Evaluation of RNA interference in developing porcine granulosa cells using fluorescence reporter genes

Gene silencing using small interfering RNA (siRNA) may be useful for functional analyses of unidentified genes expressed during cell differentiation. The present study was performed to evaluate RNA interference (RNAi) in porcine granulosa cells stimulated with bovine FSH, by using two fluorescence reporter genes: a plasmid encoding green fluorescent protein (GFP) and a plasmid encoding red fluorescent protein (RFP). The siRNA targeting GFP mRNA sequence (GFP-siRNA) with both plasmids was introduced into cultured cells by lipofection. GFP- and RFP-expressing cells were observed under fluorescence microscopy and analyzed by flow cytometry. Strong fluorescence was observed after introduction of both plasmids into cells. The intensity of green fluorescence generated by GFP was greatly suppressed by introduction of GFP-siRNA, showing an approximate 70% decrease in the ratio of green to red fluorescence. Consequently, we concluded that gene silencing by siRNA can be used to analyze the functions of genes of interest during differentiation of porcine granulosa cells.     

Genetic relationship of feed efficiency traits of bulls with growth and carcass traits of their progeny for Japanese Black (Wagyu) cattle

Genetic parameters for feed efficiency traits of 740 Wagyu bulls and growth and carcass traits of 591 of their progeny, and the genetic relationship between the traits of bulls and their progeny were estimated with the residual maximum likelihood procedure. The estimations were made for the test periods of 140 days (77 bulls), 112 days (663 bulls) and 364 days (591 steer progeny). Feed efficiency traits of bulls included feed conversion ratio (FCR), phenotypic residual feed intake (RFI_phe) and genetic residual feed intake (RFI_gen). Progeny traits were bodyweight at the start of the test (BWS), bodyweight at finish (BWF), average daily gain (ADG), rib eye area (REA), marbling score (MSR), dressing percentage (DRS) and subcutaneous fat thickness (SFT). The estimated heritability for MSR (0.52) was high and for BWS (0.35), BWF (0.40) and ADG (0.30) were moderate, whereas REA, DRS and SFT were low. Positive genetic correlations among BWS, BWF, ADG and SFT and negative genetic correlations between MSR and DRS and between REA and SFT were found. The genetic correlations between residual feed intake (RFI_phe and RFI_gen) of bulls and bodyweights (BWS and BWF) of their progeny ranged from ?0.27 to ?0.61. Residual feed intake was positively correlated with REA and DRS and negatively correlated with MSR and SFT. No responses in ADG and weakly correlated responses in REA and DRS of progeny were found to select against feed efficiency traits of bulls. The present experiment provides evidence that selection against lower RFI (higher feed efficiency) would be better than selection against lower FCR for getting better correlated responses in bodyweights.     

Fractography of shear failure surface of softwood decayed by brown-rot fungus

In order to investigate shear failure surface of decayed wood, wood pieces of ezomatsu (Picea jezoensis) were exposed to brown-rot fungus (Fomitopsis palustris), and standard shear test in radial plane was conducted. The failure surfaces were examined by scanning electron microscopy and surface roughness measurements were also conducted. Transwall failure that the crack elongated parallel to the tracheid axis was observed in the earlywood region through all phases of decay. Intrawall failure principally occurred in the latewood region on the early phase of decay. When decay progressed considerably, transwall failure that the crack elongated perpendicularly to the tracheid axis occurred. And transwall failure was also dominant failure morphology in the latewood region. Fragments of tracheids which were partly peeled out from S₂ layers were observed in some specimens. Size and appearance frequency of fragments of tracheid were smaller and lower when decay progressed. Arithmetic average roughness, which was the index of fragment size and appearance frequency, had positive correlation to shear strength ratio. Especially, line surface roughness of radial direction, which was measured across the radial files of tracheid, had the highest correlation to the shear strength ratio. The surface roughness would be a good indicator to evaluate the decay degree.     

Effects of salinity on the nail-holding power of dimension lumber used in light-frame wood building

We studied the effect of salinity on nail-hold-ing power in wood construction. In saline solution, the holding power of nails was less than in purified water. With the increase of salt concentration, t...     

Effect of Simulated Acid Rain on Deterioration of Concrete

In the study the long-term exposure tests to simulated acid rain were performed in order to clarify the effect of acid rain on deterioration of concrete. Mortar specimens with 40 mm in width, 15 mm in thickness and 160 mm in length were used for the tests. At each time after the fixed rainfall was attained, those were tested physically and analyzed chemically. Finally total rainfall of 9000mm was given to the specimens. From the test results, it was confirmed that the eroded depth of the specimen has a good linear relation to the total rainfall under simulated acid rain with various pH. Surface erosion rates of the mortar specimens with an ordinary mix proportion under simulated acid rains with pH 3.0 and 2.5 were about 1.2 and three times larger than that under pH 5.6, respectively. It was also confirmed that flexural strength of the specimens with an ordinary mix proportion hardly changed under low pH simulated acid rain even after total rainfall of 9000mm was given.     

Production and analysis of reciprocal hybrids between <Emphasis Type="Italic">Asparagus officinalis</Emphasis> L. and <Emphasis Type="Italic">A. schoberioides</Emphasis> Kunth

The genus Asparagus (Asparagaceae) encompasses 100–300 species, including the important vegetable crop, A. officinalis (garden asparagus). Previous attempts to hybridize garden asparagus with A. densiflorus (Kunth) Jessop, with the aim of introducing disease resistance, were unsuccessful because of the failure of endosperm development. In the present study, reciprocal interspecific hybrids between garden asparagus and A. schoberioides Kunth were generated by hand pollination. The F1 hybrids were analyzed by using both morphological and molecular techniques. This is the first report describing the production of an interspecific hybrid between garden asparagus at the diploid level (2n = 2x = 20) and its diploid (2n = 2x = 20) wild relative, A. schoberioides. Morphological characteristics of candidate hybrids were a mixture of those found in the parents, and cytological and RFLP analyses confirmed that morphologically intermediate plants were indeed diploid hybrids of those two species. In other words, post-zygotic isolation is not complete between phylogenetically distinct these two species. Our results suggest that other Asparagus species may be capable of hybridizing with A. officinalis and that introducing wild characters through interspecific hybridization may offer advantages for breeding for novel traits. Takuro Ito and Toshinori Ochiai authors are contributed equally to this work