Porcine MHC classical class I genes are coordinately expressed in superantigen-activated mononuclear cells

The expression of the major histocompatibility complex (MHC) classical class I genes is important for the adaptive immune response to target virus-infected cells and cancer cells. The up-regulation of the MHC is achieved by hormonal/cytokine signals including IFN-γ-inducible elements. The swine leukocyte antigen (SLA), the MHC class I region of pigs, consists of the duplicated classical class I genes, SLA-1, SLA-2 and SLA-3, but the molecular mechanisms involved in their up-regulation after T cell stimulation have not been fully elucidated. In order to better understand some of the putative regulatory mechanisms of SLA class I gene expression in activated T cells, we examined the coordinated expression of the SLA classical class I, IFN-γ and interferon regulatory factor-1 (IRF-1) genes in the peripheral blood mononuclear cells (PBMCs) of SLA homozygous Clawn miniature swine stimulated for 72h with either IFN-γ or an enterotoxin produced by Staphylococcus aureus. This enterotoxin, toxic shock syndrome-1 (TSST-1), is known to act as a superantigen (sAG) to activate the T cells in various vertebrate species. We showed by using mAbs and flow cytometry that the CD4(+)CD25(+) cell number of swine PBMCs was also increased by TSST-1 and to a lesser degree by IFN-γ. Time course analyses of the expression of the IFN-γ, IRF-1 and the three classical class I genes, SLA-1, SLA-2, and SLA-3, in PBMCs by quantitative real-time PCR revealed a transitory response to TSST-1 or IFN-γ stimulation. The IFN-γ mRNA levels in the PBMCs were continuously up-regulated over the first 48h by TSST-1 or IFN-γ. In contrast, SLA class I expression moderately increased at 24h and then decreased to a baseline level or less at 72h of IFN-γ or TSST-1 stimulation. The three classical SLA class I genes showed similar expression kinetics, although SLA-3 mRNA level was consistently lower than those of SLA-1 and -2. The expression of IRF-1, a modulator of SLA expression, showed similar kinetics to those of the three classical SLA class I genes. The expression profiles detected by flow cytometry of the SLA molecules on the cell surface of PBMCs were maintained at a consistently high level during cell stimulation with either TSST-1 or IFN-γ, which was distinct from the kinetics of mRNA expression. These results showed that miniature swine SLA class I mRNA expression was effectively and equally up-regulated among the three loci and coordinately with IRF-1 gene expression after stimulation of T cell activation by sAG or IFN-γ.     

Influence of stem rot pathogen <Emphasis Type="Italic">Fomitiporia</Emphasis> sp. on “Sanbu-sugi” cultivar of the Japanese cedar <Emphasis Type="Italic">Cryptomeria japonica</Emphasis>

An unidentified Fomitiporia sp. causes severe white-rot on stems of a cultivar “Sanbu-sugi” of the Japanese cedar, Cryptomeria japonica. The influence of the fungus on tree health and wood properties was determined. Bark from dead branches and xylem from living branches contained more glucose than bark from living branches and xylem from dead branches. Tree heights at which annual rings were disconnected were 2, 4 and 6 m at ages 37, 15 and 24 years old, respectively. The pH values of damaged parts were lower than those of non-damaged parts, and the damaged parts were clearly identified using bromocresol green solution. Weight loss of sapwood during 60 days of fungal degradation was 1.4 %, which was less than that by a saprophyte, Trametes versicolor. The amount of polyphenols in the heartwood from damaged tree stems was less than that from non-damaged stems. Degraded parts were less stiff than the non-degraded sapwood as measured with a wood-decay tester, Pilodyn. Our observations indicate that damaged stems are chemically and physically inferior to non-damaged stems.     

Insulin-like Growth Factor 1 mRNA Expression in the Uterus of Streptozotocin-treated Diabetic Mice

Reproductive functions decline with the onset of diabetes in female mice. Diabetic mice have smaller uteri with an underdeveloped endometrium, suggesting diminished estrogen-induced growth. We aimed to clarify the changes in the estrous cycle and in insulin-like growth factor 1 (IGF1) expression in the uteri of streptozotocin (STZ)-treated diabetic mice, because IGF1 is one of the main growth factors involved in estrogen-induced uterine growth. ICR female mice were intraperitoneally administered STZ (10 mg/100 g BW), and blood glucose levels were determined. Mice with blood glucose levels > 200 mg/dl were classified as diabetic mice. The onset of diabetes was associated with acyclic estrous cycles. Diabetes was also induced with STZ in ovariectomized mice. Uterine Igf1 mRNA levels were reduced in ovariectomized STZ-treated diabetic mice. Estrogen is known to stimulate Igf1 mRNA expression in the uterus, but estrogen action was abolished in the uteri of STZ-treated diabetic mice. mRNA expressions of estrogen receptor α (ERα) and steroid hormone receptor coactivators (SRC-1/Ncoa1, SRC-2/Ncoa2, SRC-3/Ncoa3 and CBP/p300/Crebbp) were reduced in the uteri of ovariectomized STZ-treated diabetic mice. The present study demonstrates that diabetes induces a decline in female reproductive functions in mice. Igf1 expression in ovariectomized diabetic female mice was decreased, and decreased responsiveness to estrogen in the uteri of diabetic mice is probably associated with a reduction in ERα and steroid receptor coactivator mRNA expression.     

Genetic variation in radish (Raphanus sativus L.) germplasm from Pakistan using morphological traits and RAPDs

Genetic diversity of 30 radish (Raphanus sativus L.) accessions was investigated at the phenotypic level with morphological characters and at the DNA level using the random amplified polymorphic DNA (RAPD) technique. Thirty-six morpho-physiological traits were recorded from seedling stage to harvest. The 31 primers used generated 202 RAPD bands, of which 158 (78.2%) were polymorphic. Multivariate procedures were used to classify the germplasm on the basis of phenotypic traits and RAPD fragments. Dendrograms were generated for the Euclidean distance from the morphological data and the Nei's genetic distance from the RAPD markers. Phenotypically, all the accessions were classified into four major groups corresponding to the different forms of cultivated radish. The morphological diversity existing within each of these groups suggested that they should be discriminated into the three botanical convarieties, sativusT (large-rooted), caudatus (pod-type) and oleifer (oilseed-type). Clustering of the accessions did not show any pattern of association between the morphological characters and the collection sites. Instead, landrace groups were associated with their morphological similarities and horticultural uses. On the other hand, the intra-specific genetic relationships of several accessions based on RAPD analysis were related primarily to their collection sites rather than to their phenotypic affinities. The level of polymorphism exhibited by the various convarieties could be exploited in genetic mapping populations to tag economically important traits. These genotypes also could serve as a useful germplasm source for root, leaf, pod and seed. This preliminary study of traditional radish landraces from Pakistan provides useful information regarding their horticultural potential.     

Keratometry in normal cats: a cross-sectional study in Japan using an automated handheld keratometer

Keratometry was performed in 73 domestic cats of varied signalment in Japan using an automated handheld keratometer. The mean corneal curvature radius was significantly lower for cats younger than 1 year than for those older than 2 years (8.04 mm vs. 8.80−8.99 mm, P<0.01). The radius was significantly greater in males than in females among the cats older than 11 years (9.22 mm vs. 8.84 mm, P=0.01), while the age distributions of the males and females were similar. Corneal astigmatism did not significantly differ across the gender and age groups. The predictability of the corneal curvature and astigmatism was approximately 41−43% and less than 3%, respectively, as a function of age and bodyweight. The results highlighted some age- and sex-related keratometric variations in domestic cats in Japan.     

Effect of fatty acids added to the milk replacer on white scour and excretion of fatty acids in Holstein calves

In order to examine the relationship between white scour and fatty acids, we added fatty acids to the milk replacer. Twenty healthy Holstein calves were divided into 4 groups, five calves per group; a control group with no fortified fatty acid, and 3 groups fed either with oleic, stearic, or palmitic acid, respectively. The calves were fed milk replacer (5% of the calf's body weight) twice a day but the fatty acids (2 wt % of milk replacer) were added only once. The fecal and blood samples were obtained at 0, 12, 24, 36, and 48 h after feeding of the acids. All five calves in the palmitic acid group, and 3 out of 5 calves each in the stealic and the oleic acid groups had whitish feces after feeding fatty acid. The stearic acid group had a significantly elevated stearic acid concentration in the feces during 24–36 h compared to the pre-feeding level. The fecal concentration of palmitic acid increased significantly at 24–36 h in the palmitic acid group. We concluded that the long-chain saturated fatty acids are one of the causes of white scour in calves.     

Effects of GH gene polymorphism and sex on carcass traits and fatty acid compositions in Japanese Black cattle

To investigate the effects of bovine growth hormone (bGH) gene polymorphism on carcass traits and fatty acid compositions in Japanese Black cattle caused by nucleotide substitution of CTG (allele A)/GTG (allele B) at codon 127 and of ACG (allele A and B)/ATG (allele C) at codon 172 of bGH, GH genotypes of 135 cattle were determined using allele specific-multiplex polymerase chain reaction (PCR). Allele A gave greater rib thickness and lower melting point of fat (MP) while allele B gave higher C18:1% ( P  < 0.05). Allele C gave higher C18:1, monounsaturated fatty acid (MUFA), unsaturated fatty acid (USFA) percentages ( P  < 0.05). It also gave lower saturated fatty acid (SFA) percentages, higher MUFA/SFA and USFA/SFA ratios, and lower MP ( P  < 0.05). Interactions of sex and GH alleles were analyzed. In heifers, allele A gave higher carcass weight, daily carcass gain, rib eye area, rib thickness, subcutaneous fat thickness, and BMS while allele B gave greater rib eye area and rib thickness ( P  < 0.05). Allele C gave higher C18:1 ( P  < 0.01), MUFA ( P  < 0.01), USFA percentages ( P  < 0.05) and MUFA/SFA and USFA/SFA ratios ( P  < 0.01), and lower C16:0 and SFA percentages ( P  < 0.05) and MP ( P  < 0.01). GH gene polymorphism affected carcass traits and fatty acid compositions although the effects were more pronounced in heifers.     

Two Types of Grapevine Yellow Speckle Viroid 1 Isolated from Commercial Grapevine Had the Nucleotide Sequence of Yellow Speckle Symptom-inducing Type

Grapevine yellow speckle viroid 1 (GYSVd 1) was first detected in Japan from commercial grapevines. The viroid seems to be spreading in Japanese commercial vineyards but without causing noticeable disease symptoms. Two Japanese GYSVd 1 isolates from the cultivars Campbell and Steuben were sequenced. The Campbell isolate is included in the type 2 variant of GYSVd 1, and the Steuben isolate is included in the type 3. Both of the isolates were grouped into the yellow speckle-inducing variants in the nucleotide sequence ; however, the etiological importance of this viroid on the viticulture of Japan has not yet been determined. Received 14 June 1999/ Accepted in revised form 25 August 1999     

Changes in the milk composition of okapi (Okapia johnstoni) during the first 6 months of lactation

We collected 17 milk samples at 1 day to 6 months post partum from one captive okapi (Okapia johnstoni). Milks were examined for nutrient composition, including water, fat, crude protein, carbohydrates, ash and five minerals. The average values of the concentrations were 79.2 ± 3.0% SD water, 8.5 ± 3.4% fat, 7.5 ± 1.1% crude protein, 3.6 ± 0.6% carbohydrate, 1.2 ± 0.1% ash, 263.7 ± 47.3 mg/100 g calcium, 183.0 ± 40.0 mg/100 g phosphorus, 95.2 ± 17.6 mg/100 g sodium, 74.4 ± 34.3 mg/100 g potassium and 28.4 ± 4.4 mg/100 g magnesium, respectively. The protein concentration increased at late lactation (P < 0.01). The contents of sodium and potassium were rather high in the early lactation period, while those of calcium and phosphorus were rather low in the early lactation period. During the course of lactation in the first week post partum and the protein concentration was high at 1 day post partum. We compare our findings with the American Association of Zoos & Aquariums's Okapi Species Survival Plan Hand‐rearing Protocol, and recommend that nutrient composition of the formula for okapi calves is developed using the present study as a guide.