in the body. Due to the specific environment of the body cavity, the exfoliated reactive mesothelial cells may show significant morphologic overlap with the morphology of cancer cells. With a focus on the differential points between reactive mesothelial cells and metastatic adenocarcinoma cells, the practical diagnostic approaches, the diagnostic clues and the pitfalls to achieve a correct diagnosis are presented in this review.

Internal neoplasia in the horse with special regard to the cytology of body cavity fluid

Internal neoplasia in the horse with special regard to the cytology of body cavity fluid

Hydration abnormalities in Nigerian patients on chronic hemodialysis

The state of hydration affects the outcomes of chronic dialysis. Bioelectrical impedance analysis (BIA) provides estimates of body water (V), extracellular volume (ECFV), and fat-free mass (FFM) that allow characterization of hydration. We compared single-frequency BIA measurements before and after 14 hemodialysis sessions in 10 Nigerian patients (6 men, 4 women; 44+/-7 years old) with clinical evaluation (weight removed during dialysis, presence of edema) and with estimates of body water obtained by the Watson, Chertow, and Chumlea anthropometric formulas. Predialysis and postdialysis values of body water did not differ between BIA and anthropometric estimates. However, only the BIA estimate of the change in body water during dialysis (-0.8+/-2.9 L) did not differ from the corresponding change in body weight (-1.3+/-3.0 kg), while anthropometric estimates of the change in body water were significantly lower, approximately one-third of the change in weight. Bioelectrical impedance analysis correctly detected the intradialytic change in body water content (the ratio V/Weight) in 79% of the cases, while anthropometric formula estimates of the same change were erroneous in each case. Compared with patients with clinical postdialysis euvolemia (n=7), those with postdialysis edema (n=5) had higher values of postdialysis BIA ratios V/FFM (0.77+/-0.01 vs. 0.72+/-0.03, p<0.01) and ECFV/V (0.53+/-0.02 vs. 0.47+/-0.06, p<0.05), respectively. Bioelectrical impedance analysis appeared to underestimate body water and extracellular volume in a patient with massive ascites and bilateral pleural effusions. Anthropometric formulas are not appropriate for evaluating the state of hydration in patients on chronic hemodialysis. In contrast, BIA provides estimates of hydration agreeing with clinical estimates in the same patients, although it tends to underestimate body water and extracellular volume in patients with large collections of fluid in central body cavities.

Comments on “Experimental determination of net protein charge, [A]tot, and Ka of nonvolatile buffers in bird plasma”

LETTERS TO THE EDITORComments on “Experimental determination of net protein charge, [A]tot, and Ka of nonvolatile buffers in bird plasma”Jeanne W. GeorgeJeanne W. GeorgePublished Online:01 Jun 2007https://doi.org/10.1152/japplphysiol.00108.2007MoreSectionsPDF (29 KB)Download PDF ToolsExport citationAdd to favoritesGet permissionsTrack citations ShareShare onFacebookTwitterLinkedInWeChat to the editor: I am writing to express some concerns about the article by Stämpfli et al. (7), “Experimental determination of net protein charge, [A]tot, and Ka of nonvolatile buffers in bird plasma.”First, it appears that the BCG dye binding technique was used to determine albumin concentration. This technique has not been validated for avian albumin and does not correlate with electrophoretic results (4). Electrophoretic determination of albumin concentration would have been a better choice of measurement. I think the authors need to qualify their statements about indexing [A]tot to albumin concentration, as their measurements may have been in error.Second, the authors use an incorrect term, “total solids,” to describe the readings obtained from a Leica Vet 360 refractometer. They were actually measuring total protein by a total solids technique. The term total solids refers to all solutes in plasma and is ∼15–20 g/l higher than the protein concentration for most mammalian samples. Refractometers measure protein by the angle of light produced by refraction caused by the total solids in the sample and then subtracting the expected amount of refraction from non-protein solids in the sample. The Leica Vet 360 includes that calculation and has protein (g/dl) listed clearly on the scales in the internal reticle (2).Refractometers have been reported to give higher results than the biuret technique for avian samples for various species of birds (4–6). That discrepancy may be due to relatively higher glucose or other non-protein solids in the plasma of many avian species (4). Pigeons have higher glucose concentrations in their plasma than most mammals, perhaps contributing to higher protein results by refractometry reported by the authors (1).Lastly, I believe Ref. 12 is incorrect (3). That article does not contain any information about comparative accuracy of refractometry in various species. Perhaps they meant to cite a review article written by me containing that information (2).REFERENCES1 Gayathri KL, Shenoy KB, Hegde SN. Blood profile of pigeons (Columba livia) during growth and breeding. Comp Biochem Physiol A Mol Integr Physiol 138: 187–192, 2004.Crossref | ISI | Google Scholar2 George JW. The usefulness and limitations of hand-held refractometers in veterinary laboratory medicine: an historical and technical review. Vet Clin Pathol 30: 201–210, 2001.Crossref | ISI | Google Scholar3 George JW, O'Neill SL. Comparison of refractometer and biuret methods for total protein measurement in body cavity fluids. Vet Clin Pathol 30: 16–18, 2001.Crossref | ISI | Google Scholar4 Harr KE. Clinical chemistry of companion avian species: a review. Vet Clin Pathol 31: 140–151, 2002.Crossref | ISI | Google Scholar5 Lumeij JT, Maclean B. Total protein determination in pigeon plasma and serum: comparison of refractometric methods with biuret method. J Avian Med Surg 10: 150–152, 1996.Google Scholar6 Spagnolo V, Crippa V, Marzia A, Sartorelli P. Reference intervals for hematologic and biochemical constituents and protein electrophoretic fractions in captive common buzzards (Buteo buteo). Vet Clin Pathol 35: 82–87, 2006.Crossref | ISI | Google Scholar7 Stämpfli H, Taylor M, McNicoll C, Gancz AY, Constable PD. Experimental determination of net protein charge, [A]tot, and Ka of nonvolatile buffers in bird plasma. J Appl Physiol 100: 1831–1836, 2006.Link | ISI | Google ScholarAUTHOR NOTESAddress for reprint requests and other correspondence: J. W. George, 1126 Haring Hall, UC-Davis, Davis, CA 95616 (e-mail: [email protected]) Download PDF Previous Back to Top Next FiguresReferencesRelatedInformationCited ByReply to GeorgeHenry Stämpfli, Michael Taylor, Carl McNicoll, Ady Y. Gancz, and Peter D. Constable1 June 2007 | Journal of Applied Physiology, Vol. 102, No. 6 More from this issue > Volume 102Issue 6June 2007Pages 2416-2416 Copyright & PermissionsCopyright © 2007 the American Physiological Societyhttps://doi.org/10.1152/japplphysiol.00108.2007PubMed17551115History Published online 1 June 2007 Published in print 1 June 2007 Metrics

Heparanase expression correlates with poor survival in metastatic ovarian carcinoma

To analyze the expression of Heparanase, an enzyme involved in cancer metastasis and angiogenesis, in ovarian and breast carcinoma cells in effusions. Heparanase protein expression was analyzed in malignant effusions from ovarian (=200) and breast (=41) carcinoma patients using immunocytochemistry. The levels of secreted heparanase were analyzed in 45 effusion supernatants using a newly established ELISA test. Heparanase expression levels were analyzed for clinical significance. Heparanase was expressed at the cell membrane in 106/200 (53%) ovarian and 22/41 (54%) breast carcinomas. Cytoplasmic expression was found in 180/200 (90%) ovarian and 26/41 (63%) breast carcinomas. Reactive mesothelial cells showed frequent cytoplasmic, but not membrane expression. ELISA showed secreted heparanase in all 45 analyzed effusions. Higher levels were detected in peritoneal compared to pleural effusions (p=0.031). In univariate survival analysis of ovarian carcinoma patients with post-chemotherapy effusions, membrane expression in >5% of tumor cells correlated with shorter overall survival (OS, p=0.013). FIGO stage (p=0.03 for all patients, p=0.045 for those with post-chemotherapy specimens) and response to first-line chemotherapy (p<0.0001 for all patients, p=0.049 for those with post-chemotherapy specimens) were the clinical parameters related to OS. In Cox analysis of this subset of patients, heparanase expression (p=0.02) and response to chemotherapy (p=0.049) were independent predictors of poor OS. Heparanase expression did not correlate with survival in breast carcinoma. Our data show that heparanase is frequently expressed in metastatic gynecologic adenocarcinomas, and that it is secreted into the effusion fluid in body cavities. The correlation between heparanase expression and poor survival in ovarian carcinoma suggests a role for this molecule in ovarian cancer metastasis and supports its role as a marker of aggressive clinical behavior at disease recurrence.