Calcium restriction for 28 days markedly and negatively influences bone mineral density of the femur and lumbar vertebrae regardless of the high-fat diet ingestion in young adult male rats.

Calcium (Ca) is necessary for bone calcification, and Ca deficiency leads to decreased bone mineral density (BMD). Epidemiological studies have reported a correlation between Ca intake and BMD. Although the influences of Ca deficiency on BMD have been reported, the effects of Ca restriction on bone during high-fat diet ingestion remain unclear. Therefore, we hypothesized that high-fat diet ingestion would potentiate the negative effects of Ca restriction on bone. Sprague-Dawley strain male rats (aged 11 weeks) were divided into 4 groups: basic control diet (Cont.) (11% lipid energy rate, 0.5% calcium), basic control diet with Ca restriction (CaR) (11% lipid energy rate, 0.02% calcium), high-fat diet (HF) (40% lipid energy rate, 0.5% calcium), and high-fat diet with Ca restriction (HFCaR) (40% lipid energy rate, 0.02% calcium). At 28 days after starting the experimental diets, body weights were higher in the high-fat diet groups (HF and HFCaR) than in the standard-fat diet groups (Cont. and CaR) on 2-way analysis of variance. The apparent Ca absorption rate in the Ca-restricted groups (CaR and HFCaR) was higher than in the Ca-sufficient groups (Cont. and HF). BMD and bone strength parameters of the femur and lumbar vertebrae in the Ca-restricted groups were markedly lower than in the Ca-sufficient groups, whereas there were no significant differences between the standard-fat diet and HF diet groups. These results suggest that 28 days of Ca restriction increases the risk of bone fracture and osteoporosis.

Coincident sequence-specific RNA degradation of linked transgenes in the plant genome.

The expression of transgenes in plant genomes can be inhibited by either transcriptional gene silencing or posttranscriptional gene silencing (PTGS). Overexpression of the chalcone synthase-A (CHS-A) transgene triggers PTGS of CHS-A and thus results in loss of flower pigmentation in petunia. We previously demonstrated that epigenetic inactivation of CHS-A transgene transcription leads to a reversion of the PTGS phenotype. Although neomycin phosphotransferase II (nptII), a marker gene co-introduced into the genome with the CHS-A transgene, is not normally silenced in petunia, even when CHS-A is silenced, here we found that nptII was silenced in a petunia line in which CHS-A PTGS was induced, but not in the revertant plants that had no PTGS of CHS-A. Transcriptional activity, accumulation of short interfering RNAs, and restoration of mRNA level after infection with viruses that had suppressor proteins of gene silencing indicated that the mechanism for nptII silencing was posttranscriptional. Read-through transcripts of the CHS-A gene toward the nptII gene were detected. Deep-sequencing analysis revealed a striking difference between the predominant size class of small RNAs produced from the read-through transcripts (22 nt) and that from the CHS-A RNAs (21 nt). These results implicate the involvement of read-through transcription and distinct phases of RNA degradation in the coincident PTGS of linked transgenes and provide new insights into the destabilization of transgene expression.