bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of Early Ileal Invasion by Salmonella enterica Serovars Choleraesuis and Typhimurium
Authors: Meyerholz D. K. , Stabel T. J.
Published: June 2003

The mechanisms of Salmonella serovar-host specificity are not well defined. Pig ileal loops were used to compare phenotypic differences in early cellular invasion between non-host-adapted Salmonella serovar Typhimurium (SsT) and host-adapted Salmonella serovar Choleraesuis (SsC). By 10 minutes postinoculation, both serovars invaded a small number of M cells, enterocytes, and goblet cells. Multiple SsC organisms (up to 6 per cell) simultaneously invaded M cells, whereas SsT often invaded as one to two organisms per M cell. Internalization of both serovars resulted in vacuoles containing a single bacterium. The follicle-associated epithelium (FAE) of SsC-inoculated loops responded with more filopodia and lamellipodia although exhibiting less cell swelling than SsT. Additionally, SsT showed an enhanced affinity for sites of cell extrusion compared with SsC at 60 minutes. These results suggest: 1) both SsC and SsT exhibit non-cell-specific invasion as early as 10 minutes postinoculation, 2) Salmonella serovars exhibit differences in early invasion of FAE and M cells, and 3) cells undergoing extrusion may provide a site for preferential adherence by SsT and SsC.
Immunohistochemical Localization of Clostridium perfringens β2-Toxin in the Gastrointestinal Tract of Horses
Authors: Bacciarini L. N. , Boerlin P. , Straub R. , Frey J. , Gröne A.
Published: June 2003

Clostridia-associated intestinal disease in horses was generally reported to be due to infection with Clostridium perfringens type A, which harbors the cpa-encoded α-toxin. A recent study demonstrated a high incidence of β2-toxigenic C. perfringens in horses suffering or dying from typhlocolitis, suggesting that this novel type of C. perfringens might play an important role in typhlocolitis and possibly other equine intestinal diseases. A retrospective study was conducted to assess the presence of the β2-toxin in tissues of the equine gastrointestinal tract. Monospecific polyclonal antibodies against recombinant β2-toxin were produced in rabbits and used to demonstrate the β2-toxin in sections of the gastrointestinal tract by immunohistochemical methods. Sections from 69 horses were stained and β2-toxin was observed immunohistochemically in 40 animals. Sections from the stomach, small intestine, and large intestine were positive. Immunopositivity for β2-toxin was significantly associated with presence of β2-toxigenic bacteria. This investigation demonstrates local production of β2-toxin and suggests that immunohistochemistry using antitoxin antibodies represents a useful diagnostic method in those cases where isolation of bacteria and polymerase chain reaction typing is not feasible. Although the association between the presence of β2-toxin and development of gastrointestinal disease in horses remains uncertain, the findings of this study indicate that the potential causal relationship warrants further investigation.
Matrix Metalloproteinase-2 and −9 Involvement in Canine Tumors
Authors: Loukopoulos P. , Mungall B. A. , Straw R. C. , Thornton J. R. , Robinson W. F.
Published: June 2003

Matrix metalloproteinases (MMPs) are a family of enzymes implicated in the degradation and remodeling of extracellular matrix and in vascularization. They are also involved in pathologic processes such as tumor invasion and metastasis in experimental cancer models and in human malignancies. We used gelatin zymography and immunohistochemistry to determine whether MMP-2 and MMP-9 are present in canine tumors and normal tissues and whether MMP production correlates with clinicopathologic parameters of prognostic importance. High levels of pro-MMP-9, pro-MMP-2, and active MMP-2 were detected in most canine tumors. Significantly higher MMP levels were measured in canine tumors than in nontumors, malignancies had higher MMP levels than benign tumors, and sarcomas had higher active MMP-2 than carcinomas. Cartilaginous tumors produced higher MMP levels than did nonsarcomatous malignancies, benign tumors, and normal tissues, and significantly greater MMP-2 than osteosarcomas and fibrosarcomas. Pro-MMP-9 production correlated with the histologic grade of osteosarcomas. The 62-kd form of active MMP-2 was detected only in high-grade, p53-positive, metastatic malignancies. Zymography proved to be a sensitive and quantitative technique for the assessment of MMP presence but has the limitation of requiring fresh tissue; immunohistochemistry is qualitative and comparatively insensitive but could be of value in archival studies. MMP presence was shown in a range of canine tumors, and their link to tumor type and grade was demonstrated for the first time. This study will allow a substantially improved evaluation of veterinary cancer patients and provides baseline information necessary for the design of clinical trials targeting MMPs.
A Morphologic and Immunohistochemical Study of the Bronchus-associated Lymphoid Tissue of Pigs Naturally Infected with Mycoplasma hyopneumoniae
Authors: Sarradell J. , Andrada M. , Ramírez A. S. , Fernández A. , Gómez-Villamandos J. C. , Jover A. , Lorenzo H. , Herráez P. , Rodríguez F.
Published: June 2003

Porcine enzootic pneumonia (PEN), caused by Mycoplasma hyopneumoniae (Mh), has been described in pigs in all geographic areas. The disease is characterized by high morbidity and low mortality rates in intensive swine production systems. A morphologic and immunohistochemical study was done to determine the cellular populations present in lung parenchyma of infected pigs, with special attention to the bronchus-associated lymphoid tissue (BALT). Polyclonal and monoclonal antibodies were used for the detection of antigens of Mh, T lymphocytes (CD3+, CD4+, and CD8+), IgG+ or IgA+ lymphocytes, and cells containing lysozyme, S-100 protein, major histocompatibility complex class II antigen or myeloid-histiocyte antigen. Findings in lung tissues associated with Mh infection were catarrhal bronchointerstitial pneumonia, with infiltration of inflammatory cells in the lamina propria of bronchi and bronchioles and alveolar septa. Hyperplasia of mononuclear cells in the BALT areas was the most significant histologic change. The BALT showed a high morphologic and cellular organization. Macrophages and B lymphocytes were the main cellular components of germinal centers. T lymphocytes were primarily located in perifollicular areas of the BALT, lamina propria and within the airway epithelium, and plasma cells containing IgG or IgA at the periphery of the BALT, in the lamina propria of bronchi and bronchioles, in alveolar septa, and around bronchial submucosal glands. The hyperplastic BALT in PEN cases consisted of macrophages, dendritic cells, T and B lymphocytes, and IgG+ and IgA+ plasma cells. CD4+ cells predominated over CD8+ cells. Local humoral immunity appears to play an important role in the infection.
Fatal Herpes simplex Infection in a Group of Common Marmosets (Callithrix jacchus)
Authors: Mätz-Rensing K. , Jentsch K. D. , Rensing S. , Langenhuyzen S. , Verschoor E. , Niphuis H. , Kaup F.-J.
Published: June 2003

An outbreak of classical herpetic infection causing vesicoulcerative stomatitis in a family group (eight animals) of Callithrix jacchus is described. In all eight infected animals, human herpesvirus 1 (HHV-1) was identified as the causative agent. This was confirmed by histologic, immunohistologic, and molecular biologic investigations, as well as by virus isolation. The clinical picture, the macroscopic appearance, and the histologic results indicated a herpes infection as the cause of mortality. Alterations of the oral mucous membranes were erosive to ulcerative with typical intranuclear inclusions. Immunohistologic and molecular biologic techniques clearly identified the HHV-1 virus and excluded other possible primate herpesviruses such as B-virus, SA8, HVP-2, and Herpes tamarinus. The significance of this herpesvirus infection for colony management is discussed.
p63: A Novel Myoepithelial Cell Marker in Canine Mammary Tissues
Authors: Gama A. , Alves A. , Gartner F. , Schmitt F.
Published: June 2003

Several immunohistochemical markers have been used to demonstrate the presence of myoepithelial cells in order to determine their role in the histogenesis of mammary tumors. p63, a recently characterized p53 homologue, is consistently expressed in myoepithelial cells of the human breast; however, no assessment of its immunoreactivity has been reported so far in canine mammary tissues. We investigated p63 immunohistochemical expression, as a novel myoepithelial cell nuclear marker, in 81 samples of normal ( n = 2), hyperplastic ( n = 11), and neoplastic ( n = 68) canine mammary tissues. Myoepithelial phenotype was confirmed by using complementary monoclonal antibodies: alpha-smooth muscle actin, cytokeratin 14, cytokeratin AE1/AE3, and vimentin. p63 expression was observed in 91.4% (74/81) of the samples evaluated. Normal mammary glands, mammary hyperplasias, and benign tumors showed 100% immunoreactivity, with p63 expression restricted to myoepithelial cell nuclei. In general, benign mixed tumors showed a basal cell compartment immunoreactive to p63, with a gradual decrease of its expression during myoepithelial transformation. p63 expression was found in 72% of malignant tumors, allowing myoepithelial or basal cell identification in spindle-cell carcinomas (2/2), tubulopapillary carcinomas (8/9), solid carcinomas (7/10), and carcinosarcomas (1/3). The osteosarcoma analyzed was p63 negative. In our series, stromal components were consistently nonreactive to p63. In conclusion, the present study reveals p63 as a sensitive and highly specific marker of myoepithelial cells in canine mammary tissues, and the authors suggest p63 as an additional marker for defining myoepithelial histogenesis.
Immunopathogenesis of Experimentally Induced Proliferative Enteropathy in Pigs
Authors: Macintyre N. , Smith D. G. E. , Shaw D. J. , Thomson J. R. , Rhind S. M.
Published: June 2003

To characterize the immune response associated with Lawsonia intracellularis infection, twenty-eight, 7-week-old pigs were dosed orally with a pure culture of L. intracellularis. Animals were killed 3, 7, 14, 21, 28, 35, and 42 days postinfection. Light microscopic studies were undertaken to immunophenotype the immunologic response using specific antibodies to T-cell subsets (CD3, CD4, and CD8), B cells, major histocompatibility complex class II, cadherin, and macrophages over the course of time. The results indicate that there is a direct association between the presence of L. intracellularis and reduced T-cell and B-cell numbers. For the first time, this provides evidence of the presence of an immunosuppressive mechanism operating in this disease. Furthermore, macrophage marker studies indicated that macrophages may play a more complex and significant role in the disease process than has been previously reported, with activated macrophages accumulating in infected hyperplastic crypts.
Follicular Stem Cell Carcinoma: Histologic, Immunohistochemical, Ultrastructural, and Clinical Characterization in 30 Dogs
Authors: Mikaelian I. , Wong V.
Published: June 2003

Diagnostic records of 30 primary and one metastatic follicular stem cell carcinomas in 30 dogs were reviewed. Neoplastic cells had a clear cytoplasm and formed lobules and nests surrounded by a basement membrane. Trichoepitheliomatous and apocrine differentiations were noted in 22 of 30 (73%) and 21 of 30 (70%) primary tumors, respectively. Glycogen was present in 20 of 20 (100%) tumors tested, suggesting tricholemmal differentiation. Antibodies against AE1/AE3 cytokeratin, vimentin, and melanA/MART1 stained 29 of 30 (97%), 29 of 30 (97%), and 12 of 27 (44%) primary tumors, respectively. Small amounts of melanin were identified in 14 primary tumors, either on the hematoxylin and eosin-stained section ( n = 6), or on the Fontana-stained section ( n = 8 of 14). Ultrastructural features of neoplastic cells included cell junction complexes, swollen mitochondria, neuroendocrine-like granules, and intracytoplasmic non-membrane-bound accumulation of proteinaceous material. Features of this neoplasm are consistent with a follicular stem cell origin. Follow-up information was available for eight dogs. Metastases developed in the draining lymph node at the time of excision of the primary tumor ( n = 1) or subsequently ( n = 3).
Parapoxvirus Infection in Harbor Seals (Phoca vitulina) from the German North Sea
Authors: Müller G. , Gröters S. , Siebert U. , Rosenberger T. , Driver J. , König M. , Becher P. , Hetzel U. , Baumgärtner W.
Published: June 2003

In the summer of 2000, proliferative lesions of the skin and oral mucosa were observed in 26 young harbor seals ( Phoca vitulina) from a rehabilitation center in Schleswig-Holstein, Germany. Verrucose, roundish nodules, approximately 1–2 cm in diameter, were presented in the oral cavity, especially on the tongue. Some animals developed similarly sized spherical dermal elevations with ulceration on flippers, chest, neck, and perineum. Necropsy of one animal showed multifocal, verrucose nodules in the oral cavity and a mild tonsillitis. Histologically, the nodules were characterized by ballooning degeneration of the outer parts of the spiny layer and stratum granulosum, with large eosinophilic cytoplasmic inclusions and a perivascular to interstitial lymphohistiocytic infiltration accompanied by fibroblastic proliferation and neovascularization. Negative staining of mucosal tissue homogenates demonstrated parapoxvirus-like particles. The presence of parapoxvirus was confirmed by polymerase chain reaction, using primers specific for parapoxvirus of ungulates. By in situ hybridization, using a parapox-specific, digoxigenin-labeled DNA probe, abundant parapoxvirus DNA-positive epithelial cells were detected in the stratum granulosum and the outer parts of the spiny layer. There was no parapoxvirus-positive signal in the adjacent submucosa. Although DNA analysis revealed that the causative agent can clearly be distinct from terrestrial parapoxviruses, lesions resembled parapoxvirus infections in other terrestrial species, and the pattern of virus DNA distribution indicated a direct effect of the virus on keratinocytes. In contrast, changes in the corium may be considered an indirect response mediated by the virus or the immune system.
Cyclooxygenase-1 and −2 Expression in Urothelial Carcinomas of the Urinary Bladder in Cows
Authors: Borzacchiello G. , Ambrosio V. , Galati P. , Perillo A. , Roperto F.
Published: June 2003

Bovine urinary bladder tumors occur frequently in animals suffering from chronic enzootic hematuria because of prolonged ingestion of bracken fern ( Pteridium spp.). Cyclooxygenase (COX) genes (COX-1 and COX-2) are known to be involved in the carcinogenesis of human and some animal urothelial tumors. The aim of the present study was to investigate COX-1 and COX-2 expression by immunohistochemical methods in 20 bovine urothelial carcinomas collected at public slaughterhouses from cows that had been suffering from chronic enzootic hematuria. COX-1 immunostaining was identified intracytoplasmically in normal urothelium and in 15 of 20 neoplastic specimens. COX-1 immunosignal in the tumor cells was either absent or weak. COX-2 was also expressed intracytoplasmically in 17 of 20 urothelial carcinomas. Moderate to intense COX-2 labeling was detected in both noninvasive and invasive urothelial carcinomas. Coexpression of both enzyme isoforms was also revealed by confocal laser scanning microscopic investigations. This study indicates that COX-2 is overexpressed in naturally occurring urothelial carcinomas of cows.
Ultrastructural and Immunologic Characteristics of Mouse × Cattle Xenogeneic Hybridomas Originating from Bovine Leukemia Virus-infected Cattle
Authors: Saini S. S. , Kaushik A. , Basrur P. K. , Yamashiro S.
Published: June 2003

Nine percent of xenogeneic hybridomas originating from a bovine leukemia virus (BLV)-infected cow secreted monoclonal IgM antibodies with multispecific reactivity. Similar reactivity was evident in some antibodies with an unusually long (>50 amino acids) third complementarity-determining region of the heavy chain. Electron microscopy of hybridomas demonstrated the presence of c-type virus particles consistent with polymerase chain reaction detection of BLV env gene. Some hybridomas contained dilated rough endoplasmic reticulum and cisternae filled with moderately electron-dense granular substance compatible with plasma cells at presecretory stage. The number of chromosomes in xenogeneic hybridomas corresponded to the sum total of mouse and bovine chromosomes. None of the hybridomas showed polyploidy. The immunochemical and genetic analysis of stable bovine immunoglobulin-secreting xenogeneic hybridomas confirms that BLV infection causes polyclonal B cell activation regardless of antigen specificity. Presence of c-type particles in hybridomas suggests that T cell-derived cytokines are not required for sustained BLV expression.
Immunohistochemical Analysis of Two Strains of Lion (Panthera leo)-adapted Canine Distemper Virus in Ferrets (Mustela putorius furo)
Authors: Stanton J. B. , Givens L. , Evermann J. F. , Brown C. C.
Published: June 2003

Canine distemper virus (CDV) caused epizootics in lions ( Panthera leo) in Tanzania's Serengeti National Park in 1994 and in captive lions and other Panthera spp. in the USA in 1991–1992. In this study, immunohistochemistry was used to compare viral distribution in tissues collected from ferrets ( Mustela putorius furo) inoculated with one of the two lion-derived CDV isolates, either from Serengeti (A94-11/13) or from California (A92-27/20). The California isolate resulted in severe morbidity in all nine ferrets, whereas the Serengeti isolate resulted in severe morbidity in five of the nine ferrets. A slightly higher proportion of infected cells was found in many tissues in the Serengeti isolate-inoculated ferrets. These findings indicate that the pathogenicity of the California isolate is not directly related to the number of infected cells.
Cardiomyopathy Associated with Angiomatous Pheochromocytoma in a Rhesus Macaque (Macaca mulatta)
Authors: Vogel P. , Fritz D.
Published: June 2003

A 24-year-old female rhesus macaque ( Macaca mulatta) presented with a clinical history of chronic heart disease and prolonged recovery from sedation and anesthesia. At necropsy, the heart was markedly enlarged, with thinned ventricular walls, dilated chambers, and severe left atrioventricular valvular insufficiency. The ventricular walls contained numerous narrow, pale, often coalescing streaks that often extended along vessels into the deeper myocardium. Histologically, bands of interstitial fibrosis in the heart were associated with areas of myofiber atrophy, myofibril degeneration and loss, and inflammation. The left adrenal gland contained a 2 × 1 × 0.5 cm tumor with an unusual angiomatous pattern consisting largely of blood-filled sinusoids lined by one to four layers of low cuboidal to polyhedral tumor cells. In most sinusoids, the tumor cells appeared to be in direct contact with blood, although endothelial cells lined some sinusoids. Ultrastructurally, the tumor cells contained abundant electron-dense cytoplasmic granules. These granules were argyrophilic by Sevier-Munger staining and intensely immunoreactive for synaptophysin, chromogranin A, neuron-specific enolase, and S-100. These findings helped confirm the diagnosis of angiomatous pheochromocytoma. The heart lesions match those reported for catecholamine cardiomyopathy in other species.
Effect of co‐administration of retinoids on rat embryo development in vitro
Authors: Ritchie Helen E. , Brown‐Woodman Patricia D. , Korabelnikoff Ann
Published: June 2003

BACKGROUND

Excess retinyl palmitate (vitamin A) induces a variety of malformations in many mammalian species, yet retinyl palmitate is not thought to be the proximate teratogen. Many metabolites are generated after oral dosing several of which are, individually teratogenic. It is not known whether these metabolites when present simultaneously interact in an additive or synergistic manner.

METHODS

Whole rat embryos were cultured with retinol, all‐trans retinoic acid (tRA), 4‐oxo all‐trans retinoic acid (4otRA) or 9‐cis retinoic acid (9cRA). Retinoids were added singly or in paired combinations at the start of culture and embryos were examined for evidence of dysmorphogenesis at the end of culture. Retinoids were added at both subthreshold and supra‐threshold concentrations.

RESULTS

Co‐administration of subthreshold concentrations of tRA and 4otRA, retinol and tRA or 4otRA and 4otRA and 9cRA resulted in a greater frequency of abnormal embryos compared to single administration of each retinoid.

CONCLUSIONS

Knowledge of the way in which retinyl palmitate metabolites interact should refine our ability to predict risk from a potentially teratogenic event e.g., accidental overdose with RP during human pregnancy. Birth Defects Research (Part A) 67444–451, 2003. © 2003 Wiley‐Liss, Inc.
Papillary microcarcinoma of the thyroid—Prognostic significance of lymph node metastasis and multifocality
Authors: Chow Sin‐Ming , Law Stephen C. K. , Chan John K. C. , Au Siu‐Kie , Yau Stephen , Lau Wai‐Hon
Published: June 2003

BACKGROUND

It is known that patients with papillary microcarcinoma (PMC) of the thyroid gland have a very favorable prognosis. The rising incidence of PMC among papillary thyroid carcinoma (PTC) necessitates the identification of prognostic factors and the formulation of treatment protocols.

METHODS

The authors conducted a retrospective analysis of 203 patients with PMC who were diagnosed on or before 1999 and were treated at the Department of Clinical Oncology, Queen Elizabeth Hospital, Hong Kong.

RESULTS

The cause specific survival, locoregional (LR) failure free survival, and distant metastases failure free survival rates at 10 years were 100%, 92.1%, and 97.1%, respectively. Five patients had lung metastases; 2 patients died of their metastases 12.9 years and 14.8 years after diagnosis, and 3 patients achieved clinical remission after radioiodine (RAI) treatment. Twelve patients had LR recurrences. Patients with LR recurrence were highly salvageable with a combination of surgery, RAI treatment, and external radiotherapy; all but one (who refused treatment) were alive without disease at last follow‐up. Multivariate analyses did not reveal any independent prognostic factor for survival. The risk of cervical lymph node (LN) recurrence increased 6.2‐fold (P= 0.01) and 5.6‐fold (P= 0.02) when LN metastases and multifocal disease were present at diagnosis. RAI ablation reduced the LN recurrence rate to 0.27 (P= 0.04). The presence of LN metastasis increased the rate of distant metastasis 11.2‐fold (P= 0.03). Age was not a significant factor in predicting disease recurrence or survival. Subdivision by tumor sizes ≤ 5 mm and > 5 mm did not affect the outcome, but no patient with tumors ≤ 5 mm had mortality related to PMC.

CONCLUSIONS

Despite the overall excellent prognosis for patients with PMC, PMC was associated with a 1.0% disease‐related mortality rate, a 5.0% LN recurrence rate, and a 2.5% distant metastasis rate. Therefore, the treatment of patients with PMC should be no different from the treatment of patients with conventional PTC: i.e., complete surgery with consideration for RAI and/or external radiation therapy if poor prognostic factors are present. Cancer 2003;98:31–40. © 2003 American Cancer Society.

DOI 10.1002/cncr.11442
Evidence of a preferred molecular pathway in patients with synchronous colorectal cancer
Authors: Dykes Sharon L. , Qui Hongming , Rothenberger David A. , García‐Aguilar Julio
Published: June 2003

BACKGROUND

A small proportion of patients with colorectal carcinoma (CRC) have synchronous tumors at the time of diagnosis. A subset of sporadic CRCs display microsatellite instability (MSI) that is associated with MLH1 silencing due to promoter methylation. In the current study, the authors investigated the proportion of tumors with MSI in patients with synchronous colorectal carcinoma (SCRC) and the concordance in MSI status among tumors in a given individual. In addition, the authors examined MLH1 and MSH2 expression and MLH1 promoter methylation in SCRCs.

METHODS

The current study included 77 patients, with a combined total of 170 invasive SCRCs, who were identified from a database of 2884 patients with CRC. Instability was determined by polymerase chain reaction (PCR) amplification using a set of five markers. Tumors that were unstable at two or more markers were considered unstable (MSI); otherwise, they were considered microsatellite stable (MSS). Expression of MLH1 and MSH2 was determined by immunohistochemistry. Methylation of the MLH1 gene promoter was determined by a methylation‐specific PCR assay. Statistical comparisons were made using the chi‐square test or the Student t test.

RESULTS

Of the 77 patients in the study, 21 (27%) had a family history of hereditary nonpolyposis colon carcinoma–related malignancy, but none fulfilled the Amsterdam II criteria. Fifty‐four of 170 tumors (32%) were found to be MSI. Patients with MSI tumors were older and more frequently female. All but 1 MSI tumor lacked expression of MLH1 (n = 44) or MSH2 (n = 8), or both (n = 1). All MLH1‐negative tumors, compared with only 3 MLH1‐positive tumors, were methylated at the MLH1 promoter. Most patients (n = 67; 87%) had either all MSS tumors (n = 48; 62%) or all MSI tumors (n = 19; 25%); 10 patients (13%) had both MSS and MSI tumors. The observed MSI/MSS distribution was significantly different from the distribution expected based on an assumption of independence (P < 0.0001).

CONCLUSIONS

There is a strong concordance in MSI/MSS status among tumors in the same individual. This finding suggests that the tumors in patients with SCRC develop along a preferred molecular pathway. Cancer 2003;98:48–54. © 2003 American Cancer Society.

DOI 10.1002/cncr.11445
Distribution of HER‐2 overexpression in ovarian carcinoma tissue and its prognostic value in patients with ovarian carcinoma
Authors: Høgdall Estrid V. S. , Christensen Lise , Kjaer Susanne K. , Blaakaer Jan , Bock Johannes E. , Glud Eva , Nørgaard‐Pedersen Bent , Høgdall Claus K.
Published: June 2003

BACKGROUND

The HER‐2 (Human Epidermal Growth factor receptor‐2, also known as c‐erb‐2/neu) protooncogene encodes a transmembrane receptor protein,Mr185,000. Studies have shown that the HER‐2 oncogene is overexpressed in approximately 25–30% of ovarian carcinoma (OC) cases, but to the authors' knowledge, to date no consensus regarding overexpression and prognosis has been possible. The objective of the current study was first to analyze the presence of HER‐2 overexpression in tissue from Danish OC patients and correlate the distribution of HER‐2 overexpression with clinical and biochemical data and second to investigate the value of HER‐2 overexpression as a prognostic marker in OC and to compare this value with the prognostic value of other biochemical markers.

METHODS

The study population was comprised of the first 181 patients diagnosed with epithelial OC who were included in the MALOVA study. The staining procedure for HER‐2 overexpression was performed using the p185 antibody.

RESULTS

HER‐2 overexpression was found in 95 of the 181 investigated cases (52.5%), in which 71 carcinomas (39.2%) were weakly positive (1+) and 24 carcinomas (13.3%) were moderately (2+) to intensely positive (3+). Increased HER‐2 expression was found to be correlated with reduced survival. Significant differences in survival between patients with (1+, 2+, and 3+) and those without HER‐2 overexpression were found for patients with International Federation of Gynecology and Obstetrics (FIGO) Stage I, Stage III, and Stage III/IV OC (Stage I:P= 0.021; Stage III:P= 0.0078; and Stage III/IV:P= 0.0054). Multivariate survival analyses including all 181 OC patients demonstrated that HER‐2 overexpression is a prognostic marker (P= 0.003) together with disease stage, serum tetranectin level, and patient age. For patients with Stage III OC, the only independent prognostic factors detected were HER‐2 overexpression (P= 0.009) and serum tetranectin level (P≤ 0.0001).

CONCLUSIONS

The results of the current study show that HER‐2 overexpression has prognostic value both in univariate and multivariate survival analyses. Therefore, the clinical relevance of this observation should be established conclusively by therapy that targets HER‐2 in a prospective Phase II clinical trial. Cancer 2003;98:66–73. © 2003 American Cancer Society.

DOI 10.1002/cncr.11476
Sudden onset of the blastic phase of chronic myelogenous leukemia
Authors: Kantarjian Hagop , O'Brien Susan , Cortes Jorge , Giles Francis , Thomas Deborah , Kornblau Steven , Shan Jianquin , Beth Rios Mary , Keating Michael , Freireich Emil , Talpaz Moshe
Published: June 2003

BACKGROUND

Therapeutic options in Philadelphia chromosome‐positive chronic myelogenous leukemia (Ph‐positive CML) in chronic phase include allogeneic stem cell transplantation (SCT) and treatments other than allogeneic SCT. The choice of therapy often depends on the mortality risk of allogeneic SCT. Delaying SCT until the disease demonstrates resistance to other strategies (e.g., imatinib mesylate or interferon‐α) entails the risk of the development of a sudden blastic phase, which may not permit enough time to perform a SCT.

METHODS

The current study was performed to define the incidence and patterns of sudden‐onset blastic‐phase CML in relation to the risk inherent in delaying allogeneic SCT. Of a study group of 1286 patients who were referred in early chronic‐phase CML between 1981–2001 and who received therapies other than allogeneic SCT, 1093 patients had regular follow‐up at the study institution. A sudden onset of blastic phase was defined as having its onset within 3 months from a previously documented complete hematologic response.

RESULTS

Among the 1093 patients, 183 patients developed blastic‐phase disease, which was of sudden onset in 46 patients (25%; 4% of the total). The incidence of sudden blastic phase was 0.4% in the first year, 1.8% in the second year, and 2.6% in the third year. Patients with sudden blastic‐phase disease were found to have more often low‐risk features at the time of presentation, including low incidences of blasts and thrombocytosis. Sudden blastic‐phase CML was found to be associated with a higher incidence of lymphoid blastic morphology (67% vs. 22%; P < 0.0001), which resulted in a higher response rate to blastic‐phase therapy (70% vs. 29%; P < 0.0001) and significantly longer survival from the onset of blastic‐phase disease (median of 12 months vs. 6 months; P < 0.01). Twenty‐four of the 42 patients with sudden blastic‐phase disease who were treated at the M. D. Anderson Cancer Center were able to undergo allogeneic SCT in second chronic phrase (n = 13) or transformed phase (n = 11); at the time of last follow‐up, 6 patients were alive without evidence of disease at a median of 18 months.

CONCLUSIONS

The low rates of sudden blastic transformation in the first 3 years of the course of disease for CML and the salvage rate of these patients with allogeneic SCT should be considered in relation to the transplantation mortality in patients with early chronic‐phase disease and the early promising results with imatinib mesylate therapy. Cancer 2003;98:81–5. © 2003 American Cancer Society.

DOI 10.1002/cncr.11477
A Phase I–II trial of polyethylene glycol‐conjugated L‐asparaginase in patients with multiple myeloma
Authors: Agrawal Neeraj R. , Bukowski Ronald M. , Rybicki Lisa A. , Kurtzberg Joanne , Cohen Lewis J. , Hussein Mohamad A.
Published: June 2003

BACKGROUND

Multiple myeloma remains an incurable disease. New agents are needed to improve therapy for patients with this disease. Previous investigators evaluated in vitro sensitivity of myeloma cells to polyethylene glycol‐conjugated L‐asparaginase (PEG‐L‐asparaginase) using the human tumor clonogenic assay. Of the 19 myeloma samples evaluated, 63% were inhibited at 0.075 IU/mL, and 74% were inhibited at 0.75 IU/mL. PEG‐L‐asparaginase is a form of Escherichia coli‐derived L‐asparaginase that is bound covalently to polyethylene glycol. Compared with the native form, it has a longer half‐life and is less likely to cause allergic reactions.

METHODS

The authors conducted a Phase I–II trial using PEG‐L‐asparaginase as a single agent in patients with recurrent and/or refractory multiple myeloma.

RESULTS

Twenty‐two patients received a median of two doses of PEG‐L‐asparaginase. In the 17 patients who are evaluable for response, a complete response was observed in one patient after four doses, and stable disease was observed in eight patients. Progression of disease was the reason for termination from study in the remaining eight patients. The median survival was 31.7 months, with four patients who were alive at 72 months after the start of therapy. Grade 3–4 toxicity was noted by the PEG‐L‐asparaginase 2000 mg/m2 level. Severe allergic reactions were noted only at the highest dose level.

CONCLUSIONS

Current data suggest that the maximal tolerated dose for single agent PEG‐L‐asparaginase in relapse/refractory multiple myeloma patients is 1000 mg/m2 every 4 weeks. We could not identify any correlation between dose, plasma level and response. In this advanced group of patients we noted stable disease and/or reponse in 52% of evaluable patients. PEG‐L‐asparaginase has lower tolerance when used in the standard dosage as a single agent in this group of patients. We therefore recommend further studying of PEG‐L‐asparaginase dose of 1000 mg/m2 on alternate weeks with steroids and/or other immune modulators. Cancer 2003;98:94–9. © 2003 American Cancer Society.
Overexpression of osteopontin is associated with intrahepatic metastasis, early recurrence, and poorer prognosis of surgically resected hepatocellular carcinoma
Authors: Pan Hung‐Wei , Ou Yueh‐Hsing , Peng Shian‐Yang , Liu Shu‐Hsian , Lai Po‐Lin , Lee Po‐Hwaung , Sheu Jin‐Chuan , Chen Chi‐Ling , Hsu Hey‐Chi
Published: June 2003

BACKGROUND

Intrahepatic metastasis via portal vein spread is an important feature and a crucial unfavorable prognostic factor of hepatocellular carcinoma (HCC). To identify the molecular factors for tumor progression, the authors used differential display (DD) to analyze aberrant gene expression in HCC. The goal of the current study was to elucidate the clinicopathologic and prognostic significance of osteopontin (OPN) in HCC progression.

METHODS

OPN mRNA levels, which were increased preferentially in HCC in a DD assay and verified with Northern blotting, were measured in 240 surgically removed, unifocal, primary HCCs using the reverse transcription–polymerase chain reaction at the exponential phase. OPN mRNA expression was correlated with clinicopathologic features, particularly portal vein invasion, early tumor recurrence, and prognosis.

RESULTS

Osteopontin mRNA was overexpressed in 133 tumors (55%). The OPN overexpression was associated closely with α‐fetoprotein elevation (P = 0.001), p53 mutation (P = 0.021), larger tumors (P = 0.002), high‐grade HCC (P < 0.001), late‐stage HCC (P < 0.001), early tumor recurrence and/or metastasis (P = 0.003), and a lower 10‐year survival rate (P = 0.00013). Multivariate analysis revealed that tumor stage and early tumor recurrence were crucial prognostic factors. In early‐stage HCC, which has no vascular invasion and a lower early tumor recurrence than late‐stage HCC, OPN mRNA overexpression predicted a higher early recurrence rate (P = 0.003).

CONCLUSIONS

OPN mRNA overexpression was correlated closely with high‐grade, late‐stage, and early tumor recurrence, which lead to poorer prognosis. Osteopontin overexpression might serve as an unfavorable prognostic factor and a useful marker for predicting early recurrence in early‐stage HCC. Cancer 2003;98:119–27. © 2003 American Cancer Society.

DOI 10.1002/cncr.11487
Cell Adhesion onto Highly Curved Surfaces: One‐Step Immobilization of Human Erythrocyte Membranes on Silica Beads
Authors: Kaufmann Stefan , Tanaka Motomu
Published: July 2003

This paper deals with single‐step, orientation‐selective immobilization of human erythrocyte membranes on bare silica beads with different topographies: 1) solid (nonporous) silica beads with a diameter of 3 μm and 2) porous silica beads with a diameter of 5 μm. Erythrocyte membranes were immobilized onto beads simply by incubation, without sonication or osmotic lysis. Membrane orientation before and after immobilization was identified with two immunofluorescence labels: 1) the extracellular part of glycophorin can be labeled with a first monoclonal antibody and a second polyclonal antibody with fluorescence dyes (outside label), while 2) the cytoplasmic domain of Band 3 can be recognized with a first monoclonal antibody and a second fluorescent polyclonal antibody (inside label). Adherent erythrocytes on the beads all ruptured, inverted the asymmetric orientation of the membrane, and selectively exposed their cytoplasmic domain. The surface topography did not influence the orientation or the amount of immobilized membrane. On the other hand, the fact that no adsorption or rupture of erythrocytes could be observed on planar quartz substrates suggests a significant influence of contact curvature on adhesion energy.
Improvement of the Kleihauer‐Betke test by automated detection of fetal erythrocytes in maternal blood
Authors: Pelikan Denise M. V. , Mesker Wilma E. , Scherjon Sicco A. , Kanhai Humphrey H. H. , Tanke Hans J.
Published: June 2003

BACKGROUND

Reliable detection and quantification of fetal red cells in maternal blood is important in routine obstetric practice. The manual Kleihauer‐Betke test (KBT) is widely used, but it is imprecise and subjective. This study investigated whether automated readout of the KBT could improve sensitivity and accuracy. Methods: Glass slides containing dilutions of fetal red cells in adult blood were prepared and stained by using acid elution. Standard manual evaluation of the KBT was performed for all slides by one investigator. In addition, automated microscopy and image analysis of the same slides were performed, whereby detected fetal cells were reviewed by two independent investigators. Results: Ten replicate measurements of fetal cell numbers showed high reproducibility and very small interobserver and intraobserver variabilities. Typical coefficients of variation were 3–4% for concentrations ranging from 0.001% to 0.1%. The automated KBT showed strong correlation between theoretical and detected concentrations of fetal cells (r2 = 0.999). In the range from 0.0001% to 0.001%, the standard KBT underestimated the fetal cell percentage, whereas the automated KBT was very precise. The correlation between methods was good (r2 = 0.999). Conclusion: Automated readout of the KBT improved accuracy of fetal cell detection in the range from 0.0001% to 1% fetomaternal hemorrhage, particularly when larger numbers of cells were analyzed. Cytometry Part B (Clin. Cytometry) 54B:1–9, 2003. © 2003 Wiley‐Liss, Inc.
Immune consequences of pediatric and adult cardiovascular surgery: Report of the 7th Leipzig workshop
Authors: Tárnok Attila , Emmrich Frank
Published: June 2003

Cardiovascular surgery in children and adults is among the most common types of interventions in the western hemisphere for innate and acquired defects. In the recent decades, the risk of cardiovascular surgery has been reduced substantially. Nevertheless, open heart surgery is risky for the patient and can lead to postoperative complications such as postpericardiotomy syndrome, capillary leak syndrome, or multiple organ failure. To gain further understanding into the response to cardiovascular surgery, it is necessary to join forces from several disciplines of medicine and natural sciences. Interdisciplinarity is the basic concept of the Leipzig Workshop. The consensus of the workshop was that cardiovascular surgery with cardiopulmonary bypass induces a systemic antiinflammatory response due to (a) elimination of activated cells, (b) compensatory reaction to a local proinflammatory responses, (c) interleukin‐10 release, (d) anesthetics and medication, and (e) leukocyte extravasation. The subsequent proinflammatory reaction is the response to surgical trauma modulating the antiinflammatory reaction. Novel therapeutic approaches include the introduction of autologous endothelial progenitor cells from the peripheral blood into the sites of injury. The analysis of immune response and outcome prediction require novel analytical tools that allow fast, accurate, and quantitative determination of the desired parameters in a multiplexed manner (i.e., cytomics), such as flow cytometric microbead array assays and slide‐based cytometry. The major goal is predictive medicine by cytomics, i.e., the individualized risk assessment by analyzing the cytome in combination with sophisticated data pattern recognition. These developments may lead to individualized therapy for the benefit of the patient and cost reduction. Cytometry Part B (Clin. Cytometry) 54B:54–57, 2003. © 2003 Wiley‐Liss, Inc.
Optimization of the chiral resolution of baclofen by capillary electrophoresis using β‐cyclodextrin as the chiral selector
Authors: Ali Imran , Aboul‐Enein Hassan Y.
Published: July 2003

The chiral resolution of baclofen was achieved by capillary electrophoresis using a fused‐silica capillary (60 cm×75 μm ID). The background electrolyte (BGE) was phosphate buffer (pH 7.0, 50 mM)‐acetonitrile (95:5 v/v) containing 10 mM β‐cyclodextrin. The applied voltage was 15 kV. The values of α and Rs were 1.06 and 1.00, respectively. The electrophoretic conditions were optimized varying the pH and the ionic strength of the BGE, concentrations of β‐cyclodextrin and acetonitrile and the applied voltage.
Analytical separations of lanthanides and actinides by capillary electrophoresis
Authors: Janoš Pavel
Published: July 2003

The separation of lanthanide and actinide elements belongs to one of the most challenging tasks of the separation science, due to a great similarity in their physical and chemical properties. The electrophoretic separation can be accomplished in the presence of suitable complex‐forming agents, from which α‐hydroxyisobutyric acid (HIBA) has been used most often. In the most effective capillary electrophoretic mode – capillary zone electrophoresis (CZE) – a complete separation of lanthanide ions can be accomplished within a few minutes. Various electrophoretic methods can be relatively easily adopted for the determinations of individual lanthanide elements in certain kinds of technical materials, concentrates, precursors, etc., where the high speed and low costs of analysis characteristics of capillary electrophoresis (CE) may be advantageously exploited. Electrophoretic techniques may also be employed for speciation studies, especially for examinations of the behavior of actinides in the environment.
Influence of a hedge surrounding bottomland on seasonal soil‐water movement
Authors: Caubel Virginie , Grimaldi Catherine , Merot Philippe , Grimaldi Michel
Published: June 2003

The influence of a hedge surrounding bottomland on soil‐water movement along the hillslope was studied on a plot scale for 28 months. The study was based on the comparison of two transects, one with a hedge, the other without, using mainly a dense grid of tensiometers. The influence of the bottomland hedge was located in the area where tree roots were developed, several metres upslope from the hedge, and could be observed both in the saturated and non‐saturated zone, from May to December. The hedge induced a high rate of soil drying, because of the high evaporative capacity of the trees. We evaluated that water uptake by the hedge during the growing season was at least 100 mm higher than without a hedge. This increased drying rate led to a delayed rewetting of the soils upslope from the hedge in autumn, of about 1 month compared with the situation without a hedge. Several consequences of this delayed rewetting are expected: a delay in the return of subsurface transfer from the hillslope to the riparian zone, a buffering effect of hedges on floods, already observed at the catchment scale, and an increased residence time of pollutants. Copyright © 2003 John Wiley & Sons, Ltd.
Environmental Stress‐Cracking Resistance of LDPE/EVA Blends
Authors: Borisova Bistra , Kressler Jörg
Published: June 2003

Blends of low‐density polyethylene with random copolymers of ethylene and vinyl acetate (PE/EVA) are studied with respect to their environmental stress‐cracking resistance (ESCR) using the Bell‐telephone test. This system shows the shortest time to failure in the ESCR test after annealing at 50 °C in a stress‐cracking agent (Igepal solution) compared with that in the tests conducted at 30 and 70 °C. The increase of the time to failure at 70 °C as compared with that at 50 °C is probably the result of the semicrystalline proportion of EVA melting. Transmission electron microscopy images (see Figure) reveal that EVA particles are molten and deformed in bending direction of the sample at 70 °C in contrast to samples annealed at 50 and 30 °C. TEM pictures of a failed sample during the test conducted at 50 °C indicate that EVA particles can stop crack propagation.

TEM image of PE/EVA‐5.4 after 1 000 h in ESCR test conditions at 70 °C.

magnified image

TEM image of PE/EVA‐5.4 after 1 000 h in ESCR test conditions at 70 °C.
Evidence for hydrochory and the deposition of viable seeds within winter flow‐deposited sediments: the River Dove, Derbyshire, UK
Authors: Goodson J. M. , Gurnell A. M. , Angold P. G. , Morrissey I. P.
Published: June 2003

Previous studies of the influence of hydrochory on plant dispersal have focused on relationships with existing vegetation patterns or litter deposits. River sediment deposition studies have assessed sediment accretion rates, particle size and quality. The link between seed and sediment transport and deposition has been overlooked and never quantified.

This paper presents observations of over‐winter viable seed and sediment deposition on three river margin sites along the River Dove, England. At these sites, 105 paired artificial turf mats were installed on the river bank top, face and toe from October 1999 to March 2000 to collect river‐deposited material. From each pair, one mat was used to determine the dry weight; median particle size; percentage sand, silt, clay and organic content of deposited sediment. The other was used in germination trials to determine the number and species of deposited viable seeds. Topographic surveys and water level measurements supported the estimation of the hydrological characteristics of each mat location.

Few seeds and little sediment were found on mats not inundated by river water. For these inundated, sediment characteristics varied primarily with elevation (relative to local mean river water level during the study period), with less marked between‐site contrasts. The species composition of viable seeds also varied with elevation. Detrended correspondence analysis (DCA) revealed associations between river/wetland‐related seed species and the bank toe samples. Regression analysis revealed significant relationships between seed number, sediment properties and mat elevation. Redundancy analysis (RDA) revealed associations between the species and abundance of viable seeds and elevation, sediment weight and organic matter content. These analyses suggest that sediment and seed deposition by rivers are closely related processes.

The implications of these results for seed remobilization and dispersal and hydrochory within riparian zones are discussed, as are the implications of changes in sediment/seed delivery mechanisms, river flow regimes, and the hydraulic properties of river margins for seeds, sediments and riparian vegetation patterns. Copyright © 2003 John Wiley & Sons, Ltd.