Computerized radiographic measurements of anatomical parameters of the elbow joint of Bernese Mountain Dogs using two different methods

Deckblatt-Impressum Inhaltsverzeichnis Abkürzungsverzeichnis Einleitung Literaturübersicht Material und Methoden Ergebnisse Diskussion Zusammenfassung Summary Literaturverzeichnis Anhang Danksagung SelbständigkeitserklärungZiel der Arbeit war es die Meßmethoden nach MUES und nach VIEHMANN zur Erfassung anatomischer Messparameter des caninen Ellbogengelenkes zu vergleichen. Eine Korrelation von Messwerten und dem gutachterlichen ED-Grad wurde berechnet. Nach der Messmethode MUES wurden der Winkel OL, der Winkel PA, der Winkel RA und der Winkel UL gemessen und nach der Messmethode nach VIEHMANN der Radius des Condylus humeri, der Öffnungswinkel beta, der Quotient Q, der Quotient Ae, die Fläche X, die Stufe 1 und die Stufe 2. Insgesamt wurden 931 Röntgenbilder von Ellbogengelenken der Rasse Berner Sennenhund, die zuvor im Rahmen der Zuchtzulassung gutachterlich mit einem ED-Grad bewertet worden waren, in jeweils drei Messdurchgängen nach beiden Methoden vermessen. Die Röntgenbilder wurden auch aufgrund der Beugungswinkel in 5 Gruppen unterteilt: 0-30°, 31-60°, 61-90°, 91-120° und größer 120°. Bei den Messparametern nach MUES waren die Mittelwerte von Winkel OL, Winkel PA und Winkel RA in den verschiedenen Beugewinkelgruppen signifikant unterschiedlich. Von den Messwerten nach VIEHMANN war die Fläche X abhängig vom Beugewinkel. Die Mittelwerte der Stufe 1 und Stufe 2 wurden signifikant kleiner, je größer der Beugewinkel war, während die Werte der Quotient Q und Ae, der Winkel beta, sowie der Radius des Condylus humeri vom Beugewinkel unabhängig waren. Die Größe des Winkels OL, des Winkels PA und des Winkel RA korrelierten signifikant mit dem ED-Grad, während der Winkel UL keine Korrelation mit dem ED-Grad aufwies. Der Radius des Condylus humeri, der Öffnungswinkel beta, der Quotient Q, der Quotient Ae sowie die Stufe 1 und die Stufe 2 korrelierten hoch signifikant mit dem ED-Grad. Die Fläche X korrelierte in einzelnen Beugewinkelgruppen signifikant mit dem ED-Grad. Die Messparameter Winkel OL, PA und RA nach MUES zeigten eine signifikante Anhängigkeit von Beugewinkel des Ellbogengelenkes. Zudem war die Korrelation zwischen den Messwerten und dem ED-Grad nur signifikant, während die Korrelation der nach VIEHMANN gemessenen Parameter mit dem ED-Grad hoch signifikant war. Im direkten Vergleich der beiden Meßmethoden war die nach VIEHMANN beim Berner Sennenhund besser geeignet, eine Ellbogengelenksdysplasie zu erfassen. Die Messparameter Öffnungswinkel beta, Quotient Q und Quotient Ae korrelierten hoch signifikant mit dem ED-Grad und waren unabhängig vom Beugewinkel. Diese beiden Voraussetzungen für eine Screeningmethode wurden in der hier vorgelegten Studie von keinem der Messparameter nach MUES erfüllt.The goal of the here presented study was the comparison of two computer- assisted measuring methods (of VIEHMANN and of MUES) for the collection of anatomical measuring parameters of the canine elbow joint. The correlation between measured values and the ED-grade score by an expert was compared. The angle OL, the angle PA, the angle RA and the angle UL were measured according to MUES as well as the radius of the humeral condyle, the opening angle beta, the quotient Q, the quotient Ae, the area X, the step 1 and the step 2 between the radius and the ulna according to the method of VIEHMANN. Altogether 931 radiographs of elbow joints of Bernese Mountain Dogs, which had been evaluated for breeding protocols, were measured. Each digital radiograph was measured three times with each method. The radiographs were also devided due to the angles of flexion of the elbow joint into 5 groups: 0-30°, 31-60°, 61-90°, 91-120° and larger than 120°. With the method according to MUES the average values of angle OL, angle Pa and angle RA were significantly different in the different flexion angle groups. From the measured values according to VIEHMANN the area X depended on the angle of flexion. The average values of the step 1 and step 2 became significantly smaller the larger the flexion angle was, while the opening angle beta, quotient Q and quotient Ae as well as the radius of the Condylus humeri showed to be independent from the degree of the elbow flexion on the radiograph. The size of the angle OL, the angle PA and the angle RA correlated significantly with the ED degree, while the angle UL did not exhibit a correlation. The radius of the Condylus humeri, the opening angle beta, the quotient Q, the quotient Ae as well as the step 1 and the step 2 correlated highly significant with the ED degree. The area X correlated in individual degrees of flexion significantly with the ED degree. The parameters angle OL, PA and RA of the method of MUES showed a significant dependence on the degree of flexion of the elbow joint. The correlation between the measured values and the ED degree was only significant, while the correlation of the parameters according to VIEHMANN was highly significant. The values of the quotient Q, the quotient Ae as well as the opening angle beta were independent from the degree of flexion. They seem to be more suitable for breeding protocols. To decide, if the VIEHMANN-method is suitable for an early breeding protocol, a prospective study using a representative population of young dogs should be accomplished including repetitive radiographs during growth

Statin and rottlerin small-molecule inhibitors restrict colon cancer progression and metastasis via MACC1

MACC1 (Metastasis Associated in Colon Cancer 1) is a key driver and prognostic biomarker for cancer progression and metastasis in a large variety of solid tumor types, particularly colorectal cancer (CRC). However, no MACC1 inhibitors have been identified yet. Therefore, we aimed to target MACC1 expression using a luciferase reporter-based high-throughput screening with the ChemBioNet library of more than 30,000 compounds. The small molecules lovastatin and rottlerin emerged as the most potent MACC1 transcriptional inhibitors. They remarkably inhibited MACC1 promoter activity and expression, resulting in reduced cell motility. Lovastatin impaired the binding of the transcription factors c-Jun and Sp1 to the MACC1 promoter, thereby inhibiting MACC1 transcription. Most importantly, in CRC-xenografted mice, lovastatin and rottlerin restricted MACC1 expression and liver metastasis. This is—to the best of our knowledge—the first identification of inhibitors restricting cancer progression and metastasis via the novel target MACC1. This drug repositioning might be of therapeutic value for CRC patients

ECRG4 is a candidate tumor suppressor gene frequently hypermethylated in colorectal carcinoma and glioma

<p>Abstract</p> <p>Background</p> <p>Cancer cells display widespread changes in DNA methylation that may lead to genetic instability by global hypomethylation and aberrant silencing of tumor suppressor genes by focal hypermethylation. In turn, altered DNA methylation patterns have been used to identify putative tumor suppressor genes.</p> <p>Methods</p> <p>In a methylation screening approach, we identified <it>ECRG4 </it>as a differentially methylated gene. We analyzed different cancer cells for <it>ECRG4 </it>promoter methylation by COBRA and bisulfite sequencing. Gene expression analysis was carried out by semi-quantitative RT-PCR. The <it>ECRG4 </it>coding region was cloned and transfected into colorectal carcinoma cells. Cell growth was assessed by MTT and BrdU assays. ECRG4 localization was analyzed by fluorescence microscopy and Western blotting after transfection of an <it>ECRG4-eGFP </it>fusion gene.</p> <p>Results</p> <p>We found a high frequency of <it>ECRG4 </it>promoter methylation in various cancer cell lines. Remarkably, aberrant methylation of <it>ECRG4 </it>was also found in primary human tumor tissues, including samples from colorectal carcinoma and from malignant gliomas. <it>ECRG4 </it>hypermethylation associated strongly with transcriptional silencing and its expression could be re-activated <it>in vitro </it>by demethylating treatment with 5-aza-2'-deoxycytidine. Overexpression of <it>ECRG4 </it>in colorectal carcinoma cells led to a significant decrease in cell growth. In transfected cells, ECRG4 protein was detectable within the Golgi secretion machinery as well as in the culture medium.</p> <p>Conclusions</p> <p><it>ECRG4 </it>is silenced via promoter hypermethylation in different types of human cancer cells. Its gene product may act as inhibitor of cell proliferation in colorectal carcinoma cells and may play a role as extracellular signaling molecule.</p

High level of conservation between genes coding for the GAMYB transcription factor in barley (Hordeum vulgare L.) and bread wheat (Triticum aestivum L.) collections

The transcription factor GAMYB is involved in gibberellin signalling in cereal aleurone cells and in plant developmental processes. Nucleotide diversity of HvGAMYB and TaGAMYB was investigated in 155 barley (Hordeum vulgare) and 42 wheat (Triticum aestivum) accessions, respectively. Polymorphisms defined 18 haplotypes in the barley collection and 1, 7 and 3 haplotypes for the A, B, and D genomes of wheat, respectively. We found that (1) Hv- and TaGAMYB genes have identical structures. (2) Both genes show a high level of nucleotide identity (>95%) in the coding sequences and the distribution of polymorphisms is similar in both collections. At the protein level the functional domain is identical in both species. (3) GAMYB genes map to a syntenic position on chromosome 3. GAMYB genes are different in both collections with respect to the Tajima D statistic and linkage disequilibrium (LD). A moderate level of LD was observed in the barley collection. In wheat, LD is absolute between polymorphic sites, mostly located in the first intron, while it decays within the gene. Differences in Tajima D values might be due to a lower selection pressure on HvGAMYB, compared to its wheat orthologue. Altogether our results provide evidence that there have been only few evolutionary changes in Hv- and TaGAMYB. This confirms the close relationship between these species and also highlights the functional importance of this transcription factor

The Effect of Micrococcal Nuclease Digestion on Nucleosome Positioning Data

Eukaryotic genomes are packed into chromatin, whose basic repeating unit is the nucleosome. Nucleosome positioning is a widely researched area. A common experimental procedure to determine nucleosome positions involves the use of micrococcal nuclease (MNase). Here, we show that the cutting preference of MNase in combination with size selection generates a sequence-dependent bias in the resulting fragments. This strongly affects nucleosome positioning data and especially sequence-dependent models for nucleosome positioning. As a consequence we see a need to re-evaluate whether the DNA sequence is a major determinant of nucleosome positioning in vivo. More generally, our results show that data generated after MNase digestion of chromatin requires a matched control experiment in order to determine nucleosome positions

Identification of Y-Box Binding Protein 1 As a Core Regulator of MEK/ERK Pathway-Dependent Gene Signatures in Colorectal Cancer Cells

Transcriptional signatures are an indispensible source of correlative information on disease-related molecular alterations on a genome-wide level. Numerous candidate genes involved in disease and in factors of predictive, as well as of prognostic, value have been deduced from such molecular portraits, e.g. in cancer. However, mechanistic insights into the regulatory principles governing global transcriptional changes are lagging behind extensive compilations of deregulated genes. To identify regulators of transcriptome alterations, we used an integrated approach combining transcriptional profiling of colorectal cancer cell lines treated with inhibitors targeting the receptor tyrosine kinase (RTK)/RAS/mitogen-activated protein kinase pathway, computational prediction of regulatory elements in promoters of co-regulated genes, chromatin-based and functional cellular assays. We identified commonly co-regulated, proliferation-associated target genes that respond to the MAPK pathway. We recognized E2F and NFY transcription factor binding sites as prevalent motifs in those pathway-responsive genes and confirmed the predicted regulatory role of Y-box binding protein 1 (YBX1) by reporter gene, gel shift, and chromatin immunoprecipitation assays. We also validated the MAPK-dependent gene signature in colorectal cancers and provided evidence for the association of YBX1 with poor prognosis in colorectal cancer patients. This suggests that MEK/ERK-dependent, YBX1-regulated target genes are involved in executing malignant properties

The effectiveness of counter-terrorism strategies

Since September 11th, there have been massive increases in personal, commercial, and governmental expenditures on anti-terrorism strategies, as well as a proliferation of programs designed to fight terrorism. These increases in spending and program development have focused attention on the most significant and central policy question related to these interventions: Do these programs work? To explore research evidence regarding this question, we conducted a Campbell systematic review on counter-terrorism strategies to determine the scope and strength of evaluation research in this area. In the course of our review, we discovered that there is an almost complete absence of evaluation research on counter-terrorism strategies. From over 20,000 studies we located on terrorism, we found only seven which contained moderately rigorous evaluations of counterterrorism programs. We conclude that there is little scientific knowledge about the effectiveness of most counter-terrorism interventions. Further, from the evidence we were able to locate, it appears that some evaluated interventions either didn’t work or sometimes increased the likelihood of terrorism and terrorism-related harm. The findings of this review dramatically emphasize the need for government leaders, policy makers, researchers, and funding agencies to include and insist on evaluations of the effectiveness of these programs in their agendas. These agendas would include identifying ways to overcome methodological and data challenges often associated with terrorism research, increasing funding to evaluate existing programs through methodologically rigorous evaluation designs, and paying attention to existing evaluations of programs when implementing them. Further, programs should be assessed to establish if they cause more harm than good or if they create unanticipated consequences