25 years International Bladder Cancer Network (IBCN): The past, the present, and the future.

In 1997 an international group of scientists organized a meeting in Barcelona, Spain, to discuss the use of biomarkers in the management of patients with bladder cancer. This meeting was the offspring of an - initially informal - group that finally resulted in the foundation and incorporation of the International Bladder Cancer Network (IBCN) e.V. in 2005. Over the years the group has supported several research initiatives and generated several recommendations on the use of biomarkers in the diagnosis and treatment of bladder cancer. Meeting quality was generated by inviting experts presenting state-of-the-art lectures or work in progress reports, interdisciplinarity and the limited number of participants supporting an open and personal exchange resulted in a format increasingly attracting participants from all over the world. The recent limitations caused by the Covid-19 pandemic were partially met by organizing several well attended webinars. The future challenge is to maintain the IBCN meeting spirit despite an increasing interest of the scientific community and industrial partners to participate. However, the integration of and interaction between increasingly more specialized disciplines is a challenge that can be better catalyzed by an international multidisciplinary network than mostly national professional associations.

Vapor Phase Exchange of Self-Assembled Monolayers for Engineering of Biofunctional Surfaces.

We show that 4'-nitro-1,1'-biphenyl-4-thiol self-assembled monolayers (NBPT SAMs) on gold can be exchanged with 11-(mercaptoundecyl)triethylene glycol (C11EG3OH) SAMs via vapor deposition (VD). The pristine and the exchanged SAMs obtained by VD as well as solution method (SM) were characterized by X-ray photoelectron spectroscopy (XPS) and polarization modulation infrared reflection-absorption spectroscopy (PM-IRRAS). Using surface plasmon resonance (SPR), it is shown that C11EG3OH SAMs on gold obtained by vapor deposition exchange (VDEx) have the same protein resistivity as SAMs obtained by the direct self-assembly process. As expected, the cross-linked NBPT SAM are found to be resistive to both exchange processes, VDEx and solution method exchange (SMEx). In this way, VDEx opens up an elegant and new approach of patterning SAM surfaces in situ at vacuum conditions without using any solvents. By combining electron irradiation-induced chemical lithography of NBPT SAMs with VDEx, biofunctional patterned substrates were engineered and used for immobilization of protein arrays.

Dermatopathology in sub-Saharan Africa: a systematic 5-year analysis of all histopathological diagnoses from the Regional Dermatology Training Centre (RDTC) in Moshi, Tanzania.

BACKGROUND: Proper diagnosis of skin diseases relies on dermatopathology, the most important diagnostic technique in dermatology. Unfortunately, there are few dermatopathology institutions in sub-Saharan Africa, where little is known about the spectrum of histopathological features observed. OBJECTIVES: To investigate the spectrum of dermatopathological diagnoses made in a sub-Saharan African reference centre of a large, mainly rural area. PATIENTS/METHODS: To retrospectively evaluate all dermatopathological diagnoses made over a period of 5 years at the Regional Dermatology Training Centre (RDTC) in Moshi, Tanzania. RESULTS: There were a total of 1554 skin biopsy specimens. In 45% of cases, there were inflammatory diseases, most frequently lichenoid conditions. Cutaneous neoplasms represented 30.4% of all diagnoses, with Kaposi's sarcoma (KS) and, less frequently, squamous cell carcinoma (SCC) being the two most common neoplastic conditions. The latter also reflected the intensive management of persons with albinism in the RDTC. The distribution of histological diagnoses seemed to correlate with the overall clinical spectrum of cutaneous diseases managed in the RDTC. CONCLUSIONS: In this African study inflammatory conditions are the main burden of skin diseases leading to a diagnostic biopsy. Our findings provide further evidence that KS, primarily related to the high prevalence of HIV infection is an epidemiological problem. Both SCC and basal cell carcinoma represent another relatively common malignant cutaneous neoplasms, reflecting the presence of specific populations at risk. The challenging spectrum of histological diagnoses observed in this specific African setting with basic working conditions shows that development of laboratory services of good standards and specific training in dermatopathology are urgently needed.

Project RATEMA--one year's experience.

The RATEMA project is an international cooperation between the University of Ulm in Germany and the Urals Research Centre for Radiation Medicine in Chelyabinsk, Russia. For one year we conducted weekly conferences between the two sites, based on a satellite link with a 384 kbit/s connection. During the videoconferences the physicians on both sides--experts in radiation medicine--discussed the health status of Russian patients who had been chronically exposed to ionizing radiation in the South Urals region. The German partners presented patients with comparable haematological and oncological diseases. The project has shown the advantages and difficulties of working in an international and interdisciplinary environment. The experience gained has been very valuable for planning new projects with similar tasks. The results and the contents of the RATEMA database are the basis for education and research for physicians involved in the management of radiation victims.

[The internet--its uses in traumatology]. / Das Internet--Nutzen für die Traumatologie.

No other words have obtained such meaning in the computer world as the catchwords "Online" and "Internet". The possibilities are innumerable, even in the medical field, and therefore we must consider whether trauma and orthopedic surgeons can also benefit from this new telecommunications medium. As a result of this inquiry, a pilot program has been initiated through an Internet connection at the computer center in the University of Ulm to test and define the possibilities of the Internet in routine clinical applications and for scientific projects. For the surgeon, there is already an abundance of information available: (1) A medical search register consisting of more than 20 million words has been comprised. the source of such searches could be clinics, individuals publications, or even an e-mail answer from a patient's "Help" inquiry. (2) The literature can be researched without difficulty on the data highway. (3) It has become possible to communicate transfer figures, texts, sound and video data within seconds to any connected physician throughout the world over the e-mail system. (4) Included in the many scientific news-letters presented in the Internet ae also several specified for the surgical field. (5) It has also become apparent that a few of the rare German-speaking institutions available on the Internet (for example, the AO Institute in Davos, some university departments and a few research institutes) offer very detailed "Home Pages". In comparison to the American institutions, however, the majority of the German surgical world is not yet connected to the Internet. The Internet offers an enormous information service that is always available and is accessible worldwide. This information can already be used in clinical and scientific applications.

Upregulation of gap junction protein connexin43 in alveolar epithelial cells of rats with radiation-induced pulmonary fibrosis.

The degree of immunoreactive connexin43 (C x 43) in rat lung was evaluated during the development of radiation-induced pulmonary fibrosis in rat by a double immunofluorescence technique using polyclonal antisera to Cx43 and monoclonal antibodies to cytokeratins on cryostat sections. In normal rat lungs, Cx43 was detected in pneumocytes type II and I, in large blood vessel endothelia, in peribronchial smooth muscle cells, and in some peribronchial and perivascular interstitial cells. As early as 1 week after irradiation, enhanced immunoreactivity for Cx43 in the epithelial cells was detected. In severely injured lungs (about 3 months after irradiation), Cx43 was found also in the cytoplasm of type II pneumocytes. These findings were confirmed by western blot data. Western blot analysis also revealed increased phosphorylation of Cx43. It remains to be investigated whether the increased content of Cx43 in irradiated rat lung may be due to an enhanced number of gap junctions between type I and II alveolar epithelial cells.

Tissue factor expression during human and mouse development.

In the adult organism the cellular distribution of tissue factor (TF) expression corresponds to biological boundary layers forming a hemostatic barrier ready to activate blood coagulation after tissue injury. Whether TF expression might also play a role in development is unknown. To determine the significance of TF in ontogenesis, we examined the pattern of TF expression in mouse development and compared it with the distribution of TF in human post-implantation embryos and fetuses of corresponding gestational age. At early embryonic periods of murine (6.5 and 7.5 pc) and human (stage 5) development, there was strong expression of TF in both ectodermal and entodermal cells. In situ hybridization and immunohistochemistry demonstrated that TF mRNA and protein were expressed widely in epithelial areas with high levels of morphogenic activity during organogenesis. Staining for TF was seen during ontogenetic development in tissues such as epidermis, myocardium, bronchial epithelium, and hepatocytes, which express TF in the adult organism. Surprisingly, during renal development and in adults, expression of TF differed between humans and mice. In humans, maturing stage glomeruli were stained for TF whereas in mice, TF was absent from glomeruli but was present in the epithelia of tubular segments. In neuroepithelial cells, there was a substantial expression of TF. Moreover, there was robust TF expression in tissues such as skeletal muscle and pancreas, which do not express it in the adult. In contrast, expression of the physiological ligand for TF, factor VII, was not detectable during early stages of human embryogenesis using immunohistochemistry. The temporal and spatial pattern of TF expression during murine and human development supports the contention that TF serves as an important morphogenic factor during embryogenesis.

Detection of circulating tissue factor and factor VII in a normal population.

The plasma tissue factor (TF) concentration was correlated to factor VII concentration (FVIIag) and factor VII activity (FVIIc) in 498 healthy volunteers ranging in age from 17 to 64 years. Immunoassays using monoclonal antibodies (mAbs) were developed for the determination of TF and FVIIag in plasma. The mAbs and the test systems were characterized. The mean value of the TF concentration was 172 +/- 135 pg/ml. TF showed no age- and gender-related differences. For the total population, FVIIc, determined by a clotting test, was 110 +/- 15% and the factor VIIag was 0.77 +/- 0.19 microgram/ml. FVII activity was significantly increased with age, whereas the concentration demonstrated no correlation to age in this population. FVII concentration is highly correlated with the activity as measured by clotting assay using rabbit thromboplastin. The ratio between FVIIc and FVIIag was not age-dependent, but demonstrated a significant difference between men and women. Between TF and FVII we could not detect a correlation.

Immunohistochemistry of new type I alveolar epithelial cell markers of the rat.

The presence of pan-cadherin and the MEP-I antigen in normal and diseased rat lung was established by employing immunoperoxidase and double label fluorescence techniques. The binding of a mouse monoclonal antibody (MEP-1) reacting specifically with type I pneumocytes was assessed on paraffin sections of normal specimens as well as those with pulmonary fibrosis induced by bleomycin or radiation treatment. In injured alveolar epithelium, a diminished type I cell and a focal type II cell immunoreactivity was found. Electron microscopy of immunogold-labelled lung tissue confirmed the type I cell specificity of MEP-1. In severely injured pulmonary parenchyma MEP-1-negative areas occurred, which were also negative with the type II pneumocyte marker Maclura pomifera lectin. Similarly, a polyclonal pan-cadherin antibody uniformly decorated luminal surfaces of alveoli except the type II pneumocytes. Furthermore, pleural mesothelial cells, bronchiolar epithelial cells, endothelial cells of large blood vessels and alveolar macrophage surfaces exhibited pan-cadherin immunoreactivity. After injury, a remarkable loss of pan-cadherin immunoreactivity in the MEP-1-positive type I epithelial cells was detectable. These findings suggest that characterization of normal alveolar epithelial cells and monitoring of the epithelial remodelling in pulmonary pathohistology are sufficiently described by the antibodies MEP-1 anti pancadherin.

Immunocytochemical distribution of E-cadherin in normal and injured lung tissue of the rat.

Affinity purified rabbit anti-mouse E-cadherin antibodies, reacting with diverse rat epithelia, were used to characterize epithelial changes in a radiation-induced fibrosis model of rat lung by immunoblotting techniques, immunoperoxidase and immunofluorescence microscopy. Immunostaining of normal rat lung tissues revealed a predominant staining of type II pneumocytes. Immunoelectron microscopy confirmed the immunohistochemical data of normal lung tissue obtained at the light microscopic level. In severely injured rat lung, we found enhanced immunoreactivity for E-cadherin at the surface of type I alveolar epithelial cells. The results suggest that E-cadherin is an adhesion molecule that is modulated after pathological alteration of the alveolar epithelium and that the antiserum may be useful for the characterization of normal and diseased rat epithelia.

Monoclonal antibodies to surfactant protein D: evaluation of immunoreactivity in normal rat lung and in a radiation-induced fibrosis model.

This report describes the development of a new panel of monoclonal antibodies established after immunization of mice with purified surfactant protein D of the rat. To enhance the detection of SP-D in formalin- or Schaffer-fixed samples, immunohistochemistry was performed by using microwave pretreatment of paraffin sections. Using these new antibodies that bind to type II epithelial cells, Clara cells, and alveolar macrophages, the responses of lung parenchymal cells were examined in a radiation-induced fibrosis model. Increased accumulation of extracellular SP-D in the alveolar space was found. Double staining with anti-surfactant protein A antibodies revealed different Clara cell populations containing one or both types of surfactant proteins.

Disseminated histoplasmosis in chronic mucocutaneous candidiasis.

Chronic mucocutaneous candidiasis (CMC) is rarely associated with disseminated Candida, but is often associated with systemic infection secondary to other organisms [1]. A 6-year-old with CMC and disseminated histoplasmosis is presented here.

An ELISA for tissue factor using monoclonal antibodies.

Whereas tissue factor, a high-affinity cell-surface receptor and essential cofactor for the serine protease factor VII is constitutively present in certain tissues such as epithelial tissue, brain and placenta, it is not normally expressed by cells within the vasculature. However, the stimulation of monocytes and endothelial cells by a variety of inflammatory and immunological reactions results in the induction of cell surface tissue factor (TF) expression. TF is also expressed on tumour cells, and may play a role in tumour growth and metastasis formation. To examine the role of TF in these processes we developed monoclonal antibodies to human tissue factor apoprotein. The antibodies were characterized by neutralization of the procoagulant activity and by immunoblotting. With two of these monoclonal antibodies a sandwich ELISA was developed for the rapid quantitation of TF. The sensitivity of the assay permits extensive studies involving the modulation of TF expression on small numbers of cells. The results are comparable to the functional clotting assay as evaluated with unpurified TF and with the tumour cell line MCF-7. For certain applications, monitoring of cellular TF expression by ELISA using anti-TF monoclonal antibodies is preferable because it is not influenced by other coagulation factors or by inhibitors of procoagulant activity on the cells.

Immunohistological detection of tissue factor in normal and abnormal human mammary glands using monoclonal antibodies.

Tissue factor (TF) is the primary cell-bound initiator of the coagulation protease cascade. The cytological distribution of TF in various tissues may be described on the basis of immunohistochemistry with epitope-defined monoclonal antibodies and the extravascular distribution of TF apparently represents a haemostatic envelope ready to activate coagulation when vascular integrity is disrupted. The present study localized TF in human breast cancer tissues when compared with normal breast gland tissues and benign disorders of the mammary gland. By use of a cocktail of three epitope-defined monoclonal antibodies, TF was detected only in the myoepithelia of the resting breast gland. In proliferating disorders like fibrocystic disease or in fibroadenomas, both myoepithelia and luminal epithelia showed TF expression. Of 115 breast cancers 93 reacted with anti-TF, in an inhomogeneous manner in terms of intensity and number of positive cells. There was a tendency for more positive and intensely stained cells to be found in well-differentiated structures such as tubules. Invasive ductal carcinomas exhibiting more positive and more strongly stained cells were less commonly metastatic to lymph nodes when compared with the tumours with no detectable or very low TF immunostaining. A semi-quantitatively recorded score of TF immunostaining correlated with the procoagulatory activity measured (7 fibroadenomas and 24 carcinomas). The results of this study suggest that proliferation and differentiation of the mammary gland is associated with enhanced TF expression in the epithelia which are negative for TF staining in the resting gland. Malignant growth is characterized by randomly expressed epithelial TF, which expression is enhanced and more frequent in well-differentiated tumour cells.