Synthetic antisense oligodeoxynucleotides as potential drugs against hepatitis C

Antisense oligodeoxynucleotides (ODNS) can be used to specifically inhibit hepatitis C viral gene expression. Due to its high degree of conservation and its important function as internal ribosomal entry site, the 5'-non-coding region of the hepatitis C virus has been the most effective target to inhibit translation so far. Inhibition of luciferase reporter gene expression of up to 96 +/- 2% has been achieved. Modifications of ODNs like phosphorothioate, methylphosphonate or benzylphosphonate modification of terminal or intramolecular internucleotide phosphates lead to altered lipophilicity and binding stability to its RNA target and resistance against serum nucleases. The mode of action of ODNs is mainly dependent on an efficient induction of RNase H activity. The uptake of ODNs occurs via receptor-mediated or absorptive and fluid-phase endocytosis. After release from the endosomes, ODNs may exert their effects by interaction with cytosolic or nuclear structures. Side effects can occur when this interaction affects intra- or extracellular targets essential for biological cell function. If these problems can be solved, antisense technology has the potential for future therapy of human disease.     

Contaminant assimilation within the water column of two newly created prairie wetlands

This research examined the influence of abiotic factors on contaminant assimilation within the water column of two wetlands (RMA3 and RMA4) located on Rocky Mountain Arsenal National Wildlife Area, north of Denver, Colorado, USA. We define contaminant assimilation as the ability of a wetland system to reduce negative impacts of contaminants on aquatic organisms. To examine the influence of abiotic factors (i.e., alkalinity, suspended solids), chronic toxicity tests were performed from February 1992 through November 1993. Ceriodaphnia dubia were exposed to water from the wetlands, which was spiked with Zn. These tests indicated higher assimilative capacity in wetland RMA4 relative to RMA3. Dissolved organic carbon (DOC) was significantly greater in RMA4 than in RMA3, suggesting that it may have influenced differences in bioavailability of Zn in wetland water. Suspended solids were also consistently higher in RMA4 than in RMA3. To test the influence of suspended solids on contaminant assimilation, C. dubia were exposed to filtered and unfiltered Zn-spiked water from RMA4. The greater toxicity of Zn to C. dubia in filtered water indicated that suspended solids were important in ameliorating impacts of Zn in RMA4. Results from this study suggest that from the suite of variables assessed, DOC and suspended solids were the most important physiochemical influences on Zn toxicity within the water column of these two wetlands. Differences in habitat characteristics of RMA3 and RMA4 demonstrated the importance of site-selection toward functional design of wetlands.     

Adenocarcinoma combined with malignant peripheral nerve sheath tumor of the gallbladder

Adenocarcinoma of the gallbladder combined with a malignant peripheral nerve sheath tumor (MPNST) in the gallbladder in an 81-year-old woman is reported. The resected gallbladder showed two distinct tumor components, the epithelioid type of MPNST and adenocarcinoma with areas of mucin production. Although the immediate postoperative course was uneventful, a pathologic fracture of her right upper femur developed 4 months after the cholecystectomy. The pathology was determined to be a feature of metastatic MPNST rather than of adenocarcinoma. A whole body bone scan revealed multiple metastases, including the left parietal skull, left ninth rib, seventh thoracic vertebra, and right upper third of the femur. Despite cholecystectomy and postoperative irradiation therapy, she died 6 months after diagnosis of the tumor. Without an autopsy the primary site of the MPNST was unknown. We found that the prognosis was very poor in patients with distal metastatic MPNST, especially in older patients.     

PCR bias toward the wild-type k-ras and p53 sequences: implications for PCR detection of mutations and cancer diagnosis

PCR-based cancer diagnosis requires detection of rare mutations in k-ras, p53 or other genes. The assumption has been that mutant and wild-type sequences amplify with near equal efficiency, so that they are eventually present in proportions representative of the starting material. Work on factor IX suggests that this assumption is invalid for one case of near-sequence identity. To test the generality of this phenomenon and its relevance to cancer diagnosis, primers distant from point mutations in p53 and k-ras were used to amplify wild-type and mutant sequences from these genes. A substantial bias against PCR amplification of mutants was observed for two regions of the p53 gene and one region of k-ras. For k-ras and p53, bias was observed when the wild-type and mutant sequences were amplified separately or when mixed in equal proportions before PCR. Bias was present with proofreading and non-proofreading polymerases. Mutant and wild-type segments of the factor V, cystic fibrosis transmembrane conductance regulator and prothrombin genes were amplified and did not exhibit PCR bias. Therefore, the assumption of equal PCR efficiency for point mutant and wild-type sequences is invalid in several systems. Quantitative or diagnostic PCR will require validation for each locus, and enrichment strategies may be needed to optimize detection of mutants.