Involvement of wild-type p53 protein in the cell cycle requires nuclear localization.

Transfection of wild-type p53 into a pre-B, p53 nonproducer cell line yielded the generation of stable clones. Although constitutively expressing the growth-suppressor wild-type p53 protein, these cells proliferate continuously in vitro. However, expression of wild-type p53 in these cells altered their cell cycle pattern and reduced their growth in vivo. When the same parental cells were transfected with a plasmid coding for a wild-type p53 lacking nuclear localization signals, a wild-type cytoplasmic p53 protein was expressed. Expression of this cytoplasmic p53 product did not exert any changes in the growth of the parental cells, suggesting that wild-type p53 affects the cell cycle only when localized in the nuclear cell compartment.     

Evidence for a major dominance component in the variation of serum pepsinogen I levels

We report here a variance component analysis of the distribution of serum pepsinogen I levels in normal individuals, using a maximum-likelihood method on entire pedigrees. The results indicate a broad heritability of 91%, with some 74% being attributed to a dominance component. This is consistent with the hypothesis that the pepsinogen I level in normals is principally determined by the action of major genes, as also seems to be the case for duodenal ulcer patients and families.     

Biocompatibility evaluation of densified bacterial nanocellulose hydrogel as an implant material for auricular cartilage regeneration

Bacterial nanocellulose (BNC), synthesized by the bacterium Gluconacetobacter xylinus, is composed of highly hydrated fibrils (99 % water) with high mechanical strength. These exceptional material properties make BNC a novel biomaterial for many potential medical and tissue engineering applications. Recently, BNC with cellulose content of 15 % has been proposed as an implant material for auricular cartilage replacement, since it matches the mechanical requirements of human auricular cartilage. This study investigates the biocompatibility of BNC with increased cellulose content (17 %) to evaluate its response in vitro and in vivo. Cylindrical BNC structures (Ø48?×?20 mm) were produced, purified in a built-in house perfusion system, and compressed to increase the cellulose content in BNC hydrogels. The reduction of endotoxicity of the material was quantified by bacterial endotoxin analysis throughout the purification process. Afterward, the biocompatibility of the purified BNC hydrogels with cellulose content of 17 % was assessed in vitro and in vivo, according to standards set forth in ISO 10993. The endotoxin content in non-purified BNC (2,390 endotoxin units (EU)/ml) was reduced to 0.10 EU/ml after the purification process, level well below the endotoxin threshold set for medical devices. Furthermore, the biocompatibility tests demonstrated that densified BNC hydrogels are non-cytotoxic and cause a minimal foreign body response. In support with our previous findings, this study concludes that BNC with increased cellulose content of 17 % is a promising non-resorbable biomaterial for auricular cartilage tissue engineering, due to its similarity with auricular cartilage in terms of mechanical strength and host tissue response.     

Infectious clones of cotton leafroll dwarf virus strains used for genotype evaluation in cotton

Cotton blue disease (CBD) and atypical-CBD are the most important viral diseases of cotton plants in the southern region of South America. Common and atypical strains of cotton leafroll dwarf virus (CLRDV and CLRDV-at, respectively) are thought to be causative agents of CBD and atypical-CBD, respectively. Inoculation of test plants via aphid vectors is difficult, as is determining strains via molecular diagnosis; accordingly, it is difficult for breeders to evaluate the effects of blue disease-associated virus infections in cotton lineages. In the present study, we attempted to circumvent these difficulties by producing six full-length cDNA infectious clones from CLRDV and CLRDV-at strains using the Gibson Assembly protocol. For inoculation of the infectious clones, a vacuum chamber-mediated agroinfiltration protocol was adapted and applied. Using this protocol, 90%–100% of cotton plants became infected with the clones, which was not possible via syringe-based agroinfiltration. A genotyping protocol based on RT-qPCR targeting a specific region of the virus P0 protein was also developed, allowing rapid differentiation of CLRDV and CLRDV-at. Applying this protocol to 68 field samples revealed that CLRDV-at was dominant (50%) over CLRDV (5.8%) in single virus infections. These preliminary results imply that CLRDV-at might occupy the ecological niche of CLRDV in the cotton fields of Brazil.