Syncytin-1 and its receptor is present in human gametes.

MAIN PURPOSE AND RESEARCH QUESTION: To determine whether the true fusogen Syncytin-1 and its receptor (ASCT-2) is present in human gametes using qRT-PCR, immunoblotting and immunofluorescence. METHODS: Donated oocytes and spermatozoa, originating from a fertility center in tertiary referral university hospital, underwent qRT-PCR, immunoblotting and immunofluorescence analyzes. RESULTS: Quantitative RT-PCR of sperm samples from sperm donors showed that syncytin-1 is present in all samples, however, protein levels varied between donors. Syncytin-1 immunoreactivity predominates in the sperm head and around the equatorial segment. The receptor ASCT-2 is expressed in the acrosomal region and in the sperm tail. Moreover, ASCT-2, but not syncytin-1, is expressed in oocytes and the mRNA level increases with increasing maturity of the oocytes. CONCLUSIONS: Syncytin and its receptor are present in human gametes and localization and temporal appearance is consistent with a possible role in fusion between oocyte and sperm.

A randomized study comparing short-time infusion of oxaliplatin in combination with capecitabine XELOX(30) and chronomodulated XELOX(30) as first-line therapy in patients with advanced colorectal cancer.

BACKGROUND: Chronotherapy is one of the several approaches to increase efficacy and reduce toxicity of chemotherapy. In a phase II study in the second-line in patients with metastatic colorectal cancer (mCRC), we found that chronomodulated XELOX (XELOX(30Chron)) was a well-tolerated regimen with potentially reduced toxicity. PATIENTS AND METHODS: One hundred and forty-one patients with unresectable mCRC were enrolled in a randomized study comparing standard XELOX (XELOX(30)), arm A, and XELOX(30Chron), arm B-both with short-time infusion of oxaliplatin-with the primary aim of reducing overall toxicity. RESULTS: Overall toxicity grade 2-4 was 90% versus 85%, P = 0.47 and grade 3-4 was 31% versus 37%, P = 0.6 in arm A and B, respectively. We found no significant differences in median overall survival (17.6 versus 15.5 months; P = 0.068) and median progression-free survival (8.9 versus 8.8 months; P = 0.7). The incidence of grade 3 neuropathy was 16% in arm A and 19% in arm B (P = 0.7) after a cumulative dose of oxaliplatin of 1000 mg/m(2). CONCLUSION: XELOX(30Chron) does not reduce toxicity or improve efficacy. A 30-min infusion of oxaliplatin is safe and does not increase the severity of chronic neuropathy.

Patient preference for oral or intravenous chemotherapy: a randomised cross-over trial comparing capecitabine and Nordic fluorouracil/leucovorin in patients with colorectal cancer.

Until recently, fluorouracil (F) and leucovorin (L) had been considered the standard therapy for patients with colorectal cancer. However, several studies have shown that oral therapy with UFT/L or capecitabine is as effective as intravenous (i.v.) therapy and in addition it is claimed that patients prefer oral to i.v. therapy as long as efficacy is not compromised. In a previous crossover study by Borner et al., it was shown that 26 out of 31 patients preferred oral therapy with UFT/L to i.v. FL (Mayo regimen) [Borner M, Schöffski P, de Wit R, et al. Patient preferences and pharmacokinetics of oral modulated UFT versus intravenous fluorouracil and leucovorin: a randomised crossover trial in advanced colorectal cancer. Eur J Cancer 2002;38:349-58]. The objective of the present study was to investigate patient preference between i.v. FL and oral capecitabine using the design described by Borner. The Nordic FL schedule is a bolus regimen with efficacy comparable to other i.v. regimens and at the same time a very tolerable and easy administered regimen. We randomised 60 patients with colorectal cancer (53 patients received adjuvant therapy and seven patients received palliative therapy) to start therapy with either oral capecitabine or Nordic bolus FL. After 6 weeks of therapy (two courses of capecitabine or three courses of Nordic FL) patients were crossed over to the other regimen. After having completed 12 weeks of therapy the patients (49 evaluable patients) were asked to choose one of the regimens for a further 12 weeks of therapy. Patients had more side-effect when treated with capecitabine and a total of 30 out of 49 (61%) preferred the Nordic FL regimen and 19 (39%) preferred capecitabine. We conclude that patients prefer the regimen with less toxicity and that it is of minor importance whether the medication is administrated orally at home or i.v. at the hospital.

Syncytin is involved in breast cancer-endothelial cell fusions.

Cancer cells can fuse spontaneously with normal host cells, including endothelial cells, and such fusions may strongly modulate the biological behaviour of tumors. However, the underlying mechanisms are unknown. We now show that human breast cancer cell lines and 63 out of 165 (38%) breast cancer specimens express syncytin, an endogenous retroviral envelope protein, previously implicated in fusions between placental trophoblast cells. Additionally, endothelial and cancer cells are shown to express ASCT-2, a receptor for syncytin. Syncytin antisense treatment decreases syncytin expression and inhibits fusions between breast cancer cells and endothelial cells. Moreover, a syncytin inhibitory peptide also inhibits fusions between cancer and endothelial cells. These results are the first to show that syncytin is expressed by human cancer cells and is involved in cancer-endothelial cell fusions.

Automated identification of cancerous smears using various competitive intelligent techniques.

In this study the performance of various intelligent methodologies is compared in the task of pap-smear diagnosis. The selected intelligent methodologies are briefly described and explained, and then, the acquired results are presented and discussed for their comprehensibility and usefulness to medical staff, either for fault diagnosis tasks, or for the construction of automated computer-assisted classification of smears. The intelligent methodologies used for the construction of pap-smear classifiers, are different clustering approaches, feature selection, neuro-fuzzy systems, inductive machine learning, genetic programming, and second order neural networks. Acquired results reveal the power of most intelligent techniques to obtain high quality solutions in this difficult problem of medical diagnosis. Some of the methods obtain almost perfect diagnostic accuracy in test data, but the outcome lacks comprehensibility. On the other hand, results scoring high in terms of comprehensibility are acquired from some methods, but with the drawback of achieving lower diagnostic accuracy. The experimental data used in this study were collected at a previous stage, for the purpose of combining intelligent diagnostic methodologies with other existing computer imaging technologies towards the construction of an automated smear cell classification device.

Regulation of ribosomal RNA gene expression in porcine oocytes.

In vitro production (IVP) of porcine embryos including in vitro maturation (IVM) of oocytes followed by in vitro fertilization (IVF) and in vitro culture (IVC) of the resultant embryos may result in live offspring, but it is still associated with great inefficiencies probably due to incomplete cytoplasmic maturation of the oocytes in vitro. Therefore, fundamental knowledge on the regulation of transcription during the oocyte growth phase when the messengers and protein synthetic machinery necessary for oocyte developmental competence are formed, is of great importance. In mammals, synthesis of RNA, up to 60-70% of which is ribosomal (rRNA), increases during oocyte growth and reaches a peak at the beginning of follicular antrum formation. In oocytes at the end of the growth phase, acquisition of full meiotic competence coincides with a markedly decreased rRNA transcriptional activity in the gametes. Our recent studies on the porcine oocyte growth phase have revealed a deeper molecular and biological insight into the complex regulation of rRNA transcription at different stages of follicular development. The data indicate that the so-called pocket protein, p130, is involved in the down-regulation of rRNA transcription at the end of the oocyte growth phase through an inhibition of the action of upstream binding factor (UBF). The latter protein is necessary for the function of RNA polymerase I (RNA Pol I), which is the actual enzyme driving rRNA gene transcription. Moreover, rRNA transcription also appears to be down-regulated by a decrease in the expression of mRNA encoding PAF53, an RNA Pol I-associated factor also required for the polymerase to exert its action. At the ultrastructural level, these molecular changes are paralleled by marginalization of the fibrillar centres of the oocyte nucleolus followed by compaction of the nucleolus into an inactive sphere of fibrils.

The most common chromosome aberration detected by high-resolution comparative genomic hybridization in vulvar intraepithelial neoplasia is not seen in vulvar squamous cell carcinoma.

We analyzed genetic changes in condylomas (four cases), vulvar intraepithelial neoplasia I-III (VIN I-III, eleven cases), and primary vulvar squamous cell carcinomas (VSCC, ten cases) by high-resolution comparative genomic hybridization (HR-CGH) and flowcytometry. All samples were also human papilloma virus (HPV)-genotyped. Gain of chromosome 1, the aberration most often seen in VIN III (67%), was not seen in HPV-positive or -negative VSCCs (0%). Both VIN III and VSCC frequently showed gain of 3q (56 and 70%, respectively). The VIN III samples often demonstrated gain of 20q (56%) and 20p (44%), and the VSCC samples gain of 8q (60%), loss of 3p (50%), and 8p (40%). None of the four most frequent changes in the VSCC samples occurred exclusively in the HPV-positive or -negative samples. As expected, we did not find any cytogenetic changes in condylomas and nearly any changes in VIN I-II.

Nucleolar ultrastructure and protein allocation in in vitro produced porcine embryos.

The nucleolus formation was studied as an indirect marker of the ribosomal RNA (rRNA) genes activation in porcine embryos following oocyte maturation, fertilization, and culture in vitro. Nucleologenesis was assessed by transmission electron microscopy (TEM), light microscopical autoradiography following 20 min of 3H-uridine incubation, and immunocytochemical localization of key nucleolar proteins involved in rRNA transcription (upstream binding factor (UBF), topoisomerase I, and RNA polymerase I) and processing (fibrillarin, nucleophosmin, nucleolin) by confocal laser scanning microscopy. During the first four post-fertilization cell cycles, TEM revealed spherical nucleolus precursor bodies (NPBs), consisting of densely packed fibrils, as the most prominent intra-nuclear entities of the blastomeres. Fibrillo-granular nucleoli were observed in some blastomeres in a single embryo during the 5th cell cycle, i.e., the tentative 16-cell stage, where formation of fibrillar centres (FC), a dense fibrillar component, and a granular component on the surface of the NPBs was seen. In this embryo, autoradiographic labeling was detected over the nucleoplasm and in particular over the nucleoli. Fibrillarin was immunocytochemically localized in the presumptive NPBs of the pronuclei. This protein was again localized to the presumptive NPBs together with nucleolin from late during the 3rd cell cycle, i.e., the four-cell stage in some embryos. UBF, RNA polymerase I, and nucleophosmin were localized to the presumptive NPBs in a proportion of the embryos at the 4th cell cycle, i.e., the tentative eight-cell stage and onwards. Toposiomerase I was not localized to intra-nuclear entities even during the 5th post-fertilization cell cycle. Moreover, a considerable proportion of the blastomere nuclei apparently did not show localization of other nucleolar proteins. In conclusion, porcine embryos produced in vitro display a substantial delay in or even lack of the development of functional nucleoli.

Osteoclast-like giant cells in leiomyomatous tumors of the uterus. A case report and review of the literature.

Osteoclast-like giant cells (OLGC) in leiomyomatous tumors of the uterus are rarely seen, and their significance is unknown. We present a case of a large leiomyomatous tumor in which OLGC were found in only few sections showing leiomyosarcoma whereas the majority of sections revealed a leiomyoma. Though radically operated, the patient died a few months later with recurrent tumor in the pelvis and metastases to the lungs.

Accumulation of the proteolytic marker peptide ubiquitin in the trophoblast of mammalian blastocysts.

Ubiquitination is a universal protein degradation pathway in which the molecules of 8.5-kDa proteolytic peptide ubiquitin are covalently attached to the epsilon-amino group of the substrate's lysine residues. Little is known about the importance of this highly conserved mechanism for protein recycling in mammalian gametogenesis and fertilization. The data obtained by the students and faculty of the international training course Window to the Zygote 2000 demonstrate the accumulation of ubiquitin-cross-reactive structures in the trophoblast, but not in the inner cell mass of the expanding bovine and mouse blastocysts. This observation suggests that a major burst of ubiquitin-dependent proteolysis occurs in the trophoblast of mammalian peri-implantation embryos. This event may be important for the success of blastocyst hatching, differentiation of embryonic stem cells into soma and germ line, and/or implantation in both naturally conceived and reconstructed mammalian embryos.

Staging uterine cervical carcinoma with low-field MR imaging.

PURPOSE: To assess the validity of low-field MR in staging cervical cancer compared to clinical staging. MATERIAL AND METHODS: A total of 95 women entered the study over a 3-year period. MR examinations with a 0.1 T resistive magnet using a body coil and clinical staging according to the FIGO recommendations (1988) were performed within 2 weeks from clinical diagnosis. T1- and T2-weighted sequences were obtained in transversal and sagittal acquisitions, and an additional T1 before and after contrast (randomisation to 0.1 or 0.3 mmol/kg b.w. gadodiamide). Treatment decisions on surgery or radiation therapy were made solely on the clinical staging. RESULTS: Sixty-one patients were found to be eligible for surgery. In 5 women, the pathological results revealed a more advanced stage of the disease than assessed by clinical staging. MR correctly staged 4 of the 5 but otherwise tended to overstate the disease. Contrast enhancement significantly reduced this trend (p<0.05) regardless of the contrast medium dose used. Divided into two groups, an operable (less than stage 2b) and an inoperable group (more than stage 2a), the clinical staging correctly classified 57 patients (accuracy 92%) compared to 52 patients with MR using contrast enhancement (accuracy 84%). The specificity was no higher than 31%, whereas the reproducibility of the MR assessment was fairly good with kappa values around 0.65 for both intra- and inter-observer variations. CONCLUSION: In the present set-up, clinical assessment was superior to low-field MR in staging cervical cancer. When using contrast enhancement, the staging accuracies of low-field MR were comparable to the ones reported for techniques with higher tesla values, whereas the specificity and reproducibility errors were lower. The method, therefore, needs to be optimised.

An empirical study of stalking victimization.

This article empirically studies the phenomenon of stalking and its victims by utilizing a random sample of college students at a large public University. The study found that 25% of the women and 11% of the men had been stalked at some point in their lives and that six percent were currently being stalked. Additionally, the study found that the majority of stalking victims are women who are stalked by male offenders. The sample reported being stalked for an average of 347 days and having engaged in a variety of actions in response. A substantial number of victims reported being threatened by their stalkers. This threat was associated with higher levels of fear among the victims and a greater chance of physical attack by the stalkers, particularly for the female victims.

Liposarcoma in association with total hip replacement.

A possible relation between joint replacement and malignancy has previously been suggested in a few cases. In the majority of these cases, the authors found either malignant fibrous histiocytomas or osteosarcomas. We hereby report the first case of a liposarcoma in the vicinity of a joint prosthesis.

Observer variation in the assessment of the histopathologic diagnosis of acute appendicitis.

BACKGROUND: The objectives of this study were to determine observer variation in the assessment of the histopathologic diagnosis of acute appendicitis. METHODS: Two consultant pathologists independently evaluated 415 appendectomy specimens (set I). After a consensus conference at which the observers unified the criteria of the diagnosis of acute appendicitis, another 396 appendectomy specimens (set II) were evaluated. To calculate the intra-observer variation, one observer evaluated set II once more. Finally, using the consensus diagnosis as the final diagnosis, the diagnostic accuracy of the surgeon, the primary pathology report, and the two observer evaluations was calculated. RESULTS: In set I no difference was found in the level of agreement on the diagnosis of acute appendicitis versus other diagnoses, and a kappa value of 0.85 was obtained. In set II a kappa value of 0.88 was obtained, which was not significantly different from the kappa value in set I. However, the consensus conference did increase the level of agreement on the diagnosis of acute appendicitis. The intra-observer variation was calculated, and a kappa value of 0.88 was obtained. The cases in which the observers disagreed on a positive diagnosis were cases of mild appendicitis. The question remains how many specimens would have shown these changes if all tissue had been developed for microscopy. The observers' diagnoses were the most reliable, and there was a significant decrease in the diagnostic accuracy compared with both the primary pathology report and the surgeon's opinion, the last one being the least accurate. CONCLUSIONS: These observations stress how important it is for the pathologist to discuss the diagnostic criteria of the diagnosis of acute appendicitis and for the surgeon to go back to the patient's record and look up the histopathologic diagnosis.

Intramuscular myxoma and fibrous dysplasia of bone--Mazabraud's syndrome. A case report.

We present a case of Mazabraud's syndrome, a rare benign disease, with multiple intramuscular myxomas of the thoracic wall associated with fibrous dysplasia of bone. CT, MR imaging and ultrasonography (US) of the thorax showed 2 well circumscribed homogeneous intramuscular tumors. A US-guided needle biopsy with a large-core needle (2.0 mm) and a fine needle (0.8 mm) showed that the tumors were intramuscular myxomas with no sign of malignancy. 99mTc bone scintigraphy showed a markedly increased uptake in the right lower skull, and multiple smaller foci. CT of the skull revealed a right-sided unilateral bone thickening of the orbit and the ethmoidal cells, and right-sided exophthalmia. This case history suggests that patients with multiple intramuscular myxomas should be preoperatively examined for osseous lesions. A postoperative follow-up should also be performed to detect other soft-tissue myxomas not as yet clinically detectable, or rare osseous complications.

[EDP registration of the types of tests, working procedures and diagnosis in a department of histopathology. Minimum dataset for assessment of the workload]. / Edb-registering af prøvetyper, arbejdsprocedurer og diagnoser på patologisk anatomisk afdeling.Minimumdatasoet for opgørelse af ressourceforbrug.

The aim of the study was to develop a computer-based system for measuring workload in a department of histo- and cytopathology using routine registration of a minimum dataset. A group with representatives from the laboratory technicians, the pathologists and the secretaries defined 18 types of specimens. By studying each step of specimen processing it was shown, that 14 items for technical details could cover all the work done in the department. This information was collected in a computer-based system connected to the hospital network. The measurement of workload is essential for the efficient management of laboratory services. The registration of specimen types and a minimum dataset for specimen processing describes the work done in a department of histo- and cytopathology.

[A time study in a department of histopathology]. / Tidsstudie på patologisk anatomisk afdeling.

In the Department of Histo- and Cytopathology, Herlev University Hospital, Copenhagen, a time study was performed according to the principles known from work sampling. From 22.11-12.12.1993 laboratory technicians, pathologists and secretaries were interviewed and an activity form was filled in. Six thousand nine hundred and fifty interviews were performed and the time study showed that for all groups of personnel 40% of the time was used on specimens sent for microscopy, 5% on autopsies (excl. microscopy), 15% on teaching and research, 20% on scientific meetings, reading and administration and 20% on other activities e.g. lunch, holiday, absence owing to illness. There was little variation for the different groups of personnel. On the other hand the time used on specific laboratory procedures, e.g. serial cutting, special-, and immunostaining, varied considerably for laboratory technicians, pathologists and secretaries respectively. Time studies, especially "work sampling" yield important information for the planning of laboratory services and provide documentation also for the time not directly related to routine work.

[A model for estimation of productivity and costs at a deparment of histopathology]. / Model til beregning af produktion og omkostninger på patologisk anatomisk afdeling.

Two cost calculation models were used. The "top down" model calculated an average cost of all investigations; this proved suitable for the calculation of the costs of autopsies and electron microscopical specimens. The "bottom up" model calculated the cost of an individual investigation, depending on the resources used in handling each particular specimen; it was necessary to adopt this model for specimens sent for microscopy. Information about the type of specimen and technical details were registered in a computer system. Production was registered in points and the costs were distributed between the clinical departments. The study showed that the cost of the histological specimens varied considerably depending on the material received from the clinical departments. A model using points for technical details in a department of cyto- and histopathology is suitable for calculating production and cost.

Computer-assisted diagnosis in gastroenterology.

During the last 20 years, Danish gastroenterologists and biostatisticians have been involved in practical and theoretical research aimed at providing computer-assisted decision support in clinical practice. On the practical side, we summarize two clinical applications, one in the acute abdomen, the other in jaundice. The former project is part of a worldwide effort which appears able to reduce considerably the perforated appendix rate and the negative laparotomy rate. In the latter project, also entwined in international activities, three methods of early statistical discrimination of jaundice causes have been tested, and it has been shown how ultrasonographic data can be incorporated into the diagnostic assessment. On the theoretical side, a Dutch-Danish group has been looking into how one best designs a trial and analyses its data in the context of measuring the quality and impact of machine-made diagnostic advice. Having pointed out the international ramifications of these activities, we outline what may lie ahead: Gastroenterologists must be prepared to join large-scale field trials of clinical advice-giving software. In the future, however, such software will often become embedded in general hospital recordkeeping systems.

Diagnostic value of cerebrospinal fluid cytology in comparison with tumor marker activity in central nervous system metastases secondary to breast cancer.

BACKGROUND: Central nervous system (CNS) metastases occur in approximately 35% of patients with breast cancer. Parenchymal brain metastases (MET) remain undetected in a large proportion of patients, and only 50% of patients with leptomeningeal carcinomatosis (MC) are diagnosed in vivo. METHODS: Cytology and activity of the tumor markers tissue polypeptide antigen (TPA) and creatine kinase-BB isoenzyme (CK-BB) were evaluated in the cerebrospinal fluid (CSF) in 71 consecutive patients with breast cancer suspected for CNS metastases. RESULTS: Forty-three patients had no CNS metastases, 12 patients had MET, 5 patients had both MET and MC, and 11 patients had MC alone. Seven of the patients with MC had an intracerebroventricular (ICV) reservoir inserted, and an additional 70 ICV CSF samples from these patients were obtained. In CSF obtained by lumbar puncture, 11% of the samples were classified as "suspicious for malignancy," but a very limited interobserver variability was demonstrated (Kappa test value, 0.81; 95% confidence limits, 0.67-0.95%). Fifty-one percent of the ICV CSF samples were classified as "suspicious for malignancy" (Kappa test value, 0.58; 95% confidence limits, 0.34-0.82%). TPA and CK-BB were both measured in 101 CSF specimens (61 from lumbar puncture and 40 ICV samples). The differences between patients with and without CNS metastases were significantly different according to TPA (P < 0.00001) and CK-BB (P < 0.00003; Mann-Whitney test). The sensitivity and predictive value of a negative test for having any CNS metastases (in case of elevated values of either TPA or CK-BB or both) were 85% (95% confidence limits, 65-96%) and 90% (95% confidence limits, 76-97%), respectively. In addition, a significant correlation between TPA and CK-BB was demonstrated in CSF from lumbar puncture (Spearmans-Rho, 0.49; P < 0.0001) and ICV (Spearmans-Rho, 0.37; P < 0.02). CONCLUSIONS: Cytologic evaluation of CSF obtained by lumbar punctures is a reliable procedure. In CSF from ICV reservoirs, cytologic evaluation is of limited use, but CK-BB and TPA is of potential value.