Quantitative genotyping by amplifying the polymorphic sequences of Pre-Melanosomal Protein (PMEL17) gene using real-time polymerase chain reaction in chickens

1. This study was conducted to develop a quantitative genotyping system of chimaeric chickens by real-time PCR. 2. The polymorphisms in exons 7 and 11 of PMEL17 gene, which is one of the major genes affecting plumage colour, were identified from White Leghorn, Barred Plymouth Rock and Rhode Island Red chickens. 3. Quantitative genotyping was successfully performed by real-time PCR using polymorphic sequence-specific TaqMan Probes. 4. This methodology can support future research of germline chimaeric chickens as well as the application of germ cell transfer technique.     

Association of tumour‐infiltrating regulatory T cells with adverse outcomes in dogs with malignant tumours

Regulatory T cells (Tregs) infiltrate into a variety of tumour tissues and associate with poor prognosis in humans. However, data on association of Treg infiltration with prognosis is limited in canine tumours. The purpose of this study was to examine the number of tumour‐infiltrating Tregs and its association with overall survival (OS) in dogs with malignant tumours. The following 168 canine tumours were included: 37 oral malignant melanomas (OMMs); 14 oral squamous cell carcinomas (OSCCs); 16 pulmonary adenocarcinomas (PAs); 37 mammary carcinomas (MCs); 36 mast cell tumours (MCTs) and 28 hepatocellular carcinomas (HCCs). Normal tissues were obtained from 8 healthy dogs as controls. The number of forkhead box P3 (Foxp3)‐positive Tregs in intratumoral and peritumoral areas was investigated by immunohistochemistry. OS was compared between high and low Treg groups. The number of intratumoral and peritumoral Foxp3‐positive Tregs was significantly higher in OMM, OSCC, PA and MC compared with each normal tissue. There were few Foxp3‐positive Tregs in MCT and HCC. With intratumoral Tregs, the OS in the high Treg group was significantly shorter than that in the low Treg group in OMM, OSCC and PA. With peritumoral Tregs, there was no significant difference for OS between the 2 groups in each tumour type. These results suggest that Tregs infiltrate into a variety of canine tumours and the abundance of Tregs are associated with poor prognosis in some solid tumour types.     

Accumulation of carbohydrates and proline in water-stressed soybean (Glycine max L.)

Abstract

A wide range of metabolites accumulates under water stress depending on certain metabolic alterations. For example, free amino acids, especially free proline, accumulate in response to water stress ((l)). Proline accumulation is closely connected with carbohydrate metabolism. Thus, a carbohydrate requirement for proline accumulation in water-stressed leaves has been reported (2, 3).     

Improved method for easy and rapid determination of root length of vegetables

We developed an improved method for the determination of the root length of crisphead lettuce (Lactuca sativa L.) and Chinese cabbage (Brassica pekinensis Rupr.). Boiling of soil containing roots and stirring to collect the roots in water reduced the contamination with debris. The time required for removing the debris was shortened by 30% as compared to washing of roots with water. Boiling had no effect on the nitrogen concentration, but resulted in changes of the phosphate, potassium and calcium concentrations of roots. Use of a crosscounter plate shortened by 30% the time required for the line intersect method. The total time required for root length determination from a 1 kg soil sample was only 15.9 min.     

The Effect of Eating Sea Cucumber Jelly on Candida Load in the Oral Cavity of Elderly Individuals in a Nursing Home

We conducted a double-blind randomized controlled study of elderly individuals in a nursing home to investigate the effect of the consumption of jelly containing sea cucumber on their oral Candida load. The jelly contained a hydrolysate of the sea cucumber Stichopus japonicus, which contained triterpene glycosides called holotoxins. The holotoxins worked as a fungicide, and their minimum inhibitory concentrations for Candida albicans were 7 µg/mL. Eight individuals in the nursing home took the sea cucumber jelly for a week and their oral Candida were counted before and after the intervention. Nine individuals took a control jelly without S. japonicus. The sea cucumber jelly showed inhibitory effects on the oral Candida. Thus, daily consumption of the S. japonicus jelly has the potential to reduce the oral Candida load in the elderly in nursing homes.     

Cell growth and nutrient uptake by cell suspensions of <Emphasis Type="Italic">Cupressus lusitanica</Emphasis>

 Conditions for cell growth of suspension cultures of Cupressus lusitanica, which has high β-thujaplicin productivity, were studied. The medium that provided the highest growth rate was IS-1 medium (pH 5.5), modified from Gamborg B5 medium containing 32 mM of total nitrogen. Its NO₃-N/NH₄-N ratio was 30 : 2. The maximum growth represented a 25-fold increase over the initial biomass on a fresh weight basis after 30 days of culture in this medium. The highest cell growth was obtained with an initial pH of 3.5–5.5, but the pH of the medium settled to about pH 4.0 from any of the initial pH values in this report. The cells cultured under this condition were able to produce a high level of β-thujaplicin. Received: September 7, 2001 / Accepted: May 7, 2002 Present address: Teijin Ltd., Ehime 791-8530, Japan Present address: Q'SAI Co. Ltd., Fukuoka 811-3422, Japan Part of this report was presented at the 10th international symposium on wood and pulp chemistry, Yokohama, Japan, June 1999 Correspondence to:K. Fujita     

Epidural spinal myelolipoma in a dog

Epidural spinal myelolipoma was diagnosed in a 13-year-old, male Siberian husky that was referred for evaluation of progressive pelvic limb paresis and urinary incontinence. An epidural mass was detected by magnetic resonance imaging and computed tomography. The mass was removed and identified histopathologically as an epidural myelolipoma. Pelvic limb paresis improved after surgery, but urinary retention associated with neurological bladder dysfunction persisted.     

Effectiveness of adjusting for heterogeneity of variance in genetic evaluation of Japanese Black cattle

Heterogeneity of variance among subclasses of an effect is a potential source of bias in genetic evaluation. The objectives of this study were to quantify the heterogeneity of variance in carcass weight in Japanese Black cattle, to develop an adjustment method to account for the heterogeneity, and to evaluate the effectiveness of the method. A total of 96,950 records were collected from steers and heifers slaughtered from 1997 to 2005. These records were grouped into 2,767 farm-market-year-sex subclasses. Fourteen log-linear models for the variances were set up to estimate the heterogeneous phenotypic variances within subclasses. Schwarz's Bayesian information criterion was used for model selection. The preadjustment of records to a baseline variance was based on maximum likelihood estimates obtained from the selected model. As a result of adjustment, the SD, the CV, and the Gini coefficient for the phenotypic variance decreased by 68.6, 69.8, and 70.1%, respectively. When the top 5% of sires and top 1% of dams were selected, Spearman's rank correlation coefficients between the adjusted and unadjusted data were 0.95 for the selected sires and 0.78 for the selected dams. The effectiveness of the adjustment was evaluated in terms of the ability to predict breeding values, using the results of the successive genetic evaluations. Mean squared error between the parent averages and actual predicted values of the genetic merit for the sires whose progeny had a carcass record only from 2003 to 2005 was significantly reduced by the adjustment (P < 0.05). The results suggest that the genetic evaluation becomes more accurate by adjusting the data using the procedure developed in this study.