Immunohistochemical analysis of expression patterns of tumor necrosis factor receptors on lymphoma cells in enzootic bovine leukosis

Tumor necrosis factor-alpha (TNF-alpha) has been reported to be associated with the progression of lymphoproliferative neoplastic diseases and retroviral infections. Hence we examined immunohistochemically the expression patterns of TNF-receptors (TNF-RI and RII) on lymphoma cells derived from the 29 cases of enzootic bovine leukosis (EBL). Lymphomas obtained in 29 animals with EBL were histopathologically classified into three types: diffuse mixed type (10 cases), diffuse large type (9 cases), and diffuse large cleaved type (10 cases). Immunohistochemically using a monoclonal antibody to a bovine lymphocyte surface antigen, the lymphomas were classified into three phenotypes: B-1a (CD5+/CD11b+), B-1b (CD5-/CD11b+) and B-2 (conventional B) (CD5-/CD11b-). Interestingly, the lymphoma cells in all animals expressed TNF-RII, but not TNF-RI. Although, in EBL, lymphoma cells of which the histopathological and immunological property differs has been formed, the expression patterns of TNF-Rs had the universality in all lymphoma cells. TNF-RII, which induces cell proliferation, was expressed but TNF-RI, which induces cell apoptosis was not expressed on all lymphoma cells, suggesting that TNF-Rs play an important role in the malignant proliferation of B cells and formation of lymphomas in EBL.     

Establishment and characterization of four canine melanoma cell lines

Four canine melanoma cell lines were established from the subcutaneous, oral gingival and mucosal melanoma tissues at the primary and metastatic sites. These cell lines were designated as CMeC-1, CMeC-2, KMeC and LMeC. The cells were spindles in shape, similar to that of primary tumor cells. The doubling times of these cells ranged from 34.1 +/- 5.61 to 57.9 +/- 3.28 hr and their chromosome number ranged from 46 to 80. When transplanted into nude mice, CMeC-1 and LMeC produced tumors, whereas CMeC-2 and KMeC did not. The morphology of the tissue formed by xenotransplantation of these cells was similar to their primary tumors.     

Relationship between serum sex hormone concentrations and histology of seminiferous tubules of captured baleen whales in the Western North Pacific during the feeding season

The present study was conducted to obtain new information on relationships among serum testosterone (T), estradiol-17 beta (E(2)), follicle-stimulating hormone (FSH), and luteinizing hormone (LH) concentrations and histology of seminiferous tubules in captured common minke and Bryde's whales during the feeding season. Blood samples and testes were collected from common minke (n=39 for blood samples, n=15 for testes) and Bryde's (n=14 for blood samples, n=7 for testes) whales captured from May 2001 to August 2001 in the Western North Pacific. Serum T concentrations, in 35.9% of the common minke and 57.1% of Bryde's whales, were below the detection limit (< 2.5 pg/ml). There were no significant differences in the serum concentrations of E(2), FSH, and LH among immature, mature common minke and Bryde's whales except that LH levels of immature Bryde's whales was higher than those of common minke whales. In most seminiferous tubules of mature whales, only a single-layer of spermatogonia was observed. However, spermatozoa were observed in seminiferous tubules in 2/13 of mature common minke and 4/4 of mature Bryde's whales with the low or undetectable T levels. These results indicate that the low serum T concentrations reflect the inactivity of spermatogenesis in both baleen whales, and that it is not possible to assess gonadal activity in either common minke or Bryde's whales using serum sex hormone concentrations during the feeding season.     

Comparative studies on lipid analysis and ultrastructure in porcine and southern minke whale (Balaenoptera bonaerensis) oocytes

The present study was conducted to clarify the difference in the color of the cytoplasm in immature follicular oocytes from prepubertal and adult minke whales. The four lipid contents (triglyceride, total cholesterol, phospholipids and non-esterified fatty acids) in vitrified immature oocytes from prepubertal and adult minke whales, and also in fresh and vitrified immature porcine oocytes, were measured. The lipid contents in vitrified-warmed minke whale oocytes were similarly high compared with those in vitrified-warmed porcine oocytes. In particular, the total cholesterol and phospholipid contents in the vitrified immature oocytes from prepubertal and adult minke whales were significantly (P<0.05) higher than those from prepubertal pigs. Furthermore, the distribution of lipid droplets in fresh and vitrified immature oocytes was observed in transmission electron microscopy. Lipid droplets in the prepubertal minke whale oocytes were distributed throughout the cytoplasm. In contrast, adult minke whales had larger lipid droplets which were distributed mainly in the central portion of the cytoplasm. The lipid droplets of immature oocytes from prepubertal pigs were larger than those in minke whale oocytes. These results indicated that the difference in the distribution of the cytoplasmic lipid droplets may result in the difference in the color tone of both prepubertal and adult whale oocyte cytoplasm.     

The pattern of ovarian development in the prepubertal antarctic minke whale (Balaenoptera bonaerensis)

This study describes the morphological and morphometrical changes associated with prepubertal ovarian development in the Antarctic minke whale (Balaenoptera bonaerensis). Ovaries were harvested from 94 immature minke whales caught in the Antarctic Ocean during the summer feeding season (December-March). Notable differences in ovarian size and morphology were found among animals. Up to 10 folds difference in ovarian weight was found among prepubertal whales of similar body size. During the prepubertal period, ovaries grew slowly and approximately doubled their weight. The morphologies of right and left ovaries were almost identical while the growth of the ovary appears to occur preferentially on the right side. The most striking morphological feature was numerous small antral follicles less than 5 mm in diameter found in ovaries of younger immature whales. The occurrence of these ovaries was highest in whales less than 6 m long and gradually decreased as body length increased. In larger whales, the occurrence of ovaries with a smaller number of follicles up to 10 mm and thick tunica albuginea increased. Thus, the ovary of the Antarctic minke whale experiences bursts of small follicular development during the early prepubertal period before becoming a more developed ovary with fewer but larger follicles, and thick tunica albuginea.     

Complete cDNA sequences and phylogenetic analyses of the Th1 and Th2 cytokines of the bactrian camel (Camelus bactrianus)

The complementary DNAs of the Th1 (IL-2, IL-12p35, and IFN-gamma) and Th2 (IL-4, IL-10 and IL-13) cytokine genes of the bactrian camel (Camelus bactrianus) were cloned, sequenced, and analyzed. IL-2, IL-4, IL-10, IL-12p35, IL-13, and IFN-gamma were found to have 465, 402, 537, 669, 411, and 501 bp length open reading frames with 154, 133, 178, 222, 136, and 166 amino acid encodings, respectively. The homology ranged from 58.8% to 100% between the nucleotide sequences of the camel cytokine genes and the published sequences of other mammalian genes, including the llama, pig, cow, horse, human, and mouse. The cDNA had highest homology with orders Artiodactyla (pigs and cattle) and Perissodactyla (horses), especially to the recently cloned llama sequences.     

Investigating phenotypic variability in Colletotrichum lindemuthianum populations

Colletotrichum lindemuthianum, causal agent of anthracnose in the common bean, has wide genetic variability. Differential bean cultivars and morphological and physiological characteristics were used to analyze 74 isolates of C. lindemuthianum collected in two counties in the state of Minas Gerais, Brazil. Six different races were found, with a predominance of race 65 at both locations. Isolates were classified according to their sensitivities to the fungicide thiophanate-methyl, normally used in the control of common bean anthracnose. In all, ≈10% of isolates were resistant to the fungicide in vitro. Characteristics such as indexes of mycelia growth rate, colony diameter, sporulation capacity, and percentage of germination demonstrated the high genetic variability of C. lindemuthianum. We also observed variation in conidial cytology. The conidia of most isolates showed septa formation after germination, in contrast to septa absence, previously reported in the literature. Sexual and asexual reproduction were evaluated for mechanisms that may contribute in the generation of variability in C. lindemuthianum. Conidial anastomosis tubes were commonly found, indicating that asexual reproduction can help increase variability in this species. Information from this study confirmed high variability in C. lindemuthianum and will guide future studies in basic knowledge and applied technologies.     

First report of fig mosaic virus infecting common fig (Ficus carica) in Japan

For the first time, fig mosaic virus (FMV) was detected in common fig (Ficus carica) trees in Shimane, Japan. These trees exhibited mosaic, ringspots, or distortion, accompanied by chlorosis on leaves and yellow spots on fruits. Some of the symptomatic trees were infested with the eriophyid mite Aceria ficus. The virus was detected based on RT-PCR, followed by sequencing. The amplified 300 base-pair fragments shared 83.5–91.5% identity with the corresponding region of RNA-dependent RNA polymerase gene of FMV isolates previously reported in Turkey, Iran, and Italy.     

Eucalyptus camaldulensis density and fiber length estimated by near-infrared spectroscopy

Density and fiber length belong to the parameters that are used by the pulping industry as indicators of wood quality for different industrial processes and final paper products. The feasibility of Fourier transform near-infrared (FT-NIR) spectroscopy for the non-destructive evaluation of fiber length and air-dry density of fast-growing E. camaldulensis from Thailand was investigated using 50 samples. NIR spectra taken from solid wood and air-dry density as well as fiber length were used for partial least squares (PLS) regression analyses. It is the first time that the fiber length of E. camaldulensis solid wood could be predicted with high accuracy and precision and that the ratios of performance to deviation (RPD) obtained are the first that fully fulfill the requirements of AACC Method 39-00 (AACC 1999) for screening in breeding programs (RPD?≥?2.5). The RPDs for cross-validation (test set validation) of the NIR-PLS-R models of 3.3 (3.8) for air-dry density and 3.5 (3.9) for fiber length allow drawing the conclusion that the models are at least applicable for screening in breeding programs as they lie in-between screening (RPD?≥?2.5) and quality control (RPD?≥?5). Even when 40% of the samples were removed in cross-validation of the air-dry density model, the RPD is 3.2, which confirms that the model is robust, stable, and well qualified for prediction. The good model statistics obtained in this study might be due to the fact that measurement sites for the measurement of NIR spectra, air-dry density, and fiber lengths were strictly coincided.