bluuebunny/crossref_metadata_2025 huggingface dataset:
Increasing incidence of testicular cancer — birth cohort effects
Authors: EKBOM ANDERS , AKRE OLOF
Published: August 2009

The incidence of testicular cancer is rising in most Western populations. A collaborative study between nine population‐based cancer registries in countries around the Baltic Sea was utilized in order to analyze in detail geographic variations and temporal trends in the occurrence of testicular cancer. There were 34,309 cases registered up until 1989 starting in Denmark in 1942 and most recently in Latvia in 1977. From the descriptive epidemiology it was obvious that there was a substantial variation in the age‐standardized incidence amounting to about a 10‐fold difference between the different countries ranging from 0.8 per 100,000 person‐years in Lithuania to 7.6 per 100,000 person‐years in Denmark. Previous studies have indicated that this increase is due to birth cohort effects. A more detailed analysis was therefore performed in those six countries with a sufficiently long period of cancer registration; Poland, former East Germany, Norway, Finland, Denmark and Sweden. This analysis showed that birth cohort is a more important determinant of testicular cancer risk than year of diagnosis. In Poland, former East Germany and Finland, there was an increasing risk for all birth cohorts. Among men born in Denmark, Norway or Sweden between 1930 and 1945, this increasing trend in risk was interrupted in these birth cohorts but followed thereafter by an uninterrupted increase by birth cohort. In conclusion, life time exposure to environmental factors which are associated with the incidence of testicular cancer appear to be more related to birth cohort than to year of diagnosis. Because testicular cancer typically occurs at an early age, major etiological factors therefore need to operate early in life, perhaps even in utero.
Negative feedback control of the spermatogenic progression by testicular oestrogen synthesis: Insights from the shark testis model
Authors: BETKA MARLIES , CALLARD GLORIA V.
Published: August 2009

The organisation of the testis of the dogfish shark is technically advantageous for stage‐by‐stage analysis of spermatogenesis in vivo and in vitro. Prior studies using this model show that total oestrogen receptors (ER) are concentrated in regions where spermatocysts (“follicle‐like” germ cell‐Sertoli cell units) are in stem cell and spermatogonial stages: respectively, germinal zone (GZ) and premeiotic (PrM) regions. By contrast, key enzymes regulating oestrogen (E) concentrations (aromatase, 17α‐hydroxylase) are maximal in meiotic (M) and postmeiotic (PoM) regions, respectively, which are upstream in the intratesticular vascular pathway. To investigate the hypothesis that E is part of a signalling mechanism between stages of development, studies were undertaken to test direct effects of oestradiol‐17β (E2) on processes in ER‐rich regions. As measured by [3H]thymidine (‐Tdr) incorporation, DNA synthesis in GZ and PrM regions was inhibited by E2 ((1000 nM) in a dose‐response fashion. The maximal response (30–40%) was significant, reproducible and observed within 72 hr of treatment. Insulin differentially affected DNA synthesis and the response to E2 in GZ and PrM regions. As measured by [3H]Tdr release after prelabelling spermatocysts of GZ regions, apoptosis progressively decreased with increasing concentrations of E2. At the maximal dose of E2 used, there was no effect on total protein synthesis or secretion in combined GZ/PrM cysts, indicating that effects on DNA synthesis and cell death were authentically physiological, not pharmacological, and consistent with a state of developmental arrest. These results support the hypothesis that E synthesised within the testis is part of a negative feedback regulatory mechanism whereby more mature stages regulate the developmental advance of less mature stages. A growth control mechanism of this type could explain the strict temporal, spatial and quantitative order of succeeding stages characteristic of normal spermatogenesis in all vertebrates. Further study is required to determine whether E signalling in this model is restricted to Sertoli cells or has a germ cell component.
Interaction of herpes simplex virus with mononuclear phagocytes is dependent on the differentiation stage of the cells
Authors: BRUUN TROND , KRISTOFFERSEN ANNE K. , ROLLAG HALVOR , DEGRÉ MIKLOS
Published: August 2009

The interaction of herpes simplex virus (HSV) with mononuclear phagocytes (MP), i.e. monocytes and macrophages, is of importance for the pathogenesis of HSV infections. MP are known to play a significant role in the cellular defence against infections with HSV, but it has also been shown that HSV‐1 affects MP. The infection of these cells at different stages of differentiation has different outcomes, and may result in the alteration of important cellular functions. HSV‐1 inhibits the morphological differentiation of human monocytes, and this inhibition occurs in spite of the fact that human monocytes are non‐permissive to HSV‐1. We have studied the effect of HSV infection of monocytes and macrophages on production of essential cytokines and related this effect to the reproduction of the virus. Blood‐derived MP were cultured in vitro and inoculated with HSV at different stages of differentiation. Replication of the virus was measured by infectivity titration, detection of HSV antigens by immunofluorescence and detection of HSV‐specific mRNA. In monocytes, no viral replication and no production of late protein was seen. HSV IE gene was transcribed in monocytes from some donors, but not from others. In macrophages, virus replicated, but less efficiently than in fully permissive fibroblast cells. The production of IL‐1β, IL‐6 and TNF‐α in both non‐permissive monocytes and permissive macrophages was assayed both at the transcriptional level, as mRNA, and as protein released from the cells. Production of cytokines by MP was affected by HSV‐1. The level of cytokine mRNA and cytokine protein did not correspond for all cytokines, which may suggest that translational regulation and/or cytokine inhibitors are important in the regulation of the cytokine response. The cytokine modulation, both at the transcriptional level and measured as biological activity, was different in monocytes and macrophages, and varied between different donors. Our results indicate a relation between permissiveness and cytokine response in mononuclear phagocytes infected with HSV‐1. Such a relation may be of importance to both intrinsic and extrinsic defence mechanisms of MP against HSV‐1. Our study also demonstrates that even the functions of non‐permissive cells such as blood‐derived monocytes may be affected by viral infections.
Microfilaremia from a Dirofilaria‐like parasite in Greece
Authors: PETROCHEILOU V. , THEODORAKIS M. , WILLIAMS J. , PRIFTI H. , GEORGILIS K. , APOSTOLOPOULOU I. , MAVRIKAKIS M.
Published: August 2009

In Europe, zoonotic filarial infections in humans are caused by two species, Dirofilaria immitis and Dirofilaria repens. These parasites are associated mainly with embolic infarcts of the pulmonary artery and subcutaneous nodules, respectively. An unusual dirofilarial infection in a Greek patient who showed marked eosinophilia and microfilaremia is presented. Although the identification of Dirofilaria species is not conclusive, this report is the first on a case of microfilaremia from a Dirofilaria infection in an immunocompetent patient.
Morphological adaptations of human liver peroxisomes in cholestasis
Authors: DE CRAEMER DIRK , PAUWELS MARINA , VAN DEN BRANDEN CHRISTIANE
Published: August 2009

Part of the bile acid synthesis takes place in peroxisomes. An altered enterohepatic circulation of bile acids might influence peroxisomal β‐oxidation enzymes and peroxisomal morphology. We performed a morphological and morphometric investigation of peroxisomes in liver biopsy samples of eight patients with cholestasis of different origin: graft versus host reaction (n=1), obstruction of the bile flow (n=3), and drug‐induced cholestatic hepatitis (n=4). Peroxisomes were identified using catalase cytochemistry. They were regularly shaped and showed individual differences in electron density. A perinuclear distribution was observed in a variable number of hepatocytes in each sample. Morphometric analysis of peroxisomes revealed an increase in numerical density and surface density in all, and a decreased mean diameter in four liver samples. Based on previously obtained data in experimental animals, we hypothesize that the observed alterations in peroxisomal morphology indicate an enhanced metabolic activity of the enzymes in the peroxisomal matrix. Among them are enzymes involved in bile acid synthesis.
Renal structural changes in insulin‐dependent diabetic patients with albuminuria
Authors: ØSTERBY R. , SCHMITZ A. , NYBERG G. , ASPLUND J.
Published: August 2009

The large interindividual variation in diabetes duration until the onset of nephropathy is partly unexplained. This study was performed to compare renal structure in insulin‐dependent (IDDM) patients who had developed signs of nephropathy after a short or long duration of diabetes. Renal biopsies were obtained from 17 IDDM patients, with albumin excretion rate 20–300 μg/min and normal blood pressure. Six patients had <25 years duration (“short‐term”, early onset of microalbuminuria) and eight patients had duration >30 years (“long‐term”, late onset of microalbuminuria). Biopsies were obtained 18 months after entry into a study testing the effect of low‐dose antihypertensives. Parameters characterizing diabetic glomerulopathy were significantly increased in IDDM patients compared with those in 17 living donors: Basement membrane thickness, mean and (CV): 591 nm (0.17) vs 320 nm (0.12), mesangial volume fraction per glomerulus 0.27 (0.19) vs 0.19 (0.10), matrix volume fraction per glomerulus 0.16 (0.20) vs 0.097 (0.22), matrix star volume 38.5 μm3 (0.43) vs 13.9 μm3 (0.31), (p<10‐4 for each). Comparison of short vs long‐term patients showed no significant differences in glomerulopathy parameters, glomerular volume or extracellular material per glomerulus, whereas the fraction of occluded glomeruli was significantly increased in long‐term patients. A close correlation obtained between fraction of occluded glomeruli and glomerular filtration rate (r=0.72, p=0.001). Glomerular occlusion occurred unrelated to the severity of diabetic glomerulopathy. It is suggested that diabetic macroangiopathy and arteriolar hyalinization may play an important role in the renal function of patients with slow development of nephropathy.
Evaluation of an immunohistochemical test with polyclonal antibodies raised against mycobacteria used in formalin‐fixed tissue compared with mycobacterial specific culture
Authors: CARABIAS E. , PALENQUE E. , SERRANO R. , AGUADO J. M. , BALLESTÍN C.
Published: August 2009

An immunohistochemical (IH) test (commercially available polyclonal antiserum rabbit anti‐Mycobacterium bovis; DAKO A/S) was used to detect the presence of mycobacteria in 65 formalin‐fixed, paraffin‐embedded tissue blocks from different organs, showing necrotizing caseous granuloma lesions on hematoxylin and eosin sections from 65 patients. These 65 samples were dyed using an acid‐fast fluorescent technique and compared using the immunohistochemical method. Both results were also compared with the mycobacterial cultures. The IH test, compared with the culture, showed a sensitivity (S) of 52%, a specificity (Sp) of 76%, a positive predictive value (PV pos) of 61% and a negative predictive value (PV neg) of 69%. We analyze these data and discuss the possible causes of false‐positive and ‐negative results of the IH test. This rapid test on paraffin embedded tissue seems valuable in the period when waiting for the culture results.
Primary mucosal melanomas of the nasal cavity and paranasal sinuses
Authors: REGAUER SIGRID , ANDERHUBER WOLFGANG , RICHTIG ERIKA , SCHACHENREITER JOCHEN , OTT ARTHUR , BEHAM ALFRED
Published: August 2009

We present 14 patients with primary sinonasal melanomas (SM) identified from 1984–1997 in our archives (11/14 lateral nose, 1/14 nasal septum, 2/14 paranasal sinuses; 8M/6F, mean age 67.7 years, range 39–88 years). Survival was poor (median 9 months) with death related to extensive local disease and/or widespread hematogenous metastases. The following histological subtypes were identified in descending order: amelanotic small blue cell, pleomorphic, epithelioid, spindle cell and myxoid. High mitotic rate and vascular invasion, absence of tumor‐infiltrating lymphocytes and regression were features shared by all SM. Negative staining of B‐ and T‐cell markers, LCA, neuroendocrine markers such as NSE, chromogranin and synaptophysin, and CK‐negativity excluded olfactory neuroblastoma, small cell undifferentiated carcinoma, and lymphoma. S‐100 protein was expressed in all SM, but demonstrated variable staining intensity with areas of complete negativity. HMB45 was strongly and uniformly (>80%) expressed in all undifferentiated small blue cell SM. The pigmented SM were predominantly HMB45‐negative. The strong HMB45 staining in amelanotic small blue cell SM is explained by the reaction of HMB45 antibody with an oncofetal antigen found in immature melanosomes. In these poorly differentiated amelanotic malignant melanomas, antibody to HMB45 proved to be a superb diagnostic marker. We therefore strongly advocate the inclusion of HMB45 antibody in the panel of antibodies for initial work‐up of undifferentiated mucosal neoplasms, since a negative S‐100 stain in small biopsy material may result in incorrect classification of these neoplasms.
Herpes simplex virus and human papillomavirus in a population‐based case‐control study of cervical intraepithelial neoplasia grade II‐III
Authors: OLSEN ANNE O. , ØRSTAVIK IVAR , DILLNER JOAKIM , VESTERGAARD BENT FABER , MAGNUS PER
Published: August 2009

In order to evaluate the association between seropositivity for herpes simplex virus (type 1 and type 2) and cervical intraepithelial neoplacia (CIN), we analysed data from a population‐based case‐control study of CIN grade II‐III which included Norwegian women aged 20 to 44 years, 94 cases and 228 controls. Our objectives were to determine if HSV‐1 and/or HSV‐2 seropositivity were independent risk factors for CIN, taking human papillomavirus exposure into account, and to elucidate the combined effect of HPV positivity and seropositivity for HSV. In logistic regression analyses, the association between HSV‐2 or HSV‐1 seropositivity and CIN II‐III was not explained by HPV (adjusted OR 3.0; 95% CI 1.3–7.2 and adjusted OR 3.3; 95% CI 1.3–8.4, respectively). In analyses restricted to HPV‐16 DNA‐positive individuals, seropositivity for HSV‐2 increased the risk of CIN (OR 11.1; 95% CI 1.2–105.7), whereas HSV‐1 seropositivity was not significantly associated with CIN. In women positive for other HPV types, only HSV‐1 seropositivity was associated with CIN (OR 8.5; 95% CI 1.3–55.8). In analyses of the HPV‐16‐seropositive individuals, neither HSV‐1 nor HSV‐2 seropositivity was associated with CIN. Compared to the reference group of jointly unexposed subjects, the HPV‐16 DNA‐positive women who were anti‐HSV‐2 negative had an increased risk of CIN (OR 29; 95% CI 12–67), whereas the risk in women who were both HPV‐16 DNA‐positive and HSV‐2 was OR = 247 (95% CI 31–1996). The estimate of interaction was strong, but did not reach significance, and our findings may suggest that the combined effect of the two viruses is of aetiological importance in cervical carcinogenesis. Furthermore, the results indicate that HPV DNA positivity is not sufficient to explain the sexual behaviour‐associated risk of cervical neoplasia and that further studies on the role of genital HSV (type 1 as well as type 2) and other STDs are warranted.
Expression of bcl‐2 in fetal thymus, thymomas and thymic carcinomas
Authors: ENGEL PETER , FRANCIS DORTHE , GRÆM NIELS
Published: August 2009

Bcl‐2 is a proto‐oncogene inhibiting apoptosis, and p53 is a tumor supressor gene which induces apoptosis. Both seem to take part in tumorigenesis. An inverse relationship between the two genes has been reported in some neoplasms, although the exact mechanism is not fully understood. We have analyzed the expression of bcl‐2 and p53 in 18 fetal thymuses and 18 clinically benign and malignant thymomas: bcl‐2 was expressed by most medullary lymphocytes and epithelial cells of the normal thymus; p53 was not expressed at all. Bcl‐2 and p53 were co‐expressed in the majority of the thymomas and the staining reaction was stronger in the clinically malignant ones. It is concluded that although co‐expression of bcl‐2 and p53 is of doubtful prognostic relevance, the staining pattern of bcl‐2 supports the histogenetic classification system of Müller‐Hermelink.
HIV infection in the first heart transplantation in Italy: fatal outcome
Authors: CALABRESE FIORELLA , ANGELINI ANNALISA , CECCHETTO ATTILIO , VALENTE MARIALUISA , LIVI UGOLINO , THIENE GAETANO
Published: August 2009

A 46‐year‐old man with alcoholic dilated cardiomyopathy underwent heart transplantation on November 14, 1985. It was the first cardiac transplant in Italy and at that time no HIV antibody screening test was available in this country. The patient remained in good health for 6 years postoperatively, with only one episode of rejection (type 3A). In June 1992 he died of fulminant complications of AIDS and severe chronic rejection. Neither the patient nor the organ donor belonged to any of the known risk groups for HIV infection; a retrospective analysis revealed that perioperative blood transfusions had been the vectors of transmission.
Immunoreactivity of pleural malignant mesotheliomas to glutathione S‐transferases
Authors: DEJMEK ANNIKA , BROCKSTEDT ULRIKA , HJERPE ANDERS
Published: August 2009

Malignant mesotheliomas show a highly variable aggressiveness, but it is difficult to predict the outcome in the individual case at the time of diagnosis. Glutathione S‐transferases are detoxification enzymes that have been correlated with the prognosis in some tumours. We have therefore assessed the value of GST expression as a prognostic parameter in mesotheliomas. The reactivities to GST‐π, ‐α and ‐μ antibodies were studied in histological sections from altogether 88 cases. Most of them showed distinct cytoplasmic reactivity to one or more of the GST antibodies tested. This high prevalence is in good agreement with the low responsiveness of mesotheliomas to chemotherapy. However, there was no prognostic value in detecting GST immunoreactivity, and it gave no information of value for distinguishing between neoplastic and reactive mesothelium.
A histomorphometric study of haematological disorders with respect to marrow fibrosis and osteosclerosis
Authors: POULSEN L. W. , MELSEN F. , BENDIX K.
Published: August 2009

A retrospective investigation of 75 EDTA‐decalcified Jamshidi biopsies collected over a 2‐year period at Aarhus University Hospital was performed. The biopsies originated from 75 patients suffering from idiopathic myelofibrosis, other chronic myeloproliferative disorders, or other conditions with known associations with bone marrow fibrosis. The relative volumes of trabecular and woven bone, as well as haematopoietic and non‐haematopoietic tissue, were estimated histomorphometrically. The degree of fibrosis was estimated semiquantitatively. Finally, the thickness of trabecular osteons was estimated from the number of lamellae by counting. Patients with idiopathic myelofibrosis had statistically significantly more bone tissue than the other groups of patients. The osteosclerosis was primarily due to woven bone. Larger cancellous osteons also suggested a positive balance in bone remodelling. The amount of bone tissue showed furthermore a statistically significant increase through the groups of polycythaemia vera, essential thrombocythaemia, chronic myelogenous leukaemia and idiopathic myelofibrosis. Parallel to the increase in the amount of bone, an increase in the degree of marrow fibrosis was detected. The positive correlation between the amount of bone and the degree of marrow fibrosis was statistically highly significant (p=0.0008).
Population genetics and molecular epidemiology of Neisseria meningitidis
Authors: CAUGANT DOMINIQUE A.
Published: August 2009

Under non‐epidemic conditions, Neisseria meningitidis causes disease primarily in children under the age of 5 and the cases are sporadic without any evident relationship between them. Occasionally, localized outbreaks of meningococcal disease occur, and sometimes epidemic waves of disease may spread to several countries or even continents and constitute a pandemic. In the past 10 years or so, population genetic analyses have provided insights into the biology of the bacterium and the epidemiology of meningococcal disease, improving our understanding of the cause of epidemics. Through the application of molecular methods, and especially multilocus enzyme electrophoresis, to N. meningitidis strains of worldwide origin, it has been possible to identify virulent clones and provide a surveillance system to warn of meningococcal epidemics. The characteristics of the predominant clones which are nowadays causing meningococcal disease in the world are summarized here and the importance of population genetics in interpreting the epidemiological data is illustrated.
A comparison of flow cytometry and immunohistochemistry in human colorectal cancers
Authors: DIEDERICHSEN AXEL COSMUS PYNDT , HANSEN TINE PLATO , NIELSEN OLE , FENGER CLAUS , JENSENIUS JENS CHRISTIAN , CHRISTENSEN PER BORUP , KRISTENSEN TOM , ZEUTHEN JESPER
Published: August 2009

In human colorectal cancer it has been reported that some tumours lack the HLA‐ABC antigens. This has been interpreted as reflecting tumour escape from the immune system. Earlier data have been obtained by immunohistochemistry. In this study, we compared the expression of HLA‐ABC, HLA‐DR, CD80 (B7–1) and CD54 (ICAM‐1) in 20 tumours using both a conventional immunohistochemistry two‐layer technique and multiparameter flow cytometry, gating on an epithelial cell marker. Colorectal cancer tissue used in flow cytometry was dissociated with collagenase, deoxyribonuclease and hyaluronidase. The intensity of expression of HLA‐ABC, HLA‐DR and CD80 was unaffected by the enzymes, but CD54 was decreased by 30%. The reproducibility of flow cytometry was good. Microscopy of sections revealed that about 5% of each tumour sample consisted of normal epithelium, but even after correction for this, flow cytometry was superior to immunohistochemistry in 33 out of 80 cases, and showed that tumours described as HLA‐ABC negative by immunohistochemistry were in fact weakly positive for HLA‐ABC. We conclude that flow cytometry and immunohistochemistry are complementary, and that flow cytometry is superior to immunohistochemistry for detecting anti‐gens/epitopes present in low amounts.
Recovery of Helicobacter pylori ATCC43504 from a viable but not culturable state: regrowth or resuscitation?
Authors: CELLINI LUIGINA , ROBUFFO IOLE , DI CAMPLI EMANUELA , DI BARTOLOMEO SORAYA , TARABORELLI TEA , DAINELLI BENEDETTO
Published: August 2009

Studies were conducted following the formation and characterization of the coccoid morphology of Helicobacter pylori. H. pylori ATCC43504 was incubated in brucella broth plus 2% fetal calf serum at three different temperatures: 37°C, room temperature and 4°C in a microaerophilic environment, and readings were taken at 2, 7, 15, 30 and 45 days. At control times, the total and the viable count, viability tests with tetrazolium salts, and ultrastructural studies were carried out. On solid media, H. pylori became nonculturable after 7 days of incubation at room temperature and 4°C, and after 15 days of incubation at 37°C. At these times of incubation, after subculturing in liquid medium under the same conditions, the growth of H. pylori was detected until the 15th day from cultures incubated at 4°C and until the 30th day from cultures stored at 37°C, and at room temperature. Ultrastructural studies showed a gradual reduction of integrity of bacterial cells that remained stable at 30 and 45 days of incubation: 30% of whole cells of bacteria incubated at 37°C and room temperature and 50% in bacteria incubated at 4°C. The viability of the VNC (viable nonculturable) state was assessed by studying the reduction of tetrazolium salts INT (p‐iodonitrophenyl tetrazolium violet) and CTC (cyanoditolyl tetrazolium chloride) to their respective formazans and this was linked to the cellular respiration. At 45 days of incubation, when bacterial regrowth was not observed in solid or in liquid medium, different resuscitation methods were applied to evaluate a possible resuscitation of VNC H. pylori. No significant growth on solid medium was observed.
Opportunistic screening for Chlamydia trachomatis cervicitis: the value of cytobrush specimens for detection by PCR compared with cell culture
Authors: WANDALL DORTE ALNOR , ØSTERGAARD LARS , OVERGAARD LENE , WORM ANNE‐MARIE , GUTSCHIK ERNÖ
Published: August 2009

Endocervical sampling for microbiological and pathological screening is laborious and expensive due to different sampling devices and techniques. The purpose of this study was to examine if the routine procedure could be simplified by using a cytobrush for concurrent cytology and sampling for Chlamydia trachomatis detection using the PCR method or cell culture. As a sampling device control we used a conventional rayon swab. Results: Culture: Out of 873 paired endocervical specimens, C. trachomatis was isolated in 68 swab specimens and in 65 cytobrush specimens (overall detection rate 8.4%). The cytobrush proved less suitable than the swab for the isolation of C. trachomatis as 31.5% of the cytobrush samples showed cytotoxicity to the cultured cells vs 0.9% of the swab samples. PCR: In a random sample of 427 paired endocervical specimens, C. trachomatis was detected in 45 pairs without any difference between the two sampling devices. The sensitivity of PCR was 93.8%vs 89.6% and 87.5% in cultured swab and cultured cytobrush specimens, respectively. The cytobrush can therefore be recommended as a cervical sampling device if a PCR assay is used for the detection of C. trachomatis, but not if the cell culture method is used, due to high cytotoxicity. Furthermore, the same cytobrush may be used for cervical cytological sampling and thereafter placed in transport medium for subsequent C. trachomatis detection if the PCR technique is used.
Encephalocele with an area mimicking giant cell fibroblastoma
Authors: HIROKAWA MITSUYOSHI , SHIMIZU MICHIO , MANABE TOSHIAKI , SHIRABE TERUO
Published: August 2009

We report a unique case of encephalocele with an area mimicking giant cell fibroblastoma. A 4‐month‐old boy had been born with a parietal midline mass of the head, measuring 3times3x1.5 cm. Microscopically, it consisted of fibrous connective tissue and brain tissue. The fibrous tissue revealed spindle cells and multinucleated giant cells lining sinusoid‐like spaces. This area was reminiscent of giant cell fibroblastoma. These cells were immunopositive for vimentin and negative for CD34. The giant cell fibroblastoma‐like lesion in our case seems to be non‐neoplastic and hamartomatous, and we think that the immunohistochemistry using CD34 is useful for differentiating the two lesions.
T‐cell activation and the dynamic world of rafts:
Authors: Viola Antonella , Lanzavecchia Antonio
Published: August 2009

Recent evidence, indicates that T‐cell receptor (TCR) triggering and T‐cell activation are dynamic processes that involve various aspects of T‐cell organization. In addition to the interaction between the TCR molecule and its ligand, T‐cell activation depends on a combination of many other events involving coreceptor molecules, actin cytoskeleton and plasma membrane lipids. Altogether, these cell structures organize the formation of a specialized junction between the T cell and the antigen‐presenting cell (APC), that plays a critical role in sustaining and amplifying TCR signalling.
Formation and structure of mixed bacterial communities
Authors: Tetz VIctor V.
Published: August 2009

Mixed bacterial communities are formed by unrelated bacteria on solid media. Mixed bacterial communities on solid media are similar to “classical” colonies and are formed after the growth of a large number of unrelated bacteria simultaneously plated onto a limited area of agar. The morphology of the mixed bacterial communities was similar for different combinations of bacteria and did not change when the bacteria were plated on different media. Different bacterial strains form zones of individual and mixed growth in the structure of mixed bacterial communities. The results of electron microscopic examination indicate that mixed bacterial communities are isolated from their external environment by a surface film. The basic part of this film is formed by an elementary membrane. The membrane of the surface film of mixed bacterial communities is a stable structure occupying a large surface area. The results of this investigation seem to indicate the existence of a special type of co‐operation between different species of bacteria. This type of co‐operation may be very important in the regulation of interactions between different bacteria and between bacteria and the environment.
Development of intestinal bacterial enzymes in infants ‐ relationship to mode of delivery and type of feeding
Authors: Grönlund Minna‐Maija , Salminen Seppo , Mykkänen Hannu , Kero Pentti , Lehtonen Olli‐Pekka
Published: August 2009

To evaluate the development of intestinal flora in young infants, and especially to estimate the influence of mode of delivery and type of feeding on the establishment of intestinal microflora, faecal flora was studied indirectly by measuring prospectively the faecal bacterial enzyme activities (β‐glucosidase, β‐glucuronidase and urease) in 29 full‐term, healthy infants during the first 6 months of life. Mode of delivery had no influence on the faecal enzyme activities. In contrast, infants receiving formula feeds were more often urease positive at 1–2 months of age (70%vs 25%, p=0.043) and had higher median activity of β‐glucuronidase at 6 months of age (0.90 and 0.19 nmoles/mg protein × min, p=0.0043) than exclusively breast‐fed infants. Through indirect methods to measure the development of a faecal microflora our results indicate that the type of milk that infants receive during the first months of life may have an important role in the development of intestinal flora.
Comparison of thymidine kinase and A‐type inclusion protein gene sequences from Norwegian and Swedish cowpox virus isolates
Authors: Hansen H. , Sandvik T. , Tryland M. , Olsvik ø. , Traavik T.
Published: August 2009

During the last decades, cowpox virus, a member of the genus Orthopoxvirus within the Poxviridae family, has appeared as a pathogen in domestic cats, zoo animal species, and humans. At the same time, vaccinia virus, another orthopoxvirus, has been used as a recombinant vaccine vector with foreign genes inserted in the thymidine kinase (TK) gene. By PCR and cycle sequencing, we have determined the nucleotide sequences of the TK gene and the A‐type inclusion protein (ATIP) gene of virus isolates from two human cowpox cases in Sweden, as well as a human and a feline case from Norway. We also obtained the corresponding sequences from ectromelia virus (strain Moscow), cow‐pox virus (strain Brighton) and vaccinia virus (strain Western Reserve). The new virus isolates differed from ectromelia virus and vaccinia virus, and were confirmed to be cowpox virus strains. Isolates originating from the same country had nearly identical TK sequences and fully identical ATIP sequences. They probably represent local geographical strains of cowpox virus.
Duodenal colonization byGeotrichum candidumin a child with transient low serum levels of IgA and IgM
Authors: Vasei Mohammad , Imanieh Mohammad Hadi
Published: August 2009

Fungal colonization was observed in the duodenal biopsy specimens of a nine‐year‐old girl who complained of anorexia and epigastric pain. Endoscopy revealed gastric redness andGeotrichum candidumwas identified in the duodenum by histopathology and tissue culture. Immunologic work‐up showed low serum levels of IgA and IgM. She was treated with ranitidine and antacids for gastritis. At follow‐up, the levels of immunoglobulins had normalized, no fungal elements were detected, and clinical symptoms had disappeared. Duodenal invasion byG. candidumhas not been described so far. Duodenal colonization byG. candidummay occur when mucosal immunity is disturbed and may be the source of dissemination in severe immunodeficiency states.
Complement haemolytic activity, circulating immune complexes and the morbidity of sickle cell anaemia
Authors: Anyaegbu C. C. , Okpala I. E. , Aken'Ova A. Y. , Salimonu L. S.
Published: August 2009

The aim of this study was to find out if the number of crises and complications of sickle cell anaemia (SCA) relate to complement function, or the levels of circulating immune complexes (CIC), complement factor B (Bf), C3 and C4. In 73 steady‐state HbSS patients and 50 HbAA control subjects, we determined the haemolytic activity of the alternative pathway of complement (AP50), of the classical pathway (CH50); and the serum concentrations of Bf, C3, C4 and CIC. By clinical examination of each patient and review of the medical records, we determined the number of complications of SCA which had occurred and the mean number of crises per year over a minimum period of 3 years. The mean±SD AP50 for the patients (14±2 U/ml) was significantly lower than the control value of 16±3 U/ml (p<0.001). AP50 had a significant inverse correlation with the number of crises (r=‐0.30, p<0.02). Mean±SD CIC in patients (0.45±0.38 g/1) was significantly higher than in controls: 0.24±0.15 g/1 (p<0.002). CIC snowed a significant direct correlation with the number of complications of SCA (r=+0.28, p<0.02). Mean±SD Bf in SCA patients (0.19±0.09) was higher than in controls (0.17±0.05). The difference reached marginal statistical significance (p=0.049). SCA patients and controls had no significant differences in CH50, C3 and C4. These parameters and Bf did not correlate with either the number of crises or complications. The mechanisms underlying the correlations observed in this study are yet to be fully elucidated.
Minute oxidative stress is sufficient to induce apoptotic death of NIT‐1 insulinoma cells
Authors: Olejnicka Beata T. , Dalen Helge , Brunk Ulf T.
Published: August 2009

When cultured NIT‐1 cells were subjected to a low level of oxidative stress (30 μM hydrogen peroxide for 15 min at 37°C) several of their lysosomes ruptured, as demonstrated by intravital staining with the lysosomotropic weak base acridine orange. Such rupture is due to intralysosomal, iron‐catalyzed oxidative reactions, since it was largely prevented by previous endocytotic uptake of desferrioxamine. The resultant limited leakage of lysosomal hydrolytic enzymes into the cytosol could be important for an apoptotic‐type degradatiodfragmentation process within initially intact plasma membranes. In contrast, extensive lysosomal rupture leads to necrosis. The development of the damage process was followed by light‐ and electron microscopy; and by the TUNEL‐reaction. As a result of the applied oxidative stress, which is comparable to that expected to occur within the microenvironment surrounding activated macrophages under oxidative burst (e.g. during autoimmune insulitis), about 90% of the cells eventually died due to post‐apoptotic secondary necrosis. The few surviving cells phagocytosed the debris from their fragmented neighbours and began to divide about 24 h after the insult. Thus the sensitivity to oxidative stress varies, perhaps as a consequence of varying amounts intralysosomal redox‐active iron, as we have found to be the case in several other cellular systems. Since the NIT‐1 cells are highly differentiated, and in many ways like beta cells, we consider our result to be of value for the understanding of beta‐cell death during the development of insulin‐dependent (Type I) diabetes mellitus (IDDM).
Expression of fibroblast growth factor‐2 transcripts in the healing of acetic acid‐induced gastric ulcers
Authors: Kimura Tsuguhiro , Noda Masao , Sugihara Hiroyuki , Kashima Kei , Hattori Takanori
Published: August 2009

Basic fibroblast growth factor‐2 (FGF‐2) has an important role in angiogenesis, and has been demonstrated to promote ulcer healing. However, the actual part played by FGF‐2 in the process of ulcer healing is not well understood. In this study, we investigated expression of FGF‐2 transcripts at each stage of gastric ulcer healing, using an acetic acid model in rats. We made ulcers in the rat stomach by direct application of acetic acid, and 3 days and 1, 2, 3, 4, 8 weeks after treatment, we examined expression of FGF‐2 transcripts by in situ hybridization. On day 3, FGF‐2 transcripts were detected in mononuclear cells infiltrating the submucosal layer around the ulcer. After 1 and 2 weeks, expression of FGF‐2 transcripts was prominent in spindle‐shaped mesenchymal cells and endothelial cells, which proliferated in the ulcerative region. Some of the spindle‐shaped cells which expressed FGF‐2 transcripts also showed immunoreactivity for α‐smooth muscle actin. After 3 and 4 weeks, FGF‐2 expression was seen mainly in endothelial cells of vessels. These results suggest that different cells produce FGF‐2 during the process of gastric ulcer healing, and some of the spindle‐shaped cells expressing FGF‐2 transcripts in the early phase are myofibroblasts.
The role of volume‐weighted mean nuclear volume in predicting disease outcome in patients with prostate cancer treated with radical prostatectomy
Authors: Fujikawa Keita , Itoh Tadahiro , Nishio Yasunori , Miyakawa Mieko , Sasaki Miharu
Published: August 2009

Background. Estimates of volume‐weighted mean nuclear volume (MNV) are the only means by which unbiased estimates of three‐dimensional parameters can be obtained from single two‐dimensional sections without any assumptions. We have reported that for prostate cancer estimates of MNV are prognostically equal or superior to morphological grading of malignancy, such as Gleason score (GS), and in particular, that MNV proved to be a meaningful predictor of prognosis for patients with clinically localized tumors. However, all previous studies were conducted on patients treated conservatively, and no authors have tested whether estimates of MNV can predict the prognosis of patients treated with radical prostatectomy. Materials and methods. A retrospective prognostic study of 52 patients with clinically localized prostate cancer diagnosed at three Hospitals in Shizuoka Prefecture, Japan (Shizuoka City Hospital, Shizuoka Prefectural Hospital and Shimada Municipal Hospital) and treated by radical prostatectomy was performed. Twenty of these patients were treated with hormone therapy before radical prostatectomy. Unbiased estimates of MNV were compared with clinical stage, histological grading according to GS and neo‐adjuvant hormone therapy with regard to the prognostic value. Results. MNV was significantly correlated with pathologcial T stage, but was not significantly correlated with the presence or absence of lymph node metastasis. Univariate analysis revealed that MNV correlated significantly with progression‐free survival (p=0.0116). Multivariate analysis revealed that MNV (p=0.0115) and GS (p=0.0275) were two significant independent predictors of progression‐free survival. Conclusions. The results of the present study suggest that MNV and GS are powerful independent predictors of prognosis for prostate cancer treated with radical prostatectomy. We recommend estimates of MNV as a supportive method to the histological grading for patients with prostate cancer.
The role of adenosine receptor engagement in murine fetal thymocyte development
Authors: Hamad Mawieh
Published: August 2009

The role of adenosine receptor engagement in murine T‐cell development was evaluated by culturing day 15–16 fetal thymic lobes in the presence of various concentrations of the adenosine receptor agonist 5′‐(N‐ethyl)‐carboxamidoadenosine (NECA) or the adenosine receptor antagonist 8‐phenyl‐theophylline (8‐PT) using the fetal thymic organ culture (FTOC) system. Before and 8 days after culture, thymocytes were isolated, counted, and analyzed for the expression of CD4 and CD8 T‐cell differentiation molecules. Analysis of fresh thymocytes prior to culture showed that the majority of cells were of the CD4 single‐positive or CD4+CD8+immature phenotype. Eight days after culture with media alone, 44% of cells were CD4+and 23% were CD8+, and the number of viable thymocytes had increased from 1.7 × 105to 2.2 × 105cells per thymic lobe. A dose‐dependent inhibition of maturation was observed in cultures with 8‐PT, with greater than 85% inhibition at 50 μM. The double‐positive thymocyte subset was most severely depleted. The number of cells obtained from cultures with NECA was also reduced, with about 65% inhibition at 50 μM, especially the CD8+subset that was most severely affected. These results suggest that adenosine receptor engagement is required for normal T‐cell differentiation and that adenosine receptor agonists and antagonists have distinct effects on thymocyte differentiation. An understanding of the cell‐type‐specific and developmental expression of adenosine receptors will help elucidate the mechanisms by which adenosine receptor engagement influences T‐cell development.
Typing ofPasteurella multocidafrom haemorrhagic septicaemia in Danish fallow deer(Dama dama)
Authors: Aalbæk Bent , Eriksen Lis , Rimler Richard B. , Leifsson Pall S. , Basse Axel , Christiansen Torben , Eriksen Erik
Published: August 2009

Isolatesof Pasteurella multocidassp.multocida(n = 31) from a Danish population of fallow deer which succumbed to haemorrhagic septicaemia during 1992–1993 and isolates from the palatine tonsils of apparently healthy fallow deer from the same area (n=6) were typed and compared withP. multocidafrom other sources. Plasmids were not observed in the fallow deer strains and one unique pattern was observed by ribotyping usingHindIIIand by pulsed‐field gel eletrophoresis usingSalIas restriction endonuclease. All Danish fallow deer isolates belonged to serotype B:3,4. On restriction endonuclease analysis usingHhaIas restriction endonuclease, all had a profile identical to that of a fallow deer isolate from the United Kingdom: profile 0033 of Wilson et al. On restriction endonuclease analysis usingHpaIIas restriction endonuclease, the Danish fallow deer isolates had a unique profile, designated 0062, which differed slightly from that of a fallow deer isolate from the United Kingdom.P. multocidafrom other animal species were genotypically different from the fallow deer isolates. It is concluded that a specific clone ofP. multocidawas responsible for the outbreak of haemorrhagic septicaemia among Danish fallow deer. A carrier rate of 27% was demonstrated among apparently normal animals from the same population.
Malformations of cranial base structures and pituitary gland in prenatal Meckel syndrome
Authors: Klær Klaus Wilbrandt , Hansen Birgit Fischer , Keeling Jean W. , Nolting Dorrit , Kjær Inger
Published: August 2009

The sella turcica region, including the clivus and the pituitary gland, was studied histologically in five human fetuses with Meckel syndrome (MS). All cases had malformed sella turcica and malformed clivus with irregularly shaped notochordal remnants. We consider that these three characteristics are constant phenotypic traits in MS. The adenohypophysis was present in three cases. In one of these, ectopia of the gland occurred with adenopituitary tissue overlying the dorsum sella, and in another remnants were found in the pharyngeal submucosa. In two fetuses the neurohypophysis was not found. The findings in the region were compared to normal findings and to findings in trisomy 18, where cranial base structures radiographically appeared similar to those in MS. We conclude that in MS specific characteristics are found in the cranial base region and that radiographic analysis needs to be supplemented by histological analysis when studying this specific region.
Immature peritoneal mast cells in neonatal rats express the CTMC phenotype, as well as functional IgE receptors
Authors: Chen Xiao‐Jun , EnerbäCk Lennart
Published: August 2009

The purpose of this study was to investigate the relationship between the differentiation and maturation of mast cells and the expression of IgE receptors on their surface in neonatal animals in vivo. Another aim was to clarify whether connective tissue mast cells (CTMC) undergo a maturation process involving a transdifferentiation from mucosal mast cells (MMC) during this period of time. Mast‐cell phenotypes were studied in terms of the profiles of proteinases and proteoglycan. In 1‐week‐old rats, the mast‐cell granules stained with Alcian blue rather than with safranin (AB+/S‐) in the Alcian blue/safranin staining sequence, normally regarded as a property of MMC. However, the AB+/S‐stained proteoglycan was degradable by nitrous acid and stained with berberine sulphate, thus indicating that it contained heparin rather than chondroitin sulphate. The mast cells expressed rat mast‐cell proteinase (RMCP) I rather than RMCP II, which is normally found in MMC. The mast cells of 1‐week‐old rats expressed functional IgE receptors, by showing a dose‐dependent IgE‐mediated histamine release of mast cells. About 70% of the IgE receptors on the mast cells were occupied by IgE. In 2‐ to 3‐week‐old rats, there was a progressive increase in mast cells stained with both Alcian blue and safranin or with safranin alone, i.e. they gradually changed towards the staining properties of CTMC (AB‐/S+). The expression and the degree of IgE occupancy of the receptors increased in 1‐ to 3‐week‐old animals. This was paralleled by an increment in cell size and in the content of heparin, histamine and serotonin in the mast cells. The findings thus indicate that the peritoneal mast cells of neonatal rats express the CTMC phenotype and undergo a maturation process at from 1 to 3 weeks of age, without involving a transdifferentiation from MMC. The maturation of the mast cells is accompanied by an increase in the expression of functional IgE receptors on the cell surface. IgE production was detectable as early as in 1‐week‐old rats.
Tyrosinase gene expression in clear cell sarcoma indicates a melanocyte origin: insight from the first reported canine case
Authors: Cangul I. Taci , Van Garderen Evert , Van Der Poel Hein J. A. , Weijer Kees , Misdorp Wim
Published: August 2009

The aim of this study was to characterize a metastasizing soft tissue tumor in a dog, which clinically, grossly and histologically showed a close resemblance to human clear cell sarcoma, a soft tissue variant of malignant melanoma. Ultrastructurally, melanosomes were found, indicating a melanocytic origin of the tumor. Using reverse‐transcription polymerase chain reaction, expression of the gene encoding tyrosinase was determined in tumor cells. With this first case of canine clear cell sarcoma, as well as the earlier report from our laboratory on amelanotic melanomas in the cat, we demonstrate that expression of the tyrosinase gene may occur in a broader range of less differentiated melanocytic tumors in different species, including man.