bluuebunny/crossref_metadata_2025 huggingface dataset:
Plumage‐based phylogenetic analyses of the Merops bee‐eaters
Authors: BURT D. BRENT
Published: December 2004

I review previous systematic work on the family Meropidae and present phylogenetic hypotheses derived from my analyses of colour, pattern and shape variation in 30 plumage regions among species and subspecies in this family. Consistent patterns are seen across shallow portions of the trees. Uncertainty remains concerning the placement of several deep branches within this group's phylogeny. In particular, the phylogenetic placement of Meropogon forsteni and Merops breweri, M. ornatus, M. hirundineus and M. boehmi remains uncertain. The biogeographical patterns in the resultant trees are similar with either a Southeast Asian or African origin for the family, with most of the early diversification occurring in Africa, and with multiple independent subsequent invasions of non‐African areas.
A new method for estimating the adult survival rate of the Corncrake Crex crex and comparison with estimates from ring‐recovery and ring‐recapture data
Authors: Green Rhys E.
Published: December 2004

The mean annual survival rate of adult Corncrakes Crex crex was estimated by three independent approaches: ring‐recovery, ring‐recapture and the shape of secondary remiges. The last method is new and uses measurements of the shape of the tips of the secondaries, which changes in the first post‐juvenile wing moult to become less pointed. The estimates obtained by the three approaches were mutually compatible and indicated that annual survival is likely to be within the range 0.2–0.3. This low survival rate is likely to make the growth rate of Corncrake populations particularly sensitive to the effects of agricultural and conservation management on breeding success and recruitment. The survival rate of adult female Corncrakes was estimated for the first time (0.259) and was found not to differ significantly from an estimate for adult males (0.298) made in the same study area by the same method. It is concluded that the method based upon population counts and measurements of the shape of remiges has potential value for studies of the demography of Corncrakes and other species.
Semen quality and sedentary work position
Authors: Støy Julie , Hjøllund Niels Henrik I. , Mortensen Jens Tølbøll , Burr Hermann , Bonde Jens Peter
Published: December 2004

Increased scrotal temperature can, in experimental settings, markedly disturb the production of semen. Sedentary work position may increase the temperature of the scrotum, but previous studies have failed to determine whether changes in scrotal temperature caused by sedentary work actually do affect semen quality. This study was carried out to elucidate the possible harmful effects of sedentary work on sperm count and other semen characteristics. In 1981–1983 a semen sample was obtained from 3119 men who attended an infertility workup in one of four Danish fertility centres. A total of 2517 men returned a postal questionnaire with information on life style, leisure time activities, occupational history and job duties. Information on job specific work position was obtained from The Danish Work Environment Cohort study 1990 (DWECS). In this analysis DWECS data for a total of 1747 men was included from men aged 18–39 years with >30 h of work per week. For all job titles represented in the DWECS, the mean proportion of sedentary work was estimated. The sperm cell concentration was 30.6 million/mL among men in the quintile with lowest job specific sedentary work compared with 40.5 million/mL in the highest quintile. The difference was, however, not statistically significant. Stratification on infertility period, educational level of the man, fertility centre, and fertility‐related disease of the spouse did not influence the results. The analyses do not suggest that sedentary work is a risk factor for abnormal semen characteristics.
Spectrophotometric application of resazurin reduction assay to evaluate boar semen quality
Authors: Zrimšek P. , Kunc J. , Kosec M. , Mrkun J.
Published: December 2004

The resazurin reduction assay depends on the ability of metabolically active cells to reduce the resazurin redox dye to resorufin. In the present study we applied and made a diagnostic evaluation of a spectrophotometric application of the resazurin reduction assay to assess the colour change of resazurin reduction in butanol extracted colour to evaluate boar semen quality. Forty‐one samples of boar semen from various breeds were included in the study. The absorption peaks for resazurin and resorufin were found to be 610 and 575 nm, respectively. Absorbance at 610 nm, where the minimum overlap of the two peaks was observed, was used in further analysis. Spearman rank correlation analysis was used to determine the correlation between the resazurin reduction assay and various semen parameters. The highest correlations were observed with the concentration of motile spermatozoa (r = −0.841; p < 0.001), sperm concentration (r = −0.833; p < 0.001), sperm index (−0.826; p < 0.001) and concentration of viable spermatozoa (r = −0.763; p < 0.001). Sensitivity and specificity, at 94.1 and 91.7%, respectively, indicate that the present test is highly accurate in discriminating between the samples according to the sperm index. When motile sperm concentration was used to distinguish between good and poor samples, high sensitivity (93.6%) was also found, whereas the test was only moderately, 80%, specific. The stability of butanol extracts in terms of A610 at different times of measurement confirmed that the resazurin reduction could be spectrophotometrically measured within 7 days from the time of assay performance, making the assay much more useful. Based on these results, the assay could be used as an additional tool for evaluating the quality of boar semen.
Use of hydrogen peroxide to assess the sperm susceptibility to oxidative stress in subjects presenting a normal semen profile
Authors: Misro M. M. , Choudhury L. , Upreti K. , Gautam D. , Chaki S. P. , Mahajan A. S. , Babbar R.
Published: December 2004

Human sperm susceptibility to oxidative stress is vital as it affects various characteristics of sperm function. In the present study, we report a simple, sensitive and quick method of assessing the capacity of the sperms to withstand increased oxidative stress. The basis for the test was derived from the fact that human sperms suspended in Ham's F‐10 medium tend to lose the forward progressive motility when co‐incubated with H2O2 (600 μm). Replacement of the medium with seminal plasma (1 : 1) was able to reduce the loss of sperm motility (40%). Retention of sperm motility in semen (0–30%) following 10 min of H2O2 (600 μm) exposure was taken as the criteria for delineating the quality of sperm as poor, moderate, good and excellent types. The protocol was tested in 87 subjects presenting a normal semen profile. On the basis of this test, 44% of the semen samples were classified as poor and the rest as moderate, good or excellent. Lipid peroxidation was found higher in the sperms from the ‘poor’ category. Activities of superoxide dismutase and catalase were also significantly elevated in the seminal plasma of these subjects as compared with combined categories of good or excellent. The test described here can be used routinely in laboratory investigations to assess sperm susceptibility to oxidative stress in subjects presenting a normal semen profile.
Use of a β‐glucan hydrocolloidal suspension in the manufacture of low‐fat Cheddar cheese: manufacture, composition, yield and microstructure
Authors: Konuklar G. , Inglett G. E. , Carriere C. J. , Felker F. C.
Published: December 2004

Low‐fat Cheddar cheese was manufactured using a β‐glucan, hydrocolloidal fat replacer denoted as Nutrim. The composition, production efficiency, microstructure, and utility of replacing fat with Nutrim were examined. Cheese samples (designated as Nutrim‐I, and Nutrim‐II) containing Nutrim were produced with mean fat levels of 6.84 and 3.47%, respectively. A low‐fat cheese was also produced as a control with a mean fat level of 11.2%. Nutrim‐II cheese had significantly higher moisture, salt, and ash contents as compared with the low‐fat control cheese. The low‐fat control cheese had a higher yield normalized for 54% moisture and 1.5% salt content as compared with the Nutrim‐II cheese. Scanning electron microscopy revealed smaller and more uniform fat droplet voids in the Nutrim cheese than the low‐fat control, and a more dense, noncontinuous background protein matrix with globular clusters suggesting a physical buffering afforded by the presence of the β‐glucan hydrocolloid or its associated water.
Effects of ultra high hydrostatic pressure on Listeria monocytogenes and natural flora in broth, milk and fruit juices
Authors: Erkmen O. , Dogan C.
Published: December 2004

Listeria monocytogenes was subjected to ultra high hydrostatic pressure (UHHP) treatments from 200 to 700 MPa at 25 °C in broth, raw milk, peach juice and orange juice. Survivor curves showed that cell death increased as pressure increased. After 10 min pressure treatment at 400 MPa reductions of about 2.09 and 2.76 log CFU mL−1 in aerobic bacteria and L. monocytogenes, respectively, were produced in raw milk, this increased to 5.09 and 6.47 log CFU mL−1, respectively, at 600 MPa. Death of bacteria at UHHP treatment was greater in orange juice than peach juice, and in peach juice than milk. Listeria monocytogenes was more sensitive to increased pressure than increased pressurization time. Injury of L. monocytogenes occurred from 0 to 100%. Factors effecting the rate of microbial inactivation are: pressure, age of cell, composition of medium, and pressurization time. UHHP inactivation can be used to extend shelf life and increase food quality during storage, and may also contribute to inactivation of L. monocytogenes.
Microfiltration of whey proteins adsorbed on yeast cells
Authors: Keskinler Bulent , Erhan Elif , Akay Galip , Kaya Mukerren , Bayguven Betul
Published: December 2004

Soluble whey proteins (WPs), adsorbed on yeast cells, were recovered by a crossflow microfiltration (MF) technique using a cellulose nitrate membrane with a pore size of 0.45 μm. The crossflow velocity was 1.5 m s−1 with a transmembrane pressure of 200 kPa at 25 °C. A series of protein rejections occured at various pH values ranging from 2 to 8. WPs adsorbed more on to yeast cells at low pH (pH < 4) than at high pH values, probably because they were positively charged at low pH. It was also shown that permeate flux increased and Modified Membrane Fouling Index values decreased at low pH levels. When the yeast concentration was 50 g L−1, the flux decreased five times compared with that in the absence of yeast. Protein recovery increased with increasing yeast concentrations. The highest protein recovery was found to be 85% at a yeast concentration of 50 g L−1 at a steady state flux rate of 10−6 m s−1 at 25 °C. When diluted solutions of whey were used, the same rejection of protein, adsorbed on yeast cells, was achieved at ten times lower amounts of yeast cells. This technique not only provides for the recovery of protein but also may give rise to the direct use of yeast cells, which are rich in protein, in the baking industry. WPs absorbed by yeast cells can be used to produce nutritionally rich products in areas where yeasts have been already used.
The effects of various salt concentrations during brine curing of cod (Gadus morhua)
Authors: Thorarinsdottir Kristin Anna , Arason Sigurjon , Bogason Sigurdur G. , Kristbergsson Kristberg
Published: December 2004

The method used for salting of cod (Gadus morhua) is believed to influence the quality and characteristics of the final product. In recent years an initial brine salting for 1–4 days has preceded dry salting; this increases both the weight yield and quality of the final product. After removal from the brine, dry salting is followed by packaging and storage. The effect of the salt concentration in the brine has been a matter of controversy, with some indication that higher weight yield and quality may be obtained by using lower salt concentrations than by using a fully saturated brine solution. Therefore to test this hypothesis, the effect of different brine salting methods was studied; traditional brine salting, maintaining a constant brine concentration and increasing the salt concentration gradually during brining. The results indicated that the effect on weight yield, chemical composition and water holding capacity of the salted or rehydrated cod were not significant. Altering salt concentration of the brine, by adding salt during brining, did not result in any significant improvements in weight yield, either after the salting process or after the rehydration. The overall quality was increased by using a lower salt concentration of 16° Bau compared with 20 and 24° Bau.
Extrauterine (tubal) placental site nodule
Authors: Campello T.R. , Fittipaldi H. , O'valle F. , Carvia R.E. , Nogales F.F.
Published: December 2004

Aims

The clinicopathological and immunohistochemical features of the second case of placental site nodule (PSN) of extrauterine, tubal location are presented.

Methods and results

The lesion was incidentally found in the right tube during a cesarean section and eventual tubal ligation in a 23‐year‐old women gesta 2 para 1, after an uneventful 39‐week intrauterine pregnancy. Grossly, the right Fallopian tube had a 1 cm dilatation filled by necrotic material. Microscopically, the lumen of the Fallopian tube was effaced and replaced by a rim of pleomorphic intermediate trophoblastic (IT) cells with pseudoinvasive parietal features which were positive for human placental lactogen, placental alkaline phosphatase, epithelial membrane antigen and CAM5.2. The Ki67 index was 3%.

Conclusion

Due to its bizarre microscopic appearance, this lesion should be included in the differential diagnosis with malignant conditions. Both origins from a previous subclinical extrauterine tubal pregnancy and a possible migration of IT from a uterine implantation are considered.
Dermatofibrosarcoma protuberans and its fibrosarcomatous variant with areas of myoid differentiation: a report of three cases
Authors: Morimitsu Y. , Hisaoka M. , Okamoto S. , Hashimoto H. , Ushijima M.
Published: December 2004

Aims

We describe three examples of dermatofibrosarcoma protuberans which demonstrated focal myofibroblastic differentiation, and discuss the nature of myofibroblastic differentiation.

Methods and results

We studied three cases of dermatofibrosarcoma protuberans, two of which had fibrosarcomatous portions containing myoid areas, and one which showed myoid areas in ordinary dermatofibrosarcoma protuberans. Myoid areas were recognized histologically as randomly distributed, scattered bundles or small nodules of spindle cells with deeply eosinophilic cytoplasm. Myofibroblastic differentiation was demonstrated immunohistochemically by positive staining for smooth muscle actin and muscle‐specific actin, and ultrastructurally by the presence of microfilament bundles with focal dense bodies and pinocytic vesicles. The myoid areas occasionally contained intraneoplastic blood vessels.

Conclusion

The presence of myoid areas may be related to reactive hyperplasia of stroma, and its relation to the histogenesis of dermatofibrosarcoma protuberans is uncertain.
Renal cell carcinoma with rhabdoid features: an aggressive neoplasm
Authors: Kuroiwa K , Kinoshita Y , Shiratsuchi H , Oshiro Y , Tamiya S , Oda Y , Naito S , Tsuneyoshi M
Published: December 2004

Aims:  Only a few reports on renal cell carcinoma with rhabdoid features have been published. This study was performed to investigate the clinicopathological characteristics of renal cell carcinomas with rhabdoid features.

Methods and results:  Among 253 cases of renal cell carcinoma in adults, eight cases with rhabdoid features were detected. Rhabdoid areas ranged from 10% to 90% of each of the cases. Seven of the eight cases were TNM stage III or IV, and four of the eight cases died within 8 months of surgery. Immunohistochemically, the rhabdoid areas were positive for CAM 5.2 (4/8), AE1/AE3 (6/8), epithelial membrane antigen (6/8) and vimentin (8/8), and negative for myogenetic markers (0/8). The mean MIB‐1 labelling index in the rhabdoid areas was higher than that in the definite carcinomatous areas. Ultrastructurally, perinuclear whorls of intermediate filaments were demonstrated in three of the eight cases using paraffin‐embedded blocks.

Conclusions:  The rhabdoid areas in renal cell carcinoma have histological, immunohistochemical and ultrastructural similarities to malignant rhabdoid tumours. Renal cell carcinoma with rhabdoid features is a highly aggressive neoplasm and its malignant behaviour may be due to the high cell‐proliferative activity of the rhabdoid areas. Rhabdoid features in renal cell carcinoma may represent the endpoint of clonal evolution of renal cell carcinoma (especially in clear cell type cases).
Clear cell sarcoma of the stomach
Authors: Pauwels P , Debiec‐Rychter M , Sciot R , Vlasveld T , Den Butter B , Hagemeijer A , Hogendoorn P C W
Published: December 2004

Aim:  Clear cell sarcoma is a high‐grade sarcoma with morphological features resembling malignant melanoma. This tumour is reported to display a characteristic distribution pattern nearly always involving the extremities. We report the first case of clear cell sarcoma of the stomach.

Methods and results:  A 30‐year‐old male developed a huge tumour of the stomach, which at first glance could be considered as a poorly differentiated carcinoma. Immunohistochemical and ultrastructural examinations were consistent with a diagnosis of metastatic melanoma. However, cytogenetic examination revealed a t(12;22) translocation, specific for clear cell sarcoma. This was confirmed by fluorescence in‐situ hybridization.

Conclusion:  Making a reliable diagnosis of clear cell sarcoma of the stomach requires cytogenetic or molecular diagnostic investigations, particularly to rule out metastatic melanoma. This diagnosis avoids an unnecessary search for a primary melanoma.
Changes in the distribution pattern of gelatin‐binding protein of 28 kDa (adiponectin) in myocardial remodelling after ischaemic injury
Authors: Ishikawa Y , Akasaka Y , Ishii T , Yoda‐Murakami M , Choi‐Miura N‐H , Tomita M , Ito K , Zhang L , Akishima Y , Ishihara M , Muramatsu M , Taniyama M
Published: December 2004

Aims:  Gelatin‐binding protein of 28 kDa (GBP28) is a collagen‐like plasma protein having a binding capacity with collagens. We investigated GBP28 role on myocardial remodelling as well as the diagnostic significance of GBP28 immunostaining in myocardial infarction.

Methods and results:  Myocardial tissues obtained from 47 autopsied hearts with infarction were immunostained with antibodies against GBP28, fibronectin, type III and IV collagens, and prolyl 4‐hydroxylase. GBP28 was distributed in interstitium of infarcted lesions at an early stage. GBP28 was linearly found both along the border with vital myocardium and at the periphery of surviving cardiomyocyte around the lesion at granulative stage. However, it was not observed in the scar. GBP28 distribution patterns were similar to those of fibronectin in infarcted lesions and those of type IV collagen at the periphery of cardiomyocyte. Type III collagen was not recognized in the early‐stage lesion but increased along with scar maturation. Prolyl 4‐hydroxylase was found in surviving cardiomyocytes around the lesion during all stages and in interstitial cells appeared in granulation tissue.

Conclusion:  GBP28 plays a role as a scaffold of newly formed collagen in myocardial remodelling after ischaemic injury, and GBP28 immunostaining may assist in a diagnosis of healing stage.
Cellular differentiation status of epithelial polyps of the colorectum: the gastric foveolar cell‐type in hyperplastic polyps
Authors: Koike M , Inada K , Nakanishi H , Matsuura A , Nakamura S , Tatematsu M
Published: December 2004

Aims:  The ‘metaplastic’ polyp of the colorectum, a synonym for the hyperplastic polyp, was named based only on features of the crypt epithelium. It is considered non‐neoplastic, but the precise cellular differentiation status remains to be proven.

Methods and results:  Forty‐eight hyperplastic polyps, 12 serrated adenomas, 45 tubular adenomas and five juvenile polyps were studied for their phenotypic expression using gastric (foveolar or pyloric gland cell), small intestinal (goblet cell), and colonic (goblet cell) cellular markers by immunohistochemical and mucin histochemical techniques. Gastric foveolar cell‐type differentiation was significantly expressed in hyperplastic polyps, while colonic differentiation was also consistently preserved. Neither gastric pyloric‐type nor small intestinal differentiation was observed. The same cell differentiation status as hyperplastic polyps was observed in serrated adenomas but not in tubular adenomas or juvenile polyps.

Conclusions:  A large proportion of hyperplastic polyps are composed of hybrid epithelium, with bidirectional differentiation to both gastric foveolar and colonic epithelial cells in the same crypt. Therefore hyperplastic polyps might be interpreted as the outcome of abnormal cell differentiation of stem cells. The same phenotypic expression suggests that hyperplastic polyps and serrated adenomas share the same cell lineage.
p53 expression in normal paraffin‐embedded tissue using different antibodies and antigen retrieval buffer systems
Authors: Pillai G , Roberts H , Gatter K , Pezzella F
Published: December 2004

Aims:  The study was undertaken to demonstrate wild‐type p53 in normal paraffin‐embedded tissues using two widely used antibodies, DO7 and 1801 and two different antigen retrieval buffer systems.

Methods and results:  Formalin‐fixed paraffin‐embedded normal tissue samples were obtained from the archives of the John Radcliffe Hospital, Oxford. Antigen retrieval was performed by microwaving using two different buffer systems: (i) the commercially available Dako target retrieval solution (Cat. no. 1699) (pH 9.8–9.9), (ii) freshly prepared buffer consisting of 0.1 m EDTA with 0.1% Tween pH 6.0, and (iii) freshly prepared buffer consisting of 0.1 m EDTA with 0.1% Tween pH 8.0. Staining was performed with DO7 and 1801 antibodies using the Dako Envision kit (peroxidase/DAB). DO7 antibody elicited strong nuclear staining in the mucosal cells of the small and large intestine, lymphoid cells, decidua, neurones such as Purkinje cells of the cerebellum, glandular epithelial cells and stromal cells of the prostate, cardiac myocytes and bronchial epithelial cells. Cytoplasmic staining was noted in Purkinje cells, glandular epithelium of prostate, exocrine pancreas and renal tubular epithelium. The 1801 antibody did not produce staining in any of these tissues.

Conclusions:  Our study demonstrates the presence of p53 in normal paraffin‐embedded tissue with nuclear and/or cytoplasmic localization in some instances. In our view, DO7 appears to be better suited for such detection.
Recent advances in the pathology of the vulva
Authors: Fox H , Wells M
Published: December 2004

This review addresses recent and important advances in our knowledge of several uncommon or rare disorders of the vulva including Paget's disease, vulva intraepithelial neoplasia, lichen sclerosus and squamous hyperplasia and their relationship to squamous carcinoma. Emphasis is placed on the two biologically different types of squamous carcinoma related and unrelated to human papillomavirus infection. Finally, the relatively recent concept of the sentinel node as applied to vulva carcinoma is discussed.
Clinicopathological features of nine cases of non‐cirrhotic portal hypertension: current definitions and criteria are inadequate
Authors: Ibarrola C , Colina F
Published: December 2004

Aims:  The clinicopathological features of nine patients with non‐cirrhotic portal hypertension were studied and an attempt was made to apply the descriptive criteria of experts to the morphological alterations of the livers in order to classify them adequately.

Methods and results:  Clinical and biochemical data and the alterations in livers resected at transplantation (n=7) or at autopsy (n=2) were gathered in five males and four females (ages 15–78 years) without aetiological factors for chronic hepatic disease who had oesophageal varices and splenomegaly in the absence of typical cirrhosis. Noting the luminal obstruction of the three hepatic vascular trees, hyperplastic nodule size and distribution, and the density of fibrosis, an attempt was made to assign each case to one of the following diagnostic categories: idiopathic portal hypertension, diffuse nodular regenerative hyperplasia, partial nodular transformation and incomplete septal cirrhosis. When a case could not be categorized into one of these groups, it was listed as non‐cirrhotic irregular architectural transformation. Only three cases could be assigned to one pure diagnostic category (two diffuse nodular regenerative hyperplasias and one incomplete septal cirrhosis). Three other cases could not be classified due to the heterogeneity of their lesions. In the remaining three cases, the hepatic morphology was a mixture of hilar partial nodular transformation combined with another abnormal architectural pattern in the peripheral parenchyma: diffuse nodular regenerative hyperplasia in two cases and idiopathic portal hypertension in the other. In seven cases, old thromboses in the hilar portal tree were observed. Stenoses were observed in some of the arterial branches in five cases and in some hepatic venous branches in four. However, no obstructions could be discovered in small or large portal veins in the two classical diffuse nodular regenerative hyperplasia cases.

Conclusions:  The hepatic morphology in this group of non‐cirrhotic portal hypertension patients was an abnormal remodelling of the liver associated with the frequent development of irregular hyperplastic nodules and frequent obstructions of the pre‐ and intrahepatic vascular lumens. It was very difficult to apply the nomenclature proposed by international experts.
The immunohistochemical expression pattern of Chk2, p53, p19INK4d, MAGE‐A4 and other selected antigens provides new evidence for the premeiotic origin of spermatocytic seminoma
Authors: Rajpert‐De Meyts E , Jacobsen G K , Bartkova J , Aubry F , Samson M , Bartek J , Skakkebæk N E
Published: December 2004

Aims:  Spermatocytic seminoma is a rare germ cell derived tumour of the testis that occurs mainly in older men. We analysed the expression of recently discovered markers for germ cell differentiation and the mitosis–meiosis transition in order to define the antigen profile for diagnostic purposes and to clarify the biology and histogenesis of spermatocytic seminoma.

Methods and results:  Twenty‐five spermatocytic seminomas were examined for immunohistochemical expression of germ cell‐specific onco‐fetal antigens and proteins involved in regulation of germ cell division, DNA repair and differentiation. The panel included Chk2, p19INK4d, p53, MAGE‐A4, KIT, TRA‐1‐60, neurone‐specific enolase and placental‐like alkaline phosphatase. Four of these proteins/antigens have never before been investigated in spermatocytic seminoma. Proteins highly expressed in gonocytes and spermatogonia, such as Chk2, MAGE‐A4 and neurone‐specific enolase, were consistently present in spermatocytic seminoma. Antigens expressed in embryonic germ cells but not in the normal adult testis, e.g. TRA‐1‐60, were undetectable, with the exception of p53 protein, which was demonstrated in 80% of cases. A proto‐oncogene p19INK4d, which is involved in the transition from mitotic to meiotic division in germ cells, was not detected in spermatocytic seminoma.

Conclusions:  The investigation provided new information concerning the expression of Chk2, MAGE‐A4, neurone‐specific enolase and p19INK4d in spermatocytic seminoma. The pattern of expression is highly consistent with the origin of spermatocytic seminoma from a premeiotic germ cell, which has lost embryonic traits and has committed to spermatogenic lineage but has not yet passed the meiotic checkpoint, most probably from the spermatogonium of the adult testis.
Differential expression of S100 calcium‐binding proteins in epidermoid cysts, branchial cysts, craniopharyngiomas and cholesteatomas
Authors: Pelc P , Vanmuylder N , Lefranc F , Heizmann C W , Hassid S , Salmon I , Kiss R , Louryan S , Decaestecker C
Published: December 2004

Aims: To investigate whether epidermoid cysts, branchial cysts, craniopharyngiomas and cholesteatomas express S100 proteins differentially by immunohistochemical assaying the presence of S100A1, S100A2, S100A3, S100A4, S100A5, S100A6 and S100B.

Methods and results: Immunopositivity/negativity was recorded for each S100 protein in a series of 52 cases consisting of 12 epidermoid cysts, 12 branchial cysts, 15 adamantinomatous craniopharyngiomas and 13 acquired cholesteatomas. Except in the case of the craniopharyngiomas, immunoreactivity was assessed independently in the basal membrane and the basal, the internal and the keratin layers. Our data show that in contrast to S100B, which was rarely expressed, S100A1, S100A2, S100A4 and S100A5 were often present in these four types of epithelial lesions. S100A3 and S100A6 and, to a lesser extent, S100A5 were the most differentially expressed proteins across the different histopathological groups analysed. These three proteins are expressed more often in craniopharyngiomas and cholesteatomas, the two more aggressive types of lesions.

Conclusions: This is the first study to report data on the expression of seven S100 proteins in different histopathological groups of epithelial head and neck lesions, whose precise embryological origins are still a matter of debate. S100 proteins could possibly be used as markers to target this embryonic origin, since our results show that S100A3 and S100A6 (and, to a lesser extent, S100A5) are expressed differentially across these different groups of epithelial lesions.
Expression of cyclin A and topoisomerase IIα of oligodendrogliomas is correlated with tumour grade, MIB‐1 labelling index and survival
Authors: Park S‐H , Suh Y‐L
Published: December 2004

Aims:  This study was designed to investigate immunoexpression of cyclin A and D1, and topoisomerase IIα in oligodendrogliomas and to evaluate the correlation with MIB‐1 (Ki67), tumour grade, and survival of the patients.

Methods and results:  Forty cases of oligodendrogliomas (20 high‐ and 20 low‐grade) were studied immunohistochemically with the above‐mentioned monoclonal antibodies.

Results:  Normal brain tissues included in tumour sections did not express any of cyclin A, MIB‐1 and topoisomerase IIα except cyclin D1, which was shown in perineuronal and interfascicular normal oligodendroglial cells. In low‐grade and high‐grade oligodendrogliomas, the mean cyclin A labelling index (LI) was 1.18 ± 0.98% versus 4.65 ± 1.99%, respectively; the mean topoisomerase IIα LI was 1.32 ± 1.04% versus 6.63 ± 4.31%, respectively; and the mean MIB‐1 LI was 1.69 ± 1.55% versus 9.46 ± 4.66%, respectively. Interestingly, cyclin D1 was not expressed in any oligodendrogliomas. Both cyclin A and topoisomerase IIα LI showed a significant positive correlation with MIB‐1 LI and histological grade of oligodendrogliomas (P < 0.01) and an inverse correlation with overall survival (P < 0.01). Univariate analysis showed that cyclin A and topoisomerase IIα LIs with a cut‐off point at 3% were a significant prognostic factor (P: cyclin A = 0.0040, topoisomerase IIα = 0.0033).

Conclusion:  Cyclin A and topoisomerase IIα expression are closely correlated with anaplastic oligodendrogliomas and worse clinical outcomes. Cyclin D1 seems not to be involved in the tumorigenesis of oligodendrogliomas.
Pathological significance of oxidative cellular damage in human alcoholic liver disease
Authors: Seki S , Kitada T , Sakaguchi H , Nakatani K , Wakasa K
Published: December 2004

Aims: To investigate the pathological significance of oxidative stress‐induced lipid peroxidation and oxidative DNA damage in alcoholic liver disease.

Methods and results: Hepatic expression of 4‐hydroxy‐2′‐nonenal (HNE) adducts and 8‐hydroxydeoxyguanosine (8‐OHdG) as reliable markers of lipid peroxidation and oxidative DNA damage, respectively, was analysed immunohistochemically and compared with histological findings in alcoholic liver disease. While no HNE adducts were observed in control livers, HNE adducts were frequently (37 of 40 cases, 92.5%) detected in alcoholic liver disease. The localization of HNE adducts was the cytoplasm of hepatocytes and sinusoidal cells in zone 3. As for 8‐OHdG, 29 of 40 cases (72.5%) with alcoholic liver disease exhibited positive immunolabelling for 8‐OHdG, while 8‐OHdG expression was not evident in control livers. The nuclear expression of 8‐OHdG was mainly detected in the hepatocytes within the areas of active inflammation. Among histological parameters, the grade of necro‐inflammation activity as well as the presence of Mallory bodies were significantly associated with the expression of HNE adducts and 8‐OHdG. In addition, the severity of steatosis also correlated with HNE adduct expression.

Conclusions: Lipid peroxidation and oxidative DNA damage occur widely and may be associated with certain pathological features in human alcoholic liver disease.
Quantifying the pathology of neurodegenerative disorders: quantitative measurements, sampling strategies and data analysis
Authors: Armstrong R A
Published: December 2004

The use of quantitative methods has become increasingly important in the study of neurodegenerative disease. Disorders such as Alzheimer's disease (AD) are characterized by the formation of discrete, microscopic, pathological lesions which play an important role in pathological diagnosis. This article reviews the advantages and limitations of the different methods of quantifying the abundance of pathological lesions in histological sections, including estimates of density, frequency, coverage, and the use of semiquantitative scores. The major sampling methods by which these quantitative measures can be obtained from histological sections, including plot or quadrat sampling, transect sampling, and point‐quarter sampling, are also described. In addition, the data analysis methods commonly used to analyse quantitative data in neuropathology, including analyses of variance (anova) and principal components analysis (PCA), are discussed. These methods are illustrated with reference to particular problems in the pathological diagnosis of AD and dementia with Lewy bodies (DLB).
How to improve microscopic images obtained with consumer‐type digital cameras
Authors: Regitnig P , Van Paasen R , Tsybrovskyy O
Published: December 2004

Aim: Digital imaging is useful in conventional photography because it immediately provides images, and the image quality can be improved afterwards by the use of computer programs. The major disadvantages of consumer‐type digital cameras mounted on microscopes are (i) unequal illumination through the image, and (ii) a coloured background. A computer program was specifically adapted and refined to improve images obtained with consumer‐type digital cameras mounted on microscopes.

Methods and results: An approach using a division operation between the specimen image and a background image leads to homogeneous illumination throughout the image, with automatically corrected brightness and white background. The correct colour spectrum is preserved by correction of the histogram. This approach was obtained from the freeware computer program ‘Image Arithmetic’. In a test, three different consumer‐type digital cameras (Sony, Nikon, Olympus) on different microscopes were used to obtain images of different types of histological specimens (cervical smear, bone marrow biopsy, and colonic biopsy). The computer program dramatically improved the quality of images obtained with all tested cameras.

Conclusion: Using this approach, even low‐cost digital cameras mounted on microscopes produce brilliant images with homogeneous illumination and a white background, the image quality being comparable with expensive cameras especially designed for microscopes.
p53 mutation and MDM2 amplification in inflammatory myofibroblastic tumours
Authors: Yamamoto H , Oda Y , Saito T , Sakamoto A , Miyajima K , Tamiya S , Tsuneyoshi M
Published: December 2004

Aims:  The pathogenic mechanism and predictive indicators of biological behaviour of inflammatory myofibroblastic tumour are poorly understood. We investigated molecular abnormalities of p53 and MDM2 in order to assess whether these play an important role in pathogenesis, and whether they also contribute to clinicopathological aggressive phenotype in inflammatory myofibroblastic tumour.

Methods and results:  We compared the immunohistochemical expression of calponin, h‐caldesmon, ALK, and p53 gene mutation and MDM2 gene amplification with clinicopathological findings in 15 cases of inflammatory myofibroblastic tumour. Histologically, cellular atypia was observed in five (33.3%) out of 15 cases. Local recurrences were observed in two (14.3%) of 14 informative cases, but no distant metastasis was observed. The expression of calponin (9/14; 64%) but not h‐caldesmon (0/14; 0%) was seen, which suggested myofibroblastic differentiation. ALK expression was seen in eight (53.3%) out of 15 cases, particularly in patients under 40 years old. Nuclear expression of p53 protein was recognized in only one (6.7%) of 15 cases, and polymerase chain reaction single‐strand conformation polymorphism followed by direct sequencing revealed p53 gene missense mutations in two (13.3%) of 15 cases. Nuclear expression of MDM2 was seen in four (26.7%) of 15 cases, and the MDM2 gene amplification was observed in two of the four cases.

Conclusion:  Inflammatory myofibroblastic tumour shows a wide spectrum of cellular atypia and biological behaviour with p53 and MDM2 expression. However, the alterations in the p53 pathway seem not to play a major role in the pathogenesis of inflammatory myofibroblastic tumour.
Gastric‐type mucin and TFF‐peptide expression in Barrett's oesophagus is disturbed during increased expression of MUC2
Authors: Van de Bovenkamp J H B , Korteland‐Van Male A M , Warson C , Büller H A , Einerhand A W C , Ectors N L E Y , Dekker J
Published: December 2004

Aims:  Barrett's oesophagus constitutes metaplastic epithelium, often diagnosed by mucin histochemistry. We determined the mucins and trefoil factor family (TFF)‐peptides that were expressed in Barrett's oesophagus, in order to study changes in protein expression in early stages of Barrett's oesophagus development.

Methods and results:  Biopsy specimens of 71 Barrett's oesophagus patients were collected, and sections were stained for secretory mucins by histochemistry. Immunohistochemistry was performed for secretory mucins (MUC2, MUC5AC, MUC5B, MUC6), TFFs (TFF1, TFF2, TFF3), and proliferation (Ki67). Protein expression in the tissue was measured semiquantitatively. MUC5AC and TFF1 showed high levels and strong colocalization in the surface epithelium, whereas MUC6, MUC5B and TFF3 were found in the deeper glandular structures. TFF2 was found in both surface and glandular epithelium. The co‐ordinate expression patterns of these six markers were similar to gastric antrum epithelium. MUC2 expression was ubiquitously associated with goblet cells within intestinal metaplasia, occurring in 68% of patients, and was correlated with increasing proliferation in the epithelium.

Conclusions:  Virtually all cells in Barrett's oesophagus epithelium displayed a secretory phenotype, demonstrating a co‐ordinate gastric‐type MUC and TFF expression. When MUC2 expression was more pronounced, the expression patterns of the other MUCs and the TFFs were increasingly disturbed. MUC2 expression may constitute a marker for early change in the phenotype of Barrett's oesophagus as a precancerous lesion.
Cutaneous breast cancer deposits show distinct growth patterns with different degrees of angiogenesis, hypoxia and fibrin deposition
Authors: Colpaert C G , Vermeulen P B , Van Beest P , Soubry A , Goovaerts G , Dirix L Y , Harris A L , Van Marck E A
Published: December 2004

Aims:  We postulated that skin metastases and cutaneous local recurrences from breast adenocarcinoma show different growth patterns with distinct angiogenic profiles.

Methods and results:  Fifty‐one surgically resected dermal breast cancer deposits were evaluated for growth pattern, E‐cadherin expression, presence of necrosis and a fibrotic focus, fibrin deposition, carbonic anhydrase IX expression (CA IX), microvessel density, endothelial cell proliferation and blood vessel immaturity. Growth patterns were infiltrative, with carcinoma cells infiltrating the dermis without significant disturbance of the pre‐existing architecture, expansive, meaning that a nodule of carcinoma cells and desmoplastic tissue pushed aside the pre‐existing dermal structures, or mixed. All lobular carcinomas showed an infiltrative growth and lacked membranous E‐cadherin expression. Different growth patterns in the ductal carcinomas were not correlated with differences in E‐cadherin expression. The presence of necrosis and/or a fibrotic focus and the expression of the hypoxia marker CA IX were significantly associated with an expansive growth. Fibrin was present in all expansive deposits and less frequently in the other growth patterns. There was a positive association between fibrin deposition, CA IX expression and microvessel density. The latter was significantly higher in the expansive and mixed growth patterns than in the infiltrative pattern. Endothelial cell proliferation was highest in the expansive growth pattern and was positively correlated with the presence of a fibrotic focus and with fibrin deposition. The maximum percentage of immature blood vessels was higher in the expansive and mixed growth patterns than in the infiltrative one.

Conclusion:  The recognition of different subgroups of cutaneous breast cancer deposits with different degrees of hypoxia‐driven angiogenesis may have important implications for the usefulness of anti‐angiogenic therapy.
Immunohistochemistry in the diagnosis of soft tissue tumours
Authors: Coindre J M
Published: December 2004

Immunohistochemistry is particularly important in the field of soft tissue tumours because of their variety and the frequent difficulty of diagnosis. The first part of this paper discusses useful or new antibodies, together with others that are no longer of use. The second part is devoted to the role of immunohistochemistry in the diagnosis of soft tissue tumours: identification of some rare or atypical benign lesions, identification of non‐mesenchymal malignant tumours, and classification of sarcomas. The respective roles of immunohistochemistry and molecular biology are underlined.
The angiogenic receptor KDR is widely distributed in human tissues and tumours and relocates intracellularly on phosphorylation. An immunohistochemical study
Authors: Stewart M , Turley H , Cook N , Pezzella F , Pillai G , Ogilvie D , Cartlidge S , Paterson D , Copley C , Kendrew J , Barnes C , Harris A L , Gatter K C
Published: December 2004

Aims:  Angiogenesis is an important factor in tumour growth and metastasis. Vascular endothelial growth factor receptor 2 (VEGFR‐2) or KDR plays a crucial role in angiogenesis. The aim of this study was to raise and characterize antibodies against phosphorylated KDR which could be used for studies on human tissues to assess KDR activation and novel inhibitors of KDR activation in clinical trials.

Methods and results:  Three monoclonal antibodies and one rabbit polyclonal antiserum were produced. The specificity of the antibodies was confirmed by ELISA. One of the mouse antibodies and the rabbit polyclonal antiserum reacted with a 200‐kDa band on a Western blot of human umbilical vein endothelial cell (HUVEC) lysates, the molecular weight of KDR. Immunohistochemical staining showed that phosphorylated KDR is present in a wide variety of normal tissues including liver, colon and placenta, and is not restricted to endothelium. It was also present in a number of human tumours including breast carcinomas, colonic carcinomas and non‐Hodgkin's lymphomas. The pattern of staining was membranous, cytoplasmic and nuclear.

Conclusions:  This study has shown that phosphorylated KDR is present in a wide variety of tumour and tissue types and is not confined to endothelium.
Subepithelial myofibroblasts and tenascin expression in microscopic colitis
Authors: Salas A , Fernández‐Bañares F , Casalots J , González C , Tarroch X , Forcada P , González G
Published: December 2004

Aims:  To assess differences in the pattern of subepithelial myofibroblasts and the expression of tenascin as a marker of extracellular matrix production in collagenous and lymphocytic colitis.

Methods and results:  Colorectal biopsies were studied from 122 patients with chronic diarrhoea and normal colonoscopy. The pathological diagnoses were collagenous colitis (n = 35), lymphocytic colitis (n = 37), mild non‐specific chronic inflammation (n = 28) and normal mucosa (n = 18). Four cases showed features of collagenous colitis but with collagen bands <10 µm thick. Normal mucosa from 14 patients without diarrhoea served as healthy control tissue. Immunohistochemical expression of α‐smooth muscle actin (myofibroblast marker) and tenascin was evaluated in well‐orientated sections. The expression of α‐smooth muscle actin was significantly increased in collagenous colitis compared with all the other groups. Strong tenascin subepithelial expression was seen in all cases of collagenous colitis, including the four without full‐blown features. The mean thickness of tenascin bands was greater than that obtained by conventional stains.

Conclusions:  There are clear differences, with respect to extracellular matrix remodelling, between collagenous and lymphocytic colitis. These results support the theory of matrix overproduction in the genesis of collagenous colitis.
Expression of apoptosis‐related markers and HER‐2/neu in thymic epithelial tumours
Authors: Pan C‐C , Chen P C‐H , Wang L‐S , Lee J‐Y , Chiang H
Published: December 2004

Aims:  To correlate the expression of a series of apoptotic and oncogene markers (including p53, Bcl‐2, BAX, Bcl‐XL, p21WAF,1/CIP1, cyclin D1, HER‐2/neu) in thymic epithelial tumours with histological type, stage and resectability and to determine whether the information on HER‐2/neu would be valuable in identifying patients who are eligible for anti‐HER‐2/neu treatment.

Methods and results:  Immunohistochemical stains were performed on 16 cases of non‐neoplastic thymus, 63 thymomas and 17 thymic carcinomas. Fluorescence in‐situ hybridization (FISH) for HER2 was performed to validate the gene amplification. Eighteen thymomas were positive for p53 and 14 of them were low‐expressors, with positive cells below 10%. All thymic carcinomas revealed over‐expression of p53 with positive cells either between 10% and 50% or >50%. The expression of p53 correlated with histological type and stage in thymoma. In both thymoma and thymic carcinoma, there was a statistically significant correlation between p53 status and resectability, with low expressors having a higher likelihood of being resectable. Thymic carcinomas, regardless of the histological subtypes, uniformly expressed Bcl‐2, while thymomas showed no or only weak cytoplasmic immunoreactivity. Most thymomas and thymic carcinomas were negative for Bcl‐XL, p21WAF,1/CIP1 and cyclin D1. The expression of BAX was inconsistent among different histological types. Nine thymic carcinomas revealed membranous positivity for HER‐2/neu, but no HER2 gene amplification could be demonstrated by FISH in any of the cases.

Conclusions:  p53 and Bcl‐2 are more implicated in the development of thymic carcinoma than thymoma. The higher level of p53 expression and the strong immunopositive pattern of Bcl‐2 in thymic carcinomas have potential value in the differential diagnosis and prediction of aggressiveness and resectability. On account of the absence of HER2 amplification, patients would probably not benefit from anti‐HER‐2/neu treatment.