Novel variations and loss of heterozygosity of BRCA2 identified in a dog with mammary tumors

OBJECTIVE: To establish novel polymorphic markers for analysis of loss of heterozygosity (LOH), so as to study the possible involvement of BRCA2 in mammary tumors obtained from dogs. SAMPLE POPULATION: Blood samples, mammary gland specimens, or mammary tumors from 3 tumor-bearing dogs and 10 tumor-free dogs. PROCEDURES: Nucleotide sequence analysis was performed with a DNA autosequencer. Loss of heterozygosity analysis was performed for markers established in the present study. The expression level of canine BRCA2 was quantified by real-time PCR analysis. RESULTS: 3 novel microsatellite markers with high heterozygosity rates (> 50%) were established, and the previously reported marker for canine BRCA2 gene locus was improved. These markers were used for the analysis of DNA from formalin-fixed and paraffin-embedded samples. By use of these markers, LOH in canine BRCA2 was identified as a result of recombination. In mammary tumor DNA that corresponded to the LOH-positive dog, the level of canine BRCA2 expression was decreased compared with that of nonneoplastic mammary gland tissue; the open reading frame contained 4 missense variations, 1 insertion variation, and 1 silent variation, some of which were localized to functional domains. CONCLUSIONS AND CLINICAL RELEVANCE: 3 novel polymorphic markers were developed for LOH analysis of canine BRCA2 and identified a dog with LOH with some variations in the functional domains. These markers could be useful for assessing the relevance of BRCA2 variation in mammary tumors of dogs.     

A pre-harvest method for predicting understory light levels created by artificial canopy gaps

A new method for predicting understory light levels around an artificial canopy gap prior to the selective cutting is presented. In two even-aged coppice stands, hemispherical photographs were taken at a height of 1.3 m, and crowns of each canopy stem on the photographs were identified by field observations. To predict light levels after cutting, we used computerized images of the hemispherical photographs, in which we digitally erased all crowns to be cut (named ‘erasing method’). Light levels were calculated by an analyzing program; HEMIPHOT. Light levels predicted by the ‘erasing method’ were compared with that calculated by the ordinary photograph-analyzing method using photographs taken after the actual cutting. The correlations between the two methods were highly significant in the two stands. The ‘erasing method’ is eminently effective for the prediction of understory light levels in an artificial canopy gap.     

Isolation of avian bornaviruses from psittacine birds using QT6 quail cells in Japan

Avian bornaviruses (ABVs) were recently discovered as the causative agents of proventricular dilatation disease (PDD). Although molecular epidemiological studies revealed that ABVs exist in Japan, no Japanese isolate has been reported thus far. In this study, we isolated four strains of Psittaciform 1 bornavirus from psittacine birds affected by PDD using QT6 quail cells. To our knowledge, this is the first report to isolate ABVs in Japan and to show that QT6 cells are available for ABV isolation. These isolates and QT6 cells would be powerful tools for elucidating the fundamental biology and pathogenicity of ABVs.     

Complementary process to response bias in the centromedian nucleus of the thalamus

Activity in several areas of the human brain and the monkey brain increases when a subject anticipates events associated with a reward, implicating a role for bias of decision and action. However, in real life, events do not always appear as expected, and we must choose an undesirable action. More than half of the neurons in the monkey centromedian (CM) thalamus were selectively activated when a small-reward action was required but a large-reward option was anticipated. Electrical stimulation of the CM after a large-reward action request substituted a brisk performance with a sluggish performance. These results suggest involvement of the CM in a mechanism complementary to decision and action bias.     

Effects of in utero and lactational exposure to the no-observed-adverse-effect level (NOAEL) dose of the neonicotinoid clothianidin on the reproductive organs of female mice

Recently, developmental exposure to clothianidin (CLO) has been shown to cause reproductive toxicity in male mice, but the effects in female mice remain to be clarified. Pregnant C57BL/6N mice were given a no-observed-adverse-effect-level (NOAEL) dose of CLO until weaning. We then examined ovaries of 3- or 10-week-old female offspring. In the CLO-administered group, morphological changes, a decrease in the immunoreactivity of the antioxidant enzyme glutathione peroxidase 4 (GPx4), and activation of genes in the steroid hormone biosynthesis pathway were observed in 3-week-old mice, and decreases of GPx4 immunoreactivity, 17OH-progesterone and corticosterone levels were observed in 10-week-old mice, along with high rates of infanticide and severe neglect, providing new evidence that developmental exposure to CLO affects juvenile and adult mice differently.     

Effects of low and high levels of maternal nutrition consumed for the entirety of gestation on the development of muscle,adipose tissue,bone, and the organs of Wagyu cattle fetuses

This study aimed to investigate the effects of high and low levels of energy intake during the entire gestation period on the skeletal muscle development, organ development, and adipose tissue accumulation in fetuses of Wagyu (Japanese Black) cows, a breed with highly marbled beef. Cows were allocated to a high-nutrition (n = 6) group (fed 120% of the nutritional requirement) or low-nutrition (n = 6) group (fed 60% of the nutritional requirement). The cows were artificially inseminated with semen from the same sire, and the fetuses were removed by cesarean section at 260 ± 8.3 days of fetal age and slaughtered. The whole-body, total muscle, adipose, and bone masses of the fetal half-carcasses were significantly higher in the high-nutrition group than the low-nutrition group (p = 0.0018, 0.009, 0.0004, and 0.0362, respectively). Fifteen of 20 individual muscles, five of six fat depots, nine of 17 organs, and seven of 12 bones that were investigated had significantly higher masses in the high-nutrition group than the low-nutrition group. The crude components and amino acid composition of the longissimus muscle significantly differed between the low- and high-nutrition groups. These data indicate that maternal nutrition during gestation has a marked effect on the muscle, bone, and adipose tissue development of Wagyu cattle fetuses.     

Effects of Ozone and/or Soil Water Stress on Growth and Photosynthesis of Fagaus Crenata Seedlings

The effects of ozone (O₃) and soil water stress, singly and in combination, on the growth and photosynthesis of Fagus crenata seedlings were investigated. Four-year-old seedlings were exposed to charcoal-filtered air (< 5 nmol mol^?1 O₃) or 60 nmol mol^?1 O₃, 7 hours per day (11:00–18:00), for 156 days from 10 May to 11 October 1999 in naturally-lit growth chambers at 20/15 °C (6:00–18:00/18:00–6:00). During the same period, half of the seedlings in each gas treatment received 250 mL of water at the 3-day intervals (well-watered treatment), while the rest received 175 mL of water at the 3-day intervals (water-stressed treatment). The exposure of the seedlings to O₃ caused reductions in the leaf, stem, root and whole-plant dry weights. The net photosynthetic rate at 350 µmol mol^?1 CO₂, the maximum net photosynthetic rate at saturated CO₂-concentration, carboxylation efficiency of photosynthesis and Rubisco content were significantly reduced by the exposure to O₃. The soil water stress induced reductions in the stem, bud and whole-plant dry weights, transpiration rate and leaf water potential during the midday. The additive effects of O₃ and soil water stress were observed on the dry matter production, leaf gas exchange rates and leaf water potential. As a result, the whole-plant dry weight of the seedlings exposed to both stresses was markedly reduced compared with that of the seedlings exposed to charcoal-filtered air and grown in the well-watered treatment.