CD38 gene disruption inhibits the contraction induced by alpha-adrenoceptor stimulation in mouse aorta

CD38 is an ectoenzyme with ADP-ribosyl cyclase and hydrolase activities, which synthesizes cyclic ADP-ribose from NAD and hydrolyzes cyclic ADP-ribose to ADP-ribose. It has been shown that cyclic ADP-ribose is a potent Ca(2+) mobilizing messenger in many cells. To know the physiological role of cyclic ADP-ribose in vascular smooth muscle, we examined the effects of various agonists in the aorta isolated from CD38 knockout (CD38(-/-)) mouse. Western blot analysis showed that CD38 protein was detected in the aorta isolated from wild-type (CD38(+/+)) mouse, but not from CD38(-/-) mouse. In the aortae isolated from both CD38(+/+) and CD38(-/-) mice, KCl, phenylephrine and norepinephrine induced concentration-dependent contraction. KCl produced similar concentration-dependent responses in the aortae from both CD38(+/+) and CD38(-/-) mice. Maximum force of contraction induced by KCl (65 mM) was same in the size. Phenylephrine- and norepinephrine-induced contractions were, however, significantly smaller in the aortae from CD38(-/-) mice than in those from CD38(+/+) mice. 5-Hydroxytryptamine, endothelin-1, caffeine and thapsigargin-induced contractions were not significantly different in these two aortae. These results suggest that CD38 gene disruption inhibits alpha-adrenoceptor-induced vascular contractions and cyclic ADP-ribose-mediated signal transduction system is committed in these responses.     

Immunohistochemical evaluation of a malignant intestinal carcinoid in a dog

An intestinal carcinoid with multiple metastases was identified in a 5-year-old male Shih Tzu with a clinical history of anemia, fatigue, anorexia, vomiting, intermittent diarrhea, intestinal bleeding, and progressive emaciation. There was a yellowish-white mass 15 mm in diameter in the anterior jejunum and white nodules consistent with metastases in many organs. Histopathologically, the mass consisted of neoplastic cells arranged in lobules, trabeculae, or closely interdigitating islands of cells. Neoplastic cells were generally polygonal with round hyperchromatic nuclei, modest amounts of eosinophilic cytoplasm, and eosinophilic cytoplasmic granules. Mitoses were common. Rosette formations of tumor cells were apparent in metastatic tumors. Immunohistochemically, tumor cells stained positive for cytokeratin 13, synaptophysin, protein gene product 9.5, neuron-specific enolase, chromogranin A, calcitonin gene-related peptide, serotonin (5-HT), and Leu-7. Serum 5-HT concentrations for this dog were increased 10-fold compared with those of normal dogs. All findings were consistent with a diagnosis of a malignant intestinal carcinoid.     

Haematogenous spread of Sporothrix schenckii in cats with naturally acquired sporotrichosis

The recovery of Sporothrix schenckii from blood samples is rare, and the diagnosis of systemic sporotrichosis is usually made at necropsy. In this report, S schenckii was isolated from two or more internal organs of nine necropsied cats with naturally acquired sporotrichosis. Haematogenous spread was demonstrated in vivo by the isolation of S schenckii from the peripheral blood of 17 (n = 49, 34.4 per cent) cats. Feline leukaemia virus (FeLV) was not detected, and co-infection with feline immunodeficiency virus (FIV), observed in nine cases (n = 43, 20.9 per cent), apparently did not affect the isolation of S schenckii from peripheral blood or from the internal organs.     

Light quality of continuous illuminating at night to induce floral initiation of Fragaria chiloensis L. CHI-24-1

A wild strawberry strain, Fragaria chiloensis CHI-24-1, produced inflorescences from both parent and asexually propagated daughter plants linked with runners when grown at 23 °C/20 °C (day/night) under a 24 h day-length (DL) of daylight plus nightly lighting by an incandescent lamp, but not under 8 or 16 h DLs. In the present study, the effect of light quality for continuous illuminating at night on floral initiation of CHI-24-1 plants grown under a 24 h DL was examined. The CHI-24-1 plants were grown under a 24 h DL consisting of natural daylight and continuous lighting at night by an incandescent, a blue fluorescent, a red fluorescent or a far-red fluorescent lamp for 40 days in summer and autumn. Also, the CHI-24-1 plants were grown for 40 days in a growth chamber at 25 °C/20 °C (day/night) with natural daylight and continuous lighting at night by red- and four types of far-red light-emitting-diodes (LEDs with peak wavelengths of 660, 700, 735, 780 and 830 nm). In both experiments, floral initiation of the parent and daughter plants was observed under a stereomicroscope. Although more than 50% of the parent and daughter plants initiated flower buds under the incandescent and far-red fluorescent lamps, about 15% and 0% of those initiated flower buds under blue and red fluorescent lamps, respectively. Floral initiation of the parent and daughter plants occurred under the far-red LED light source whose peak wavelength was 735 nm, but not under the red or the other far-red LEDs. From these results, it can be concluded that the effective light wavelength range of nightly continuous illuminating for floral induction in the CHI-24-1 plants is 735 nm in the far-red light region. Hence, the induction of floral initiation by nightly continuous far-red light (735 nm) appeared to be a response mediated by phytochrome.     

Eimeria organisms develop in the epithelial cells of equine small intestine

Histopathologic and immunohistochemical examinations were performed to determine the origin of host cells parasitized by Eimeria in the small intestines collected from five foals. Eimeria organisms at various stages (mainly microgametes and macrogametes) were frequently found in the cytoplasm of hypertrophied host cells in the lamina propria at the tips of villi of the jejunum and ileum. The cytoplasm of the host cell was immunohistochemically positive for cytokeratin AE1/AE3 and cytokeratin 13 and was negative for vimentin, desmin, alpha-smooth muscle actin, chromogranin A, neuron-specific enolase, and factor VIII. The host cells parasitized by Eimeria species had the immunostaining characteristics of epithelial cells but not of mesenchymal cells, endothelial cells of lacteals or capillaries, smooth muscle cells or neuroendocrine cells. These results suggest that the host cell of Eimeria species is possibly derived from intestinal epithelial cells and then displaced into the lamina propria of the small intestine.     

Myofiber expression of class I major histocompatibility complex accompanied by CD8+ T-cell-associated myofiber injury in a case of canine polymyositis

A 7-year-old female Labrador Retriever dog showed extreme muscular weakness, muscle wasting, dysbasia, and mild dysphagia. An elevated value of creatine kinase (335 IU/liter) in the serum was detected. Electromyographic findings included increased insertional activity, fibrillation potentials, and bizarre high-frequency repetitive potentials. Histopathologic examination of skeletal muscles revealed myofiber necrosis and phagocytosis, regeneration of myofibers, and perivascular, perimysial, and endomysial infiltrations of lymphocytes, macrophages and plasma cells. Immunohistochemical evaluation demonstrated that infiltrative cells in the early stage of myositis were CD8+ T-cells and that an increased expression of major histocompatibility complex (MHC) class I was apparent on the surface of nonnecrotic muscle fibers. In contrast, many CD3+ cells (T cells) and HLA-DR-positive macrophages and B lymphocytes were found in the severely affected areas. These results suggest that both expression of MHC class I and CD8+ T-cell infiltration may play an important role in initiation of myositis. These histopathologic findings resemble those reported in naturally occurring polymyositis in humans.     

Availability of NMR microscopic observation of mouse embryo disorder: examination in malformations induced by maternal administration of retinoic acid

Nuclear magnetic resonance (NMR) microscopy is a magnetic resonance imaging method with enhanced spatial resolution due to the use of a high static magnetic field and high magnetic field gradients. It is considered to be a useful tool for non-invasive and continuous investigation of tissue and organs at the histological level. In this study, we applied NMR microscopy to assessment of morphology in mouse embryos using a developmental disorder model induced by retinoic acid administration. Pregnant mice were given 50 mg/kg all-trans retinoic acid at 8.5 dpc. Embryos were collected at several time points after treatment and examined by NMR microscopy after fixation. Two-dimensional and three-dimensional spin echo sequences were used. Tissue contrast on two-dimensional images changed according to length of repetition time and echo time, and also to developmental stage of embryos. Two-dimensional and three-dimensional images nondestructively demonstrated defects in development of the skeleton and soft tissue, e.g. hypoplasia of vertebrae in the lumbar and tail regions and dysplasia of the spinal cord, in embryos exposed to retinoic acid. These morphological abnormalities were confirmed by conventional assessment after imaging. Although further improvements are required, NMR microscopy will provide a new approach for multi-parameter assessment of embryonic development under physiological and pathological conditions.     

Primary malignant peripheral nerve sheath tumor with eosinophilic cytoplasmic globules arising from the greater omentum in a dog

A 10-year-old Golden Retriever dog had a solitary tumor mass arising from the greater omentum. Histologically, the tumor showed varying cellularity and patterns of cellular arrangement. In dense cellular areas, spindle-shaped cells were arranged in interlacing bundles. The sparse cellular area was characterized by loosely arranged fusiform cells. The neoplastic cells frequently contained PAS-positive eosinophilic globules in the cytoplasm, and mitotic figures were frequently observed. The tumor cells were positive to vimentin, S-100 protein, glial fibrillary acidic protein, myelin basic protein, neuron-specific enolase and myoglobin. The present tumor was diagnosed as a malignant peripheral nerve sheath tumor (MPNST) with eosinophilic cytoplasmic globules arising from the greater omentum. To our knowledge, this may be the first case of primary omental MPNST in domestic animals.     

Anticoccidial activity of 8-aminoquinolines, pamaquine, primaquine and several molecular complexes and salts of pamaquine, against Eimeria tenella, E. necatrix, E. acervulina, E. maxima, and E. brunetti in battery experiments.

Pamaquine and primaquine, the well known antimalarial 8-aminoquinolines, have not been reported for their anticoccidial activity. A series of battery experiments was conducted to investigate their activity against a laboratory strain of Eimeria tenella, E. necatrix, E. acervulina, E. maxima, or E. brunetti and revealed that both drugs were effective against E. tenella and E. necatrix, but not against the other three species. Pamaquine suppressed the symptoms of E. tenella induced coccidiosis at concentrations above 125 ppm in feed and primaquine controlled the clinical signs as well at levels above 31.2 ppm. The activity against E. necatrix was observed with pamaquine at 250 ppm and with primaquine at levels above 125 ppm. Pamaquine showed a tendency apparently to reduce body weight gain at 125-500 ppm, whereas primaquine showed the same tendency at 500 ppm. In a concomitantly conducted experiment, this adverse effect of pamaquine was averted in its molecular complexes with benzophenone, nitropyrazole, dinitrobenzoic acids and quinoline, and in its salts of sulfate or zinc chloride, and yet these compounds retained the same anticoccidial activity as of pamaquine. This suggests that these compounds had the broadened safety margin. Judging from their susceptibility to these compounds. E. tenella and E. necatrix will have similar metabolic functions to those of blood cell parasitizing protozoa like plasmodia and prioplasma, which are easily suppressed by this class of compound.     

Histological and Immunohistochemical Evaluation of Canine Ovary

In the present study, 15 canine ovaries without morphological lesions were examined histologically and immunohistochemically by using a large number of proteins including AE1/AE3, cytokeratin7 (CK7), CK13, CK20, vimentin, desmin, alpha smooth muscle actin (alphaSMA), calponin, S100, Neurofilaments, Inhibinalpha, placental alkaline phosphatase (PLAP) and neuron-specific enolase. Ovarian structures observed in this study included surface epithelium (SE), cortical tubules (CT), tunica albuginea (TA), stromal cells (SC), internal endocrine cells (IE), rete ovarii (RO) and fallopian tubes (FT). SE, CT, RO and FT were broadly immunoreactive for desmin. Besides AE1/AE3 and vimentin, desmin was also closely linked to these structures. Rete ovarii forming a reticular structure showed a positive reaction to S100. Surface epithelium was immunoreactive for PLAP at a significantly high level. In conclusion, these results indicate a specific segment of immunoreactivity as well as the broad range of immunoreactivity in canine ovary. The distinct patterns of immunoreactive for various kinds of proteins will play an important role in facilitating their identification and discrimination even in a normal canine ovary with a complex structure.