Establishment of a syngeneic orthotopic model of prostate cancer in immunocompetent rats

We previously established 3 cell lines (PLS10, PLS20 and PLS30) from a chemically-induced prostate carcinoma in F344 rats, and demonstrated high potential for metastasis in nude mice. In the present study, we investigated the feasibility of establishing an orthotopic model using the 3 rat prostate cancer cell lines in immunocompetent rats with the aim of resolving species-mismatch problems and defects of immune systems. The PLS10, PLS20 and PLS30 cell lines were injected into the ventral prostates of 6-week-old rats, which were then sacrificed at experimental weeks 4 and 8. Tumor mass formation was found in rats with PLS10, but not in those with PLS20 or PLS30. Additionally, metastatic carcinomas could be detected in lymph nodes and lungs of PLS10-inoculated rats. Genetic analysis demonstrated K-ras gene mutations in PLS10 and PLS20, but not in PLS30 cells. There were no mutations in p53 and KLF6. In conclusion, we established a syngeneic orthotopic model for prostate cancer in immunocompetent rats simulating human castration-resistant prostate cancer (CRPC), which should prove useful for development and validation of therapeutic agents, especially with immunotherapy.     

Genetic diversity and similarity of pear (<Emphasis Type="Italic">Pyrus</Emphasis> L.) cultivars native to East Asia revealed by SSR (simple sequence repeat) markers

Simple sequence repeat (SSR) markers were used to assess genetic diversity and relationship of Pyrus L. cultivars native mainly to East Asia. A total of 168 putative alleles were generated from six primer-pairs (BGA35, KU10, BGT23b, NH004a, NH011b and NH015a). All the SSR markers showed a high level of genetic polymorphism with a mean of 28 putative alleles per locus and the heterozygosity of 0.63. The Dice’s similarity coefficient between cultivars ranged from 0.02 to 0.98 and Occidental pears generally had low affinities to Asian pears. Ten major groups were generated from all the accessions by UPGMA clusters analysis. Chinese sand pears consisted of four groups with Chinese white pears and Japanese pears, of which Chinese sand pears occurred in all four groups, presenting a large genetic diversity, Chinese white pears were included in three groups, and Japanese pears only fell into one group. In the dendrogram, Chinese sand pears and Chinese white pears did not form discrete group, even subgroups. Some Japanese pear cultivars had high affinities to Chinese sand pear cultivars. These findings supports the authors’ previous viewpoints of Chinese white pears as a variety or an ecotype of Chinese sand pears (P. pyrifolia var. sinensis (Lindley) Y. Teng et K. Tanabe) and the progenitor of Japanese pears coming from China. Cultivars of P. ussuriensis Maxim. were clustered together with one clone of P. hondoensis Nakai et Kikuchi, a relative species of P. ussuriensis. Cultivars of P. communis L. and other Occidental species formed three independent groups and were distant from most Asian pears, except for P. betulaefolia Bge.     

Leaf margin phenotype-specific restriction-site-associated DNA-derived markers for pineapple (Ananas comosus L.)

To explore genome-wide DNA polymorphisms and identify DNA markers for leaf margin phenotypes, a restriction-site-associated DNA sequencing analysis was employed to analyze three bulked DNAs of F1 progeny from a cross between a ‘piping-leaf-type’ cultivar, ‘Yugafu’, and a ‘spiny-tip-leaf-type’ variety, ‘Yonekura’. The parents were both Ananas comosus var. comosus. From the analysis, piping-leaf and spiny-tip-leaf gene-specific restriction-site-associated DNA sequencing tags were obtained and designated as PLSTs and STLSTs, respectively. The five PLSTs and two STSLTs were successfully converted to cleaved amplified polymorphic sequence (CAPS) or simple sequence repeat (SSR) markers using the sequence differences between alleles. Based on the genotyping of the F1 with two SSR and three CAPS markers, the five PLST markers were mapped in the vicinity of the P locus, with the closest marker, PLST1_SSR, being located 1.5 cM from the P locus. The two CAPS markers from STLST1 and STLST3 perfectly assessed the ‘spiny-leaf type’ as homozygotes of the recessive s allele of the S gene. The recombination value between the S locus and STLST loci was 2.4, and STLSTs were located 2.2 cM from the S locus. SSR and CAPS markers are applicable to marker-assisted selection of leaf margin phenotypes in pineapple breeding.     

A practical method for apple cultivar identification and parent-offspring analysis using simple sequence repeat markers

Differentiation of cultivars with simple sequence repeat (SSR) markers is a very useful technique for the true-to-type characterization of cultivars and clarification of parent-offspring relationships. We developed an SSR marker set for cultivar identification comprising 15 markers that were screened from 46 previously published SSRs. This marker set could be used for apple varieties including Malus × domestica and/or other Malus species. These SSRs successfully characterized 95 apples, including the leading and major founding cultivars used worldwide for modern apple breeding. Therefore, this marker set could be applied to almost all apple cultivars. We also analyzed the parent-offspring relationships of 69 cultivars by considering allele transmissions. This analysis revealed the true parentage of the following seven cultivars: ‘Kizashi’, ‘Chinatsu’, ‘Honey Queen’, ‘Haruka’, ‘Seirin’, ‘Ozenokurenai’, and Morioka #48. This analysis also revealed a parentage discrepancy for ‘Hacnine’. From the parent-offspring analysis, two microsatellite mutation events at alleles inherited from pollen parents were observed.     

Tree mortality in a natural mixed forest affected by stand fragmentation and by a strong typhoon in northern Japan

Windthrow is recognized as an extremely significant disturbance in many forests. Its effects are increased by stand fragmentation, which exposes the stand to strong winds. In this study, we investigated the change in tree mortality in fragmented stands with the distance from the stand edge, in a conifer–broadleaved mixed forest in northern Japan. We set out experimental plots having three stand sizes (400, 1,600, 6,400 m²) and examined the stand dynamics over 10 years. Tree mortality tended to be higher in smaller stands, although an effect of stand size was found only in the first 5 years of the study. Distance-dependent individual mortality was obvious in Abies sachalinensis, the most vulnerable major tree species, again in the first 5 years, suggesting that wind-risk management should emphasize the area within ca. 20 m from the stand edge. No distant-dependent effect was found, however, in the latter 5 years, in which there was a strong typhoon; tree deaths occurred throughout the stand (irrespective of distance within) as a result of this event. We conclude that the severity of wind can cause the features of wind-induced damage to differ; stand edge effects peculiar to a small forest are unlikely to occur with particularly strong winds, and the effect of fragmentation might therefore be clear only in weaker disturbances.     

Does BDNF have pre- or postsynaptic targets?

A type of synaptic plasticity in the brain called long-term potentiation (LTP) is thought to form the molecular basis of learning and memory. In a Perspective, Manabe discusses new findings (Kovalchuk et al.) showing brain-derived neurotropic factor modulates LTP by binding to TrkB receptors on the postsynaptic neuron.     

Spatial and temporal variations of sediment quality in and around fish cage farms: A case study of aquaculture in the Seto Inland Sea, Japan

ABSTRACT: Fish cage farming generates large amounts of organic waste in the form of unconsumed feed and fecal matter, resulting in sediment deterioration and a threat to its own sustainability. Field studies analysed the scale of the impact of fish cage farming on sediment quality, variability of impact and spread of sediment enrichment in the vicinity of a fish farm. Two fish culture sites near the Fukuyama area of the Seto Inland Sea of Japan were chosen for the study. There was a significant difference between the quality of the sediment in aquaculture and non-aquaculture areas. The sediment underlying the fish cage farms was found to be extremely reducing, acidic and sulfidic. Sediment ignition loss did not vary markedly with the season. However, the redox potential and acid volatile sulfide content of aquaculture areas did show some seasonal variation. Sediment encircling a 165 m radius from the center of a farm in Yokota was judged as organically enriched by fish farming. A variation in sediment quality within the two sites and also within the stations was observed. Based on sediment quality, the stations could be categorized as less affected, highly affected or moderately affected.     

Increase in muscle endurance in mice by dietary Yamabushitake mushroom (Hericium erinaceus) possibly via activation of PPARδ

Skeletal muscle fiber is largely classified into two types: type 1 (slow‐twitch) and type 2 (fast‐twitch) fibers. Meat quality and composition of fiber types are thought to be closely related. Previous research showed that overexpression of constitutively active peroxisome proliferator‐activated receptor (PPAR)δ, a nuclear receptor present in skeletal muscle, increased type 1 fibers in mice. In this study, we found that hexane extracts of Yamabushitake mushroom (Hericium erinaceus) showed PPARδ agonistic activity in vitro. Eight‐week‐old C57BL/6J mice were fed a diet supplemented with 5% (w/w) freeze‐dried Yamabushitake mushroom for 24 hr. After the treatment period, the extensor digitorum longus (EDL) muscles were excised. The Yamabushitake‐supplemented diet up‐regulated the PPARδ target genes Pdk4 and Ucp3 in mouse skeletal muscles in vivo. Furthermore, feeding the Yamabushitake‐supplemented diet to mice for 8 weeks resulted in a significant increase in muscle endurance. These results indicate that Yamabushitake mushroom contains PPARδ agonistic ligands and that dietary intake of Yamabushitake mushroom could activate PPARδ in skeletal muscle of mice. Unexpectedly, we observed no significant alterations in composition of muscle fiber types between the mice fed control and Yamabushitake‐supplemented diets.     

Identification of tightly linked SSR markers for flower type in carnation (Dianthus caryophyllus L.)

Single or double flower type is one of the most important breeding targets in carnation (Dianthus caryophyllus L.). We mapped the D ₈₅ locus, which controls flower type, to LG 85P_15–2 using a simple sequence repeat (SSR)-based genetic linkage map constructed using 91 F₂ progeny derived from a cross between line 85–11 (double flower) and ‘Pretty Favvare’ (single flower). A positional comparison using SSR markers as anchor loci revealed that the map positions of the D ₈₅ locus corresponded to the single locus controlling the single flower type derived from wild D. capitatus ssp. andrzejowskianus. We identified four co-segregating SSR markers on the D ₈₅ locus. Verification of the SSR markers in commercial cultivars revealed that two of the four SSR markers (CES0212 and CES1982) were tightly linked to the D ₈₅ locus, and amplified a 176-bp and 269-bp allele, respectively, which were common and unique to double flower cultivars. The map positions of the D ₈₅ locus and the tightly linked SSR markers will be useful for determining the genetic basis of flower type and for marker-assisted breeding of carnations.