Multiple Conserved Elements Structuring Inverted Repeats in the Mammalian Coat Color-Related Gene Asip.

In the agouti signaling gene protein (Asip) of the house mouse (Mus musculus), inverted repeat (IR) arrays are known to exist in a non-coding region adjacent to the ventral-specific promoter region and the accompanying two exons (exons 1A and 1A'), which are around 100 kb upstream from the amino acid coding regions of exons 2, 3, and 4. To determine the gene structure of mammalian Asip and to elucidate trends in its evolution, non-coding sequences of six rodent (mouse, rat, Chinese hamster, squirrel, guinea pig, and naked mole rat) and three non-rodent (rabbit, human, and cow) species were retrieved from databases and compared. Our homology search analyses revealed the presence of three to five highly conserved non-coding elements (CNE). These CNEs were found to form IRs in rodents and lagomorphs. Combinations of IRs were further shown to build symmetric, long IR arrays. Intra- and inter-specific comparisons of the sequences of three universal CNEs showed homogeneity between CNE pairs within species. This implies that certain evolutionary constraints maintained the IR structure in the rodent and rabbit species.

Whole-genome sequencing analysis of wild house mice (Mus musculus) captured in Madagascar.

In Madagascar, the house mouse (Mus musculus) is widely believed to have colonized with human activities and is now one of the most abundant rodents on the island. However, its genetic background at the genomic level remains unclear, and clarifying this would help us to infer the timing of introduction and route of migration. In this study, we determined the whole-genome sequences of five Madagascar house mice captured from an inland location in Madagascar. We examined the genetic background of samples by analyzing the mitochondrial and autosomal genomes. We confirmed that the mitochondrial genome lineages of collected samples formed a single clade placed at one of the most basal positions in the Mus musculus species. Autosomal genomic sequences revealed that these samples are most closely related to the subspecies M. m. castaneus (CAS), but also contain a genetic component of the subspecies M. m. domesticus (DOM). The signature of a strong population bottleneck 1,000-3,000 years ago was observed in both mitochondrial and autosomal genomic data. In a comparison with global samples of M. musculus, the Madagascar samples showed strong genetic affinity to many CAS samples across a wide range of Indian Ocean coastal and insular regions, with divergence time estimated as around 4,000 years ago. These findings support the proposition that the ancestors of these animals started to colonize the island with human agricultural activity and experienced a complex history during their establishment.

[Effect of oral adsorbent AST-120 under standard treatment of care in patients with chronic renal failure].

BACKGROUND: Oral adsorbent AST-120 reduces uremic toxins, such as indoxyl sulfate, and retards the progression of chronic kidney disease (CKD). The present study was conducted to elucidate the association between the progression of CKD and the combined effect of AST-120 and standard care based on diet therapy and medications such as ACE inhibitor and ARB. METHOD: Retrospective analysis was performed using forty-four CKD patients (chronic glomerular nephritis 16, diabetic nephropathy 9, nephrosclerosis 13, and others 6) who were treated by AST-120 during the period from October, 2001 through December, 2004 and followed up for more than 6 months. The selection criteria were an estimated creatinine clearance (eCCr) of 30 mL/min or under at the initiation of treatment and a negative change in eCCr over time (DeltaeCCr (mL/min/year)) before AST-120 treatment. The eCCr was calculated using the Cockcroft-Gault equation and the DeltaeCCr was evaluated as a marker for the progression of chronic renal failure. RESULT: Overall DeltaeCCr before and after AST-120 treatment significantly improved from -7.28 +/- 6.33 to -4.29 +/- 5.09 (paired Wilcoxon test, p < 0.05). AST-120 led to greater improvement of DeltaeCCr in patients with an DeltaeCCr of less than -10 mL/min/year before AST-120 treatment and who had higher blood uric acid, urinary protein, and urinary specific gravity valves. CONCLUSION: This finding suggests that AST-120 treatment is effective in slowing the progression of chronic kidney disease, especially, in patients who exhibit a poor response to standard care.

Establishment of a Nipah virus rescue system.

Nipah virus (NiV), a paramyxovirus, was first discovered in Malaysia in 1998 in an outbreak of infection in pigs and humans and incurred a high fatality rate in humans. Fruit bats, living in vast areas extending from India to the western Pacific, were identified as the natural reservoir of the virus. However, the mechanisms that resulted in severe pathogenicity in humans (up to 70% mortality) and that enabled crossing the species barrier were not known. In this study, we established a system that enabled the rescue of replicating NiVs from a cloned DNA by cotransfection of a constructed full-length cDNA clone and supporting plasmids coding virus nucleoprotein, phosphoprotein, and polymerase with the infection of the recombinant vaccinia virus, MVAGKT7, expressing T7 RNA polymerase. The rescued NiV (rNiV), by using the newly developed reverse genetics system, showed properties in vitro that were similar to the parent virus and retained the severe pathogenicity in a previously established animal model by experimental infection. A recombinant NiV was also developed, expressing enhanced green fluorescent protein (rNiV-EGFP). Using the virus, permissibility of NiV was compared with the presence of a known cellular receptor, ephrin B2, in a number of cell lines of different origins. Interestingly, two cell lines expressing ephrin B2 were not susceptible for rNiV-EGFP, indicating that additional factors are clearly required for full NiV replication. The reverse genetics for NiV will provide a powerful tool for the analysis of the molecular mechanisms of pathogenicity and cross-species infection.

Dynamic changes in the expression of protein tyrosine phosphatases during preimplantation mouse development: semi-quantification by real-time PCR.

Protein tyrosine phosphatase (PTP) expression was examined in preimplantation mouse embryos. We previously reported that SHP-2, LAR, PTPT9, SHP-1, and mRPTPB were expressed in preimplantation mouse embryos. Here, we examined changes in the expression levels of these PTPs during preimplantation development. cDNA was obtained by reverse transcription of embryo mRNA, amplified with 10 PCR cycles, and then subjected to real-time fluorescence-monitored PCR. Experiments with an mRNA dilution series revealed that the data obtained matched the quantities of mRNA used. The measurements obtained with real-time fluorescence-monitored PCR showed that the expression of each PTP mRNA changed dynamically, and that each had a different expression pattern. This suggests that PTPs are involved in the regulation of growth and differentiation during preimplantation development.