Cost-effective development of highly polymorphic microsatellite in Japanese quail facilitated by next-generation sequencing.

Next-generation sequencing technologies permit rapid and cost-effective identification of numerous putative microsatellite loci. Here, from the genome sequences of Japanese quail, we developed microsatellite markers containing dinucleotide repeats and employed these for characterisation of genetic diversity and population structure. A total of 385 individuals from 12 experimental and one wild-derived Japanese quail lines were genotyped with newly developed autosomal markers. The maximum number of alleles, expected heterozygosity and polymorphic information content (PIC) per locus were 10, 0.80 and 0.77 respectively. Approximately half of the markers were highly informative (PIC ≥ 0.50). The mean number of alleles per locus and observed heterozygosity within a line were in the range of 1.3-4.1 and 0.11-0.53 respectively. Compared with the wild-derived line, genetic diversity levels were low in the experimental lines. Genetic differentiation (FST ) between all pairs of the lines ranged from 0.13 to 0.83. Genetic clustering analyses based on multilocus genotypes of individuals showed that most individuals formed clearly defined clusters corresponding to the origins of the lines. These results suggest that Japanese quail experimental lines are highly structured. Microsatellite markers developed in this study may be effective for future genetic studies of Japanese quail.

Evolutionary and dispersal history of Eurasian house mice Mus musculus clarified by more extensive geographic sampling of mitochondrial DNA.

We examined the sequence variation of mitochondrial DNA control region and cytochrome b gene of the house mouse (Mus musculus sensu lato) drawn from ca. 200 localities, with 286 new samples drawn primarily from previously unsampled portions of their Eurasian distribution and with the objective of further clarifying evolutionary episodes of this species before and after the onset of human-mediated long-distance dispersals. Phylogenetic analysis of the expanded data detected five equally distinct clades, with geographic ranges of northern Eurasia (musculus, MUS), India and Southeast Asia (castaneus, CAS), Nepal (unspecified, NEP), western Europe (domesticus, DOM) and Yemen (gentilulus). Our results confirm previous suggestions of Southwestern Asia as the likely place of origin of M. musculus and the region of Iran, Afghanistan, Pakistan, and northern India, specifically as the ancestral homeland of CAS. The divergence of the subspecies lineages and of internal sublineage differentiation within CAS were estimated to be 0.37-0.47 and 0.14-0.23 million years ago (mya), respectively, assuming a split of M. musculus and Mus spretus at 1.7 mya. Of the four CAS sublineages detected, only one extends to eastern parts of India, Southeast Asia, Indonesia, Philippines, South China, Northeast China, Primorye, Sakhalin and Japan, implying a dramatic range expansion of CAS out of its homeland during an evolutionary short time, perhaps associated with the spread of agricultural practices. Multiple and non-coincident eastward dispersal events of MUS sublineages to distant geographic areas, such as northern China, Russia and Korea, are inferred, with the possibility of several different routes.

Prognosis after hepatic resection in patients with hepatocellular carcinoma, estimated on the basis of the morphometric indices.

To determine whether the morphometric indices of hepatocellular carcinoma (HCC) correlated with the prognoses, the microscopic morphometric values for 84 HCC cases treated by hepatic resection were studied using an image analyzer in relation to the survival rate and the gross classification. The mean survival time (MST) was 58 months in cases with a nucleocytoplasmic area ratio (N/C) of less than 0.28; this was significantly longer than the 38-month MST in cases with an N/C of more than 0.28 (P < 0.05). In stage III disease, the MST for cases with an N/C of less than 0.28 was 63 months, which was significantly longer than the MST of 13 months for cases with an N/C of more than 0.28. After relatively noncurative hepatic resection, the MST for cases with an N/C of less than 0.28 was 49 months, and this was significantly longer than the MST of 8 months for cases with an N/C of more than 0.28. The MST was 71 months for cases with a coefficient of variance of the nuclear form factor (NCV) of less than 5.5%, which was significantly longer than the MST of 33 months for cases with an NCV of more than 5.5% (P < 0.05). In stage III disease, the MST was 69 months for cases with an NCV of less than 5.5%, and this was significantly longer than the MST of 29 months for cases with an NCV of more than 5.5% (P < 0.05). In cases with an N/C of less than 0.28, 18% had vascular invasion and 38% had intrahepatic metastases, whereas in those with an N/C of more than 0.28, 62% had vascular invasion and 67% had intrahepatic metastases (P < 0.01, P < 0.05). Based on the results of these morphometric studies on HCC cases treated by hepatic resection, N/C and NCV may be useful as prognostic factors.

[Penetration into bone and joint tissues of rabbits and in vitro antibacterial activity of tosufloxacin].

The penetration of tosufloxacin (TFLX) into the bone and joint tissues of rabbits and its in vitro antibacterial activity were compared with those of lomefloxacin (LFLX) which is a efficacious drug for orthopedic infections. Serum levels of TFLX at 1, 2, 4 and 6 hours after oral administration (100 mg/kg) were 0.41, 0.65, 0.62 and 0.42 micrograms/ml, respectively. Except in synovial fluid and femur, TFLX concentrations in bone and joint tissues were higher than those in serum (0.69 approximately 1.92 micrograms/g in bone marrow of sternum, 0.55 approximately 1.53 micrograms/g in bone marrow of femur). TFLX concentrations in synovial fluid at 4 and 6 hours after the administration were equal to those in serum, which were lower than those of LFLX, but the ratio of tissue level/serum level of TFLX was similar to that of LFLX. TFLX was 8- to 64-fold more active than LFLX against Staphylococcus aureus (including methicillin-resistant S. aureus), Streptococcus pyogenes, Haemophilus influenzae, which are major pathogens of purulent osteomyelitis and arthritis. TFLX inhibited the growth of these bacteria at less than 0.39 micrograms/ml. These results indicate that TFLX is a useful drug for orthopedic infection.