Superficial necrolytic dermatitis associated with extrapancreatic glucagonoma in a dog

An 11-year-old Shih Tzu presented with crusting and erythema, mainly on the abdomen and the root of the tail. Based on histopathological findings, blood examinations and necropsy findings, the condition was diagnosed as superficial necrolytic dermatitis associated with a glucagon-secreting extrapancreatic neuroendocrine tumour. Gross necropsy revealed tumour invasion into the spleen, liver, adrenal glands and mesenteric lymph nodes. Immunohistochemical analysis of the neoplastic cells revealed that the tumour was a glucagonoma, consistent with earlier findings of persistent glucagonaemia and hypoaminoacidaemia.     

Brazilian isolates of Trypanosoma (Megatrypanum) theileri: diagnosis and differentiation of isolates from cattle and water buffalo based on biological characteristics and randomly amplified DNA sequences

We detected and cultivated isolates of Trypanosoma (Megatrypanum) theileri from cattle and water buffaloes in S?o Paulo state, southeastern Brazil, which were characterized by comparing morphological, growth and molecular features. Although isolates from cattle and water buffalo were morphologically indistinguishable, they differed in their growth characteristics. A dendrogram based on randomly amplified polymorphic DNA (RAPD) patterns indicated close-genetic relationships among all isolates from both species, which were all tightly clustered together and distant from Megatrypanum species from wild mammals. In addition, isolates within the T. theileri-cluster were clearly segregated into two host-associated groups. The sequence of a synapomorphic RAPD-derived DNA fragment (Tth625), which was shared by all T. theileri trypanosomes from cattle and buffalo but not detected in any of 13 other trypanosome species, was used as target for a conventional T. theileri-specific PCR assay. We also defined RAPD fragments (Tthc606 and Tthb606) that distinguished cattle from buffalo isolates. Thus, distinct growth features and genetic variability distinguished between isolates from cattle and water buffaloes of the same geographic origin, suggesting an association of these isolates with their host species. The trypanosomes from water buffalo reported here are the first T. theileri-like isolates from the Asiatic buffalo (Bubalus bubalis) to be continuously cultured and compared with cattle isolates using biological and molecular methods.     

Alterations of gene expression in adult male rat testis and pituitary shortly after subacute administration of the antiandrogen flutamide

In the course of profiling alterations of gene expression in the male reproductive system induced by anti-androgenic agents, 28 genes expressed in the testis or pituitary of adult rats were examined shortly after subacute administration of the well-known anti-androgen, flutamide (FM). FM (25 mg/kg/day) was orally administered to male rats for six days. On day 8 (D8) after the first dose of FM, intratesticular testosterone (T) levels had dramatically increased, but daily sperm production on D36 was significantly decreased. The mRNA levels of testicular and pituitary genes on D8 were measured by semiquantitative RT-PCR. Among the six testicular steroidogenic enzyme genes, the mRNAs of the P450 side chain cleavage, P450 17 alpha/C(17-20) lyase, and 3beta-hydroxysteroid dehydrogenase type I (3betaHSD) genes significantly increased, whereas 17beta-hydroxysteroid dehydrogenase type III slightly decreased. Among the three steroid receptors examined, androgen receptor (AR) and glucocorticoid receptor (GR) mRNAs were significantly down-regulated (29% and 35%, respectively) in the testis, but there was no change in estrogen receptor alpha. There were no clear changes in expression of the gonadotropin receptors and Sertoli cell specific genes, but a slight increase was observed in expression of the lactose dehydrogenase-c mRNA, a germ cell specific gene. Among the three immediate early genes, c-myc mRNA was increased approximately 1.4-fold. In the pituitary, on the other hand, mRNAs for LHbeta and FSHbeta subunits and gonadotropin releasing hormone receptor had increased significantly. These results show that subacute FM administration first affected hypothalamus/pituitary hormone gene expression, then altered gonadotropin secretion, and subsequently induced over-expression of testicular steroidogenic enzyme genes. However, the significant up-regulation of 3betaHSD and down-regulation of AR mRNAs, despite the higher level of intratesticular T, might be explained by an antagonistic action of hydroxyflutamide retained in the testis. The profiles of alterations in gene expression observed will provide important information for the screening of adult male animals for anti-androgenic chemicals.     

A binding protein for fungal signal molecules in the cell wall of Pisum sativum

In the plant cell wall of Pisum sativum seedlings, we found an NTPase (E.C. 3.6.1.5.) with ATP-hydrolyzing activity that was regulated by an elicitor and suppressors of defense from pea pathogen Mycosphaerella pinodes. The ATPase-rich fraction was purified from pea cell walls by NaCl solubilization, ammonium sulfate precipitation, and chromatography with an ATP-conjugated agarose column and an anion-exchange column. The specific activity of the final ATPase-rich fraction increased 600-fold over that of the initial NaCl-solubilized fraction. The purified ATPase-rich fraction also had peroxidase activity and generated superoxide, both of which were regulated by the M. pinodes elicitor and suppressor (supprescins). Active staining and Western blot analysis also showed that the ATPase was copurified along with peroxidases. In this fraction, a biotinylated elicitor and the supprescins were bound primarily and specifically to ca. 55-kDa protein (CWP-55) with an N-terminal amino acid sequence of QEEISSYAVVFDA. The cDNA clone of CWP-55 contained five ACR domains, which are conserved in the apyrases (NTPases), and the protein is identical to a pea NTPase cDNA (GenBank accession AB071369). Based on these results, we discuss a role for the plant cell wall in recognizing exogenous signal molecules.     

Interspecies nuclear transfer embryos reconstructed from cat somatic cells and bovine ooplasm

This study was conducted to investigate the developmental capacity of domestic cat-bovine reconstructed embryos via interspecies somatic cell nuclear transfer (iSCNT) and to observe the mitochondrial DNA (mtDNA) content of the iSCNT embryos. The iSCNT embryos were generated using mixed-breed domestic cat fibroblasts as donor cells and enucleated bovine oocytes as the recipient cytoplasm. When the developmental capacities of iSCNT embryos and parthenogenic bovine embryos were compared, there was no difference (P>0.05) in the rates of cleavage and development to the 8-cell stage (86.6 vs. 84.0% and 32.2 vs. 36.2%, respectively). However, in contrast to development of parthenogenic embryos to the morula and blastocyst stages, no iSCNT embryos (0/202) developed beyond the 8-cell stage. For mtDNA analysis, iSCNT embryos at the 1-cell, 2-cell, 4-cell and 8-cell stages were randomly selected. Both cat and bovine mtDNA quantification analysis were performed using quantitative PCR. The levels of both cat and bovine mtDNA in cat-bovine iSCNT embryos varied at each stage of development. The cat mtDNA concentration in the iSCNT embryos was stable from the 1-cell to 8-cell stages. The bovine mtDNA in the iSCNT embryos at the 8-cell stage was significantly lower than that at the 4-cell stage (P<0.05). No difference in the proportions of cat mtDNA in the iSCNT embryos was found in any of the observed developmental stages (1- through 8-cell stages). In conclusion, bovine cytoplasm supports domestic cat nucleus development through the 8-cell stage. The mtDNA genotype of domestic cat-bovine iSCNT embryos illustrates persistence of heteroplasmy, and the reduction in mtDNA content might reflect a developmental block at the 8-cell stage.     

Pedigree analysis of factor XI deficiency in Japanese black cattle

Using a DNA-based diagnostic test for factor XI deficiency in Japanese black cattle, we surveyed 123 cattle (42 sires and 81 dams) in Gifu and Hyogo prefectures, and calculated gene frequencies. In sires, we drew up the pedigree network of the cattle with the factor XI deficiency. Results showed that the mutated allele of factor XI deficiency was retroactive in at least 6 or more generations of sires. Frequencies of the mutant gene were higher at 26.4% in total, and at 33.3% in sires. All 7 cattle with the homozygote of mutated allele were clinically normal, and showed no bleeding episodes. The mutated allele of factor XI deficiency might be widespread among Japanese black cattle.     

Some Physicochemical Properties of Small‐, Medium‐, and Large‐Granule Starches in Fractions of Waxy Barley Grain

Barley grain was divided into eight fractions from the surface layer to the center with a machine used to polish brewers' rice. Small‐, medium‐, and large‐granule starches were isolated from classified barley flour, and their physicochemical properties were investigated. The starch granules were oval to round with a median size of 2 μm for small, 10 μm for medium, and 12–19 μm for large granules. From the surface layer to the center, both the median sizes and the ratio of large granules decreased, and the ratio of medium‐ and small‐granules increased. The starches had A‐type X‐ray diffraction patterns typical of cereal starches. The moisture sorption showed a negative correlation to the granule size. The gelatinization temperatures of starch granules in each layer were approximately the same, but the enthalpies decreased in the order of large, medium, and small granules.     

Dehydroeffusol Rescues Amyloid β25–35-Induced Spatial Working Memory Deficit

Plant Foods for Human Nutrition - Amyloid β (Aβ) peptides produced from the amyloid precursor protein, a transmembrane protein, are neurotoxic and blocking the neurotoxicity may lead to...     

Genetic resources of salt tolerance in wild Hordeum species

Salt tolerance was evaluated in 340 accessions of Hordeum, consisting of 41 brittle-rachis forms of Hordeum vulgare L. subsp. vulgare (H. agriocrithon) accessions, 154 H. vulgare L. subsp. spontaneum (H. spontaneum) accessions, and 145 accessions of ten other species or subspecies of wild Hordeum. Germination was carried out at concentrations of 171, 257, and 342 mM NaCl. The levels of salt tolerance for seed germination in wild Hordeum species were generally lower than those found by Mano et al. (1996) in cultivated barley; the NaCl tolerance level of the different species were as follows: H. agriocrithon > H. spontaneum > other wild Hordeum species. In addition, leaf injury index was used to assess tolerance at the seedling stage after treatment with 500 mM NaCl solution for four weeks. The levels of salt tolerance at the seedling stage in wild Hordeum species were generally higher than those found by Mano & Takeda (1995) in cultivated barley. Most wild Hordeum species showed high NaCl tolerance at the seedling stage and are considered good sources of germplasm for salt tolerance breeding. This revised version was published online in August 2006 with corrections to the Cover Date.     

Physicochemical Properties and Molecular Structures of Starches from Millet (Pennisetum typhoides) and Sorghum (Sorghum bicolor L. Moench) Cultivars in Nigeria

Some physicochemical properties and molecular structures of starches from millet (Pennisetum typhoides, Doro and Gero) and sorghum (Sorghum bicolor, red and white) in Nigeria were examined. Starch granules of millet and sorghum were 3–14 μm and 4–26 μm in diameter, respectively. Millet cultivars had similar peak viscosities (204–205 RVU) on pasting, while sorghum showed similar minimum viscosities (155–156 RVU). The actual amylose content (%) calculated from iodine affinity (IA, g/100 g) was 20.1 and 21.4 for sorghum and 21.3 for millet. The IA of amylopectin was high (1.27–1.42) and its average chain lengths were 20–21 with β‐amylolysis limit of 56%. Amylopectins showed a polymodal molecular weight distribution on a molar basis. The distributions differed among the samples with a higher amount of larger molecules in Doro and red sorghum. Weight‐ and molar‐based distributions of debranched amylopectins on HPSEC were polymodal with weight‐based distribution showing presence of long chains. Peak DP values for A+B₁ and B₂+B₃ chain fractions were 13–16 and 42–43, respectively. The (A+B₁)/(B₂+B₃) ratio on molar basis (9.0–11.5) was similar to maize and rice amylopectins. Peak DP on molar‐based distribution for white sorghum and millet amyloses were similar (490–540) and the DP_n range was narrow (1,060–1,300), but weight‐based distribution profiles differed. The average chain lengths were 260–270 with 3.9–4.8 chains per molecule.