In vivo and in vitro tests showing sensitization to Japanese cedar (Cryptomeria japonica) pollen allergen in atopic dogs

Using both in vivo and in vitro tests, dogs with atopic dermatitis were examined for sensitization with Japanese cedar (Cryptomeria japonica, CJ) pollen allergen. Ten dogs with clinical manifestation of atopic dermatitis were shown to be sensitized to CJ pollen based on the results of intradermal skin test and serum antigen-specific IgE test. In vitro lymphocyte stimulation test showed blastogenic response after stimulation with crude antigen of CJ pollen in all of the 5 cases examined. The peripheral leukocytes showed increased histamine release after stimulation with crude antigen of CJ pollen in 2 cases examined. These data indicate that a proportion of dogs with atopic dermatitis is sensitized to CJ pollen in a cell-mediated manner and show immediate phase reaction of type I hypersensitivity.     

Immunohistochemical and ultrastructural detection of intestinal spirochetes in Thoroughbred horses.

Studies of equine intestinal spirochetes have long focused on intestinal contents alone, but intestinal spirochetosis has been reported recently in a 21-month-old Thoroughbred colt in Japan. To define the clinical and pathological significances of intestinal spirochetosis in several horses, an epizootiologic survey with histologic, immunohistochemical, and ultrastructural methods was conducted for Brachyspira antigen-containing intestinal spirochetes in 12 diseased or injured Thoroughbred horses, aged from 35 days to 17 years. Brachyspira antigen-containing spirochetes were found in 7 of 12 horses (58.3%) and were more frequent in the cecum than in other parts of the bowel. It was not clear whether the infection was clinically related to diarrhea or dysentery, but histopathology revealed a close association between the bacterial infection and epithelial hyperplasia. Crypt epithelium consisted mainly of goblet cells and showed frequent mitosis throughout its length. Inflammatory cells and congestion were also present. There were numerous spirochetes in the crypts, and some invaded the cecal and colonic epithelia and underlying lamina propria. Ultrastructurally, the spirochetes were divided into 4 types. Three types were identified in degenerative epithelial cells or intracellularly. Brachyspira antigen-containing intestinal spirochetes invading the mucosa were capable of causing epithelial hyperplasia in the cecum and colon in the horses. The findings in this study will increase awareness of the importance of intestinal spirochetosis and may also be helpful for diagnosis and treatment of this condition.     

Association of p53 Gene Mutation with the Prognosis of Canine Lymphoid Tumors

Introduction:  Many dogs with lymphoid tumors develop resistance to chemotherapy. As a mechanism of drug resistance in canine lymphoma, ATP‐dependent drug efflux by P‐glycoprotein was reported, however, inhibition of apoptosis mediated by P53 inactivation has not been investigated. In this study, we investigated the relationship between p53 gene mutation and clinical drug resistance in canine lymphoid tumors.
Methods:  Tumor specimens were obtained from 44 dogs with lymphoid tumors. Mutations of p53 gene at exon 4–8 of these tumor tissues were examined by PCR‐SSCP (single strand conformational polymorphism) analysis, followed by nucleotide sequencing of the abnormal bands. The cases were treated with UW‐Madison protocol, and its response was evaluated by the tumor size or the number of peripheral leukemic cells.
Results:  Of the 44 dogs, 15 dogs (34%) had p53 mutation, whereas 29 dogs (66%) were devoid of p53 mutation, before or during the chemotherapeutic protocol. Rate of good response (CR and PR) to chemotherapy was significantly lower in the dogs with p53 mutation (20%) than those without p53 mutation (55%)(p = 0.022). Median overall survival duration after examination of p53 mutation was significantly shorter in dogs with p53 mutation (101days) than those without p53 mutation (223days)(p = 0.008).
Conclusions:  Lymphoid tumors with p53 mutations were shown to have worse prognosis than those without p53 mutation.     

Biology of the prolactin family in bovine placenta. II. Bovine prolactin-related proteins: Their expression, structure and proposed roles

The placenta produces various peptides and steroid hormones that regulate placental function and fetal growth. Prolactin‐related proteins are peptides that are produced by the placenta and belong to the growth hormone/prolactin family, and have structural similarity to prolactin and placental lactogen. Although several prolactin‐related protein genes have been detected in bovine placenta, their expression profiles and functions are not clear. The main difficulties in examining their biological function is the similarity between their genes and the lack of information about their proteins. Recently, molecular biology methods have been used to detect some new bovine prolactin‐related proteins, and elucidate their biological functions. This review focuses on the structures, expression profiles and conceivable functions of prolactin‐related proteins in bovine placenta. With respect to their expression profiles, bovine prolactin‐related proteins fall into four groups: (i) those expressed around the implantation period; (ii) those that reach peak expression in the middle of gestation; (iii) those that increase with the progress of gestation, reaching a peak in late gestation; and (iv) those that reach a plateau in early gestation and are maintained at that level throughout gestation. Data indicate that bovine prolactin‐related proteins have different biological roles in different periods of gestation. In situ monitoring suggests that bovine prolactin‐related protein‐I has a role in the attachment of trophoblast cells to endometrium during the early implantation period.     

Molecular cloning of feline interferon-gamma-inducing factor (interleukin-18) and its expression in various tissues

Interleukin-18 (IL-18) is a cytokine with potent interferon-gamma-inducing activity, and plays an important biologic role in the enhancement of the activity of natural killer cells and cytotoxic T-lymphocytes. In this study, feline IL-18 cDNA was cloned and characterized to establish a basis for the prospective cytokine therapy in small animal practice. The nucleotide sequence of feline IL-18 cDNA obtained in this study was 712bp long and contained its entire open reading frame encoding 192 amino acid residues. The predicted amino acid sequence of feline IL-18 cDNA showed 77.2, 84.8, 60.2 and 62.6% similarity with those of human, dog, rat and mouse counterparts, respectively. The feline IL-18 cDNA included a putative cleavage site of IL-1beta-converting enzyme (ICE) and IL-1 signature-like sequences identified in human and mouse IL-18 cDNAs. Expression of IL-18 mRNA was detected in various tissues including spleen, liver and cerebrum in the cat.     

Seasonal rhinitis in a cat sensitized to Japanese cedar (Cryptomeria japonica) pollen

A cat showing seasonal allergic symptoms of rhinitis was examined for reactivities to Japanese cedar (Cryptomeria japonica, CJ) pollen allergen by intradermal skin test (IDST), Prausnitz-Kustner (P-K) test, and lymphocyte blastogenic response. In IDST for 26 common allergens. the cat showed a positive reaction to CJ pollen allergen. P-K test using CJ pollen allergen also showed a positive reaction, indicating the presence of serum IgE specific to CJ pollen. In the lymphocyte blastogenic response, the stimulation index in the presence of CJ pollen allergen was 2.4. These data suggested that the seasonal rhinitis observed in the cat was caused by the sensitization to CJ pollen allergen.     

Morphology and heredity of tepal spots in Asiatic and Oriental hybrid lilies (Lilium spp.)

Petal spots constitute an important trait for ornamental flowers, because they generate variation in flower colour and pigmentation patterning. However, only a few reports have described the morphologies and inheritances of petal spots. Two types of spots—raised spots and splatters—appear on the flower tepals of lily (Lilium spp.). Here, we microscopically analysed the morphologies of raised spots and splatters, and compared them with those of petal spots in other species. The raised spots of lily showed an increase in the numbers of parenchymal and epidermal cells, and accumulation of anthocyanin pigments in these cells. We did not observe this type of morphology in the petal spots of rhododendron or Tricyrtis macropoda, indicating that the morphology observed in the raised spots is unique to lily. In the splatters of lily, anthocyanin pigments accumulated only in epidermal cells, and the shape of pigmented cells did not differ from that of unpigmented cells. These features were also observed in spots of T. macropoda. We subsequently investigated the inheritance of raised spots and splatters in an F₁ segregating population. We revealed that F₁ plants with raised spots, splatters, raised spots and splatters, or no spots showed a 1:1:1:1 segregation ratio, indicating that the two types of spots are genetically independent. Such knowledge will facilitate efficient selection during breeding programs focused on the development of cultivars with (or without) spots.     

The prognostic significance of minimal residual disease in the early phases of chemotherapy in dogs with high-grade B-cell lymphoma

The prognostic significance of minimal residual disease (MRD) in the early phases of chemotherapy was examined in 36 dogs with multicentric high-grade B-cell lymphoma. Sequences of immunoglobulin heavy chain (IgH) gene fragments from lymphoma cells were amplified and used to design allele-specific primers and probes for real-time PCR. The dogs were treated with a 6-month modified version of the University of Wisconsin–Madison chemotherapy protocol (UW-25) and evaluated for the MRD level at weeks 6 and 11 of UW-25.Of the 31 dogs that remained on the protocol at week 11, 14 were found to be MRD negative (<10 tumour cells/10⁵ peripheral blood mononuclear cells [PBMCs]), whereas the other 17 were MRD positive (?10 tumour cells/10⁵ PBMCs). The progression-free survival of the dogs with MRD-negative status at week 11 (median, 337 days) was significantly longer than that of the MRD-positive dogs at the same time point (median, 196 days) (P = 0.0002). These results indicate the clinical significance of MRD as a prognostic marker in the early phase of chemotherapy.     

Food preservative potential of gassericin A‐containing concentrate prepared from cheese whey culture supernatant of Lactobacillus gasseri LA39

Gassericin A (GA) is a circular bacteriocin produced by Lactobacillus gasseri LA39. In this study, GA‐containing concentrate was prepared using a cross‐flow membrane filtration device (30 kDa cut‐off) from the culture supernatant of Lb. gasseri LA39 cultivated in a cheese whey‐based food‐grade medium. The bacteriocin activity titer in the concentrate was 16 times as high as that of the culture supernatant and was completely maintained through each incubation at 4°C for 3 months, 37°C for 2 months, 60°C for 5 h, and 100°C for 30 min. The GA‐containing concentrate was used with glycine powder to make custard creams, and then four representative strains of custard cream spoilage bacteria (Bacillus cereus, Lactococcus lactis subsp. lactis, Achromobacter denitrificans and Pseudomonas fluorescens) were individually inoculated at c. 10³ colony forming units/g in the custard creams. Throughout 30 days of incubation at 30°C, all of the inoculated bacteria were completely inhibited by the combination of 5% (w/w) of the GA‐containing concentrate and 0.5% (w/w) glycine. This is the first highly practical application of GA to foods as a biopreservative, and the concentration method and the bacteriocin concentrate would contribute to biopreservation of several foods.     

Reaction behavior of cellulose in an ionic liquid, 1-ethyl-3-methylimidazolium chloride

We investigated the reaction behavior of cellulose in an ionic liquid, 1-ethyl-3-methylimidazolium chloride ([C2mim][Cl]), which can dissolve cellulose. The cellulose samples were treated with [C2mim][Cl] at 100, 120 and 140 °C. At the beginning of the treatment, the solubilized cellulose in [C2mim][Cl] is depolymerized into various low molecular weight compounds such as cellobiose, cellobiosan, glucose, levoglucosan and 5-hydroxymethylfurfural. As the treatment continued, some of the low molecular weight compounds reacted with the ionic liquid to form new polymers, which were black and contained nitrogen. [C2mim][Cl] is, therefore, not only a solvent for cellulose, but also a reagent for both depolymerization to produce various low molecular weight compounds, and subsequent polymerization of those compounds.