bluuebunny/crossref_metadata_2025 huggingface dataset:
A species‐specific satellite DNA family in the genome of the coffee root‐knot nematode Meloidogyne exigua: application to molecular diagnostics of the parasite
Authors: Randig Onivaldo , Bongiovanni Michel , Carneiro Regina M. D. G. , Sarah Jean‐Louis , Castagnone‐Sereno Philippe
Published: March 2003

A new BglII satellite DNA has been isolated, cloned and sequenced from the coffee root‐knot nematode, Meloidogyne exigua (Nematoda: Tylenchida). It is represented as tandemly repeated sequences with a monomeric unit of 277 bp. The monomers are present at approximately 17 900 copies per haploid genome, and represent about 9.7% of the total genomic DNA. Twenty randomly chosen monomers have been sequenced. The deduced unambiguous consensus sequence is 277 bp long, and displays an A + T content of 54.2%. The monomers are very homogenous in sequence, showing on average 2.4% divergence from their consensus. Therefore, it is hypothesized that this repeated family may have recently appeared in the genome of the nematode, through some extensive amplification burst. Using a cloned monomer as a probe, dot‐blot experiments demonstrated the species‐specific distribution of the BglII satellite DNA. Moreover, squash‐blot assays allowed us to detect single M. exigua individuals, at any developmental stage, and even within root tissues, without the need for preliminary DNA purification. From these results, it is concluded that the procedure described, using the satellite DNA as a sensitive species‐specific probe, should constitute an improved and accurate diagnosis method for the detection and identification of the nematode, which would contribute to the implementation of targeted pest management strategies in all coffee growing countries of South and Central America.
Effect of packaging under vacuum or under nitrogen on the keeping quality of Danedar khoa
Authors: Sharma H K , Singhal R S , Kulkarni P R
Published: March 2003

A khoa variety popularly known as Danedar khoa, which has a very short life under ambient conditions, was studied for its shelf life at room and refrigerated temperatures under controlled conditions after packaging in a flexible material (poster paper/aluminium foil/low‐density polyethylene [LDPE]). The samples were observed periodically for characteristics such as moisture content, peroxide value, titratable acidity and overall acceptability. Samples packed under nitrogen and stored under refrigeration were found to be distinctly superior in all respects.
Effects of starter culture and its exopolysaccharides on the gelation of glucono‐δ‐lactone‐acidified bovine and caprine milk
Authors: Vlahopoulou Ioanna , Bell Alan E , Wilbey R Andrew
Published: March 2003

The effects of the starter culture biomass material and its exopolysaccharides (EPS) on the structure of bovine and caprine acid gels were investigated. Inactivated cell material and/or extracellular polysaccharide, which had been separated from yogurt starter cultures, were added to both types of milk. The milk systems were acidified using glucono‐δ‐lactone (GDL) and the gelation processes were monitored using dynamic (oscillatory) rheological testing. The addition of cell material had no effect on the bovine acid gel, whereas the addition of polysaccharide produced a weaker gel structure. The addition of cell material (> 5 × 108 cells/mL) and polysaccharide (0.35% w/v) weakened the caprine acid gel system. The treated gels were some three to eight times weaker than the untreated samples, suggesting that the starter culture metabolites and especially the EPS may act by interfering with the gel structures, particularly in the cases of milk systems such as caprine yogurt or low protein content bovine yogurt.
Sleeping giants: towards an understanding of the needs of deaf people
Authors: ANDERSON W.
Published: March 2003

Joan* was pre‐lingually deaf, from three generations of deaf people. Today she is outgoing and successful in her job, and a ready communicator with everyone, deaf and hearing alike. Her communicating idioms, with both groups, are rich. She benefited from the twin forces in her childhood of family (all her family were highly attuned to the non‐verbal idioms of communication) and one grandmother (who was hearning, and who provided what came to be seen as teh key intervening role at that time in Joan's development of a second language, the idiom of spoken English).

Her own intellectual drive was the final component in a combination of motivating influences which led her to respond to both the financial and social necessities to gain verbal codes, leading to an ability to read and to understand what she read. She remains totally deaf and she knows that her speaking voice is not understood by strangers.

Frank* is a regular reader of newspapers, at least, that's what you would think if you saw him regularly. He does know certain words (or rather parts of words) and he constructs a story from his understanding of whatever catches his eye.

At his job, which he does with skill, he is confident and earns the trust of colleagues and superiors. He is, however, illiterate. He knows that if he is seen to be ‘reading’ he will sustain the image he has created that he can read: he acts like a literate person.

He is confident and competent in what he knows, which is enough for the regular and recurring nature of the tasks he is asked to perform in his job. He is knowledgeable and defensive about that knowledge but as a accepted at work, but any change or new demand, print‐based, would sweep away his precarious confidence and, probably along with that, his job.
A case for local current‐awareness publications
Authors: MAYHEW JANICE
Published: March 2003

An analysis of two current‐awareness services designed to exploit the journal stock of a small specialist library. The benefits of locally produced publications are identified and some of the omissions and bias in the major indexes and abstracts noted. An attempt is made to identify the workload involved.
The questionnaire—a quality control method for online searching?
Authors: LYON ELIZABETH
Published: March 2003

The quality of the online search service in a university medical school/large hospital library was investigated by user survey. The use of a questionnaire as a tool to measure quality was assessed. The survey indicated that most users were satisfied with the quality of the service. User ratings for all search performance criteria were high and most readers found online searches to be good value for money. Information about user groups, frequency of searching, search applications, pre‐search work and user expectations was obtained. The most important aspect of a computer search to the reader was ‘savings in your time’. Three types of user were described based on search output analyses and reading levels. Some areas where the existing service could be improved were identified. The introduction of a standard questionnaire for monitoring the quality of computer searches and other library services was proposed.
The quality movement in higher education in the United States
Authors: SHIPP BUCHANAN HOLLY
Published: March 2003

Continuous quality improvement (CQI), often implemented as part of an integrated management system called total quality management (TQM), has been institutionalized within many manufacturing, military and service organizations in the USA as a response to declining market share, low productivity and customer complaints about poor quality. Signs and symptoms suggest that higher education has similar problems which are systemic and relate to the quality of higher education, financing, facilities, curriculum and graduates. In the 1990s, the quality movement has begun to spread to the field of education as a means of diagnosing and treating the problems widely recognized as residing in US educational institutions, especially in colleges and universities.

Many business leaders and authors believe that ‘quality is the most important strategic issue facing top management in the 1990s’.1 This belief arises partly due to the fact that managers are beginning to understand the relationship between healthy, high quality organizations and healthy profits. This article traces the recent US quality movement from its roots in manufacturing and the military, its adoption by service institutions, and its more recent application by higher education institutions.
Quality assurance in health sciences literature searching: applying the ISO 9000 quality standard
Authors: CULLEN ROWENA , MASON DAVID
Published: March 2003

Medicine is a literature‐based discipline. Ensuring that the literature review which precedes a significant piece of medical research has met predetermined standards is essential. A list of items reviewed carries no guarantees that all appropriate items have been included in the survey of the literature, or that appropriate sources have been efficiently searched. This would be a matter for concern in any discipline. In medicine it is a matter of life and death. Quality assurance procedures that offer guarantees of the standards built into the process, rather than quality control which measures only outputs, can provide the necessary reassurance. The ISO 9000 quality standard offers a much needed quality assurance process. A methodology for applying the ISO 9000 standard to the task of searching the medical literature is outlined in this paper. A new role for medical librarians in promoting a rigorous methodology in the literature review equal to that of the research it supports is defined.
The value of information supplied to clinicians by health libraries: devising an outcomes–based assessment of the contribution of libraries to clinical decision–making
Authors: URQUHART CHRISTINE , HEPWORTH JOHN
Published: March 2003

The Value project, a study funded by the British Library Research and Development Department and a group of postgraduate medical deans, has examined the effectiveness of NHS library and information services. Clinicians at 11 hospital library sites were asked to describe the purposes for which they needed information, the steps they took to obtain it and to assess the value of the information obtained from interlibrary loan requests, database searches and end‐user searches. A sample of non‐library users and of users of the BMA Dial‐Up MEDLINE service were also asked similar questions. Results show that information did, or would in the future, assist in personal clinical decision‐making and which kinds of decisions were most supported. Information new to clinicians was often provided. Follow‐up interviews provided illustrations of the ways in which developments aimed at more cost‐effective patient care were supported by information obtained from the library. Results are being used to guide the production of a quality assurance toolkit to help libraries audit their services and improve the delivery of information to clinicians.
Information in primary health care
Authors: WOOD FRANCES , PALMER JUDITH , ELLIS DAVID , SIMPSON SHARON , BACIGALUPO RUTH
Published: March 2003

The results of two recently completed studies, one in the Trent Region and the other in the Anglia and Oxford Region, of the information‐related behaviour of general medical practitioners (GPs) and of information activities within the general practices illustrating the information seeking behaviour of GPs, their generation of information, and communication within, to and from the practice. GPs stated that there is a need for more information to enable practices to function within the changing NHS environment. The increasing emphasis on evidence‐based medicine also creates the need for appropriate information sources. The Trent study resulted in guidelines for best information practice, especially the need for each practice to have an information strategy. The Anglia and Oxford Study examined the role which the library and information services could play in supporting primary health care.
Human development: competencies for the 21st century. IFLA CPERT 3rd International Conference on Continuing Professional Education
Authors: BRYANT SUE LACEY
Published: March 2003

In 1996 the Health Libraries Group, wishing to enhance knowledge and support the development of the profession, agreed to provide funds for an annual scholarship for travel to a conference, for research or for a study visit. The Committee also felt it important to encourage applicants to take an active role in securing funds from other sources in order to demonstrate the degree of their commitment to the profession. The scholarship was advertised and the response was encouraging. The first award was made to Sue Lacey Bryant for her application to attend and present a paper at the 3rd International Conference on Continuing Professional Education, held in Copenhagen in August 1997. Part of the requirement is that the successful candidate writes a report, Sue's report follows.

JUDY PALMER Chair: Health Libraries Group 1994–96.
Cutaneous lung tissue heterotopia
Authors: Singer  , Haag  , Anabitarte
Published: March 2003

Aims:

We describe the clinical and pathological findings of a previously unreported cutaneous lung tissue heterotopia in a child.

Case details:

The 3‐year‐old female patient developed a 8‐mm secreting papule over her left scapula. Pathological examination revealed a lesion composed of bronchioles and alveoli within the subcutis and the dermis, with bronchiolar connection to the epidermis. Alveolar type II cells indicating full pulmonary differentiation were detected with a monoclonal antibody (AMH 152).

Conclusions:

The described features suggest that this lesion is a unique variant of rarely observed bronchus‐like entities of the skin, designated as cutaneous bronchogenic cysts or cutaneous branchial cleft cysts.
Detection of P‐glycoprotein in frozen and paraffin‐embedded gastric adenocarcinoma tissues using a panel of monoclonal antibodies
Authors: Lacueva  , Teruel  , Calpena  , Medrano  , Mayol  , Perez‐Vazquez  , Rufete  , Camarasa  , Ferragut
Published: March 2003

Aims:

Most chemotherapeutic regimens used against gastric carcinoma include anthracyclines whose effectiveness can be impaired by the presence of P‐glycoprotein. In order to obtain a reliable pattern of P‐glycoprotein expression in these tumours an immunohistochemical study using a panel of anti‐P‐glycoprotein antibodies was performed in frozen and paraffinized tissues.

Methods and results:

Frozen and paraffinized samples from 25 gastric carcinomas were immunohistochemically analysed using a panel of four anti‐P‐glycoprotein monoclonal antibodies including C219, MRK16, JSB‐1 and C494. Semiquantitative analysis indicated that moderate or high P‐glycoprotein levels were detected in 40% to 76% of gastric adenocarcinomas, depending on the anti‐P‐glycoprotein antibody used. The antibody C494 was the most sensitive in detecting P‐glycoprotein in both frozen and paraffinized gastric carcinoma samples. Moreover, C494 showed a pattern of staining exclusively associated with the plasma membrane, in contrast to the cytoplasmic with reinforcement of plasma membrane pattern displayed by the other three antibodies. Significant differences in P‐glycoprotein levels were obtained when C494 and MRK16 were used in frozen tissues. Finally, detection of P‐glycoprotein in frozen samples did not improve when compared to paraffinized ones.

Conclusions:

It appears that P‐glycoprotein is frequently expressed in gastric adenocarcinomas, and the use of C494 complemented by JSB‐1 is recommended for reliable detection of P‐glycoprotein in this neoplasm.
Intrahepatic bile duct loss in primary sclerosing cholangitis: a quantitative study
Authors: Casali  , Carbone  , Cavalli
Published: March 2003

Aims

Intrahepatic bile duct volume density has been evaluated in four cases of primary sclerosing cholangitis (PSC) by applying a semiautomatic image analysis system to histological sections of surgical biopsies. Comparison was made between normal livers and cases of primary biliary cirrhosis.

Methods and results

A marked decrease in both bile duct volume density in liver (up to 50% of normal value) and bile duct volume density in portal tracts (up to 21% of normal value) was found. The analysis of correlations between the portal tract size and the portal tract volume fraction constituted by bile ducts confirms that the destruction mainly affects small and medium‐sized ducts. The ratio of bile duct to arterial component volume fractions in portal tracts turned out to be inverted with respect to that determined in normal liver, showing a decrease of up to 30% of normal value.

Conclusions

The features observed in primary sclerosing cholangitis, as well as the extent of bile duct loss proved to be quite similar to what we had previously described in primary biliary cirrhosis. Thus, regardless of the causes and mechanisms leading to the bile duct loss, the quantification of ductopenia produced results which are alike for the two distinct cholestatic diseases.
Apoptosis and expression of bcl‐2 protein are inverse factors influencing tumour cell turnover in primary carcinoid tumours of the lung
Authors: Zirbes T K , Lorenzen J , Baldus S E , Moenig S P , Wolters U , Ottlik A , Thiele J , Hölscher A H , Dienes H P
Published: March 2003

Aims

This study evaluates potential regulating factors in primary pulmonary carcinoid tumours, 16 typical and four atypical samples, with special emphasis on apoptosis and the bcl‐2 gene family. Furthermore, p53‐related oncogenes were analysed in a search for associated biological parameters.

Methods and results

The in‐situ end‐labelling technique (ISEL) was used to determine apoptotic cells, in addition to immunohistochemical methods, which were used to investigate the expression of the Ki67 antigen (avidin–biotin complex (ABC) method) and bcl‐2, bcl‐x, p53, p21/waf1, p27 and mdm‐2 proteins (catalysed reporter deposition (CARD) technique). The incidence of apoptotic tumour cells was significantly enhanced in typical carcinoids. The bcl‐2 protein was expressed to a higher degree in atypical carcinoids, which displayed a higher proliferative capacity as well. In contrast, bcl‐x was observed predominantly in so‐called typical carcinoids. The tumour cell turnover index was the most distinguishing parameter between both entities. All carcinoid tumours failed to show a staining for p53, p21/waf, p27 and mdm‐2 proteins.

Conclusions

The different biological behaviour of the carcinoid tumours under study seems to be influenced by the bcl‐2 gene family preventing programmed cell death. We speculate that this results in a more aggressive course in atypical carcinoid tumours.
Distinct clonal origin of low‐grade MALT‐type and high‐grade lesions of a multifocal gastric lymphoma
Authors: Matolcsy  , Nagy  , Kisfaludy  , Kelényi
Published: March 2003

Aims

Low‐grade mucosa‐associated lymphoid tissue (MALT) lymphoma and high‐grade B‐cell non‐Hodgkin's lymphoma (NHL) of the stomach may occur simultaneously. To determine the clonal relationship between these tumours, we compared the immunoglobulin heavy chain gene (IgH) rearrangements of low and high‐grade components of a multifocal gastric NHL.

Methods and results

The complementary determining region 3 (CDR3) of the IgH gene rearrangements were polymerase chain reaction (PCR) amplified, cloned and sequenced. The analysis of the CDR3 sequences rearranged by tumour cells of low‐grade MALT and the high‐grade NHL revealed different nucleic acid sequences.

Conclusion

These findings suggest that low‐grade MALT and high‐grade B‐cell components of multifocal gastric NHL may represent unrelated clones.
Phenotypic and cytotoxic characteristics of peripheral T‐cell and NK‐cell lymphomas in relation to Epstein–Barr virus association
Authors: Chan  , W Y Ho  , Chiang  , Srivastava
Published: March 2003

Aims

We investigated the phenotypic and cytotoxic characteristics of different types of peripheral T‐cell and NK‐cell lymphomas and correlated the findings of cytotoxic phenotype with Epstein–Barr virus (EBV) association.

Methods and results

Eighty cases of peripheral T‐cell and NK‐cell lymphomas, classified according to the REAL classification, were investigated for cytotoxic phenotype (by studying T‐cell intracellular antigen‐1 (TIA‐1) expression immunohistochemically) and EBV association (by in situ hybridization for EBV‐encoded small non‐polyadenylated RNAs), and the results were correlated with the specific clinicopathological types and the immunophenotype with special emphasis on CD56 expression and CD4/CD8 status. Overall, 39/80 cases (49%) expressed TIA‐1. Angiocentric lymphoma (23/24 cases; 96%), aggressive NK‐cell leukaemia (‘large granular lymphocyte (LGL) leukaemia’) (3/3 cases; 100%), intestinal T‐cell lymphoma (5/6 cases; 83%) and anaplastic large cell lymphoma (4/6 cases; 67%) were the major subtypes showing a cytotoxic phenotype. Only four of the 27 cases (15%) of peripheral T‐cell lymphoma, unspecified, were TIA‐1+ , while all the seven cases of angioimmunoblastic T‐cell lymphoma, six cases of mycosis fungoides and one case of adult T‐cell lymphoma/leukaemia were TIA‐1−.

Conclusions

Within the group of peripheral T‐cell and NK‐cell lymphomas, angiocentric lymphoma, aggressive NK‐cell leukaemia (‘LGL leukaemia’), intestinal T‐cell lymphoma and anaplastic large cell lymphoma are the major subtypes displaying a cytotoxic phenotype. The relationships between the cytotoxic phenotype and EBV association, CD56 expression or CD4/CD8 status are secondary to the relationship between cytotoxic phenotype and specific lymphoma subtype.
Papillary renal cell carcinoma with clear cell cytomorphology and chromosomal loss of 3p
Authors: Füzesi L , Gunawan B , Bergmann F , Tack S , Braun S , Jakse G
Published: March 2003

Aims

Cytogenetic studies on renal cell carcinomas (RCCs) have disclosed a correlation between chromosome aberrations and histomorphological features. Nevertheless, it is still controversial whether the cytomorphology of the tumour cells (clear cell, chromophilic, chromophobe) or their growth pattern (nonpapillary, papillary) is more discriminative for the combined histomorphological–cytogenetic classification of RCCs.

Methods and results

Three RCCs with papillary growth pattern and clear cell cytomorphology were analysed by classical cytogenetics using standard G‐banding techniques. Each tumour displayed clonal aberrations leading to loss of terminal 3p chromosomal segments. Monosomy 14 was also consistently found. Trisomy 17 was not observed in any of the tumours.

Conclusions

This series of three RCCs consisting of clear cells with papillary architecture revealed chromosomal aberrations characteristic for the conventional (clear cell) RCC. Irrespective of the predominant papillary growth pattern, none of the cases were characterized by trisomy of chromosomes 3q, 7, 8, 12, 16, 17 and 20 and loss of Y chromosome which are widely regarded as the most consistent genetic alterations for papillary RCC. Therefore, our cytogenetic findings provide evidence that papillary clear cell RCCs should be classified according to their cytomorphology rather than their growth pattern even when papillary architecture is prominent.
Molecular detection of minimal residual disease in colorectal and breast cancer
Authors: Liefers  , Tollenaar  , Cleton‐Jansen
Published: March 2003

Recent developments in the field of molecular biology enable us to detect tumour cells at a submicroscopical level. In colorectal and breast cancer the most important prognostic factor is dissemination of malignant cells to locoregional lymph nodes. An important issue is whether molecular ‘super’‐staging augments the accuracy by which the prognosis of individual patients can be assessed. Over the past few years numerous studies have reported the use of different PCR‐based techniques in various types of cancer. The reported incidence of micrometastases and specificity of different assays varies tremendously. This clearly indicates the need for uniformity in protocols. For colorectal cancer the use of molecular techniques may improve staging and guide clinical decisions. For breast cancer there is still need to prove the clinical implication of finding occult metastatic disease. Nevertheless, PCR‐based techniques are a powerful tool in the staging of common solid tumours and are likely to find their way into the daily practice of diagnostic histopathologists in the near future.
Enhanced cell kinetics, p53 accumulation and high p21WAF1 expression in chronic cholecystitis: comparison with background mucosa of gallbladder carcinomas
Authors: Yanagisawa  , Mikami  , Koike  , Okayasu
Published: March 2003

Aims

Since neoplasia resulting from chronic inflammation has recently attracted increasing attention, we have investigated surgically removed gallbladders to examine the relationship between chronic cholecystitis and carcinogenesis.

Methods and results

The mucosa of 108 cholecystectomy specimens without gallbladder cancer and 54 surgically resected gallbladder carcinomas were classified into three groups according to the degree of lymphocytic infiltration, and assessed immunohistochemically for Ki67, p53, p21WAF1 and apoptosis. In gallbladder mucosa without carcinoma, all four parameters tended to increase with the inflammation score (IS). Significantly positive correlations were revealed between Ki67 and p53, Ki67 and p21WAF1, and p53 and p21WAF1. However, in gallbladder carcinoma cases, values of p53 and p21WAF1 for background mucosa were elevated as compared to the mucosa of cholecystitis with low IS, but there was no correlation between their expression and IS, except for Ki67.

Conclusions

Severe chronic cholecystitis is associated with acceleration of epithelial cell turnover, damaged cells being eliminated by apoptosis. The background mucosa of gallbladder carcinomas showed similar cell proliferative activity (Ki67) to that in cholecystitis, with no parallel changes of p53 and p21WAF1 expression, suggesting the possibility of unknown cofactors causing genomic damage.
Immunohistochemical evidence for mesothelial origin of paratesticular adenomatoid tumour
Authors: Delahunt  , Eble  , King  , Bethwaite  , Nacey  , Thornton
Published: March 2003

Aims

To investigate the histogenesis of paratesticular adenomatoid tumour by use of immunohistochemical markers for a variety of carcinomas and mesothelioma.

Methods and results

Immunohistochemical staining of sections from 12 cases of paratesticular adenomatoid tumour was undertaken using primary antibodies to antigens expressed by benign epithelial cells and carcinoma (cytokeratin AE1/AE3, cytokeratin 34ßE12, epithelial membrane antigen, MOC‐31, Ber‐EP4, CEA, B72.3, LEA.135, Leu M1), stromal and vascular markers (vimentin, CD34, factor VIII), and mesothelioma‐associated antigens (thrombomodulin, HBME‐1, OC 125) and p53 protein. There was absence of immunohistochemical expression of epithelial/carcinoma markers MOC‐31, Ber‐EP4, CEA, B72.3, LEA.135, Leu M1 and to factor VIII and CD34. All tumours expressed cytokeratin AE1/AE3, epithelial membrane antigen and vimentin, with weak expression of cytokeratin 34ßE12 in 25% of tumours. Each tumour showed expression of thrombomodulin, HBME‐1 and OC 125 in a membranous distribution. p53 protein expression was not detected.

Conclusions

The immunohistochemical profile of paratesticular adenomatoid tumour is strongly supportive of a mesothelial cell origin.
Immunoreactivity for cadherins, HGF/SF, met, and erbB‐2 in pleural malignant mesotheliomas
Authors: Thirkettle I , Harvey P , Hasleton P S , Ball R Y , Warn R M
Published: March 2003

Aims

To assess the immunoreactivity of malignant mesotheliomas for N‐ and E‐cadherins, hepatocyte growth factor/scatter factor (HGF/SF) and the tyrosine kinase receptors, met and erbB‐2.

Methods and results

Pleural malignant mesotheliomas were stained using a standard indirect immunoperoxidase method applied to paraffin sections. Malignant mesotheliomas were immunoreactive for N‐cadherin (26/29; 90%), met (29/29; 100%) and erbB‐2 (28/29; 97%). Focal immunoreactivity was present for E‐cadherin in epithelioid or mixed tumours (14/25; 56%), and for HGF/SF (9/24; 38%).

Conclusions

Expression of N‐cadherin supports the diagnosis of malignant mesothelioma and use of appropriate antibodies would be a useful addition to a diagnostic antibody panel. Focal staining for E‐cadherin does not exclude mesothelioma. Signalling pathways mediated via met and erbB‐2 may play a role in the growth and spread of malignant mesotheliomas.
Merkel cell tumour with leiomyosarcomatous differentiation
Authors: Cooper L , DeBono R , Alsanjari N , Al‐Nafussi A
Published: March 2003

Aims: To report and confirm the identity of tumour cells showing leiomyosarcomatous differentiation in a regional lymph node containing metastatic Merkel cell tumour.

Methods and results

A 79‐year‐old woman was found to have metastatic Merkel cell tumour within an axillary lymph node 4 months after excision of the primary tumour from the forearm. The lymph node was effaced by tumour identical to that of the primary tumour but there was an additional focus of loose spindle cells. Immunocytochemical staining showed the coexpression of cytokeratin 20, neurofilament and desmin in these sarcomatous cells.

Conclusion

This, to the best of our knowledge, is the first report of Merkel cell carcinoma showing sarcomatous differentiation, and adds to the expanding morphological spectrum of malignant biphasic tumours.
Gastric adenomas and superficial adenocarcinomas display distinct patterns of mucin carbohydrate and core protein expression
Authors: Wang Y‐Z , Mitomi H , Kurihara M , Ishihara K , Hotta K , Tanigawa H , Okayasu I
Published: March 2003

Aims

To investigate the histogenetic relationship between gastric epithelial neoplasms we studied differences in expression of mucin carbohydrate antigens and mucin core protein, in normal and metaplastic gastric mucosa, and in gastric adenomas and superficial adenocarcinomas.

Methods and results

We generated four monoclonal antibodies, including HGM72/75 against human gastric mucin and HCM14/21 against human colonic mucin, and investigated immunoreactivities of these antibodies and MUC2 protein expression in normal and metaplastic gastric mucosa, adenomas (15 samples) and superficial adenocarcinomas (intestinal‐type, 77; diffuse‐type, 59 samples). HGM72 reacted with mucous neck cells of the fundic glands and with pyloric glandular cells whereas HGM75 stained foveolar cells and metaplastic goblet cells. Weak binding of HCM14/21 and strong staining with MUC2 were found in metaplastic goblet cells. Binding of HGM75, HCM14, MUC2, but not HGM72 was high in adenomas. An equivalent staining with HGM72 and HGM75 with low expression of MUC2 and HCM14 was shown in intestinal‐type carcinomas while the diffuse‐type demonstrated more strong reactivity with HGM75 than with HGM72, MUC2 and HCM14. Little binding of HCM21 was observed in any specimens.

Conclusions

This study demonstrates that adenomas predominantly have a intestinal phenotype, but both types of adenocarcinomas retain some cells with a gastric phenotype during the early steps of neoplastic development.
HSP70 is selectively over‐expressed in the blast cells of the germinal centres and paracortex in reactive lymph nodes
Authors: Leopardi O , Naughten W , Giannulis I , Mirra M , Frigo B
Published: March 2003

HSP70 is selectively over‐expressed in the blast cells of the germinal centres and paracortex in reactive lymph nodes

Aims: We evaluated by immunohistochemistry HSP70 expression in reactive lymph nodes since its morphological expression and location have not been previously described and correlated with lymphocyte kinetics.

Methods and Results: Ninety‐six cases of non‐specific lymphadenitis were immunostained for HSP70, CD20, CD3, Ki67, Bcl‐2, CD21. The type and the location of HSP70‐positive cells were determined. Their number out of 2000 cells in each germinal centre and in each paracortical area was counted at 60× magnification with the help of a quantitative grid. Seventeen percent of germinal centre cells and 7.6% of the paracortex cells were positive. This difference was highly significant. The positively reacting cells were B‐cells and had a blast (centroblast or immunoblast) morphology, with negative mantle and marginal lymphocytes and T‐cells. Lymphoplasmacytoid cells and plasma cells reacted only weakly or were negative. Germinal centre antigen‐presenting cells and interdigitating dendritic cells reacted from lightly to moderately.

Conclusions: HSP70 was selectively over‐expressed by B‐blasts mainly located within germinal centres with a lower number in the paracortex. The difference in the mean number between the two sites was statistically highly significant. No correlation was found with bcl‐2 and Ki67 expression. Mantle, marginal and T‐lymphocytes were always negative. The biological meaning and role of this over‐expression in centroblasts and immunoblasts remain to be elucidated.
Enhanced expression of decay‐accelerating factor and CD59/homologous restriction factor 20 in intestinal metaplasia, gastric adenomas and intestinal‐type gastric carcinomas but not in diffuse‐type carcinomas
Authors: Kiso T , Mizuno M , Nasu J , Shimo K , Uesu T , Yamamoto K , Okada H , Fujita T , Tsuji T
Published: March 2003

Enhanced expression of decay‐accelerating factor and CD59/homologous restriction factor 20 in intestinal metaplasia, gastric adenomas and intestinal‐type gastric carcinomas but not in diffuse‐type carcinomas

Aims: Variable expression of the complement regulatory proteins, decay‐accelerating factor, CD59/homologous restriction factor 20 (HRF20) and membrane cofactor protein has been shown in human gastrointestinal malignancies, but their expression in gastric cancer has not been fully described. Thus, we immunohistochemically defined the distribution of these proteins in human normal gastric mucosa, intestinal metaplasia, adenomas and gastric cancers.

Methods and results: Gastric tissues were obtained by endoscopic biopsy or surgical resection and stained with mouse monoclonal antibodies to decay‐accelerating factor, CD59/HRF20, and membrane cofactor protein. In the normal gastric mucosa, membrane cofactor protein was diffusely stained on the basolateral surface of epithelial cells, whereas the expression of decay‐accelerating factor and CD59/HRF20 was inconspicuous. In intestinal metaplasia, adenoma and intestinal‐type gastric carcinoma cells, decay‐accelerating factor and HRF20 were intensely stained on the apical surface; membrane cofactor protein retained its location on the basolateral surface. In diffuse‐type gastric carcinomas, the expression of decay‐accelerating factor, CD59/HRF20 was lost, but membrane cofactor protein was present on the tumour cell surface.

Conclusions: These findings suggest that membrane cofactor protein plays a primary role in the regulation of complement activation in normal and neoplastic gastric cells and that the expression pattern of the complement regulatory proteins is closely related to gastric carcinoma development.
Telepathology: current status and future prospects in diagnostic histopathology
Authors: Cross S S , Dennis T , Start R D
Published: March 2003

Telepathology: current status and future prospects in diagnostic histopathology

Telepathology is the process of diagnostic histopathology performed on digital images viewed on a display screen rather than by conventional glass slide light microscopy. The technology of telepathology has radically improved over the past 5 years so that it is no longer the limiting factor in the diagnostic process. This review looks at the resources needed for dynamic and static telepathology, including image quality, computers and software interfaces, means of transmission and human resources. It critically analyses 32 published trials of telepathology, including some large prospective studies, in all areas of diagnostic histopathology including intraoperative frozen sections, routine and referral cases. New developments, including internet solutions and virtual microscopy, are described and there is analysis of the economics of telepathology within health care systems. The review concludes that all the necessary technology for telepathology is available, there is strong published evidence for a diagnostic accuracy comparable with glass slide diagnosis, in many contexts there is a clear‐cut economic argument in favour of telepathology, and that the technique should now be integrated into mainstream diagnostic histopathology.
Intranasal immunization of mice with herpes simplex virus type 2 recombinant gD2: the effect of adjuvants on mucosal and serum antibody responses
Authors: UGOZZOLI  , O’HAGAN  , OTT
Published: March 2003

Mucosal immunization offers the potential for inducing IgA antibody responses in the vagina, the site of infection for many viruses, including herpes simplex type 2 (HSV‐2). To investigate this possibility, mice were immunized intranasally with 10 μg glycoprotein D2 (gD2) from HSV combined with a series of adjuvants of proven efficacy; the oil in water emulsion MF59, poly(d,l‐lactide‐co‐glycolide) microparticles (PLG) (encapsulated or co‐administered), immune‐stimulating complexes (iscoms) (incorporated or co‐administered with iscomatrix) and the genetically detoxified enterotoxin from Escherichia coli, LT‐K63. Encapsulation of gD2 into PLG microparticles, incorporation of gD2 into iscoms and co‐administration of gD2 with LT‐K63 induced mucosal IgA antibody responses (nasal wash, saliva and vaginal wash) which were greater than those induced by intramuscular administration of gD2 with MF59. Intranasal immunization with these formulations also induced substantial levels of serum IgG and neutralizing antibodies. These studies demonstrated that intranasal immunization with potent adjuvants is an effective means to induce mucosal antibody responses, even in the lower genital tract.
The plasma levels of conglutinin are heritable in cattle and low levels predispose to infection
Authors: HOLMSKOV  , JENSENIUS  , TORNØE  , LØVENDAHL
Published: March 2003

Conglutinin, like mannan‐binding lectin (MBL) and CL‐43, is a serum collectin involved in the innate immune defence system. In man, low serum MBL concentrations, resulting from mutations in the collagen region, are associated with a common opsonic defect. Plasma levels of conglutinin in cattle were assayed by rocket immunoelectrophoresis to examine whether they were genetically determined. Samples were collected from calves (309 bull‐calves and 260 heifers with complex pedigree relationships). The number of respiratory infections from the 42nd to 336th day of life was recorded. The number of infections was found to be genetically determined (heritability: h2=0·31±0·07). A wide concentration range of conglutinin was found in plasma (<1·25–35 μg/ml for females, geometric mean 8·1 μg/ml, and <1·25–47 μg/ml for males, geometric mean 15·5 μg/ml), and the concentration was found to be genetically determined (heritability, h2=0·52±0·07). The analysis revealed a negative association between disease frequency and the conglutinin levels (−0·56±0·18 for female; −0·50±0·18 for male). Levels of conglutinin below the detection limit of the assay (1·25 μg/ml) were found in 2% of the animals. If these animals are assumed to be homozygous for a single recessive allele causing low concentrations a gene frequency of 0·15 could be calculated. These findings suggests that selection for resistance against infectious disease is possible in cattle and that the level of plasma conglutinin may be a helpful trait in such a breeding scheme.
Interleukin‐10 differentially modulates MHC class II expression by mesangial cells and macrophages in vitro and in vivo
Authors: CHADBAN  , TESCH  , FOTI  , LAN  , ATKINS  , NIKOLIC‐PATERSON
Published: March 2003

Inhibition of major histocompatibility complex (MHC) class II expression by macrophages is the primary mechanism by which interleukin‐10 (IL‐10) exerts imm_une suppression. Little, however, is known of the effects of IL‐10 on other types of cells which can be induced to express MHC class II during an inflammatory response. We therefore studied the effects of IL‐10 treatment on the expression of MHC class II molecules in a rat model of imm_unologically induced glomerulonephritis. MHC class II mRNA levels in whole kidney were increased in saline‐treated (control) animals with glomerulonephritis (2·6‐fold increase versus normal, P=0·028) and this was partially inhibited by treatment with IL‐10 (P=NS). Double imm_unostaining of tissue sections was used to compare MHC class II expression by infiltrating macrophages and resident glomerular cells. IL‐10 treatment reduced the proportion of glomerular macrophages which expressed detectable MHC class II (70% reduction, P=0·03). In contrast, IL‐10 treatment was associated with an increase in the number of resident glomerular cells expressing MHC class II, particularly within mesangial areas. Therefore, the effects of IL‐10 on macrophages and mesangial cells were compared in vitro. IL‐10 reduced constitutive MHC class II mRNA and cell surface expression by peritoneal macrophages. In contrast, IFN‐γ‐stimulated mesangial cells (1097 cell line) cultured with IL‐10 for 24 hr showed increased MHC class II mRNA (26% increase) and surface expression (72% increase in percentage MHC II+ by flow cytometry, P=0·04) as compared with cells stimulated with IFN‐γ alone. IL‐10 also directly up‐regulated expression of ICAM‐1 by 1097 cells. In conclusion, IL‐10 was found to have contrasting effects on the production and cell surface expression of MHC class II molecules by mesangial cells and by macrophages, both in vitro and in vivo. The implications of these findings for IL‐10‐mediated imm_unosuppression are discussed.
Down‐regulation of CD28 via Fas (CD95): influence of CD28 on T‐cell apoptosis
Authors: WALKER  , MCLEOD  , BOULOUGOURIS  , PATEL  , HALL  , SANSOM
Published: March 2003

Following antigen engagement of the T‐cell receptor (TCR), T‐cell survival is largely dictated by the provision of additional signals, such as those from costimulatory receptors and cytokine receptors. Whilst CD28‐mediated signalling is increasingly associated with survival, ligation of alternative T‐cell antigens, such as Fas (CD95), can trigger apoptosis. The T‐cell response following antigen engagement may therefore be influenced by the relative expression levels of these coreceptors as well as by the availability of their ligands (CD80/86 and Fas‐L). In this study we demonstrate functional interplay between the death receptor Fas and the costimulatory receptor CD28 in human T cells. In Jurkat T cells, we show that Fas signalling leads to rapid and selective CD28 down‐regulation, and that this is associated with a specific decrease in mRNA for CD28, indicating that mechanisms exist which target CD28 at a transcriptional level. Moreover, cells that down‐regulate CD28 also undergo apoptosis. Studies on activated human peripheral blood T cells demonstrate that cells expressing high levels of CD28 are resistant to Fas‐mediated apoptosis whereas cells expressing low levels are more susceptible, implicating CD28 in the provision of anti‐apoptotic signals. Consistent with this hypothesis, direct ligation of CD28 using B7 transfectants concomitant with anti‐Fas challenge protects from apoptosis. Since antigen‐presenting cells may express Fas‐L under certain circumstances, the maintenance of T‐cell CD28 expression may be crucial for the prevention of Fas‐mediated apoptosis during the course of antigen engagement.
Morphological alterations in rat peritoneal mast cells by stem cell factor
Authors: Kim H. M. , Shin H. Y. , Lee E. H.
Published: March 2003

Stem cell factor (SCF) stimulates mast cell adhesion and, because SCF is produced normally in tissues, it may be a major factor responsible for the adhesion of mast cells to connective tissue matrix. We found that the morphology of rat peritoneal mast cells (RPMC) altered after the addition of recombinant murine SCF (rmSCF) in vitro. The ability of rmSCF to enhance morphological alteration was dose dependent and completely abolished by anti‐c‐kit ACK2 monoclonal antibody. Exposure of RPMC to transforming growth factor‐β1, wortmannin, genistein, herbimycin A, staurosporine, indomethacin and cytochalasin D before the addition of rmSCF antagonized rmSCF‐induced morphological alteration. However, nordihydroguiaretic acid had no effect. Many RPMC appeared to respond also to nerve growth factor (NGF) but the total number of cells with altered morphology was much greater when the culture was stimulated by rmSCF than by NGF. We suggest that morphological alterations of mast cells by rmSCF is an important step for the participation in adhesion to tissue under resident physiological conditions.

NDGA, nordihydroguiaretic acid NGF, nerve growth factor PI3‐kinase, posphatidylinositol 3‐kinase rmSCF, recombinant murine stem cell factor RPMC, rat peritoneal mast cells SCF, stem cell factor.
Low CD3+CD28‐induced interleukin‐2 production correlates with decreased reactive oxygen intermediate formation in neonatal T cells
Authors: Kilpinen S. , Hurme M.
Published: March 2003

The capacity of neonatal T cells to secrete interleukin‐2 (IL‐2) has been reported to be variable. We analysed IL‐2 production in purified neonatal and adult T cells using polyclonal activator phorbol ester+calcium ionophore (PDBu+iono) or receptor‐mediated anti‐CD3/anti‐CD3+anti‐CD28 stimulation. PDBu+iono induced equally high IL‐2 levels in both groups and, when stimulated with plate‐bound anti‐CD3 monoclonal antibody (mAb), the IL‐2 secretion by neonatal cells was undetectable and adult cells produced low amounts of IL‐2 (mean 331±86 pg/ml). The addition of anti‐CD28 mAb to anti‐CD3‐stimulated cells markedly increased IL‐2 production in both cell types, but levels of IL‐2 in neonatal T cells remained clearly lower than those of adult T cells (respective mean values: 385±109 pg/ml and 4494±1199 pg/ml). As NF‐κB is a critical transcription factor in the control of IL‐2 expression, we next analysed its nuclear translocation in neonatal and adult T cells using the electrophoretic mobility shift assay and, because induction of reactive oxygen intermediates (ROI) is required for the activation of NF‐κB, we also analysed levels of intracellular ROI in these cells using the ROI‐reactive fluorochrome DCFH‐DA and flow cytometry. In neonatal T cells NF‐κB activation and ROI formation after anti‐CD3 stimulation were low compared with adult T cells and, although addition of anti‐CD28 mAb increased induction of NF‐κB and ROI formation, levels similar to those of adults were not achieved. After PDBu+iono stimulation, the cells showed similar ROI formation and IL‐2 secretion. Our results suggest that reduced IL‐2 production by neonatal T cells is specific for anti‐CD3 and anti‐CD3+anti‐CD28‐mediated stimulation and that these activators cannot effectively activate the ROI–NF‐κB signalling pathway in neonatal T cells.
Immunity to vaginal HSV‐2 infection in immunoglobulin A knockout mice
Authors: PARR  , HARRIMAN  , PARR
Published: March 2003

An immunoglobulin A (IgA) knockout (KO) mouse was used to study the role of IgA in protective immunity against vaginal infection with herpes simplex virus‐type 2 (HSV‐2). Intact and KO mice were immunized intravaginally (IVAG) with attenuated HSV‐2, challenged IVAG with wild‐type virus 6 weeks later and evaluated for vaginal infection and neurological disease. Non‐immunized/challenged intact and KO mice showed vaginal infection and succumbed to neurological disease, while immunized/challenged mice exhibited reduced or no vaginal infection and no neurological disease. Log 2·5 enzyme‐linked immunoassay (ELISA) titres of viral IgA, immunoglobulin G (IgG) and immunoglobulin M (IgM) in vaginal secretions collected from intact immune mice before challenge were 0·6±0·3, 6·4±0·32 and 0·0, while those in KO immune mice were 0·0, 6·7±0·19 and 3·0±0·29, respectively. Twenty‐four hours after challenge, the percentage of vaginal epithelium that was infected in non‐immune intact and KO mice was 2·0±0·6 and 2·4±0·6, which was reduced to 0·2±0·1 and 0·1±0·06 in immune intact and KO mice, respectively. No shed virus protein was detected in vaginal secretions 3 days after challenge in any immune mouse, whereas titres were 1400 and 1700 in the two groups of non‐immune mice. Thus, immune protection against vaginal HSV‐2 infection was similar in both KO and intact mice, indicating that this mucosal immunity does not depend mainly on IgA.