Interspecies nuclear transfer embryos reconstructed from cat somatic cells and bovine ooplasm

This study was conducted to investigate the developmental capacity of domestic cat-bovine reconstructed embryos via interspecies somatic cell nuclear transfer (iSCNT) and to observe the mitochondrial DNA (mtDNA) content of the iSCNT embryos. The iSCNT embryos were generated using mixed-breed domestic cat fibroblasts as donor cells and enucleated bovine oocytes as the recipient cytoplasm. When the developmental capacities of iSCNT embryos and parthenogenic bovine embryos were compared, there was no difference (P>0.05) in the rates of cleavage and development to the 8-cell stage (86.6 vs. 84.0% and 32.2 vs. 36.2%, respectively). However, in contrast to development of parthenogenic embryos to the morula and blastocyst stages, no iSCNT embryos (0/202) developed beyond the 8-cell stage. For mtDNA analysis, iSCNT embryos at the 1-cell, 2-cell, 4-cell and 8-cell stages were randomly selected. Both cat and bovine mtDNA quantification analysis were performed using quantitative PCR. The levels of both cat and bovine mtDNA in cat-bovine iSCNT embryos varied at each stage of development. The cat mtDNA concentration in the iSCNT embryos was stable from the 1-cell to 8-cell stages. The bovine mtDNA in the iSCNT embryos at the 8-cell stage was significantly lower than that at the 4-cell stage (P<0.05). No difference in the proportions of cat mtDNA in the iSCNT embryos was found in any of the observed developmental stages (1- through 8-cell stages). In conclusion, bovine cytoplasm supports domestic cat nucleus development through the 8-cell stage. The mtDNA genotype of domestic cat-bovine iSCNT embryos illustrates persistence of heteroplasmy, and the reduction in mtDNA content might reflect a developmental block at the 8-cell stage.     

Trans-endothelial and trans-epithelial transfer of lactoferrin into the brain through BBB and BCSFB in adult rats

The transportation of intravenously administered bovine lactoferrin (bLF) into the cerebrospinal fluid (CSF) was immunohistochemically investigated in adult rats. Administered bLF was detected in the vesicular membranes of endothelial cells in cerebral blood vessels 10 min after the infusion. Numerous immunoreactive small vesicles were also detected at the ependymal cells in the choroid plexus. Moreover, the bLF concentration in the CSF was significantly increased at 1-2 hr after the intravenous infusion of bLF (10 or 30 mg/kg). These findings clearly demonstrate that LF is possibly transported into the brain matter even in adult animals.     

Antiproliferative effects of the readily extractable fractions prepared from various citrus juices on several cancer cell lines.

To eliminate the masking effect by flavonoid glycosides, which comprise approximately 70% of conventionally prepared sample, the readily extractable fraction from Citrus juice, which was prepared by adsorbing on HP-20 resin and eluting with ethanol and acetone from the resin, was subjected to antiproliferative tests against several cancer cell lines. Screening of 34 Citrus juices indicated that King (Citrus nobilis) strongly inhibited proliferation of all cancer cell lines examined. Sweet lime and Kabuchi inhibited three of the four cancer cell lines. In contrast, these samples were substantially less cytotoxic toward normal human cell lines.     

Accumulation of paralytic shellfish poisoning toxin by the swimming crab <Emphasis Type="Italic">Charybdis japonica</Emphasis> in Kure Bay,Hiroshima Prefecture

The accumulation of paralytic shellfish poisoning (PSP) toxin in the crab Charybdis japonica was investigated in Kure Bay, when a bivalve Mytilus galloprovincialis, in the same water, showed toxicity caused by PSP toxin. In 2005, 17 specimens of the crab had PSP toxin over 4 MU/g in the hepatopancreas, and the highest toxicity was at 37.4 MU/g. Since the regulation limit of PSP for crab hepatopancreas was set in 2004, this is the first observation of regulatory level of PSP toxin in C. japonica. Unlike in the hepatopancreas of the crab, the hazardous level of the toxin was not detected in the muscle of the cephalothorax and the appendage. The toxin accumulation in the crab was also investigated by feeding toxic mussels to the crab. The crab retained the toxin mainly in the hepatopancreas, and the ratio of retention in the crab was from 12.9 to 24.6%. The toxin profiles, shown in the feeding experiments, suggest that the transformation of PSP toxin occurs in the crab because dcGTX2 and dcGTX3 was detected in all crab specimens despite the lack of these analogs in the mussels used as feed.     

Fatigue of structural plywood under cyclic shear through thickness II: a new method for fatigue life prediction

For plywood specimens under shear through the thickness, a fatigue life prediction method based on strain energy has been newly developed with the fatigue process and failure criterion applicable to various loading conditions. Once the fatigue process and failure criterion of the plywood specimen were determined by the fatigue data measured under a loading condition other than the square loading waveform, the fatigue life of a specimen under various loading conditions could be predicted easily and accurately by the first cycle loading test. The relationship between stress level and the predicted fatigue life was also similar to that between stress level and the experimentally determined fatigue life. The fatigue life prediction method proposed may be widely applicable to the prediction of the fatigue life of solid wood and wood composites.     

Heat stress decreases plasma vitamin C concentration in lactating cows

Heat stress causes oxidative stress, which decreases plasma levels of vitamin C, an antioxidative vitamin in pigs and poultry. We investigated the plasma vitamin C concentration in lactating cows exposed to a hot environment temperature. In the first experiment, plasma vitamin C concentrations of 8 lactating cows were measured in summer and autumn. The concentration was found to be significantly lower in summer. The second experiment was performed using 4 lactating cows in a crossover design. The ambient temperature was set at 18 °C in the control treatment and at 28 °C in the heat-stressing treatment. Milk yields and feed intake tended to be lower in the stressing than control treatment. Rectal temperature and respiration rate were significantly (P < 0.05) higher in the heat-stressing treatment. The plasma vitamin C concentration was significantly (P < 0.05) lower in the heat-stressing period. These results indicate that heat stress decreases the plasma vitamin C concentration. Dietary supplementation with vitamin C may be beneficial for lactating cows in hot weather.     

Plasma concentration of vitamin C in dogs with a portosystemic shunt

Most mammals, including dogs, synthesize vitamin C in the liver. We measured the plasma concentration of vitamin C to assess the body vitamin C status in 15 dogs with a portosystemic shunt (PSS). The plasma biochemical parameters indicated liver abnormalities in all the dogs. In contrast, the plasma concentration of vitamin C ranged from 2.21 to 9.03 mg/L in the 15 dogs and was below the reference range (3.2 to 8.9 mg/L) in only 2 dogs. These findings suggest that vitamin C status is not impaired in dogs with PSS.     

Natural remission of nephrotic syndrome in a dog with immune-complex glomerular disease.

A nephrotic syndrome caused by immune-complex glomerular disease was diagnosed in a 4-year-old male Great Dane. The syndrome was characterized by proteinuria, hypoproteinemia, hypoalbuminemia, hypercholesterolemia, and subcutaneous edema. Renal biopsy revealed segmental membranous glomerular disease. The edema underwent complete remission 18 days after admission. Two months after admission, there was no clinical or laboratory evidence of glomerular disease. Periodic reevaluation of the dog during the next 2 years revealed recurrence of proteinuria, but no other clinical or laboratory abnormalities. Serial renal biopsies revealed persistence, but no appreciable increase, in the severity of the segmental membranous glomerular disease. The natural course of the nephrotic syndrome and immune-complex glomerular disease has been associated with unpredictable variability. It was concluded that the widespread use of corticosteroid or immunosuppressant therapy in dogs with immune complex glomerular disease should be withheld until the natural course of the disease has been evaluated in a significant number of patients and until the results of well-controlled clinical studies confirm or deny their therapeutic value.