[Geometry and function of the dog nose: how does function change when form of the nose is changed?]. / Geometrie und Funktion der Hundenase: Wie ändert sich die Funktion, wenn die Form verändert wird?

Nasal airflow resistance in brachycephalic dogs is significantly elevated compared to normal dogs. LaserAssisted TurbinEctomy (LATE)-surgery as well as xylometazolin were shown to reduce pathologically increased intranasal airway resistance in brachycephalic dogs by approximately 50 %. Impulse oscillometry provides a reliable and sensitive method to examine intranasal stenoses in the canine nose. Acoustic rhinometry allows assessment of changes in cross sectional area and volume of the canine nasal cavity.

[Brachycephaly in dog and cat: a "human induced" obstruction of the upper airways]. / Brachyzephalie bei Hund und Katze: eine "menschengemachte" Obstruktion der oberen Atemwege.

Selective breeding for exaggerated features caused in many brachycephalic dog and cat breeds virtually a loss of the nose, with serious anatomical and functional consequences. In addition to respiratory and olfactory tasks, in dogs the nose is of vital importance for thermoregulation. As obligatory nose breathers, dogs suffer far more than humans when their nasal ventilation is restricted. An open discussion in the broad public has to motivate authorities and kennel clubs to recognize extreme brachycephalic breeding as seriously affecting animal health and welfare.

Sequential chemotherapy with carboplatin followed by weekly paclitaxel in advanced ovarian cancer: results of a multicenter phase II study of the northeastern German society of gynecological oncology.

BACKGROUND: For the adjuvant setting of advanced ovarian cancer (AOC) after primary radical surgery the combination of paclitaxel and platinum in a 3-week schedule has emerged as the current standard. In preclinical studies additional anti-angiogenic effects of low dose paclitaxel infusion were demonstrated. A sequential schedule of carboplatin and paclitaxel has the potential to improve the therapeutic index. METHODS: In this multicenter phase II trial four cycles of carboplatin at a dose of AUC 5 (d1/q21d) followed by 12 cycles of weekly paclitaxel at a dose of 80 mg/m(2) (d1/q7d) were applied after primary radical surgery. Eligible were all optimally or sub-optimally debulked patients with FIGO IA-IV ovarian cancer. All patients with hemoglobin levels <12 mg/dl received erythropoietin additionally. RESULTS: Between July 2003 and May 2005, 105 patients from 27 institutions were enrolled. The median age was 60 years (range: 23-80 years). A median number of 16 courses (range 1-16) were applied. The incidence of non-hematological toxicities was very low. Only 41% of patients experienced alopecia (grade 1-2). Neurotoxicity (grade 3-4) was not observed. Grade 3-4 hematological toxicity (43% of all patients) included thrombocytopenia (17%), anemia (3%), leucopenia (23%), and neutropenic fever (0%). Ninety-seven percent received erythropoietin. Thromboembolic events (4%) were not increased in patients who received erythropoietin. After a median time of 23 months (range: 1-42 months) 32 patients had died, and the median overall survival was not reached. The progression-free survival was 25.4 months (95% CI: 18.8-40+). CONCLUSION: These results suggest that this sequential regimen using weekly paclitaxel represents an efficacious and well-tolerated regimen. A randomized study comparing this new schedule with the conventional 3-week protocol is warranted.

Pegylated liposomal doxorubicin (CAELYX) in patients with advanced ovarian cancer: results of a German multicenter observational study.

PURPOSE: Pegylated liposomal doxorubicin (PLD, CAELYX) has demonstrated activity in several phase-III trials and has been approved for the therapy of relapsed ovarian cancer after platinum treatment. Aim of this observational study was to analyze the efficacy and toxicity profile of PLD under routine clinical conditions and without the general restrictions of defined inclusion and exclusion criteria of clinical trials. METHODS: Between 2003 and 2005, a total of 190 patients with relapsed ovarian cancer were enrolled. 183 patients were available for evaluation; dose-intensity, modifications, treatment duration, toxicities and response were systematically analyzed. RESULTS: The median patient age was 62 years (range 23-86 years). 45.4% of the patients received PLD as second-line therapy and a median of four courses per patient were administered. The median dose of PLD was 40 mg/m(2), most frequently used every 4 weeks (68.8%). Grade 3 Leucopenia (1.6%) and grade 3 and 4 thrombocytopenia (0.5%) were the most frequent hematological toxicities. The most frequent non-hematological toxicities were skin toxicity, pain and nausea, which were observed in 38.8, 41 and 45.9% of the patients, respectively. Twenty-seven percent of the patients showed a response to therapy with 6.9% achieving complete remission and 20.1% achieving partial remission. 37.7% achieved a stable disease. The median duration of response for all patients was 4.8 months (range 0-51.8 months). Median progression-free interval and overall survival were 5.8 months (95% CI 5.1-6.6 months) and 16.6 months (95% CI 13.9-22.6 months), respectively. CONCLUSIONS: PLD is safe and effective in patients with relapsed ovarian cancer, even after numerous previous treatment regimens. A dose of 40 mg/m(2) every 28 days seems to be an effective and well-tolerated therapeutic option in advanced ovarian cancer with a low incidence of hematological toxicities and acceptable non-hematological toxicities.

[Health-related quality of life in children with spina bifida]. / Gesundheitsbezogene Lebensqualität bei Kindern mit Spina bifida.

BACKGROUND: There are no studies on the health-related quality of life (HRQoL) in German children with a myelomeningocele (MMC). This study aims to obtain generalizable epidemiological data on the HRQoL of such children. PATIENTS AND METHODS: KINDL-R questionnaires were filled out and clinical findings on typical MMC disabilities were also documented. Of the 115 families contacted, 70 MMC families responded (response rate 61%). Normative KINDL-R data from a sample of healthy children served as reference. RESULTS: No differences in clinical data were found when comparing responders and non-responders. KINDL total scores as well as scores across all scales were highly concordant for parental reporting and self-reporting. CONCLUSIONS: Despite the fact that children with MMC often suffer from severe physical limitations, their HRQoL is not necessarily lower than that of healthy children.

"Very delayed" graft function in a patient after living related kidney transplantation: a case report.

We report the case of a patient who experienced anuric renal transplant failure for 44 days after living related kidney transplantation. Immunosuppressive and other therapies were carefully adapted to the findings of frequent renal transplant biopsies, which ultimately led to excellent graft function.

Genomic organization, sequence analysis and transcriptional regulation of the human MCP-4 chemokine gene (SCYA13) in dermal fibroblasts: a comparison to other eosinophilic beta-chemokines.

The eosinophil chemotactic beta-chemokine MCP-4 is assumed to be involved in the accumulation of eosinophils characteristic for eosinophilic inflammatory diseases. We here describe the genomic organisation (3 exons of 138, 115 and 578 bp, 2 introns of 867 and 437 bp and 1.4 kb of regulatory sequences from the immediate 5' upstream region), sequence (genomic and transcribed) and mRNA expression of the human MCP-4 gene in dermal fibroblasts. Among the promoter elements potentially regulating MCP-4 gene expression and/or mediating the effects of anti-inflammatory drugs we identified consensus sequences known to interact with nuclear factors like NF-IL6, AP-2, a NF-kappaB like consensus sequence, gamma-interferon- response and YY-1 elements as well as glucocorticoid response elements. Like MCP-3, MCP-4 mRNA expression in dermal fibroblasts is upregulated by TNF-alpha, IL-1alpha, IFN-gamma or IL-4 and differs from RANTES and eotaxin mRNA expression in its response to IFN-gamma and/or IL-4.

Ventricular tachycardia during follow-up in patients resuscitated from ventricular fibrillation: experience from stored electrograms of implantable cardioverter-defibrillators.

OBJECTIVE: The purpose of this study was to use the electrogram storage capabilities of the implantable cardioverter-defibrillator (ICD) to categorize any arrhythmic event during follow-up in a group of patients who had survived an episode of ventricular fibrillation (VF) and to possibly identify clinical predictors of future arrhythmic events. BACKGROUND: Little is known about the electrophysiologic characteristics of ventricular arrhythmias recurring during follow-up in survivors of VF as the sole documented arrhythmia at the time of resuscitation. METHODS: Forty patients (58+/-10 years; 73% men; left ventricular ejection fraction 42+/-18%; 70% with coronary artery disease) who had survived an episode of VF and subsequently received an ICD capable of intracardiac electrogram recording and storage were followed for 23+/-11 months. In all patients, the arrhythmogenic substrate was investigated by means of programmed electrical stimulation (PES). RESULTS: Among the 40 patients, 41 episodes of ventricular arrhythmias were documented in 13 patients (33%): 36 episodes of ventricular tachycardias (VT) were recorded in 11 patients (28%) and 5 episodes of VF were recorded in the remaining 2 patients (5%). Age, gender, cardiac disease and left ventricular ejection fraction failed to distinguish between patients with clinical recurrences and patients without. The sensitivity, specificity and positive accuracy of PES were 29%, 63% and 46%, respectively, for prediction of ventricular arrhythmia recurrence; 45%, 70% and 36%, respectively, for prediction of VT; and 50%, 98% and 50%, respectively, for prediction of VF during follow-up. CONCLUSIONS: In survivors of VF receiving ICD therapy, VT is the most common ventricular arrhythmia recorded on device-incorporated electrograms during follow-up. This finding, associated with the relatively well-preserved ventricular function, may account for the ability of these patients to survive at time of the index arrhythmia; the use of antitachycardia pacing as a modality to treat arrhythmia recurrences may contribute to reduce the incidence of shock during follow-up in these patients.

The CXC receptor 2 is overexpressed in psoriatic epidermis.

The CXC chemokines interleukin-8 and GRO/melanoma growth-stimulatory activity (GRO-alpha) are potent activators of neutrophils and lymphocytes, but also stimulate growth and differentiation of nonhematopoietic cells like keratinocytes, fibroblasts, and melanocytes. High mRNA and protein levels have been detected in psoriatic epidermis. Chemokine activation of target cells is mediated by specific receptors and two CXC receptors have been described with similar affinity for interleukin-8 but different affinities for GRO-alpha. In this study, we examined the expression of both CXCR1 and CXCR2 in psoriatic tissue, identifying the target cells of chemokine activation in psoriasis. By immunohistochemistry and in situ hybridization, as confirmed by northern blot analysis and reverse transcriptase polymerase chain reaction, we could detect expression of the CXCR2 in suprabasal lesional psoriatic keratinocytes but not in healthy skin. The CXCR1 could not be localized in psoriatic keratinocytes with immunohistochemistry and in situ hybridization, but infiltrating cells in the dermal compartment expressed both types of receptors. These data suggest that in addition to neutrophil activation by both CXCR1 and CXCR2, activation of keratinocytes mediated by CXCR2 could contribute to the characteristic epidermal changes observed in psoriasis.

Genomic organization, sequence, and transcriptional regulation of the human eotaxin gene.

Eotaxin is an eosinophil specific beta-chemokine assumed to be involved in eosinophilic inflammatory diseases such as atopic dermatitis, allergic rhinitis, asthma and parasitic infections. Its expression is stimulus- and cell-specific. We here describe the genomic organisation (3 exons of 132, 112 and 542 bp and 2 introns of 1211 and 378 bp) and sequence including 3 kb of DNA from the immediate 5' upstream region of the human eotaxin gene. Among the regulatory promoter elements potentially regulating eotaxin gene expression and/or mediating the effects of anti-inflammatory drugs we identified consensus sequences known to interact with nuclear factors like NF-IL6, AP-1, a NF-kappa-B like consensus sequence and gamma-interferon- as well as glucocorticoid response elements.

[Anatomic distribution, conduction properties and recurrences after ablation of multiple in comparison with single accessory conduction pathways]. / Anatomische Verteilung, Leitungseigenschaften und Rezidivverhalten nach Ablation von multiplen im Vergleich zu singulären akzessorischen Leitungsbahnen.

In 1076 consecutive patients referred for radiofrequency current catheter ablation, the anatomical distribution and conduction properties of accessory pathways (APs) as well as the mode of recurrence after ablation were retrospectively analyzed and compared in patients with multiple and single APs. Except for 17 patients with Ebstein's anomaly, the prevalence of patients of multiple APs in this cohort was 5.4%. Patients with multiple APs. as opposed to patients with a single AP, had significantly more often APs located on the right free wall (23% versus 10%) and--since the prevalence of septal APs was identical in both groups--less frequently APs located on the left free wall (44% versus 56%). Also, concealed APs were significantly more often encountered in patients with multiple APs (45% versus 24%). Recurrence of conduction across an AP which had presumably been ablated was observed in both groups with statistically equal incidence of < 5%. In 11 patients with multiple APs, the additional AP was only found at the repeat session. These "new" APs were mostly concealed (9 out of 11) and necessitated an intervention predominantly late after the initial ablation session. Intermittent concealed conduction appears to be a likely explanation for this phenomenon. Patients with multiple APs exhibit a higher incidence of right free-wall and concealed APs, yet they stand the same, approximately 95%, chance of cure as do patients with a single AP. Nearly 25% percent of repeat sessions in patients initially thought to have a single AP are caused by the late manifestation of an additional AP.

Clinical recurrences after successful accessory pathway ablation: the role of "dormant" accessory pathways.

INTRODUCTION: Recurrence of clinical symptoms after radiofrequency catheter ablation of an accessory atrioventricular pathway (AP) may be due to the late manifestation of an additional AP that was not detected during the initial ablation session. It was the purpose of this study to elucidate the phenomenon of these "dormant" APs. METHODS AND RESULTS: Of 1280 consecutive patients who underwent radiofrequency catheter ablation of an AP, 54 patients (4.2%) developed clinical symptoms postablation, necessitating a repeat ablation session. Recurrence of conduction over the AP targeted at the initial ablation session was found in 45 patients, whereas in the other 9 patients (0.7%) the manifestation of a previously unnoticed AP had caused symptom recurrence. Retrospective analysis of the data from these patients' ablation sessions revealed that the late manifesting AP was ablated at a site clearly different from that of the initially targeted AP, and that the manifestation of conduction over a previously "dormant" AP occurred significantly later than the recovery of a presumably ablated AP. Seven (78%) of the 9 "dormant" APs were concealed, and none exhibited decremental conduction properties. CONCLUSION: The incidence of clinical recurrences mediated by the late manifestation of conduction over a previously "dormant" AP is low. The lack of an anatomic vicinity of these predominantly concealed APs with the initially targeted AP and the lack of evidence for their presence during the initial ablation session suggest intermittent conduction as the most likely explanation for their late manifestation.

Increased eotaxin-mRNA expression in non-atopic and atopic nasal polyps: comparison to RANTES and MCP-3 expression.

Eosinophilic tissue infiltration of nasal mucosa typical for allergic rhinitis and chronic polypous sinusitis may be due to chemotactic activity of chemokines specific for eosinophils. The CC-chemokines eotaxin, RANTES and MCP-3 have been postulated to be involved in the recruitment of eosinophils to certain inflamed tissues. To explore their possible role in chronic polypous sinusitis we examined eotaxin-, RANTES- and MCP-3-gene expression in human nasal polyps and normal human nasal mucosa of patients undergoing endonasal surgery for treatment of chronic polypous sinusitis. Using gene-specific primers in semi-quantitative reverse-transcriptase polymerase-chain-reaction experiments we found elevated expression of eotaxin- and RANTES-mRNA but no MCP-3-mRNA in non-atopic and atopic nasal polyps when compared to normal nasal mucosa.

Human dermal fibroblasts express eotaxin: molecular cloning, mRNA expression, and identification of eotaxin sequence variants.

Recently we discovered and purified a novel beta-chemokine with eosinophil specific chemotactic activity from supernatants to long-term TNF-alpha stimulated dermal fibroblasts. Using degenerated specific oligonucleotides based on partial amino acid sequence data and a PCR protocol, we obtained different clones sharing high sequence homology with this novel chemokine and with human eotaxin cDNA. Semi-quantitative RT-PCR experiments using eotaxin-specific primers indicate low constitutive eotaxin mRNA expression in human dermal fibroblasts which is upregulated by IL-1 alpha and TNF-alpha within 6 hrs and modulated by IFN-gamma. While IL-1 alpha-induced eotaxin mRNA accumulation is transient, long-term stimulation with TNF-alpha resulted in a further increase of eotaxin mRNA.

The use of reverse transcription polymerase chain reaction to analyse large numbers of mRNA species from a single cell.

A PCR method is described for determining the expression of multiple heterogeneous mRNAs from single cells. The total mRNA pool of a single selected cell is subjected to reverse transcription and subsequent tailing with poly(dA). This cDNA is preamplified by a sequence non-specific PCR protocol using oligo(dT)-containing primers. The single cell cDNA library obtained permits the analysis of virtually unlimited numbers of mRNA species per cell using sequence-specific PCR. This procedure of multiple mRNA analysis enables phenotyping of any cell for its mRNA composition and could be used to study the cytokine mRNA expression of individual human T cells ex vivo. The method should greatly facilitate the analysis of combinatorial expression of known genes in any cell.

Molecular characterization of the gene locus of the human cell proliferation-associated nuclear protein defined by monoclonal antibody Ki-67.

The human antigen defined by the monoclonal antibody Ki-67, the 'Ki-67 protein', is an ubiquitously expressed human nuclear protein strictly associated with cell proliferation and is widely used in routine pathology as a 'proliferation marker' to measure the growth fraction in human tumours. In immunoblots of proteins from proliferating cells, Ki-67 detects two bands with the apparent molecular weights of 345 and 395 kDa. Recently we reported on the cloning and sequencing of the complete cDNA of the Ki-67 protein. We found two isoforms of cDNA with full lengths of 11.4 and 12.5 kb, respectively, likely formed by the alternative splicing of exon 7. The remarkable exon 13 at the 'centre' of this gene contains 16 homologous segments of 366 bp (Ki-67 repeats), each including a highly conserved new motif of 66 bp (Ki-67 motif). Computer analyses confirmed that the cDNAs encode for a new class of nuclear proteins. The complete gene locus of the Ki-67 protein, comprising a 74 bp 5' region and a 264 bp 3' region, has been sequenced and aligned to a continuous sequence of 29,965 bp length located on chromosome 10q25-ter. The gene is organized in 15 exons with sizes from 67 to 6845 bp and in 14 introns with sizes from 87 to 3569 bp. Three introns contain homologue copies of 'Alu-repeats'. Interestingly, the introns flanking the alternative spliced exon 7 are free of any consensus donor and acceptor splicing signal. All other intron-exon transitions contained a potential branch site. The complete 5' region including the first two exons represents a CpG-rich island. We found the transcription initiation site in exon two adjacent to the consensus sequence of a cap site. Upstream of this cap site no TATA- or CCAAT-box could be located, but downstream we found two remarkable directly repeated elements of 24 bp lengths each containing a TATA box in inverse orientation.

Assessment of mycobacterial DNA in cells and tissues of mycobacterial and sarcoid lesions.

In this study we applied a polymerase chain reaction (PCR) assay for the detection and species-specific identification of mycobacteria to samples from patients with sarcoidosis and mycobacterial infections and from control patients. The PCR-technique is based on the amplification of mycobacterial DNA coding for 16S rRNA, which is present in all mycobacterial species, and on the additional sequencing of the PCR fragment to determine the species. Mycobacterial DNA could be detected in lung tissues and bronchoalveolar lavage cells from cases of tuberculosis and infections with atypical mycobacteria. On the other hand, mycobacterial DNA was amplified only in lung tissue from one patient with sarcoidosis. Twenty-three samples from patients with sarcoidosis were negative for mycobacterial DNA. From our results we conclude that the granulomatous lesions in sarcoidosis may not be due to mycobacterial infections.

Evidence for a hepatocyte membrane fatty acid transport protein using rat liver mRNA expression in Xenopus laevis oocytes.

The aim of the present study was to directly demonstrate that hepatocellular uptake of long-chain fatty acids represents a non-diffusional uptake mechanism. Xenopus laevis oocytes were used for expression of rat liver mRNA to identify the liver fatty acid uptake system. Injection of total rat liver poly(A)+ RNA into oocytes resulted in a dose-dependent increase in fatty acid uptake. The most active mRNA was found in the 1.1-2.1 kb subfraction. In contrast, expression of the liver cytosolic fatty acid binding protein (L-FABP) or the previously suggested candidate carrier protein, mitochondrial aspartate aminotransferase (mGOT), did not induce fatty acid uptake. It is concluded that in rat liver, fatty acid transport represents a protein-mediated transport system.

Endotoxin activates human vascular smooth muscle cells despite lack of expression of CD14 mRNA or endogenous membrane CD14.

During infection or inflammation, cells of the blood vessel wall, such as endothelial cells (EC) and smooth muscle cells (SMC), contribute to the regulation of the immune response by production of cytokines or expression of adhesion molecules. Little is known about the mechanism(s) involved in the stimulation of vascular cells by endotoxin (lipopolysaccharide [LPS]). As reported previously, LPS antagonists reduce LPS-induced cytokine production or adhesion in vitro specifically, suggesting a specific LPS recognition mechanism. We thus investigated the role of CD14 for stimulation of vascular SMC by LPS. Complement-fixing antibodies directed against CD14 (LeuM3, RoMo I, or Mo2) lysed monocytes but failed to mediate lysis of EC or SMC, indicating the lack of endogenous membrane CD14 in vascular cells. In addition, we did not detect expression of CD14 protein on EC and SMC in cell sorting analysis or cell immunoassay experiments. These observations are in line with our finding that a CD14 probe did not hybridize with mRNA or EC or SMC in Northern (RNA) blot experiments, although it hybridized well with monocyte-derived mRNA. We obtained the same results with the much more sensitive reverse transcription-PCR. Since the vascular SMC did not express endogenous CD14, we investigated the role of human serum-derived soluble CD14 (sCD14) for activation of SMC by LPS. In medium containing human serum, anti-CD14 antibodies inhibited activation of SMC by LPS. In contrast, the same antibodies did not inhibit activation of cells cultured in medium containing fetal calf serum. SMC cultured in sCD14-depleted medium responded 1,000-fold less to LPS than cells cultured in presence of sCD14. Reconstitution of sCD14-depleted serum or supplementation of serum-free medium with recombinant CD14 restored the capacity of the cells to respond to LPS. These results show that specific activation of vascular SMC by LPS does not involve binding to endogenous membrane CD14, but that the activation of vascular SMC by LPS is mediated to a great extent by serum-derived sCD14.

Cell proliferation-associated nuclear antigen defined by antibody Ki-67: a new kind of cell cycle-maintaining proteins.

A decade of studies on the human nuclear antigen defined by monoclonal antibody Ki-67 (the "Ki-67 protein") has made it abundantly clear that this structure is strictly associated with human cell proliferation and that the expression of this protein can be used to assess the growth fraction of a given cell population. Until recently the Ki-67 protein was described as a nonhistone protein that is highly susceptible to protease treatment. We have isolated and sequenced cDNAs encoding for this antigen and found two isoforms of the full length cDNA of 11.5 and 12.5 kb, respectively, sequence and structure of which are thus far unique. The gene encoding the Ki-67 protein is organized in 15 exons and is localized on chromosome 10. The center of this gene is formed by an extraordinary 6845 bp exon containing 16 successively repeated homologous segments of 366 bp ("Ki-67 repeats"), each containing a highly conserved new motif of 66 bp ("Ki-67 motif"). The deduced peptide sequence of this central exon possess 10 ProGluSerThr (PEST) motifs which are associated with high turnover proteins such as other cell cycle-related proteins, oncogenes and transcription factors, etc. Like the latter proteins the Ki-67 antigen plays a pivotal role in maintaining cell proliferation because Ki-67 protein antisense oligonucleotides significantly inhibit 3H-thymidine incorporation in permanent human tumor cell lines in a dose-dependent manner.