bluuebunny/crossref_metadata_2025 huggingface dataset:
Serological diagnosis of experimental Enterococcus faecalis endocarditis
Authors: KJERULF ANNE , ESPERSEN FRANK , GUTSCHIK ERNÖ , MAJCHERCZYK PAUL ANTHONY , HOUGEN HANS PETTER , RYGAARD JØRGEN , HØIBY NIELS
Published: August 2009

A modified rat model of endocarditis with catheterization for 2 days was established in female Lewis rats using different inocula of Enterococcus faecalis (strain no. EF 19) in order to measure IgG antibodies in serum during the course of infection. Increasing the inocula intravenously resulted in an increase in the CFU/g vegetation and the CFU/g spleen, the ID50 being about 10 CFU/ml and the ID90 about 1 × 102 CFU/ml. The lowest bacterial inoculum infecting 100% of the rats was 3 × 103 CFU/ml, and for further investigations we used this inoculum size. Rats were sacrificed on day 2, 5, 7, 9, 11 and 28 after infection. The CFU/g vegetation and the CFU/g spleen increased until day 7 and then decreased. Serum samples were collected from 129 rats at different times after challenge. Three different ELISA systems were established to measure the IgG antibody responses: E. faecalis sonicate ELISA (a pool of four sonicates of strain no. EF 10, EF 11, EF 19 and EF 48), E. faecalis whole cell ELISA (strain no. EF 19) and E. faecalis purified cell wall ELISA (strain no. EF 19). An IgG antibody response was detected already on day 2, and except for a minor decrease on day 6/7 the antibody response continued to increase until day 14 (whole cell ELISA and sonicate ELISA) and day 21 (purified cell wall ELISA) when a plateau was reached. Significant increases in IgG antibody responses (p<0.05) were found between groups of rats from days 0–2, 2–8/9 and 8/9–14 in the E. faecalis whole cell and sonicate ELISAs and from days 0–2, 2–10/11 and 10/11–21 in the E. faecalis purified cell wall ELISA. In conclusion, we established a model of endocarditis in rats with catheterization for 2 days and were able to demonstrate an increase in IgG antibodies during the course of infection.
Effect of intrathymic administration of mycobacterial heat shock protein 65 and peptide p277 on the development of diabetes in NOD mice: caution required in vaccination studies
Authors: FUNDA DAVID P. , HARTOFT‐NIELSEN MARIE‐LOUISE , KAAS ANNE , BUSCHARD KARSTEN
Published: August 2009

Heat shock protein 65 (hsp65) and a derived peptide, p277, are autoantigens reported in IDDM. I.p. injection of hsp65 reduced diabetes incidence in NOD mice and administration of p277 cured already diabetic mice. Also, intrathymic (i.t.) administration of whole islets or GAD65 prevented diabetes in NOD mice. The aim of this study was to evaluate whether i.t. injection of mycobacterial hsp65 or p277 can prevent diabetes in NOD mice. Three‐week‐old NOD female mice were injected intrathymically with 50 μg of hsp65 (n=30), 5 μg of p277 (n=30), and PBS (n=29). Diabetes incidence was observed for the following 300 days. Pancreas was then used for histological and immunohistological evaluation. No significant differences in diabetes incidence were observed among the three groups of mice. Interestingly, hsp65‐treated mice developed diabetes slightly faster at 177±6 days compared to 202±8 days (p=0.015) for the p277‐treated group and 197±7 days (p=0.033) for controls. The insulitis score and average islet size did not differ significantly among the three groups of diabetic mice. Scattered TCR‐gamma/delta positive cells were found in the pancreas of all groups of mice. In contrast, a huge infiltrate of TCR‐gamma/delta positive cells was detected in four out of eight (50%) p277‐diabetic NOD mice. Thus, our data show an earlier onset of diabetes in hsp65‐treated mice and no improvement in the incidence with either hsp65 or p277, suggesting that hsp65 acts in a different way from what was reported with GAD65. Caution is advised in future vaccination studies as hsp65 poses a potential danger.
Morphological studies of baseline needle biopsies from living donor kidneys: Light microscopic, immunohistochemical and ultrastructural findings
Authors: SUND STÅLE , REISÆTER ANNA V. , SCOTT HELGE , FAUCHALD PER , BENTDAL ØYSTEIN , SØDAL GUNNAR , HOVIG TORSTEIN
Published: August 2009

Fifty‐seven consecutive living donors (31 women and 26 men aged 20.7–72.3 years, mean 50.6 years) were subjected to needle biopsy during nephrectomy, immediately before removal of the kidney. By light microscopy, semiquantitative scoring (0–3) was performed for arteriosclerosis, arteriolar hyalinosis (hyalin arteriolosclerosis), glomerulosclerosis, interstitial mononuclear cell infiltration, and interstitial fibrosis/tubular atrophy. Whereas vascular changes were striking in many biopsies (arteriosclerosis grades 2–3: 28/54 cases, arteriolar hyalinosis grades 2–3: 15/55 cases), glomerular and tubulo‐interstitial changes were mostly low grade. The morphological changes tended to be more pronounced in middle‐aged and older individuals, but, in particular, vascular changes were seen also in the younger age group. Immunofluorescence microscopy revealed glomerular granular staining for IgM in 52.7% of the cases, IgA in 9.1%, IgG in 1.8%, and C3 in 12.7%. The main ultrastructural finding was glomerulosclerosis; one case with diffuse glomerular IgA showed distinct dense deposits, and one case showed similar dense deposits without IgA deposition. Arteriolar wall deposition of C3 was found in 58.2% of the cases, and IgM in 10.9%. Especially C3 occurred both with arteriolar hyalinosis and in arterioles without light microscopic alterations. The observation of significant vascular changes in baseline biopsies is relevant especially in the differential diagnosis of chronic rejection and cyclosporine nephropathy. The immunohistochemical findings strongly indicate the occurrence of immunoglobulins and complement factor C3 in both glomeruli and arterioles without clinical or morphological signs of renal disease. The possible pathophysiological significance of such deposits remains, however, uncertain.
Immunohistochemical analysis of Fas ligand expression in sarcomas. Sarcomas express high level of FasL in vivo
Authors: LEE SUG HYUNG , JANG JA JUNE , LEE JUNG YOUNG , KIM SOO YOUNG , PARK WON SANG , KIM CHOO SOUNG , KIM SANG HO , YOO NAM JIN
Published: August 2009

Fas ligand (FasL) and its receptor, Fas, play a key role in the regulation of apoptosis within the immune system. Several prior experimental studies of Fas ligand expression in tumors have suggested a mechanism that enables tumors to evade immune destruction by inducing apoptosis in activated lymphocytes near the tumor cells. Many types of carcinomas have been shown to express FasL, but at present nothing is known about the range of sarcomas capable of expressing FasL in vivo. The aim of this study was to determine the in vivo patterns of FasL expression in human sarcomas. Archival paraffin‐embedded tissues of 57 sarcomas and 30 carcinomas were analyzed by immunohistochemistry for the expression of FasL. FasL immunoreactivity was seen in 39 of 57 (68%) sarcomas, including 10 of 10 rhabdomyosarcomas, 5 of 5 malignant schwannomas, 2 of 2 Ewing's sarcomas, 8 of 11 malignant fibrous histiocytomas, 4 of 5 angiosarcomas, 2 of 5 synovial sarcomas, 2 of 5 liposarcomas, 3 of 5 leiomyosarcomas, 2 of 6 osteosarcomas, and 1 of 3 chondrosarcomas. All carcinomas tested (10 gastric adenocarcinomas, 10 hepatocellular carcinomas, and 10 renal cell carcinomas) were positive for FasL. These results demonstrate that FasL expression in sarcomas, although less frequent than in carcinomas, is widespread among the sarcoma types, and suggest that FasL might contribute to the immune escape of sarcomas through killing Fas‐bearing lymphocytes.
Quantification of Bordetella pertussis in clinical samples by colorimetric detection of competitive PCR products
Authors: ERLANDSSON ANN , BÄCKMAN ANDERS , NYGREN MALIN , LUNDEBERG JOAKIM , OLCÉN PER
Published: August 2009

Quantification of microorganisms is an important part of the normal diagnostic work of a clinical microbiology laboratory. Traditionally the diagnosis of pertussis is subject to a yes or no approach with no quantitative dimension. This can, however, be of interest as a factor when judging the risk of a patient spreading the bacterium and as a research tool. The aim of the present study was to develop a PCR‐based quantitative assay for Bordetella pertussis DNA in clinical nasopharyngeal aspirates by combining a quantitative PCR with a colorimetric detection principle, DIANA (detection of immobilised amplified nucleic acid). A competitor to the PCR target sequence in IS‐481, containing a lac‐operator, was constructed and calibrated, and a test protocol prepared. A total of 46 clinical nasopharyngeal aspirates, previously diagnosed using a standard nested PCR assay and quantified by culture, were analysed by the quantitative PCR. The method showed acceptable precision and accuracy considering that it estimates the total number of bacterial genomes while culture detects viable bacteria. Recognised advantages were the simple colorimetric detection, the inborn indication of a working PCR assay, and the possibility of obtaining results even when partial inhibition of the PCR assay was seen. In addition, the quantitative PCR result can be obtained within one day compared to 3–10 days for culture. The present results and the qualities of the quantitative PCR suggest that this assay will be a useful complement in routine diagnostics and in research.
Establishment of an adrenocortical carcinoma xenograft with normotensive hyperaldosteronism in vivo
Authors: YAMAZAKI HITOSHI , ABE YOSHIYUKI , KATOH YUKO , SAWA NOBUKO , OHNISHI YASUYUKI , TANAKA YUJI , SASANO HIRONOBU , OSHIKA YOSHIRO , TOKUNAGA TESTUJI , KIJIMA HIROSHI , TAMAOKI NORIKAZU , NAKAMURA MASATO , UEYAMA YOSHITO
Published: August 2009

We established a xenograft line of human adrenocortical carcinoma (ADR‐1), and analyzed the hyper‐aldosteronism induced by the xenograft in vivo. Adrenocortical carcinoma specimens from a 25‐year‐old woman were subcutaneously inoculated into nude mice (BALB/c‐nu/nu) followed by serial passages in vivo. ADR‐1 retained the histopathological features (trabecular and sinusoid nests) seen in the primary carcinoma. The patient showed hyperaldosteronism (serum aldosterone >4000 pg/ml) and hypokalemia (serum K 2.1 mEq/1), but did not show hypertension. The nude rat (F344‐rnu/rnu) bearing ADR‐1 showed hyperaldosteronism (serum aldosterone 3320 ± 1420 pg/ml; control 191 ± 130 pg/ml) and hypokalemia (serum K 3.4 ± 0.4 mEq/1; control 5.2 ± 1.0 mEq/1) in vivo, and hypertension was not obvious. ADR‐1 was shown immunohistochemically to retain production of human‐specific corticosteroid synthetase. The xenograft ADR‐1 will be useful to elucidate the regulatory mechanism of normotensive hyperaldosteronism.
Association ofSerpulina hyodysenteriaewith the colonic mucosa in experimental swine dysentery studied by fluorescentin situhybridization
Authors: JENSEN T. K. , BOYE M. , MØLLER K. , LESER T. D. , JORSAL S. E.
Published: August 2009

The localization ofSerpulina hyodysenteriaein experimental swine dysentery was studied by fluorescentin situhybridization (FISH) using an oligonucleotide probe targeting the 23S rRNA ofS. hyodysenteriae.Nine 8‐week‐old pigs were challenged. Seven of the pigs were intragastrically dosed with 1 times 109cfuS. hyodysenteriaefor 3 consecutive days, whereas two pigs were infected by contact. Six non‐challenged pigs served as negative controls. The challenged pigs developed clinical swine dysentery from 8 to 14 days postinfection with typical gross lesions. By FISHS. hyodysenteriaecells in huge numbers were found colonizing the mucus layer, the luminal epithelium and the crypts of the large intestinal mucosa. The orientation of the spirochaetes to the epithelium appeared to be random. Spirochaetes in clusters or as single cells were invading the epithelium and were occasionally detected in the adjacent lamina propria. The distribution of spirochaetes in the mucosa provides further evidence thatS. hyodysenteriaeis intimately associated with the mucus layer and the epithelium in a random pattern. Furthermore, the results demonstrate the applicability of FISH for specific detection ofS. hyodysenteriaecells in clusters or as single cells in formalin‐fixed tissue samples.
Pattern of thyroid lesions and role of fine needle aspiration cytology (FNA) in the management of thyroid enlargement: a retrospective study from a teaching hospital in Riyadh
Authors: AL‐RIKABI A. C. , AL‐OMRAN M. , CHEEMA M. , EL‐KHWSKY F. , AL‐NUAIM A.
Published: August 2009

The study aims to determine the cytological pattern of thyroid lesions, in addition to the utility and accuracy of fine needle aspiration (FNA) cytology as an initial diagnostic method in the investigation of these lesions among Saudi patients. Four hundred and seventy‐nine (479) FNAs were performed on patients presenting with diffuse or nodular thyroid enlargement to the endocrinology clinic at King Khalid University Hospital in Riyadh during the period September 1993 to September 1996. The results of the FNA and the 125 histological diagnoses obtained from the subsequent partial and subtotal thyroidectomies were retrospectively and independently reviewed and compared by two cytopathologists. The results of the FNA cytological diagnosis showed that 372 patients (77.7%) had benign lesions, 24 (5%) had lesions which were suspected of malignancy, 25 (5.2%) had malignant neoplasms, and 58 (12.1%) had FNAs which were inadequate for cytological assessment. Colloid and adenomatous nodules were the most common benign lesions reported (47.8%). Papillary carcinoma was the most common malignant neoplasm (4.2%). Despite the relatively limited number of patients who underwent surgery, cytohistopathology combined with statistical analysis of the results showed that our FNA accuracy rate is in the region of 94.4% with a sensitivity of 78% and a specificity of 100%. In addition, the negative predictive value (NPV) and the positive predictive value (PPV) of FNA thyroid cytology were 93% and 100% respectively. Our findings indicate that FNA cytology can be used effectively in the evaluation of both nodular and diffuse thyroid lesions whilst cytopathologist should be aware of the potential diagnostic pitfalls and the limitations of the procedure in the diagnosis of follicular, cystic, and small neoplasms, the positive identification of thyroiditis and most cases of neoplasia by itself provides justification for FNA.
Immunohistochemical expression of c‐erbB‐2 oncoprotein and EGF‐R in pre‐ and postmenopausal breast cancer
Authors: RATNAKAR NITESH , SHARMA BHUVNESH KUMAR , CHATURVEDI K. UMA , KAUR SUDERSHAN , NAIK S. L. DAYALU , RAY AMITABHA
Published: August 2009

Tissues from 40 cases each of premenopausal and postmenopausal breast cancer were studied immunohistochemically for epidermal growth factor receptor (EGF‐R) and c‐erbB‐2 oncoprotein. In the premenopausal group, immunopositivity for c‐erbB‐2 was 15% and for EGF‐R 22.5%, whereas in the postmenopausal group, 45% of cases were positive for c‐erbB‐2 and 42.5% for EGF‐R. The difference in immunoexpression of c‐erbB‐2 between the two groups was significant. A significant correlation was observed between the concomitant expression of c‐erbB‐2 as well as EGF‐R and lymph node involvement. Furthermore, an association was found between c‐erbB‐2 positivity and histological grading of the tumour. It is interesting that the pattern of the investigated parameters indicates the difference in the pathological events of pre‐ and postmenopausal breast cancer.
Accumulation of immunoreactivity to ubiquitin carboxyl‐terminal hydrolase PGP 9.5 in axons of human cases with spinal cord lesions
Authors: YU WEN‐RU , OLSSON YNGVE
Published: August 2009

The protein gene product PGP 9.5 is one of the major polypeptides in neurons. It can act as a ubiquitin carboxyl‐terminal hydrolase in ubiquitin‐mediated degradation of proteins. The present study was performed to find out if human cases with spinal cord trauma present immunohistochemical signs of PGP 9.5 accumulation in injured axons known to accumulate ubiquitin. For comparison, we used six autopsy cases without spinal cord pathology, one case with syringomyelia, one case with ischaemic injury of the cord, and six ALS cases. Controls presented PGP 9.5‐immunostained axons of weak to moderate intensity in the longitudinal tracts. Immunoreactivity was not detected in nerve cell bodies, glial cells or axons of the grey matter. All nine trauma cases showed axonal swellings, but their numbers varied. Intensely immunostained axonal swellings were particularly abundant in cases with a survival period up to 1 month after trauma. Strongly immunoreactive axons were present also in the cases with infarct and syringomyelia. In conclusion, human cases with spinal cord trauma and other focal injuries present signs of PGP 9.5 accumulation in severed axons possibly resulting from disturbed axonal transport. PGP 9.5 thus seems to be present and may take part in ubiquitin‐mediated degradation of proteins in injured axons of the spinal cord.
Inhibition of Entamoeba histolytica proteolytic activity by human salivary IgA antibodies
Authors: GUERRERO‐MANRÍQUEZ GLORIA G. , SÁNCHEZ‐IBARRA FRANCISCO , AVILA EVA E.
Published: August 2009

Entamoeba histolytica is a protozoan parasite that causes amoebiasis in humans; as the infection occurs mainly in the intestinal epithelium, the secretory immune response of the host could have an influence on the outcome. Secretory IgA antibodies against E. histolytica have been detected in asymptomatic and symptomatic patients, but little is known about their protective role. E. histolytica cysteine proteases seem to be involved in the pathogenesis of amoebiasis; therefore, it is important to evaluate the human IgA response against these proteases and its effect on their enzymatic activity. When human saliva samples with and without antibodies against E. histolytica were tested by Western blot against one purified 70 kDa amoebic cysteine protease, 84% of anti‐amoeba‐positive samples recognized it. The secretory IgA purified from a pool of anti‐protease‐positive samples had a strong in vitro inhibitory effect on the E. histolytica proteolytic activity. These results suggest that this effect, if it occurs in vivo, could be an important protective factor against this parasite.
Neuroendocrine carcinoma of the breast: A case report of pure type
Authors: FUKUNAGA MASAHARU
Published: August 2009

A case of pure neuroendocrine carcinoma of the breast occurring in a 51‐year‐old female is described. The tumor located in the upper outer quadrant was a relatively well‐circumscribed, uncapsulated, yellowish white, rubbery mass measuring 2.5 times 2.0 times 2.0 cm. Microscopically, the tumor was characterized by solid nest, lobular, trabecular, and insular arrangements of small to medium‐sized cells with uniform round to oval hyperchromatic nuclei and pale to eosinophilic cytoplasm. It showed focal stromal invasion. The stroma was fibrous and rich in vascularity. The tumor cells were argyrophilic and positive for cytokeratin CAM 5.2, Leu‐7, neuron‐specific enolase, and chromogranin A. Electron microscopy showed neurosecretory granules. The tumor was DNA diploid with a low S‐phase fraction by flow cytometry. The axillary lymph nodes were negative for metastasis. The patient was well and without disease 16 months after mastectomy.
Abnormal mitochondria and sarcoplasmic changes in rabbit skeletal muscle induced by immobilization
Authors: LEIVO ILMO , KAUHANEN SUSANNA , MICHELSSON JARL‐ERIK
Published: August 2009

Immobilization of the rabbit knee in extended position results in damage to the vastus intermedius profundus (VIP) muscle. To examine the mechanisms involved in initiation of the injury, we studied the light and electron microscopic morphology of the VIP muscle, as well as the activity and distribution of NADH tetrazolium reductase (NADH‐TR) in the affected muscle, and determined serum total creatine kinase (CK) activity in immobilized rabbits. The VIP muscle of the immobilized right hindlimb was removed at various time points (10 h, 24 h, 36 h and 48–72 h, n=5 for each time point). The nonimmobilized left hindlimb and five nonimmobilized animals served as controls. No morphological changes were observed by light microscopy within 48–72 h in routine stainings. Transient ultrastructural abnormalities, including abnormal cristae, matrix lucencies and mild swelling of mitochondria, were observed between 10 h and 36 h of immobilization, subsiding by 48–72 h. On the other hand, progressive disorganization of myofibrils with breaking‐up of Z‐bands and an increase in the number and size of sarcoplasmic lipid vacuoles was seen with increasing duration of immobilization. NADH‐TR activity at subsarcolemmal locations had decreased by 10 h and disappeared by 24 h of immobilization, while the intermyofibrillar mitochondria remained unaltered. Serum total CK activity began to increase by 2 h of immobilization and reached a peak by 24 h. The results indicate that already a few hours of immobilization of the rabbit knee in extension leads to signs of metabolic disturbance of the VIP muscle and sarcolemmal leakage. The simultaneous occurrence of transient mitochondrial abnormalities, transient CK efflux and progressive myofibrillar damage suggests the operation of multiple adverse mechanisms already at the onset of disuse muscle atrophy.
Ossifying fibromyxoid tumour of soft parts: report of four cases including one mediastinal and one infantile
Authors: EKFORS T. O. , KULJU T. , AALTONEN M. , KALLAJOKI M.
Published: August 2009

Four cases of ossifying fibromyxoid tumour of soft parts are described. One of them was in the mediastinum, a hitherto unreported location of this rare neoplasm. Another was removed from the subcutaneous tissue of the head of a two‐year‐old girl, the youngest patient so far described. A peculiar feature of this tumour was haphazard spindle cell groups showing smooth muscle differentiation. One tumour was remarkably proliferative with 20 mitotic figures per 10 high power fields and 50% of cells positive for Ki‐67 antigen. Immunohistochemical analysis revealed that all the tumours were diffusely positive for vimentin, and focally for S‐100‐protein. In addition to this the infantile tumour expressed focal α‐smooth muscle actin, desmin and glial fibrillary acidic protein, while the mediastinal tumour expressed only α‐smooth muscle actin and the highly proliferative one expressed none of these antigens. Background cells, including histiocytes, lymphocytes and mast cells were numerous. DNA cytometry analysis using both static and flow methods showed that the mediastinal tumour contained two cell clones, while the others were diploid. The proliferative fraction of cells (S plus G2 phases) was prominent in the proliferative and infantile tumours.
A rapid method for generating cystic forms of Borrelia burgdorferi, and their reversal to mobile spirochetes
Authors: BRORSON ØYSTEIN , BRORSON SVERRE‐HENNING
Published: August 2009

Mobile Borrelia burgdorferi were transferred to distilled water (106 per ml). The cultures were observed by dark field microscopy (DFM), interference contrast microscopy (ICM) and transmission electron microscopy (TEM). 95% of the spirochetes were converted to cysts after 1 min, and after 4 h no normal mobile borreliae were observed. When transferred to growth medium (BSK‐H), the cysts became smaller and more irregular, and were filled with organic substances. After 1 day, 1–5 thin structures sprouted from the cysts. They continued to grow in both length and thickness until they attained a normal spirochetal structure. Finally, these new‐born spirochetes detached from the cysts, by which time their mobility had become normal. The present method for producing large amounts of cystic forms of B. burgdorferi is well suited for further studies of this unique microbe.
Nitric oxide suppresses bFGF‐ and IL‐1‐alpha‐mediated but not VEGF165‐mediated angiogenesis in natively vascularized mammalian tissue*
Authors: NORRBY KLAS
Published: August 2009

Using the rat mesenteric window angiogenesis assay, we studied the systemic effect of the nitric oxide synthase inhibitor Nw‐nitro‐L‐arginine methyl ester, L‐NAME, on angiogenesis induced by basic fibroblast growth factor (bFGF), interleukin‐1‐alpha (IL‐1) or vascular endothelial growth factor (VEGF165). Using technically independent morphometric and image analysis methods, the angiogenic response was quantified in variables related to (i) microvascular spatial extension and (ii) microvascular density. The test tissue, which is natively vascularized and lacks significant angiogenesis physiologically, was unaffected by surgical intervention. Two daily intraperitoneal injections of bFGF (2.20 nmole), IL‐1 (1.18 nmole) and VEGF165 (480 pmole) for 5 days elicited significant angiogenesis in the mesenteric windows. L‐NAME (0.5 g/L in drinking water) caused further enhancement of the angiogenic response produced by human recombinant bFGF (p<0.001), bovine purified bFGF (p<0.05) or murine recombinant IL‐1 (p<0.05). In contrast, the L‐NAME treatment did not affect the angiogenic response produced by human and murine recombinant VEGF165. These data suggest that nitric oxide can act as an endogenous suppressor of mammalian de novo angiogenesis, which is a new finding, and, moreover, that angiogenesis induced by VEGF165 on the one hand and by bFGF and IL‐1 on the other in the rat mesenteric window depends on different pathways.
Complement activation and regulator expression after anoxic injury of human endothelial cells
Authors: VÄKEVÄ A. , MERI S.
Published: August 2009

Complement activation is involved in the ischemia‐reperfusion injury of various organs, but the mechanisms leading to activation of the complement system are incompletely understood. In this study we show that EA. hy 926 human endothelial cells cultured under anoxic conditions (24 or 48 h) become activators of the homologous complement system. Flow cytometric analysis indicated that C1q, C3c, C3d, C4, C5, C9 components of complement are deposited on anoxic but not on normoxic cells after incubation with normal human serum. Cell membrane‐associated regulators of complement, membrane cofactor protein (CD46), decay‐accelerating factor (CD55) and protectin (CD59) were expressed on EA. hy 926 cells grown under normal oxygen tension. Under anoxic conditions the expression of protectin was clearly decreased, whereas the expression of CD46 and CD55 diminished only slightly. Our results suggest that anoxia can convert human endothelial cells to activators of the complement system. The diminished expression of protectin, CD46 and CD55 can sensitize the cells to complement‐mediated damage. Activation of the complement system due to the anoxic injury of human endothelial cells might be an important triggering mechanism in the pathogenesis of ischemia‐reperfusion injury of human heart.
Placental candidiasis: report of four cases, one with villitis
Authors: RIVASI FRANCESCO , GASSER BERNARD , BAGNI ALBERTO , FICARRA GUIDO , NEGRO ROSA MARIA , PHILIPPE EMILE
Published: August 2009

Four cases of placental candidiasis, an uncommon complication of rupture of the membranes, are presented. In addition to chorioamnionitis, in one of these cases villitis was also observed. Villitis is a rare occurrence in Candida infection and this represents only the second case in the literature. The involvement of villi may be suggestive of blood‐borne infection. However, since neither the mother nor the foetus presented any signs of systemic dissemination, the authors suggest a hypothesis of contamination of the villi from foci of chorioamnionitis.
Can alterations in integrin and laminin‐5 expression be used as markers of malignancy?
Authors: THORUP ALIS KARABULUT , REIBEL JESPER , SCHIØDT MORTEN , STENERSEN TOM C. , THERKILDSEN MARIANNE H. , CARTER WILLIAM G. , DABELSTEEN ERIK
Published: August 2009

Development of squamous cell carcinomas (SCCs) involves alterations in the adhesive interactions in the epithelium and invasion through the basement membrane. Therefore, changes in the expression of receptors and ligands involved in cell‐cell and cell‐matrix adhesion may be essential for the transformation of a premalignant into a malignant lesion. The aim of this study was to examine if expression of specific cell adhesion molecules can be used as markers of malignant development. By immunohistochemistry, we examined the expression pattern of integrins α2β1, α3β1, α6β4 and laminin‐5 in biopsies from SCCs (n=18), premalignant lesions (leukoplakias, n=21) and non‐premalignant tissue with chronic inflammation (n=11). In poorly differentiated SCCs, patchy loss of α3β1, α6β4 and laminin‐5 expression was pronounced at the invasion front, whereas there was a tendency to increased expression of α2β1. Analogous to the SCCs, biopsies from the leukoplakias and the non‐premalignant inflammatory tissue showed alterations of the expression of α3β1 and α6β4 in the basal cell layers and of laminin‐5. However, a characteristic finding in biopsies from leukoplakias was loss of α2β1 and α3β1 in the suprabasal cells. There was no unequivocal expression of the adhesion molecules distinguishing between inflammatory tissue, premalignant, and malignant lesions.
Production, isolation and purification of bacteriocins expressed by two strains of Neisseria meningitidis
Authors: ALLUNANS JURIS , BJØRÅS MAGNAR , SEEBERG ERLING , BØVRE KJELL
Published: August 2009

The systemic Neisseria meningitidis strain P241 and the healthy pharyngeal carrier strain BT878 produce bacteriocin‐like substances during growth. A method has been devised for obtaining the active substances in solution. The activity was recovered by freeze‐thaw extraction of dialyzed Todd‐Hewitt agar medium into which the bacteriocins had diffused during growth of the producer strains. The bacteriocins were purified more than 50‐fold by ammonium‐sulphate precipitation and hydrophobic interaction chromatography. They are quite stable to heat and remain active 100% after 30 min at 100°C. However, the protein nature of the bacteriocins has been confirmed by their sensitivity to α‐chymotrypsin. Gel filtration indicated an Mr of 100–110 kDa, whereas SDS‐polyacrylamide gel electrophoresis produced a common band by Coomassie staining corresponding to an Mr of 47–48 kDa, suggesting a dimer form of the active protein component.
Recurrent fetal polycystic kidneys associated with glutaric aciduria type II
Authors: KJAERGAARD SUSANNE , GRAEM NIELS , LARSEN TORBEN , SKOVBY FLEMMING
Published: August 2009

A woman had two pregnancies terminated in the 20th and 21st weeks of gestation after ultrasonographic detection of enlarged hyperechoic kidneys in both fetuses. The combination of polycystic kidneys and steatotic liver found at autopsy suggested glutaric aciduria type II (GA II), which was confirmed by biochemical investigation. GA II or multiple acyl‐CoA dehydrogenase deficiency is an autosomal recessively inherited defect of mitochondrial energy metabolism, which usually results in neonatal death. When pregnancy is terminated because of enlarged hyperechoic kidneys in the fetus, autopsy is crucial for establishing the correct diagnosis. The combination of polycystic kidneys and steatotic liver should bring GA II to mind, and prompt appropriate biochemical investigations so that genetic counselling and first trimester diagnosis can be offered in future pregnancies.
The 1997 list Proposed new bacterial taxa and proposed changes of bacterial names published during 1997 and considered to be of interest to medical or veterinary bacteriology
Authors: FREDERIKSEN WILHELM , MAGEE JOHN , URSING JAN
Published: August 2009

A list of names published or validated in 1997 is presented. We also comment on the tendency to base names of new taxa on a single bacterial strain, and the consequences for reliable descriptions that this tendency implies.
The value of the biopsy of the contralateral testis in patients with testicular germ cell cancer: The recent German experience
Authors: DIECKMANN KLAUS‐PETER , LOY VOLKER
Published: August 2009

Purpose: Testicular intraepithelial neoplasia (TIN; so‐called carcinomain situof the testis), the precursor of testicular germ cell neoplasms can be detected by testicular biopsy many years before the clinical manifestation of the tumour. This study looked at the prevalence of contralateral TIN in patients with testicular germ cell cancer. The purpose was to evaluate this new approach of early detection of testicular cancer and to evaluate the current management strategies. Patients, methods: 1954 consecutive patients with unilateral testicular germ cell tumour underwent contralateral biopsy. All specimens were examined immunohistologically with staining for placental alkaline phosphatase. Patients with TIN were usually submitted to low‐dose radiotherapy of the testis. A rebiopsy was performed after 3 months. Endocrinological evaluations were done before, during and after treatment. Results: TIN was observed in 4.9% (95% confidence intervals 3.95%– 5.91%). Testicular atrophy constitutes a 4.3 fold increased risk of having contralateral TIN. 64% of the cases with TIN were found in clinically normal testes. Patients with TIN were significantly younger than those without (p<0.017). No case with TIN was found in patients older than 50 years. Three patients developed a second testicular tumour during follow‐up despite a negative biopsy. After radiotherapy, all of 23 patients had complete disappearance of TIN in the rebiopsy. After chemotherapy, 3 of 10 patients had persistent TIN histologically. After radiotherapy, 12 of 41 patients required testosterone replacement. Conclusion: The prevalence of contralateral TIN accords well with the known prevalence of bilateral testicular tumours. Testicular atrophy is a strong indicator for the presence of TIN but about 60% of TIN‐cases occur without atrophy. Local radiotherapy to the testis with 18 – 20 Gy is efficaceous in eradicating TIN, but it causes significant damage to almost one quarter of these patients. Chemotherapy is an unsafe treatment for TIN. This study shows the feasibility of early detection of testicular cancer in a high‐risk population by means of searching for TIN. Although the management of the condition still needs refinement, the TIN‐concept offers an avenue for the early detection of testicular cancer and early conservative management.
Hormonal stimulation of the recovery of spermatogenesis following chemo‐ or radiotherapy
Authors: MEISTRICH MARVIN L.
Published: August 2009

Radiation and chemotherapeutic drugs produce prolonged depression of sperm counts in rodents and humans. Previously, three approaches have been developed in experimental animals that have had some success in preventing or reversing this toxicity. These approaches included pretreatment with hormones that suppress spermatogenesis, stimulation of stem cell number, and supplementation with testosterone. A different rationale for the ability of particular hormonal treatments to reverse prolonged azoospermia is presented in this review. In many cases prolonged azoospermia occurs even though the stem spermatogonia survive the toxic insult, but the differentiation of these spermatogonia to produce sperm fails. In the rat, the block appears to be at the differentiation of the A spermatogonia. Hormone treatments with testosterone or with GnRH agonists, which suppress intratesticular testosterone levels, relieve this block and result in the production of differentiating cells. When the hormone treatment is stopped the production of differentiating cells continues, mature sperm are produced, and fertility is restored. If a similar mechanism can be demonstrated to hold in humans, the fertility of men who have been rendered infertile by treatments for testicular and other cancers could be improved.
Different fate of primordial germ cells and gonocytes following transplantation
Authors: JIANG FANG‐XU , SHORT ROGER V.
Published: August 2009

We have previously demonstrated that both donor primordial germ cells (PGCs) and gonocytes are capable of establishing spermatogenesis in the lumen of the seminiferous tubules of an adult host following transplantation in rats. Here we show that the PGCs, either in crude suspensions or after purification, undergo spermatogenesis only in the intraluminal compartment of the host's seminiferous tubules, while 4–5 days postpartum gonocytes also interdigitate with the host's seminiferous epithelium. The donor seminiferous epithelium was always in synchrony with the cycles of the host's spermatogenesis. It seems that the pattern of spermatogenesis of donor germ cells following transplantation in terms of its spacial location and the connection with the host's seminiferous epithelium depends on their developmental stages at transfer.
Candidate regions for testicular cancer susceptibility genes
Authors: Bishop D. Timothy ,   THE INTERNATIONAL TESTICULAR CANCER LINKAGE CONSORTIUM
Published: August 2009

Although the aetiology of testicular cancer is unknown, a clear familial component has been identified in a number of studies. In an effort to identify susceptibility genes for testicular cancer, the International Testis Cancer Linkage Consortium has been collecting families with multiple cases of testicular cancer. These families have been genotyped for a set of markers across the genome in two separate studies. The family information has also been pooled across the two studies so that the combined evidence can be assessed. While no genomic region gives overwhelming support for a testicular cancer predisposition gene, several regions, notably on chromosomes 3, 5, 12 and 18 show suggestive evidence worthy of further examination. The Consortium is now attempting to identify further families in an attempt to confirm or deny these leads.
Towards the isolation of a human malignant extragonadal germ cell tumour‐associated breakpoint in chromosome 11q13
Authors: SINKE R. J. , VAN ASSELDONK M. , DE BRUIJN D. , STRIJK J. A. , MERKX G. , WEGHUIS D. OLDE , DE JONG B. , OOSTERHUIS J. W. , VAN KESSEL A. GEURTS
Published: August 2009

In a previous study we have defined a subgroup of human malignant extragonadal germ cell tumours that is characterized by complex translocations involving chromosomes 6 and 11 (Echten et al. 1995). Here we report (i) the use of fluorescent in situ hybridization, pulsed field gel electrophoresis and direct visual hybridization techniques to localize the tumour‐associated breakpoint within band 11q13, and (ii) the construction of a phage library enriched for this region to facilitate genomic walks towards the breakpoint. Extensive breakpoint‐flanking contigs were generated and within these contigs six candidate genes could be identified.
The cytogenetic theory of the pathogenesis of human adult male germ cell tumors
Authors: CHAGANTI R. S. K. , HOULDSWORTH JANE
Published: August 2009

Human male germ cell tumors (GCTs) represent a biological paradox because, in order to develop into a pluripotential tumor, a germ cell destined to a path of limited or no proliferation must acquire the potential for unlimited proliferation. In addition, it must acquire the ability to elicit embryonal differentiation patterns without the reciprocal inputs from fertilization and the imprinting‐associated genomic changes which are a part of normal embryonal development. Although much speculated about, the genetic mechanisms underlying these properties of male GCTs remain enigmatic. Recent cytogenetic and molecular genetic analyses of these tumors are providing new insights and new testable hypotheses. Based on our recent work, we propose two such hypotheses. One relates to the mechanism of germ cell transformation and germ cell tumor development. We suggest that the invariable 12p amplification noted as early as in carcinoma in situlintratubular germ cell neoplasia (CIS/ITGCN) lesions leads to deregulated overexpression of cyclin D2, a cell cycle G1/S checkpoint regulator with oncogeneic potential. Such overexpression reinitiates the cell cycle. We visualize this happening during the pachytene stage of meiosis through aberrant recombinational events which lead to 12p amplification. The other hypothesis relates to the origin of primary extragonadal GCTs. By comparing cytogenetic changes in primary mediastinal versus gonadal lesions, we propose that, in contrast to long‐standing speculation that primary extra‐gonadal tumors arise from embryonally misplaced primordial germ cells, these lesions arise from migration of transformed gonadal germ cells.
P53 tumour suppressor gene and germ cell neoplasia
Authors: LUTZKER STUART G.
Published: August 2009

P53 tumour suppressor gene mutations occur in approximately 50% of common solid tumours such as breast, colon and lung. In contrast, p53 gene mutations occur infrequently (<3%) in germ cell tumours, even though p53 protein is expressed at high levels in the vast majority of tumour samples. P53‐regulated genes are not correspondingly over‐expressed in germ cell tumour samples and cell lines, indicating that p53 functions poorly as a transcription factor in this tumour type. High levels of wild‐type p53 may contribute, in part, to the chemosensitivity of germ cell tumours.
Immunohistochemical and mutational analysis of the p53 tumour suppressor gene and the bcl‐2 oncogene in primary testicular germ cell tumours
Authors: HEIDENREICH AXEL , SCHENKMAN NOAH S. , SESTERHENN I. A. , MOSTOFI KASH F. , MOUL JUDD W. , SRIVASTAVA SHIV , ENGELMANN UDO H.
Published: August 2009

The role of p53 in testicular germ cell tumours is still contradictory based on the finding of immunohistochemical overexpression at the protein level, but lack of mutations at the DNA level. In addition, p53 wild‐type activity has been demonstrated in cell culture experiments. Overexpression of the proto‐oncogene bcl‐2 might block p53‐induced apoptosis and might inhibit p53 functional activity. To clarify the apparent paradox with respect to p53 overexpression and lack of mutations, an immunohistochemical and mutational analysis of p53 and bcl‐2 in TGCT was performed. Ten normal testes, 52 CIS and 151 clinical stage I nonseminomatous GCTs were included in our study. A commercially available anti‐p53 polyclonal rabbit antibody and an anti‐bcl‐2‐mouse monoclonal antibody were used to stain the 5pm sections. Staining was assessed by counting at least 500 cells from the area of the most intense staining in each tumour cell type, and this was scored semiquantitatively for intensity of staining on a 4 point scale. In addition, 30 primary GCTs were included in the mutational analysis: areas with p53 overexpression were identified and microdissected prior to DNA extraction. p53 exons 5–8 were amplified by polymerase chain reaction (PCR) followed by single strand conformation polymorphism analysis. Templates demonstrating band shifts on SSCP were subjected to direct DNA sequence analysis. None of the normal testes, 32/52 (62%) CIS, and 142/151 (94%) germ cell tumours exhibited p53 overexpression. p53 expression was significantly lower in mature teratomas (0.8 ± 0.2) than in other germ cell tumour components (2.8 ± 1.2, p>0.001). PCR‐SSCP did not reveal any missense mutations or deletions for the p53 gene. Bcl‐2 protein expression was observed in none of the normal testes, in none of the CIS, and in 14/151 (9.3%) germ cell tumours. 13/14 germ cell tumours demonstrated bcl‐2 expression only in the glandular and stromal elements of their teratomatous components whereas all other components were negative for bcl‐2. Our results ‐ p53 overexpression, lack of p53 mutations, undetectable bcl‐2 ‐ are consistent with recent in vitro studies. High susceptibilty of testicular cancer to drug‐induced apoptosis appears to be the result of wild‐type p53 and lack of bcl‐2. Radiation and chemotherapeutic insensitivity of mature teratomas might be the result of bcl‐2 overexpression and lack of p53 overexpression. Therefore, chemoresistance to DNA damaging agents might be reflected by the expression of p53 and bcl‐2 and it might be useful to evaluate p53 and bcl‐2 in primary tumours and metastatic lesions in order to identify patients early with primary or secondary chemo‐resistance.
Inhibin gene expression in a large cell calcifying Sertoli cell tumour and serum inhibin and activin levels
Authors: TOPPARI JORMA , KAIPIA ANTTI , KALEVA MARKO , LAATO MATTI , DE KRETSER DAVID M. , KRUMMEN LYNNE A. , MATHER JENNIE P. , SALMI TOIVO T.
Published: August 2009

Inhibin is a potential tumour suppressor gene product in the gonads. While inhibin gene products may have a role in tumourigenesis, serum inhibin levels can be used as a marker for ovarian tumours derived from granulosa cells. Tumours derived from Sertoli cells, testicular counterparts of granulosa cells, are rare. To assess whether inhibin could be used as a human Sertoli cell tumour marker, serum inhibin and activin levels and inhibin subunit mRNA expression in the testis were studied. Northern blot and in situ hybridization revealed abundant expression of inhibin α, βA, and βB subunit mRNAs in large cell calcifying Sertoli cell tumours found in a 12‐year old boy with Carney complex. The tumours were multifocal and bilateral. Serum inhibin levels were clearly elevated at the time of the diagnosis, decreased by 50% after one of the testes was removed, and were low or undetectable after the second orchidectomy six weeks later. Activin was undetectable before the orchidectomies, while a low concentration of activin‐A was measured after them. Follicle stimulating hormone (FSH) concentration increased from normal pubertal value to castration level as expected. Normal seminiferous tubules also showed inhibin subunit α and βB mRNA expression, whereas inhibin βA mRNA was expressed in normal Leydig cells. These data suggest that serum inhibin reflects Sertoli cell activity and can be used as a human tumour marker.
Oct‐4: more than just a POUerful marker of the mammalian germline?
Authors: BREHM ALEXANDER , OVITT CATHERINE E. , SCHÖLER HANS R.
Published: August 2009

Mammals lack visible cytoplasmic components in the oocyte that could account for ‘germline determinants’ as identified in various non‐mammalian species. Actually, mammals might not define the germline autonomously by localized ‘germline determinants’ but conditionally depending on the position of cells within the embryo. The Oct‐4 gene encodes a transcription factor that is specifically expressed in the toti‐ and pluripotential stem cells of the mouse embryo and so far has only been found in mammalian species. Oct‐4‐expressing embryonal cells retain the capacity to differentiate along multiple lineages and they have been suggested to be part of a ‘totipotent germline cycle’ that links one generation to the next.
Early stages in male germ cell differentiation in the mouse
Authors: ANDERSON ROBERT , GARCIA‐CASTRO MARTIN , HEASMAN JANET , WYLIE CHRISTOPHER
Published: August 2009

Primordial germ cells arise during gastrulation and migrate from the hindgut into the gonad primordium during early organogenesis. In this article, we discuss factors that control migration, proliferation and targeting of the PGCs. In particular we discuss how changes in adhesiveness control germ cell positioning in the gonad, and the molecules involved.
Regulation of germ cell death in mammalian gonads
Authors: MATSUI YASUHISA
Published: August 2009

A large number of primordial germ cells (PGCs), as well as spermatogonia, undergo programmed cell death or apoptosis in the physiological context. In this process, environmental, cytoplasmic and nuclear factors are involved. Bcl‐2 and its related molecules are known as general regulators of cell death, and some are important for survival of PGCs and spermatogonia. Steel factor, a ligand for c‐Kit, also supports growth and survival of these cells. In addition, bone morphogenetic protein (BMP)8B and Desert Hedgehog (Dhh), which are secreted proteins, and a nuclear factor, c‐Myc, play a role in spermatocyte survival. This suggests that germ cell survival or death at each stage of differentiation is precisely controlled by specific signalling pathways which consist of a number of molecules.