bluuebunny/crossref_metadata_2025 huggingface dataset:
The reproductive biology of Polytrichum formosum: clonal structure and paternity revealed by microsatellites
Authors: Van der Velde M. , During H. J. , Van de Zande L. , Bijlsma R.
Published: March 2003

Using highly polymorphic microsatellite markers, we assessed clonal structure and paternity in a population of the bryophyte species Polytrichum formosum. Identical multilocus genotypes of individual shoots were almost never observed in spatially separated cushions, but were found to be highly clustered within moss cushions. Therefore, asexual reproduction through dispersal of gametophyte fragments is not very important in P. formosum. However, asexual reproduction on a very localized scale through vegetative growth of genets (branching of gametophytes via clonal growth of rhizomes) is very extensive. The patchy spatial distribution of genets and the absence of intermingling among genets suggest that this species follows a ‘phalanx’ clonal growth strategy. Vegetative proliferation of genets will increase their size, and, consequently, will have considerable fitness consequences for individuals in terms of increased genet longevity and reproductive output. Although paternity analysis of sporophytes confirmed male genet size, i.e. gamete production, to be an important determinant of male reproductive fitness, it also showed that the spatial distance to female genets is the predominant factor that governs male reproductive success. Moreover, we showed that male gamete dispersal distances in P. formosum are much further than generally assumed, and are in the order of metres rather than centimetres. Combining the findings, we conclude that the high genotypic diversity observed for this facultatively clonal species is most likely explained by a preponderance of sexual reproduction over clonal reproduction.
High within‐population mitochondrial DNA variation due to microvicariance and population mixing in the land snailEuhadra quaesita(Pulmonata: Bradybaenidae)
Authors: Watanabe Y. , Chiba S.
Published: March 2003

A high level of geographical variation at an exceptionally fine scale was detected in the mitochondrial 16Sribosomal RNA genes of a land snail speciesEuhadraquaesitafrom the Kanto region of Japan. In total, 50 haplotypes were detected from 27 populations, with most sample sites possessing private alleles. In some individual populations the different haplotypes do not fall as a monophyletic group, so that some of the haplotypes are phylogenetically distant, differing from each other by > 10%. In contrast, phylogenetically similar haplotypes were found in separate sites at long distances from their main distribution. Together, these findings strongly suggest that contraction and expansion of populations has occurred repeatedly in the past. The subsequent expansion of populations and migration from different areas may have mixed distant populations. This repetition of isolation and mixing has resulted in an exceptionally fine scale of geographical variation, and the accumulation of high genetic diversity within and between populations of this species.
A genetic analogue of ‘mark–recapture’ methods for estimating population size: an approach based on molecular parentage assessments
Authors: Pearse D. E. , Eckerman C. M. , Janzen F. J. , Avise J. C.
Published: March 2003

Molecular polymorphisms have been used in a variety of ways to estimate both effective and local census population sizes in nature. A related approach for estimating the current size of a breeding population, explored here for the first time, is the use of genetic ‘marks’ reconstructed for otherwise unknown parents in paternity or maternity analyses of progeny arrays. This method provides interesting similarities and contrasts to traditional mark–recapture methods based on physical tags. To illustrate, this genetic method is applied to a population of painted turtles on the Mississippi River to estimate the number of successfully breeding males. Non‐genetic mark–recapture approaches were also applied to animals trapped at this location. Results demonstrate that such genetic data on parentage can be helpful not only in estimating contemporary population sizes, but also in providing additional information, not present in customary mark–recapture data, about possible extended movements of breeding individuals and the size of the pool of mates which they encounter.
Variation with age in the timing and duration of spawning in American plaice
Authors: Morgan M. J.
Published: March 2003

Spawning time of female American plaice Hippoglossoides platessoides varied significantly with age. This effect, however, was not consistent across years. Generally, younger fish spawned earlier than older fish, but in the 1990s they spawned later than older fish. Spawning duration also varied with age, with younger fish generally having a longer spawning period than older fish. Changes in depth distribution were related to changes in both spawning time and duration. As depth increased, spawning time became later and duration became shorter. Fish <11 years old showed the largest change in both spawning time and depth distribution.
A case of successful management of maternal septic shock with multiple organ failure following amniocentesis at midgestation
Authors: Hamanishi Junzo , Itoh Hiroaki , Sagawa Norimasa , Nakayama Takahiro , Yamada Shigetoshi , Nakamura Kaoru , Saito Ayaka , Kumakura Erika , Yura Shigeo , Fujii Shingo
Published: March 2003

Maternal sepsis is an unusual but catastrophic complication of amniocentesis. We report a case of successful treatment of maternal septic shock and multiple organ failure following amniocentesis at midgestation, possibly due to needle puncture of the sigmoid colon, which was tightly adherent to the anterior surface of the pregnant uterus.
‘Partnership’: a co‐operative inquiry between Community Mental Health Nurses and their clients. 2. The nurse–client relationship
Authors: Hostick T. , Mcclelland F.
Published: March 2003

This paper describes the output of a co‐operative enquiry between Community Mental Health Nurses and their clients. Two nurses and two clients volunteered to participate as co‐researchers and co‐subjects with two facilitators in a co‐operative inquiry group. The subject of the inquiry, agreed by the group, was the relationship between the nurse and client. The description of the nurse–client relationship and the influences on it is extremely rich and potentially useful with implications for clinical and managerial practice. Despite the limitations of the study, co‐operative inquiry appears to be a useful vehicle for contributing to an increased understanding of the nurse–client relationship with potential for further research and development. Unsurprisingly, when both the relationship characteristics and the health care context are appropriate there seems to be more positive outcomes for both nurse and client. Within the current health policy context there are increased external demands with the potential to disrupt this relationship. Nurses need to be aware of the potential effects of these so that the relationship and the client do not suffer and health service managers need to be aware of the contribution their behaviour may have on the nurse–client relationship.
A dsbA mutant of Pseudomonas syringae exhibits reduced virulence and partial impairment of type III secretion
Authors: Kloek Andrew P. , Brooks David M. , Kunkel Barbara N.
Published: March 2003

To identify virulence genes of P. syringae pv. tomato strain DC3000 we screened for mutants with reduced virulence on its plant hosts, Arabidopsis thaliana and tomato. We isolated a Tn5‐insertion mutant that exhibited reduced virulence on both hosts. Further characterization showed that this mutant carried a single Tn5 insertion in the dsbA gene, which encodes a periplasmic disulphide bond‐forming protein. In addition to reduced virulence, the dsbA mutant exhibits mucoid colony morphology, loss of fluorescence, decreased motility, and a reduced growth rate in culture. The dsbA mutant is able to multiply in A. thaliana and tomato plants, trigger the hypersensitive response on tobacco and elicit Pto‐mediated resistance in tomato, indicating that type III secretion occurs in this background. However, type III secretion appears to function with reduced efficiency in the dsbA mutant, as type III‐dependent secretion of HrpZ and AvrRpt2 is impaired. These findings indicate that while the dsbA gene is required for multiple cellular functions in P. syringae, type III secretion in P. syringae is only partially dependent on dsbA.
Mutations in Ror1 and Ror2 genes cause modification of hydrogen peroxide accumulation in mlo‐barley under attack from the powdery mildew fungus
Authors: Hückelhoven Ralph , Trujillo Marco , Kogel Karl‐Heinz
Published: March 2003

Race nonspecific resistance of barley against the barley powdery mildew fungus (Blumeria Graminis f.sp. Hordei, Speer, Bgh) is mediated by recessive mlo alleles and is controlled by at least two additional genes ‘required for ml
o‐specified disease
resistance’ (Ror1 and Ror2). The pathogenesis‐related accumulation of hydrogen peroxide (H2O2) was comparatively analysed in a susceptible barley line (Hordeum vulgare L. Cv Ingrid, genotype Mlo Ror1, Ror2), a resistant Ingrid backcross line carrying the mutant allele mlo5 (BCIngrid‐mlo5, genotype mlo5 Ror1 Ror2), and in the moderately susceptible mutants A44 and A89 (genotypes mlo5 Ror1 ror2 and mlo5 ror1‐2 Ror2, respectively). In situ localization of H2O2 was performed by microscopic detection of 3,3‐diaminobenzidine (DAB) polymerization. In BCIngrid‐mlo5, penetration resistance against Bgh attack was closely correlated to H2O2 accumulation in cytoplasmic aggregates and cell wall appositions beneath the appressorium. In contrast, H2O2 accumulation was almost completely absent in susceptible Ingrid. Lines with mutations in Ror genes showed less H2O2 accumulation beneath appressoria, but more interaction sites with whole cell H2O2 accumulation and hypersensitive cell death response than resistant BCIngrid‐mlo5. Thus, mutations in Ror1 or Ror2 genes influence the cellular pattern of H2O2 accumulation in mlo plants attacked by Bgh. The data support the hypothesis that H2O2 accumulation is involved in resistance to fungal penetration.
DNA microarrays: new tools in the analysis of plant defence responses
Authors: Kazan Kemal , Schenk Peer M. , Wilson Iain , Manners John M.
Published: March 2003

Large‐scale DNA sequencing is providing information on the number and organization of genes and genomes of plant species and their pathogens. The next phase is to identify gene functions and gene networks with key roles in compatible and incompatible plant–pathogen interactions. DNA microarrays can provide information on the expression patterns of thousands of genes in parallel. The application of this technology is already revealing new features of plant–pathogen interactions and will be a key tool for a wide range of experiments in molecular plant pathology.
Pto update: recent progress on an ancient plant defence response signalling pathway
Authors: Bogdanove Adam J.
Published: March 2003

The Pto resistance gene in a gene‐for‐gene interaction with the avrPto avirulence gene governs resistance to bacterial speck of tomato. A member of a small gene family in tomato, Pto encodes a serine/threonine kinase that interacts in the yeast two‐hybrid system with the product of avrPto, an 18‐kDa hydrophilic protein. Over the past decade, studies of these genes, their products, and the defence response signalling pathway they govern have led to significant advances in our understanding of the biochemistry of Pto, the bacterial delivery and Pto recognition specificity for AvrPto, and candidate components in the pathway and their potential functions. This article provides an update of recent advances, which include the discovery of AvrPto structure–function relationships in disease and resistance, discovery of a second avirulence protein (AvrPtoB) recognized by Pto and its limited similarity to AvrPto, expression analysis and functional characterization of transcription factors Pti4, Pti5, and Pti6 that interact with Pto, analyses of Pto over‐expression that activates defence responses independent of AvrPto, and comparisons of Pto gene family members and homologues in tomato and other Solanaceae, as well as other plant species. These comparisons, in particular, have provided exciting new insight into the antiquity of the Pto gene family and of the capacity for specific recognition of AvrPto that activates plant defence.
Mining the enzymes involved in the detoxification of reactive oxygen species (ROS) in sugarcane
Authors: Kurama Eiko E. , Fenille Roseli C. , Rosa Vicente E. , Rosa Daniel D. , Ulian Eugenio C.
Published: March 2003

Adopting the sequencing of expressed sequence tags (ESTs) of a sugarcane database derived from libraries induced and not induced by pathogens, we identified EST clusters homologous to genes corresponding to enzymes involved in the detoxification of reactive oxygen species. The predicted amino acids of these enzymes are superoxide dismutases (SODs), glutathione‐S‐transferase (GST), glutathione peroxidase (GPX), and catalases. Three MnSOD mitochondrial precursors and 10 CuZnSOD were identified in sugarcane: the MnSOD mitochondrial precursor is 96% similar to the maize MnSOD mitochondrial precursor and, of the 10 CuZnSOD identified, seven were 98% identical to maize cytosolic CuZnSOD4 and one was 67% identical to putative peroxisomal CuZnSOD from Arabidopsis. Three homologues to class Phi GST were 87–88% identical to GST III from maize. Five GPX homologues were identified: three were homologous to cytosolic GPX from barley, one was 88% identical to phospholipid hydroperoxide glutathione peroxidase (PHGPX) from rice, and the last was 71% identical to GPX from A. thaliana. Three enzymes similar to maize catalase were identified in sugarcane: two were similar to catalase isozyme 3 and catalase chain 3 from maize, which are mitochondrial, and one was similar to catalase isozyme 1 from maize, whose location is peroxisomal subcellular. All enzymes were induced in all sugarcane libraries (flower, seed, root, callus, leaves) and also in the pathogen‐induced libraries, except for CuZnSOD whose cDNA was detected in none of the libraries induced by pathogens (Acetobacter diazotroficans and Herbaspirillum rubrisubalbicans). The expression of the enzymes SOD, GST, GPX, and catalases involved in the detoxification was examined using reverse transcriptase‐polymerase chain reaction in cDNA from leaves of sugarcane under biotic stress conditions, inoculated with Puccinia melanocephala, the causal agent of sugarcane rust disease.
In planta induced genes of Puccinia triticina
Authors: Thara Venkatappa K. , Fellers John P. , Zhou Jian‐Min
Published: March 2003

Wheat leaf rust disease, caused by the biotrophic fungus Puccinia triticina, is a result of complex interactions requiring the coordinated activities of the two organisms involved. In an effort to understand the molecular basis of wheat–rust interactions, we isolated and characterized cDNA corresponding to in planta induced fungal genes (PIGs) from susceptible wheat leaves infected with P. triticina by using suppression subtractive hybridization to construct a cDNA library. 350 clones were sequenced, of which 104 were unique. Forty‐four cDNA clones encode ribosomal proteins, comprising the single largest category of clones isolated. Twenty‐five of these ribosomal protein genes are likely to be of fungal origin, as was suggested by sequence homology. Hybridization of 56 selected non‐ribosomal protein clones to rust germling cDNA or genomic DNA probes showed that at least 44 were of fungal origin, demonstrating that the library was highly enriched for fungal cDNA. Differential expression analysis identified 26 non‐ribosomal protein genes that were induced in rust‐infected leaves. At least 21 of the induced genes were from the rust fungus, indicating that the majority of the induced genes were rust PIGs that are likely to play a role in parasitism. Some of the induced genes share homology to known PIGs or virulence genes in other fungi, suggesting similarities in parasitism among different fungi. Eight clones correspond to novel PIGs that have not been reported in any organism. This paper reports, for the first time, the isolation of P. triticina PIGs and discusses the use of total rust genomic DNA to identify the source of genes.
Detecting and differentiating Acremonium implicatum: developing a PCR‐based method for an endophytic fungus associated with the genus Brachiaria
Authors: Kelemu Segenet , Dongyi Huang , Guixiu Huang , Takayama Yuka
Published: March 2003

Brachiaria
is a pan‐tropical genus of grasses with about 100 species. The fungus
Acremonium implicatum
can develop an endophytic association that is mutually beneficial with
Brachiaria
species. We developed a polymerase chain reaction (PCR)‐based method by first amplifying DNA from
A. implicatum
isolates using the Random amplified polymorphic DNA (RAPD) technique with arbitrary 10‐mer primers. A 500‐bp PCR product, amplified with primer OPAK‐10 and common to
A. implicatum
isolates, was selected for further evaluation. The fragment was digoxygenin‐labelled and used to probe a dot blot containing genomic DNA from isolates of
A. implicatum
and non‐endophytic fungi, and from
Brachiaria
species free of endophytes. Strong signals were obtained only for DNA from
A. implicatum
isolates. This fragment was cloned and subsequently sequenced. Based on the sequence data, two primers were selected and synthesized: P1 (5′‐TTCGAATGATAAGGCAGATC‐3′) and P4 (5′‐ACGCATCCACTGTATGCTAC‐3′). The primer pair amplified a single fragment of about 500 bp from DNA of
A. implicatum
isolates, whether from pure culture or in association with
Brachiaria
plants. No amplification product was detected in DNA from endophyte‐free plants, pathogenic fungi, the bacterium
Xanthomonas campestris
pv.
graminis
, or non‐pathogenic fungi associated with
Brachiaria
. This assay thus allows the precise and rapid detection of endophytes in
Brachiaria plants and permits a differentiation between endophytic and non‐endophytic fungi.
Predicting safety and quality parameters for UHT‐processed milks
Authors: Browning Emma , Lewis Mike , MacdougalL Douglas
Published: March 2003

A spreadsheet was developed to evaluate safety and quality parameters for milks subjected to different heating and cooling profiles and holding time conditions in a continuous heat exchanger. Safety parameters evaluated were lethality ( Fo) and microbial inactivation (

B*) values; quality parameters were chemical change (

C*), thiamin loss, lactulose formation, Maillard browning and hydroxymethylfurfural (HMF) formation. The spreadsheet was used to predict these parameters for a pilot plant, heating milks at 120–150°C for 4, 27 and 58 s, using its heating and cooling profiles. Milks were processed in the same plant at these conditions and a selection of the milks were analysed to establish the validity of the spreadsheet. All samples with a

C* value < 3 gave a positive Aschaffenburg turbidity. The activation energy that gave the best agreement between the predicted and experimental results for lactulose was found to be 122 kJ/mol. One set of optimized conditions for this product was 135°C for 10 s, giving

B* = 1.24 and

C* = 0.55. This milk was compared with two others and its sensory characteristics were similar to that with a

C* of 1.5 but different to that with a

C* value of 6. Browning was only found to be noticeable in the most severely heat‐treated sample immediately after processing, but would become apparent on storage at 30°C and above. The spreadsheet is useful for comparing the characteristics of products from plants with different heating profiles or as a means for optimizing UHT plant performance where the heating and cooling rates are fixed.
Confidentiality and the Data Protection Act*
Authors: WILLS CHRIS
Published: March 2003

The provisions of the Data Protection Act are outlined and its implications discussed from the viewpoint of the health professional and the patient. The rights and expectations of the latter are emphasized together with the need for the collection of clear and accurate data within the NHS.
Community access to health information in Ireland
Authors: Macdougall Jennifer
Published: March 2003

This paper is based on a research project conducted on consumer health information (CHI) in the Republic of Ireland, the results of which were published in a report entitled Well Read: Developing Consumer Health Information in Ireland. The paper describes the research methodology and the Irish experience in relation to CHI followed by a discussion of access problems, illustrated with examples from the special needs and primary care sectors. The role of information providers in relation to primary healthcare and libraries is examined briefly, and finally the main research conclusions and recommendations are highlighted.
Up‐regulation of lysozyme production in colonic adenomas and adenocarcinomas
Authors: Yuen  , Wong  , Chung  , Chan  , Cheung  , HO  , Leung
Published: March 2003

Aims:

The presence of lysozyme protein in some gastric adenomas and adenocarcinomas has been well documented. There have been relatively few studies investigating the presence of lysozyme in tumours of the large intestine and they show contrasting results. We aim to investigate the cellular source and expression of lysozyme in colonic adenomas and adenocarcinomas.

Methods and results:

We randomly selected 29 and 27 colonic adenomas and adenocarcinomas, respectively. Using in‐situ hybridization (ISH) and immunohistochemistry (IHC), we found an up‐regulation of lysozyme in the dysplastic epithelium of all the adenomas studied, with more than 80% of cases expressing moderate to strong signals. Although the up‐regulation of lysozyme was also observed in adenocarcinomas, only 30% of the cases showed moderate to strong signals, mostly with an uneven distribution. Down‐regulation of lysozyme in the severely dysplastic and invasive foci were noted in some cases of adenoma with malignant transformation. Normal colonic glands were consistently negative for lysozyme at both the mRNA and the protein level, but inflamed and immature regenerative colonic epithelium at the crypt base showed positive signals in a similar pattern to those observed in the dysplastic epithelium of the adenomas.

Conclusions:

Our results confirm that colonic epithelium can produce lysozyme and its expression is up‐regulated in the dysplastic epithelium in adenomas and in invasive cancer cells. It is interesting that regenerative colonic epithelium showed a similar pattern of lysozyme expression as in adenomas. The loss of lysozyme secreting phenotype in most of the invasive tumours suggests that lysozyme may not confer an advantage to tumour progression.
Analysis of CD44 expression in serous and mucinous borderline tumours of the ovary: comparison with cystadenomas and overt carcinomas
Authors: Darai  , Walker‐Combrouze  , Fauconnier  , Madelenat  , Potet  , Scoazec
Published: March 2003

Aims:

To determine the diagnostic and prognostic value of the immunohistochemical analysis of CD44 variants in benign borderline and malignant tumours of the ovary.

Methods and results:

The reactivity of tumour cells with three monoclonal antibodies, respectively, directed to all CD44 variants, CD44‐v3 isoform and CD44‐v6 isoform, was assessed by using an indirect immunoperoxidase technique applied to formalin‐fixed, paraffin‐embedded samples of 36 cases of borderline, as compared to 20 cases of benign tumours and 20 cases of carcinomas. CD44 variants were detected in 97% of borderline tumours, as compared to 60% of benign tumours and 100% of carcinomas. CD44‐v3 was detected in 25% of borderline tumours, as compared to 0% of benign tumours (P = 0.003) and 55% of carcinomas (P = 0.065). The expression of CD44‐v6 was detected in 28% of borderline tumours, as compared to 20% of benign tumours and 30% of carcinomas. In borderline tumours, as in carcinomas, CD44‐v6, but not CD44‐v3, expression was correlated with an increased proliferative index and with a higher incidence of p53 expression.

Conclusion:

Borderline tumours of the ovary present frequent quantitative and qualitative alterations in the pattern of expression of CD44 proteins. However, these alterations are unlikely to represent useful diagnostic or prognostic markers.
Alpha‐1‐antichymotrypsin immunoreactivity in papillary carcinoma of the thyroid gland
Authors: Lai M L , Rizzo N , Liguori C , Zucca G , Faa G
Published: March 2003

Aim

Papillary thyroid carcinoma (PTC) is the most common malignant tumour of the thyroid gland. The immunohistochemical profile of PTC is characterized by immunoreactivity of tumour cells for cytokeratins, thyroglobulin, vimentin, EMA and S100 protein. Recently, the presence of a serum protease inhibitor, alpha‐1‐antitrypsin (A1AT), has been demonstrated in tumour cells of PTC. The aim of our study was to test immunoreactivity of PTC for another inhibitor of proteases, alpha‐1‐antichymotrypsin (A1ACT).

Methods and results

Serial paraffin sections of nine consecutive cases of PTC were tested with anti‐A1AT and anti‐A1ACT antibodies. No immunoreactivity for A1AT and A1ACT was found in the normal thyroid tissue surrounding each tumour. In seven out of nine cases, tumour cells of PTC showed cytoplasmic immunoreactivity for A1ACT. In two cases, A1ACT was detected even in the nuclei. Immunoreactivity for A1AT was found only in three cases. Two cases of PTC showed no staining for both A1ACT and A1AT. No significant correlation of A1ACT staining was found with various prognostic indices (age of patients, histological pattern, tumour size, presence of regional lymph node metastases). The two cases showing a lack of staining for both A1ACT and A1AT showed a more aggressive clinical behaviour.

Conclusions

Our preliminary study shows that A1ACT is expressed by tumour cells in a large proportion of papillary carcinomas of the thyroid gland. Its significance remains, to the best of our knowledge, still unknown. The observation of a more aggressive behaviour in the two cases characterized by the absence of immunoreactivity for both A1ACT and A1AT suggests that the presence or absence of protease inhibitors could play a role in controlling tumour progression in PTC.
Endolymphatic sac tumours
Authors: Kempermann G , Neumann H P H , Volk B
Published: March 2003

This review article surveys clinical and pathological literature on endolymphatic sac tumours (ELST) and summarizes characteristics that describe the entity.

ELST are rare neuroectodermal neoplasms in the petrous bone, originating from inner ear structures. They can be encountered sporadically or in von Hippel–Lindau disease. The most prominent symptom is sensorineural deafness. Historically, nomenclature of invasive adenoid tumours in the petrous bone has been divergent, the term papillary adenocarcinoma used most frequently. Histologically, they have a follicular or papillary and adenoid pattern that can be easily confused with various other neoplastic conditions including metastatic carcinoma. It remains to be verified whether similar tumours (papillary adenocarcinomas) can originate from the middle ear. Middle ear adenomas have a similar appearance but probably originate from neural crest cells in the middle ear. ELST can express a variety of epitopes (including cytokeratin and neuro‐ectodermal markers) which can be detected immunohistochemically.

In cases of von Hippel–Lindau disease the cerebello‐pontine angle should be included in routine radiological examinations to detect ELST before the tumours lead to deafness. In apparently sporadic cases of ELST, genetic testing for von Hippel–Lindau disease should be considered. Correct distinction of ELST from metastatic carcinoma prevents futile searches for unknown primary tumours.
Direct sequencing of the p53 gene shows absence of mutations in endometrioid endometrial adenocarcinomas expressing p53 protein
Authors: Stewart R L , Royds J A , Burton J L , Heatley M K , Wells M
Published: March 2003

Aims

To investigate the mutational status of the p53 gene (Tp53) in endometrial carcinomas expressing p53 protein by direct gene sequencing.

Methods and results

Eighteen cases of endometrial carcinoma, selected on the basis of p53 protein expression as detected immunohistochemically by the monoclonal antibody DO7, were microdissected in the p53 positive areas. DNA was extracted and subjected to polymerase chain reaction (PCR) for exons 5–8 which code for conserved areas containing mutational hot‐spots. The PCR amplified material was then sequenced using an ABI automated sequencer and analysed using BLAST software at the NCBI web site. All sequences analysed were wild‐type.

Conclusion

The results confirm that expression of p53 protein may occur in endometrial adenocarcinoma without mutation and may be due to stabilization of the protein during its normal function, possibly by an mdm2 mediated process.
p34cdc2 in invasive breast cancer: relationship to DNA content, Ki67 index and c‐erbB‐2 expression
Authors: Wiesener  , Hauser‐Kronberger  , Zipperer  , Dietze  , Menzel Ch , Hacker
Published: March 2003

Aims: One‐hundred and eighty‐eight cases of human mammary carcinoma were examined immunohistochemically for their expression of Ki67, p34cdc2 and c‐erbB‐2. DNA image cytometry was performed to evaluate DNA ploidy, Auer type, S‐phase fraction (SPF), 5c exceeding rate (5cER) and 2c deviation index (2cDI).

Methods and results

One‐hundred and sixty‐eight cases were invasive ductal carcinomas, 20 were of invasive lobular type. Routinely assessed oestrogen and progesterone receptor scores were available. The results were analysed statistically in comparison to tumour type, histopathological grade, lymph node status, menopausal status, patient age and overall survival. Ki67 (P < 0.002) and c‐erbB‐2 (P < 0.0001) correlated well with overall survival (P < 0.0008) and grade (P < 0.038) but not with lymph node status and tumour type. p34cdc2 showed a trend towards a positive correlation with Ki67 (P < 0.058) and a significant negative correlation with receptor status (P < 0.008) but with none of the other parameters examined.

Conclusions

No association between the DNA measured parameters (Auer type, SPF, 5cER and 2cDI) and survival was found. Our results suggest that c‐erbB‐2 and Ki67 are parameters which might, in combination with receptor status, help to define subgroups with different outcomes.
Interobserver agreement in the assessment of gastritis reversibility after Helicobacter pylori eradication
Authors: Tepeš  , Ferlan‐Marolt  , Juteršek  , Kavčič  , Zaletel‐Kragelj
Published: March 2003

Aim

Our aim was to determine interobserver agreement in the application of the Sydney system to assess reversibility of gastritis after Helicobacter pylori eradication.

Methods and results

Forty‐three patients with a Helicobacter pylori‐positive duodenal ulcer disease were included in the study. All patients included had successful H. pylori eradication after different antimicrobial drug combinations. Biopsy samples were collected from antrum and body, according to the Sydney recommendations, before antimicrobial therapy, 2 months after and at yearly intervals during 2–4 years of follow up. Three pathologists, who were blind to clinical data, evaluated histological changes in 221 antral and 219 body specimens stained with haematoxylin and eosin and with Warthin Starry. The percentage of pairwise agreement, kappa and weighted kappa statistic were used. Agreement in recognizing the presence of H. pylori colonization of the gastric mucosa, activity of inflammation and intestinal metaplasia was over 90%. Agreement in recognizing chronic inflammation in the body and atrophy in the antrum was between 78 and 89% respectively. The kappa values were excellent (more than 0.75) for the grade of H. pylori in the body, good (between 0.50 and 0.75) for the grade of H. pylori in the antrum, grade of inflammatory activity and intestinal metaplasia in the antrum and moderate to good (0.38–0.53) for the grade of chronic inflammation. Kappa values were poor to good (from 0.17 to 0.57) only in evaluation of the grade of atrophy.

Conclusion

Interobserver agreement in the application of the Sydney system to reversibility of gastritis after H. pylori was good. More strict criteria should be used for atrophy and to differentiate normal and mild chronic inflammation.
Expression of MUC1 and MUC2 mucin gene products in Barrett's metaplasia, dysplasia and adenocarcinoma: an immunopathological study with clinical correlation
Authors: Chinyama  , Marshall  , Owen  , Mason  , Kothari  , Wilkinson  , Sanderson
Published: March 2003

Aims

Changes in the histochemical characteristics of the surface epithelial mucins is the hallmark of Barrett's metaplasia. The study investigated the pattern of expression of MUC1 and MUC2 mucin gene products in Barrett's metaplasia, dysplasia and adenocarcinoma as possible indicators of increased malignant potential.

Methods and results

Tissue sections from 51 patients with Barrett's intestinal metaplasia, nine with dysplasia (three indefinite) and 28 resected adenocarcinomas were stained with monoclonal antibodies to MUC1 and MUC2. The majority of the patients were men (70/88, 80%) who were treated over a period of 3 years. None of the patients with dysplasia or carcinoma were under surveillance at the time of presentation. All 51 biopsies with Barrett's metaplasia expressed MUC2 and MUC1 was consistently absent. Neither MUC1 or MUC2 were expressed in the dysplastic epithelium whether in its pure form (6/6) or when associated with carcinoma (26/28) (P < 0.005). Three biopsies which were initially classified as high‐grade dysplasia expressed MUC1 and these turned out to be carcinomas on further investigations. MUC1 was also expressed in 12/28 (43%) of the adenocarcinomas and majority of these were poorly differentiated stage 3 tumours (P < 0.05). MUC2 was only positive in mucin‐secreting carcinomas (4/28; 14%) irrespective of the tumour stage.

Conclusion

Despite the large number of patients with Barrett's metaplasia and carcinoma, very few patients presented with dysplasia, implying that Barrett's oesophagus is a silent disease in the community presenting late as carcinoma. The study has demonstrated aberrant expression of MUC2 (an intestinal mucin) in Barrett's metaplasia and this expression is lost when the cells become dysplastic. The lack of MUC1 in dysplastic epithelium and its expression in carcinoma could be utilized as a marker which could differentiate dysplasia from carcinoma in mucosal biopsies. Furthermore, expression of MUC1 in advanced stage oesophageal cancers (as in breast cancer) suggests an unfavourable prognosis.
Microinvasive carcinoma of the breast: can it be diagnosed reliably and is it clinically significant?
Authors: Hoda  , Prasad  , Moore  , Hoda  , Giri
Published: March 2003

Deciding whether in‐situ breast carcinoma is associated with microinvasion is a common problem. Histological features resembling invasion can be simulated by in‐situ carcinoma distorted by inflammatory and reparative changes. Having expended the effort to diagnose genuine microinvasion, just how useful is this diagnosis in planning further treatment and follow‐up? In the following articles, Hoda et al. comment on the utility of immunohistochemistry in resolving uncertainty about the presence of microinvasion, and Ellis et al. critically appraise the definition of microinvasion and its clinical significance.
Varied B‐cell immunophenotypes of Hodgkin/Reed–Sternberg cells in classic Hodgkin's disease
Authors: Watanabe  , Yamashita  , Nakayama  , Hasegawa  , Kojima  , Nagasawa  , Mori
Published: March 2003

Aims

Recent analyses have suggested that Hodgkin's or Reed–Sternberg (HRS) cells in most cases of classic Hodgkin's disease are derived from germinal centre B‐cells. However, there is controversy over which B‐cell antigens are expressed in HRS cells.

Methods and results

We studied 51 cases of classic Hodgkin's disease to immunohistochemically characterize HRS cells for pan‐B‐cell markers and specific markers for plasma cells. HRS cells expressed CD20 (L26) in 18 cases (35%), CD19 (B4) in nine (18%) and CD79a (mb‐1) in 13 (25%). Furthermore, HRS cells were positive for CD138 (B‐B4) in 24 cases (45%) and for PCA‐1 in 24 (45%). In 41 (80%) cases, HRS cells expressed more than one B‐cell marker. We then subclassified cases into those positive for plasma cell markers (n = 27) (group 1) and those negative for them (n = 24) (group 2). The average age in group 1 (40 years) was younger than in group 2 (54 years) (P < 0.05). The percentage of nodular sclerosis (NS) subtype in group 1 (52%) was 1.5 times greater than in group 2 (33%) (P < 0.05). With regard to Epstein–Barr virus encoded small RNA (EBER) in‐situ hybridization, 14 cases (64%) were positive in group 2, but only seven cases (31%) were positive in group 1 (P < 0.025).

Conclusion

In most cases of classic Hodgkin's disease, HRS cells expressed later stage of B‐cell development. We consider that two different clinicopathological groups may correlate with the two different expressions of B‐cell markers.
Rhabdoid tumour of the lung is a dedifferentiated phenotype of pulmonary adenocarcinoma
Authors: Miyagi J , Tsuhako K , Kinjo T , Iwamasa T , Hashimoto H , Ishikawa S
Published: March 2003

Aims

Primary rhabdoid tumour of the lung is rare, and histological and biological characteristics have not been fully documented. We describe three cases of primary lung rhabdoid tumour, all associated with adenocarcinoma, and investigate the histological features and biological characteristics.

Methods and results

Three cases were obtained from a total 902 cases of surgically removed primary lung tumours between 1986 and 1998. The rhabdoid cells were found to occupy about 50–90% of each tumour. All of the tumours had nonrhabdoid adenocarcinoma foci in the centre of the tumours. Transition between the adenocarcinomatous and rhabdoid components was demonstrated. Detailed immunohistochemical studies were carried out. The epithelial markers, cytokeratins and epithelial membrane antigen (EMA), were strongly expressed in rhabdoid and adenocarcinomatous components. Furthermore, surfactant apoprotein A was positive in both components in one case, but myoglobin, MyoD and HHF35 were not expressed. Vimentin was strongly and diffusely stained in all cases. The neuroendocrine markers, chromogranin A (all cases), neuron‐specific antigen (NSE) (two cases) and CD56 (one case) were occasionally positive in only a small number of the rhabdoid tumour cells. GM‐CSF was positively stained in one case, and the dedifferentiated characteristics of the rhabdoid cells was suggested. Proliferative cell nuclear antigen (PCNA) was strongly demonstrated in the rhabdoid tumour cells (all cases). To gain better understanding the highly proliferative characteristics of the tumours, p53 gene (exons 5–8) mutation was examined by DNA sequencing analysis; mutation of the p53 DNA was not detected. Overexpression of p53 protein was also not demonstrated in all cases. HPV6 was demonstrated in one case by PCR method and also non‐isotopic in‐situ hybridization (NISH). Two cases died in a short period of time (3 years and 4 months, respectively).

Conclusion

The rhabdoid cells in these three cases were considered to represent the dedifferentiated components of the accompanying adenocarcinoma. Dedifferentiated characteristics (neuroendocrine markers, GM‐CSF, vimentin, and the aggressive behaviour) were evident.
Expression of VCAM‐1, ICAM‐1, E‐ and P‐selectin and tumour‐associated macrophages in renal cell carcinoma
Authors: Hemmerlein B , Scherbening J , Kugler A , Radzun H‐J
Published: March 2003

Aims

Neoangiogenesis is accompanied by an increase in endothelial surface, which can support infiltration by immune cells depending on adhesion molecule expression. Therefore, the expression of cell adhesion molecules on microvessels and epithelial cells was analysed in renal cell carcinomas as compared to tumour‐free tissue.

Methods and results

PECAM‐1, CD34, ICAM‐1, VCAM‐1, VLA‐4, P‐ and E‐selectin, the macrophage antigens Ki‐M1P and Mac‐1, and lymphocyte function antigen LFA‐1 were identified immunohistochemically. VCAM‐1, ICAM‐1, and E‐selectin were equally or less expressed, whereas P‐selectin was increased on microvessels in tumour tissue. The density of VCAM‐1‐positive tumour microvessels correlated positively with an advanced tumour stage and E‐ and P‐selectin‐positive tumour microvessels with the amount of associated macrophages. The expression of ICAM‐1 and VCAM‐1 on neoplastic epithelia correlated with an increased density of macrophages and a minor degree of tumour differentiation.

Conclusions

The positive correlation of macrophage infiltration and expression of cell adhesion molecules on tumour microvessels and epithelia with minor tumour differentiation and an advanced stage indicates that adhesion molecule expression is not associated with an effective antitumour function of macrophages
Microcystic adenoma of the pancreas (glycogen‐rich cystadenoma) with stromal amyloid deposits
Authors: Tripodi S A , Civitelli S , Schürfeld K , Cintorino M
Published: March 2003

Aims

We report a case of a pancreatic glycogen‐rich microcystic serous adenoma with stromal amyloid deposits, focusing on the significance of isolated amyloid deposits in tumours.

Methods and results

The architectural pattern was characterized by thin‐walled cysts lined by a single layer of flat or cuboidal epithelial cells intensely stained by the PAS‐reaction only before diastase digestion, suggesting the presence of glycogen. Tumour stroma was composed of broad fibrocollagenous tissue with lamellar hyalinized areas which were positively stained by Congo red and showed green birefringence and dichroism with polarized light. For amyloid protein characterization, immunohistochemical studies were performed with anti‐β amyloid protein and anti‐amyloid precursor pre‐A4695. The former antibody diffusely stained tumour stroma, while the latter stained only scattered stroma cells.

Conclusions

This is the first documented case of amyloid deposition in pancreatic serous adenoma. We indicate that the source of amyloid is an APP‐like precursor secreted by stromal myofibroblasts.
Cotyledonoid hydropic intravenous leiomyomatosis: a new variant leiomyoma
Authors: Jordan L B , Al‐Nafussi A , Beattie G
Published: March 2003

Cotyledonoid hydropic intravenous leiomyomatosis: a new variant leiomyoma

Aims: We present the histopathological findings of a series of six cases of a benign uterine smooth muscle tumour with an unusual growth pattern.

Methods and results: All cases have the appearances of the recently described dissecting (cotyledonoid) leiomyoma. In addition, three of these lesions demonstrate the features of intravenous leiomyomatosis with varying degrees of hydropic degeneration.

Conclusions: This combination of phenotypes has not previously been described within the literature; therefore we propose that these are classified as examples of `cotyledonoid hydropic intravenous leiomyomatosis', a new variant of unconventional leiomyoma.
Ultrastructural and immunohistochemical characterization of the so‐called giant multinucleate cells in cutaneous collagenomas
Authors: Ramos D , Monteagudo C , Carda C , Ramón D , González‐Devesa M , Llombart‐Bosch A
Published: March 2003

Ultrastructural and immunohistochemical characterization of the so‐called giant multinucleate cells in cutaneous collagenomas

Aims:  This study was undertaken to compare the histopathological, immunohistochemical and ultrastructural features of the so‐called giant multinucleatecells in cutaneous collagenomas: giant‐cell collagenoma and solitary sclerotic fibroma.

Methods and results:  We studied four collagenomas: one giant‐cell collagenoma and three solitary sclerotic fibromas. All the cases showed an indolent clinical presentation and were histologically constituted by a well‐demarcated dome‐shaped proliferation of coarse collagen bundles with a varying number of interspersed giant multinucleate cells and stellate mononuclear cells. The immunohistochemical study on paraffin sections revealed that neoplastic cells in both collagenomas were vimentin and CD34‐positive, whereas FXIIIa was only expressed in solitary sclerotic fibromas. In regard to the so‐called giant multinucleate cells, we have ultrastructurally found that these cells were ‘real’ multinucleate cells in giant‐cell collagenoma, whereas in solitary sclerotic fibromas they consisted of closely packed aggregates of individual stellate mononuclear cells. Moreover, perinuclear cisternae focally containing finely textured material of moderate density were unexpectedly found in giant cells of giant‐cell collagenoma, a finding which was not observed in solitary sclerotic fibromas. Additionally, a characteristic cell–cell interaction between tumour cells and mast cells was encountered in all collagenomas.

Conclusions:  This study supports a distinctive immunohistochemical and overall ultrastructural profile of giantmultinucleate cells in giant‐cell collagenoma and solitary sclerotic fibroma, which suggests a different pattern of differentiation for these two related cutaneouslesions.
Polymorphous low‐grade adenocarcinoma of the salivary glands with transformation to high‐grade carcinoma
Authors: Simpson R H W , Reis‐Filho J S , Pereira E M , Ribeiro A C , Abdulkadir A
Published: March 2003

Aims:  Polymorphous low‐grade adenocarcinoma of the minor salivary glands is an infiltrative neoplasm characterized by bland‐looking tumour cells arranged in diverse architectural patterns. It is considered to be of low‐grade malignant potential in that nodal metastases are seen in only a minority, and distant spread is rare. Even more unusual is the transformation of polymorphous low‐grade adenocarcinoma to a histologi‐cally high‐grade carcinoma, i.e. dedifferentiation. In this paper, we describe the clinicopathological and immunohistochemical findings in two further examples.

Methods and results:  Two patients presented each with a tumour of the palate. Histopathological examination showed the typical morphological, cytological and immunohistochemical features of a polymorphous low‐grade adenocarcinoma. In one case there was a second component of high‐grade carcinoma showing nuclear atypia, markedly increased mitotic activity and MIB1 index, as well as prominent zones of necrosis. It expressed epithelial markers and androgen receptors, and thus resembled salivary duct carcinoma. Similar tumour tissue was observed in one of the cervical nodal metastases, which was biopsied at the same time as the palate. In the second patient, a high‐grade component was discovered when the tumour recurred in the palate 13 years after the initial biopsy. Whilst morphologically similar to that in first case, there were significant immunohistochemical differences such as retention of some of the polymorphous low‐grade adenocarcinoma profile and absence of androgen receptor expression.

Conclusions:  Polymorphous low‐grade adenocarcinoma was first described relatively recently, and as experience with it continues to accumulate, it is becoming clear that late recurrences and metastases, whilst still infrequent, may not be quite as rare as previously thought. Reports of histological transformation are even scarcer, and most occurred at least 13 years after the polymorphous low‐grade adenocarcinoma was initially recognized. It is a real possibility that this phenomenon, like clinical progression, may also be encountered more often as time passes. Therefore, we believe that, whilst polymorphous low‐grade adenocarcinoma is certainly far less aggressive than, for example, adenoid cystic carcinoma, it nevertheless remains a true malignancy with a potential to prove fatal in a minority of patients.