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.     

Thermal stability of the synthetic peptides with the sequence of fish fast skeletal muscle tropomyosin

Tropomyosins from fish skeletal muscle show high amino acid sequence homology, although their thermal stability is clearly different among species. In order to determine the regions that are responsible for the stability of this protein, five synthetic peptides of 30mer were synthesized by Fmoc method, based on the sequence of walleye pollack Theragra chalcogramma fast skeletal muscle tropomyosin, namely, N terminal Met¹-Lys³⁰, the variable region Asp⁸⁴-Leu¹¹³, the middle region Val¹²⁸-Ala¹⁵⁷, the region containing the conservative Cys (Leu¹⁷⁶-Lys²⁰⁵), and C terminal Asp²⁵⁵-Ile²⁸⁴. The thermal stability of these peptides was measured by circular dichroism and differential scanning calorimetry. The helical contents of these peptides were decreased in a temperature-dependent manner, although they showed no clear melting temperature, suggesting that the enthalpy necessary for the complete denaturation of these peptides was low. Peptides Asp²⁵⁵-Ile²⁸⁴ and Asp⁸⁴-Leu¹¹³ showed the highest and second highest α-helical contents, respectively, and the other peptides gave rise to lower α-helical contents.     

Immunohistochemical localization of steroidogenic enzymes in the corpus luteum and the placenta of the ribbon seal (Phoca fasciata) and steller sea lion (Eumetopias jubatus)

To study the luteal and placental function of pinnipeds, we analyzed the localization of steroidogenic enzymes (P450scc, 3 beta HSD and P450arom) in the corpus luteum and the placenta of ribbon seals (Phoca fasciata) and Steller sea lions (Eumetopias jubatus) immunohistochemically. P450scc and 3 beta HSD were present in all luteal cells of both species. Almost all of the luteal cells were immunostained for P450arom, while P450scc and 3 beta HSD were negatively immunostained in placentae and P450arom was present in the syncytiotrophoblast of placentae. These findings suggest that 1) corpora lutea of both species synthesize pregnenolone, progesterone and estrogen during the entire pregnancy period, and 2) like other terrestrial carnivores in the suborder Caniformia, placentae of both species do not have the capability for synthesizing progesterone in the latter half of active pregnancy period.     

Purulent rhinitis and otitis caused by Pseudomonas aeruginosa in sheep showered with contaminated 'shower wash'

Three crossbred lowland ewes developed a severe purulent rhinitis and another three ewes developed a severe purulent otitis externa/media after being showered with a wash that had been used 24 to 48 hours before on a separate group of Cheviot ewes with lesions of dermatitis. Pseudomonas aeruginosa was isolated in pure growth from the aural and nasal abscesses and also from the dermatitis lesions. Extended antibiotic susceptibility testing and the random amplification of polymorphic DNA indicated that a single clonal type was associated with the rhinitis and otitis and with the dermatitis, providing strong evidence of an epidemiological link between the lesions of dermatitis and the aural and nasal abscesses through the use of the contaminated 'shower wash'.     

Isolation of microsatellite markers by in silico screening implicated for genetic linkage mapping in Japanese pufferfish Takifugu rubripes

ABSTRACT:   The search for dinucleotide repeat microsatellites within scaffolds 1–25 of genome database JGI Fugu v3.0 for the pufferfish Takifugu rubripes revealed that 80% of microsatellite loci consisted of five to 13-fold repeats with locus-specific differences in density. Eleven out of 15 microsatellite loci isolated from the database with which genotyping using wild pufferfish was successfully performed showed polymorphism; that is, the means of the number of alleles and expected and observed heterozygosities at these 11 loci were 21.8, 0.915 and 0.829, respectively. It was confirmed that eight out of the 11 polymorphic loci were inherited through the Mendelian law and one pair of microsatellite loci derived from the same scaffold was linked. These results demonstrated that these loci are useful for constructing a linkage map in the pufferfish as DNA markers.     

Thermal tolerance of a rainbow trout <Emphasis Type="Italic">Oncorhynchus mykiss</Emphasis> strain selected by high-temperature breeding

ABSTRACT:   Thermal tolerance was studied in a rainbow trout strain successively selected through high temperature breeding at 20–27°C since 1966 in Miyazaki Prefecture, Japan. The hatching rate and fry mortality at high temperatures were examined in the selected strain along with normal strains cultured at water temperature of 9–17°C. The hatching rate of embryos fertilized at either 10 or 14°C and subsequently subjected to high temperatures in the blastula or neurula stage of the selected strain, was marginally higher than that of the normal strain counterparts. The upper 50% lethal temperatures (LT₅₀) for embryos in the early segmentation, blastula and neurula stages of the selected strain were also higher than those of the normal strain counterparts. Death temperatures and LT₅₀ of fries acclimated to 20°C of the selected strain were significantly higher than those of the normal strains. However, no difference in the critical thermal maximum was detected between the different strains. These results suggest that the selected strain of rainbow trout established by selecting successively for many generations at high temperatures acquired a degree of thermal tolerance.