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Biomedical subjects

R Kühn

Publications and source records attributed to R Kühn.

At least 37 records · Page 2Linked to original sources
PubMed1998

Radiotherapy alone or radiochemotherapy with platin derivatives following transurethral resection of the bladder. Organ preservation and survival after treatment of bladder cancer.

PURPOSE: Multivariate analysis of prognostic factors influencing survival and bladder preservation after radiochemotherapy for bladder cancer following transurethral resection of the bladder (TURB). PATIENTS AND METHODS: At the University Hospital of Erlangen 333 patients with bladder cancer were treated with either radiotherapy alone (RT, n = 128) or platin based radiochemotherapy (RCT, n = 205) after TURB between 5/1982 and 5/1996. Two-hundred and eighty-two curative patients, with either muscle invasive or T1-high risk cancer, were analyzed. Median age was 66 years, median follow-up is 7.5 years. Uni- and multivariate analysis was performed for age, grade, R-status after initial TURB, T-category and treatment modality relevant to the endpoints initial response, survival and bladder preservation. RESULTS: Treatment related mortality was below 1%. Complete remissions were achieved at 57%, 70%, and 85% after RT or RCT with carboplatin or cisplatin. This difference was multivariately significant. Further significant prognostic factors were pT-category and R-status. For all patients survival was 59% and 43% after 5 and 10 years. 79% of survivors could keep their own bladder. Five-year survival rates after RT alone, RCT with carboplatin or cisplatin were 47%, 57%, and 69%, respectively. This was univariately significant. The only multivariately significant factor for survival and bladder preservation was the R-status after initial TURB. CONCLUSIONS: Treatment of bladder cancer by TURB and RT/RCT is an alternative to primary cystectomy. The addition of chemotherapy leads to significantly more complete remissions and better survival. Initial TURB is recommended to be as radical as possible.

Adult↗
PubMed1998

Effect of testosterone replacement after neonatal castration on craniofacial growth in rats.

Neonatal castration precludes the pubertal increase in serum testosterone and reduces general and craniofacial growth in the male Wistar rat. This study aimed to determine whether exogenous testosterone given at an age beyond the normal pubertal peak restores general and craniofacial growth in male, neonatally castrated rats. The design was a randomized, double-controlled, cross-sectional trial. Male Wistar rats were assigned by weighted randomization to be either castrated early after birth (n = 35) or not (n = 15). On day 57, a 1.5-cm Silastic tube with testosterone was implanted in 18 of the castrated rats. On day 70 and day 110, body length, weight, and craniofacial growth were measured together with the weight of the prostate, and blood samples taken. The exogenous testosterone resulted in a significant increase in serum testosterone and prostate weight. All measures of general and craniofacial growth had higher mean values in the non-castrated control group than in the castrated group, while in the testosterone-implant group the mean values lay between these of the castrated animals and the non-castrated controls. Two-way ANOVA indicated a significant effect of the testosterone administration on lower-incisor growth and the size of the total skull vault.

Analysis of Variance↗
PubMed1998

Rhythmic variation in beta1-adrenergic receptor mRNA levels in the rat pineal gland: circadian and developmental regulation.

In the rat pineal gland noradrenaline is released in large quantities from sympathetic nerve endings at the onset of darkness, thereby driving rhythmic melatonin synthesis with elevated levels at night-time. Upon release, noradrenaline interacts with postsynaptic beta1-adrenergic receptors to activate the cyclic AMP signalling pathway. Well characterized third messengers of this signalling cascade affect cyclic AMP-inducible genes that are crucially involved in initiation, maintenance and termination of hormone production. Among these third messengers are CREB (cyclic AMP responsive element binding protein) as an activating and ICER (inducible cyclic AMP early repressor) as an inhibitory transcription factor. Because a cyclic AMP-inducible promoter element is present on the beta1-adrenergic receptor gene, the expression of the receptor itself may be under control of the cyclic AMP-signalling pathway. By in situ hybridization, Northern blot analysis and RT-PCR we demonstrate a day/night rhythm in beta1-adrenergic receptor mRNA in the rat pineal gland with elevated levels during the dark period. As this rhythm persists, under constant darkness but is abolished upon removal of the sympathetic innervation, it is truly circadian. A marked day/night difference in the levels of beta1-adrenergic receptor mRNA becomes evident only after postnatal day 10, coinciding with the appearance of a functional cyclic AMP signalling pathway in the rat pineal gland. Furthermore, targeting ICER expression by transfection of pinealocytes with an antisense ICER construct, clearly indicates that the levels of the beta1-adrenergic receptor mRNA are regulated by the cyclic AMP-signalling pathway in a feedback mechanism.

Animals↗
PubMed1997

Analysis of granuloma formation in double cytokine-deficient mice reveals a central role for IL-10 in polarizing both T helper cell 1- and T helper cell 2-type cytokine responses in vivo.

In response to i.v.-injected eggs of Schistosoma mansoni, normal mice develop a dominant type 2 response, whereas IL-10-deficient animals generate a mixed type 1/type 2 cytokine profile and show reduced pulmonary granuloma formation. IL-4-deficient mice, while displaying diminished type 2 responses and granulomatous inflammation, also do not fully default to a type 1 cytokine profile. Strikingly, mice doubly deficient in IL-4 and IL-10 are completely defective in pulmonary granuloma formation and develop a highly polarized type 1 cytokine pattern. In analogous fashion, mice deficient in both IL-12 and IL-10 generate highly exacerbated type 2 cytokine responses, whereas in wild-type animals, IL-12 depletion minimally effects egg-induced cytokine production. Together, these results argue first that IL-10 is an important endogenous down-regulator of type 2 as well as type 1 cytokine synthesis, and second, that its induction is critical for type 2 response polarization in vivo.

Animals↗
PubMed1997

Generation of Cre recombinase-specific monoclonal antibodies, able to characterize the pattern of Cre expression in cre-transgenic mouse strains.

Transgene-encoded Cre recombinase can target alteration of loxP-tagged genes to specific cell types and developmental stages in mice, depending on the pattern of transgene expression. To facilitate determination of the latter, we have generated monoclonal anti-Cre antibodies which are specific for distinct epitopes on the recombinase and detect Cre both on immunoblots and intracellularly by immunofluorescence. We demonstrate the usefulness of these antibodies by an analysis of Cre expression in mice carrying a cre-transgene under B cell-specific control.

Animals↗
PubMed1997

In vivo ablation of surface immunoglobulin on mature B cells by inducible gene targeting results in rapid cell death.

Gene targeting experiments have demonstrated that the expression of immunoglobulin heavy chain in the pre-B cell receptor (pBCR) and of heavy and light chains in the B cell antigen receptor (BCR) marks checkpoints in early B cell development that the cells have to pass to survive. To investigate whether the persistence of mature B cells in the peripheral immune system also depends on BCR expression, we have generated a transgenic mouse in which the BCR can be inducibly ablated through V region gene deletion. Ablation leads to rapid death of mature B lymphocytes, which is preceded by down-regulation of MHC antigens and up-regulation of CD95 (Fas) and can be delayed by constitutive bcl-2 expression.

Animals↗
PubMed1997

[Advantages and limitations of intraoperative mechanical autotransfusion in al prostatectomies].

UNLABELLED: Intraoperative autotransfusion (MAT), preoperative autologous blood donation, and preoperative normovolaemic haemodilution are three different methods to avoid homologous blood transfusion during surgical procedures. The controversial use of MAT via cell saver in tumour surgery as well as contamination of the operative field with urine illustrate the particular difficulties of autologous blood transfusion in connection with radical prostatectomy. We investigated changes in the osmotic resistance of the retransfused red blood cells (RBC), bacterial contamination, changes in coagulation parameters, and the presence of tumour cells. PATIENTS AND METHODS: After written informed consent, 24 patients who presented for radical prostatectomy were randomly allocated to either a group that used MAT or a group that used homologous transfusion. The patients received "balanced anaesthesia" with midazolam, fentanyl, atracurium, and nitrous oxide/oxygen. The analysed parameters from the preoperative period to the 3rd postoperative day are shown in Table 1. The Haemonetics 3 Plus Cell Saver was used for autotransfusion. RESULTS: Our results showed that the haematologic parameters, coagulation factors, and serum chemistry did not differ between the two groups (Tables 2-4). However, there were significant differences during the investigated period. The osmotic resistance of the salvaged RBCs was higher than that preoperatively. Furthermore, there were no tumour cells in the autologous salvaged RBCs. CONCLUSION: Our results showed no decrease in the quality of the autotransfused RBCs, urine was not retransfused; and there were no significant differences between the groups postoperatively. Although there were no tumour cells in the salvaged blood, the possibility of blood irradiation is discussed. We concluded that because of the risk of infection of homologous blood products, MAT is a safe possibility to reduce the amount of homologous blood transfusion required in connection with radical prostatectomy.

Aged↗
PubMed1997

A comparative histopathological study of the influence of chemo- and radiotherapy on human prostatic carcinoma grown in nude mice.

Histological investigations were carried out to determine the effects of various kinds of treatment on the heterotransplanted prostatic carcinoma PC EW in athymic nude mice. Morphological changes after chemotherapy, radiotherapy, and combined radiochemotherapy were analyzed qualitatively and quantitatively. Effects of chemo- and radiotherapy like reduction of mitotic activity and increase of necrotic resp. fibrotic areas cumulated under the combined therapy. Results after radiotherapy or combined therapy corresponded to those reported for human prostatic carcinoma. Our observations indicate that the heterotransplanted tumor line PC EW represents a well suited preclinical test system for treatment of human prostatic carcinoma.

Adenocarcinoma↗
PubMed1997

Advances in gene targeting methods.

Gene targeting in embryonic stem cells is commonly used for gene inactivation and the generation of mouse mutants. The combined use of methods for site-specific and homologous DNA recombination expands the potential of gene targeting in embryonic stem cells considerably and offers the opportunity of conditional gene targeting in mice.

Animals↗
PubMed1996

Interleukin (IL)-4-independent immunoglobulin class switch to immunoglobulin (Ig)E in the mouse.

Immunoglobulin (Ig) class switching in B cells is regulated by stimuli transduced by cytokines and cell-cell contact. Among these stimuli, interleukin (IL)-4 has been considered an absolute prerequisite for class switching to IgE in the mouse. Here we report that IL-4-deficient (IL-4-/-) and wildtype mice had comparably elevated serum IgE levels during the course of a murine retrovirus-induced immunodeficiency syndrome, MAIDS. IgE switching in IL-4-/- mice was also induced by injection of anti-IgD antibody. Treatment with anti-IgD induced germline epsilon (g epsilon) transcripts with comparable efficiency in IL-4-/- mice and controls, but the levels of productive epsilon transcripts (p epsilon) were lower by a factor of 200 and serum IgE levels were lower by a factor of 300 in IL-4-/- mice as compared with controls. Induction of g epsilon after anti-IgD treatment of IL-4-/- mice was unaffected by simultaneous treatment with monoclonal antibodies to IL-4 and IL-4 receptor alpha chain. Infection of IL-4-/- mice with Nippostrongylus brasiliensis, a potent stimulus for IgE production, resulted in induction of g epsilon transcripts; however, p epsilon transcripts were barely detectable and serum IgE was not detected. These findings establish a novel IL-4-independent pathway for IgE switching in the mouse that is strongly activated in retroviral infection but weakly in nematode infection. This pathway appears to be dependent on distinct factors that separately control induction of g epsilon transcription and switch recombination to p epsilon.

Animals↗
PubMed1996

Light-induced expression of transcription factor ICER (inducible cAMP early repressor) in rat suprachiasmatic nucleus is phase-restricted.

The mammalian hypothalamic suprachiasmatic nucleus (SCN) harbors an intrinsic circadian oscillator driving a variety of endogenous rhythms. SCN activity is entrained to environmental lighting conditions by photic information from the retina. Light-induced phase shifts involve cAMP and Ca2+ as second messengers and are linked to transcriptional and translational processes. Using in situ hybridization with a cDNA fragment of the cAMP responsive element modulator (CREM) isoform inducible cAMP early repressor (ICER) we demonstrate a light-induced upregulation of CREM mRNA in rat SCN during the second half of the night. Notably, ICER is the only member of the CREM family which is transcriptionally inducible. We suggest that a stimulus-induced upregulation of ICER expression can inhibit cAMP-inducible genes in rat SCN via its trans-repressing potency.

Animals↗
PubMed1996

Enterocolitis and colon cancer in interleukin-10-deficient mice are associated with aberrant cytokine production and CD4(+) TH1-like responses.

We have characterized the progressive stages of chronic intestinal inflammation that develops spontaneously in specific pathogen-free (SPF) mice with a targeted disruption in the IL-10 gene (IL-10-/-). Our longitudinal studies showed that inflammatory changes first appear in the cecum, ascending and transverse colon of 3-wk-old mutants. As the disease progressed, lesions appeared in the remainder of the colon and in the rectum. Some aged IL-10-/- mice also developed inflammation in the small intestine. Prolonged disease with transmural lesions and a high incidence of colorectal adenocarcinomas (60%) was observed in 6-mo-old mutants. Mechanistic studies have associated uncontrolled cytokine production by activated macrophages and CD4+ Th1-like T cells with the enterocolitis exhibited by IL-10-/- mice. A major role for a pathogenic Th1 response was further suggested by showing that anti-IFNgamma antibody (Ab) treatment significantly attenuated intestinal inflammation in young IL-10-/- mice. When weanlings were treated with IL-10, they failed to develop any signs of intestinal inflammation. Interestingly, IL-10 treatment of adults was not curative but did ameliorate disease progression. Our studies have also shown that inheritable factors strongly influence the disease susceptibility of IL-10-/- mice. In 3-mo-old mutants, intestinal lesions were most severe in IL-10-/- 129/SvEv and IL-10-/- BALB/c strains, of intermediate severity in the IL-10-/- 129 x C57BL/6J outbreds, and least severe in the IL-10-/- C57BL/6J strain.

Aging↗
PubMed1996

In the absence of endogenous IL-10, mice acutely infected with Toxoplasma gondii succumb to a lethal immune response dependent on CD4+ T cells and accompanied by overproduction of IL-12, IFN-gamma and TNF-alpha.

To examine the function of IL-10 synthesis during early infection with the intracellular protozoan Toxoplasma gondii, IL-10 knockout (KO) mice were inoculated with an avirulent parasite strain (ME-49). In contrast to control littermates that displayed 100% survival, the IL-10-deficient animals succumbed within the first 2 wk of the infection, with no evidence of enhanced parasite proliferation. The mortality in the IL-10 KO mice was associated with enhanced liver pathology characterized by increased cellular infiltration and intense necrosis. Levels of IL-12 and IFN-gamma in sera of infected IL-10-deficient animals were four- to sixfold higher than those in sera from control mice, as were mRNA levels for IFN-gamma, IL-1 beta, TNF-alpha, and IL-12 in lung tissue. Similarly, macrophages from IL-10 KO mice activated in vitro or in vivo with T. gondii produced higher levels of TNF-alpha and IL-12 than macrophages from control animals. Moreover, spleen cells from IL-10 KO mice infected with T. gondii secreted more IFN-gamma than splenocytes from nondeficient animals. In vitro depletion experiments indicated that CD4+ lymphocytes are the major source of the latter cytokine in the spleen cell populations, and in vivo depletion with anti-CD4 Abs protected the IL-10 KO mice from parasite-induced mortality. Together the data suggest that endogenous IL-10 synthesis plays an important role in vivo in down-regulating monokine and IFN-gamma responses to acute intracellular infection, thereby preventing host immunopathology.

Acute Disease↗
PubMed1996

T helper cell 1-type CD4+ T cells, but not B cells, mediate colitis in interleukin 10-deficient mice.

Mice rendered deficient in the production of interleukin 10 (IL-10-/-) develop a chronic inflammatory bowel disease (IBD) that predominates in the colon and shares histopathological features with human IBD. Our aim was to identify which cell type(s) can mediate colitis in IL-10-/- mice. We detected an influx of immunoglobulin-positive cells into the colon and the presence of colon-reactive antibodies in the serum of IL-10-/- mice. To assess a pathogenic role for B cells, we generated a B cell-deficient (B-/-) strain of IL-10-/- mice. B-/-IL-10-/- mice acquired a severe colitis analogous to that IL-10-/- mice, implying that B cells were not the primary mediator of IBD in this model. A series of cell transfer experiments was performed to assess a pathogenic role for T cells. When IL-10-/- T cell-enriched lamina propria lymphocytes (LPL) or intraepithelial lymphocytes (IEL) were transferred into immunodeficient recombinase-activating gene (RAG)-2-/- recipients, a mild to severe colitis developed, depending on the cell number transferred. Lymphocytes recovered from the colon of transplanted RAG-2-/- mice with colitis were predominantly alpha beta TCR+CD4+, including a large proportion of CD4+CD8 alpha + cells. These cells were also CD45RB-/low and CD44+, indicative of an activated/memory population. Individual populations of CD4+CD8 alpha-, CD4+CD8 alpha + and CD4-CD8 alpha + T cells were then isolated from the lamina propria compartment of IL-10-/- mice and transferred into RAG-2-/- recipients. Only IL-10-/- CD4-expressing LPL, including both the CD4+CD8 alpha- and CD4+CD8 alpha + populations, induced colitis in recipient mice. Interferon-gamma, but little to no IL-4, was produced by CD4+CD8 alpha- and CD4+CD8 alpha + LPL recovered from the inflamed colons of RAG-2-/- recipients implicating alpha T helper cell 1 (TH1)-mediated response. We thus conclude that colitis in IL-10-/- mice is predominantly mediated by TH1-type alpha beta TCR+ T cells expressing CD4 alone, or in combination with the CD8 alpha molecule.

Animals↗
PubMed1996

Requirement of mammalian DNA polymerase-beta in base-excision repair.

Synthesis of DNA by DNA polymerase-beta is distributive on single-stranded DNA templates, but short DNA gaps with a 5' PO4 in the gap are filled processively to completion. In vitro studies have suggested a role of beta-polymerase in different types of DNA repair. However, the significance of these studies to the in vivo role of beta-polymerase has remained unclear. Because genetic studies are essential for determining the physiological role of a gene, we established embryonic fibroblast cell lines homozygous for a deletion mutation in the gene encoding DNA polymerase-beta. Extracts from these cell lines were found to be defective in uracil-initiated base-excision repair. The beta-polymerase-deleted cells are normal in viability and growth characteristics, although they exhibit increased sensitivity to monofunctional DNA-alkylating agents, but not to other DNA-damaging agents. Both the deficiency in base-excision repair and hypersensitivity to DNA-alkylating agents are rescued following stable transfection with a wild-type beta-polymerase minitransgene. These studies demonstrate that beta-polymerase functions specifically in base-excision repair in vivo.

Animals↗
PubMed1996

Prognostic value of ploidy and proliferation markers in renal cell carcinoma.

BACKGROUND: The prognosis for patients with renal cell carcinoma depends mainly on pathological stage and grade of the tumor at the time of surgery. Cellular proliferation may prove to be another measure for predicting biologic aggressiveness and, therefore, the prognosis. METHODS: The authors compared four different methods to assess proliferation in a series of 87 curatively resected (R0) renal cell carcinomas: flow cytometry analysis (FCM), silver-stained nucleolar organizer regions (AgNOR), and immunohistochemical assessment of the MIB-1 (Ki-67) antigen, and proliferating cell nuclear antigen (PCNA). The results obtained were compared with pathologic stage (according to the International Union Against Cancer [UICC]) and grade with disease-related survival rate; finally, we assessed whether the methods led to similar results. RESULTS: In each carcinoma examined, we could demonstrate MIB-1, PCNA, and AgNOR dots in varied proportions. Statistical correlations were seen between the tumor grade, the rate of nuclear positivity for MIB-1 and PCNA, and the number of AgNOR dots. Additionally, the MIB-1 index was significantly higher in more advanced tumor stages. A good correlation between MIB-1 and AgNOR as well as for PCNA was found. In univariate survival analysis, tumor stage and grade, MIB-1 and PCNA index, and mean AgNOR number were related significantly to patient survival. On multivariate Cox disease-related survival analysis, stage of disease and MIB-1 were significant independent prognostic factors. Flow cytometry was not related to prognosis nor to other examined parameters. CONCLUSIONS: These results indicated that MIB-1 immunostaining is an additional prognostic parameter for patient outcome. MIB-1 and PCNA immunostaining, as well as AgNOR, demonstrated good correlations among themselves. We failed to establish flow cytometry as a method to predict proliferative capacity or prognosis in renal cell carcinoma patients.

Antigens, Nuclear↗