Craniofacial fibrous dysplasia (CFD) of the maxilla in an 11-year old boy: A case report

We present the case of a surgically treated 11-year old boy with a diagnosis of craniomaxillofacial fibrous dysplasia (CFD) in the maxillary sinus. When first seen in the outpatient clinic of our department he had minimal symptoms. After initial radiological diagnostics by computed tomography scans (CT-scans) the patient was treated operatively by radical excision of the tumor. The radiographs showed the typical intramedullary located and well-defined lesions, which eroded the cortical bone with the typical appearance of fibrous dysplasia. The histopathology showed the typical curved extending fibrous trabeculae in C, O and Y-shape which were embedded in a moderately cellular morphologically inconspicuous stroma, confirming the initial suspicion of fibrous dysplasia of the maxillary bone. Cone beam tomography was a valuable tool in determining the re-ossification of bone at the affected side. Local resection can be curative in limited disease.     

Low doses of 2,3,7,8-tetrachlorodibenzo- p-dioxin increase transforming growth factor beta and cause myocardial fibrosis in marmosets ( Callithrix jacchus)

Epidemiological studies have suggested an association between exposure to dioxins and cardiovascular morbidity and mortality. However, cardiotoxic effects of low doses of 2,3,7,8-tetrachlorodibenzo- p-dioxin (TCDD) in animals have not been reported so far. We studied the hearts of male marmosets ( Callithrix jacchus)after treatment with single subcutaneous doses of 1, 10 or 100 ng TCDD/kg body weight or vehicle (toluene/DMSO 1+2 v/v, 100 microl/kg body weight). The animals were killed 2 or 4 weeks after treatment. Tissue samples of left ventricular myocardium were stained with picrosirius red and examined histologically along with quantitative image analysis. Extracellular matrix proteins were additionally analysed by western blotting. Monkeys showed no overt signs of toxicity nor did their relative heart weights differ significantly depending on treatment. Histology revealed an increase of picrosirius red-positive area above control values in 2 of 4 (1 ng TCDD/kg body weight), 6 of 12 (10 ng/kg) and 6 of 10 (100 ng/kg) marmosets. Western blotting confirmed these histological findings showing an increase of collagen, fibronectin and laminin in the hearts of TCDD-treated animals. Western blotting additionally showed an increased concentration of transforming growth factor beta1 (TGF-beta1) as well as TGF-beta receptor type I which could be a functional link to the effects on extracellular matrix. Our findings might explain the association of TCDD exposure with increased cardiovascular mortality observed in epidemiological studies and should stimulate further research on the role of changes in the extracellular matrix in the toxic effects of dioxins and related substances on other organs.     

Reliable and sample saving gene expression analysis approach for diagnostic tool development

This work answers the question of whether it is necessary to hybridize individual instead of pooled RNA samples on microarrays for screening gene targets suitable as diagnostic tools for radiation exposure scenarios, while at the same time meeting comparable microarray quality criteria. For developing new clinical diagnostic tools, a two-stage study design was employed in five projects. At first, pooled and not individual RNA samples were hybridized on microarrays for screening purposes. Potential gene candidates were selected based on their fold-change only. This was followed by a validation/quantification step using individual RNA samples and quantitative RT-PCR. Quality criteria from the screening approach with pooled RNA samples were compared with published data from the MicroArray Quality Control (MAQC) consortium that hybridized each reference RNA sample separately and established quality criteria for microarrays. When comparing both approaches, only insignificant differences for quality criteria such as false positives, sensitivity, specificity, and overall agreement were found. However, material, costs, and time were drastically reduced when hybridizing pooled RNA and gene targets applicable for clinical diagnostic purposes could be successfully selected. In search of new diagnostic tools for radiation exposure scenarios, the two stage study design using either pooled or individual RNA samples on microarrays shows comparable quality criteria, but the RNA pooling approach saves unique material, costs, and efforts and successfully selects gene targets that can be used for the desired diagnostic purposes.     

The role of prostaglandin E receptor-dependent signaling via cAMP in Mdr1b gene activation in primary rat hepatocyte cultures

Multidrug resistance (mdr) proteins of the mdr1 type function as multispecific xenobiotic transporters in hepatocytes. In the liver, mdr1 overexpression occurs during regeneration, cirrhosis, and hepatocarcinogenesis and may contribute to primary chemotherapy resistance. Cultured rat hepatocytes exhibit a time-dependent "intrinsic" increase in functional mdr1b expression, which depends on cyclooxygenase-catalyzed prostaglandin E(2) release. In the present study, the prostaglandin E (EP) receptor agonist misoprostol (1-10 microg/ml) further enhanced intrinsic mdr1b mRNA expression in primary rat hepatocytes. On the other hand, [1alpha(z),2beta,5alpha]-(+)-7-[5-[1,1'-(biphenyl)-4-yl]methoxy]-2-(4-morpholinyl)-3-oxocyclopentyl]-4-heptenoic acid (AH23848B) (30 microM), an antagonist of the cAMP-coupled EP4 receptor, and the protein kinase A (PKA) inhibitor, N-(2-[bromocinnamylamino]ethyl)-5-isoquinolinesulfonamide (H89) (10 nM), repressed intrinsic mdr1b mRNA up-regulation, whereas the stable cAMP analog 8-bromo-cAMP (10 microM) and the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX) (100 microM) further enhanced intrinsic mdr1b expression. Primary rat hepatocytes, transiently transfected with reporter gene constructs controlled by mdr1b 5'-gene-flanking regions [-1074 to +154 base pairs (bp) or -250 to +154 bp], demonstrated pronounced mdr1b promoter activity, already without the addition of exogenous modulators. Nevertheless, activity was further stimulated by misoprostol, 8-bromo-cAMP, or IBMX. Cotransfection with expression vectors for PKI, an inhibitor protein of cAMP-dependent PKA, or KCREB, a dominant-negative mutant of the cAMP-responsive element-binding protein (CREB), decreased high-intrinsic mdr1b promoter activity. KCREB also counteracted misoprostol-induced mdr1b promoter activation. In conclusion, these data provide evidence for a pivotal role of EP receptor-stimulated, cAMP-dependent activation of PKA and CREB or CREB-related proteins in mdr1b gene activation in primary rat hepatocytes. Thus, these data might offer potential new target structures for the reversal of primary drug resistance, for example, of liver tumors.