Preparation of anti-bovine and porcine mu chain sera

Absorption of anti-L chain antibodies contained in anti-IgM sera immunized with purified bovine and porcine IgM was undertaken by affinity chromatography on a column of Sepharose 4B, coupled with the normal follicular fluids of the respective species as an immunosorbent. Anti-L chain antibodies in both anti-IgM sera were completely absorbed without incurring reduction of the antibody titer, and anti-bovine and anti-porcine mu chain sera were prepared.     

FIV-infected cats respond to short-term rHuG-CSF treatment which results in anti-G-CSF neutralizing antibody production that inactivates drug activity

The hematological and virological effects of recombinant human granulocyte colony-stimulating factor (rHuG-CSF) were evaluated in feline immunodeficiency virus (FIV)-infected cats. Six age-matched, FIV-infected cats used in this cross-over study were injected subcutaneously with 5 microg/kg of rHuG-CSF daily for 3 weeks, while six control cats received a placebo. Five of six rHuG-CSF-treated cats had significant increases in neutrophil counts that peaked on days 11-21 of treatment. All rHuG-CSF-treated cats exhibited an increase in myeloid:erythroid ratios of the bone marrow cells without significant changes in lymphocyte, CD4 counts, CD4/CD8 ratios, RBC counts, FIV antibody titers, and FIV loads in peripheral blood, and without clinical and hematological toxicities. Five of six rHuG-CSF-treated cats developed antibodies to rHuG-CSF by 14-21 days of treatment, which correlated with decreasing neutrophil counts and increasing neutralizing antibodies to rHuG-CSF. Three cats re-treated with rHuG-CSF rapidly developed neutralizing antibodies to rHuG-CSF, while one cat also developed neutralizing antibodies to recombinant feline G-CSF (rFeG-CSF). Overall, rHuG-CSF treatment increased neutrophil counts in FIV-infected cats without affecting the infection status of cats. However, long-term use of rHuG-CSF is not recommended in cats because of the neutralizing antibody production to rHuG-CSF that affects the drug activity. In addition, a preliminary finding suggests that repeated treatment cycle can also induce cross-neutralizing antibodies to rFeG-CSF, which may potentially affect the homeostasis of endogenous FeG-CSF.     

Control of the collagen fibril diameter in the equine superficial digital flexor tendon in horses by decorin

The distribution pattern of collagen fibril diameter in the equine superficial digital flexor tendon (SDFT) is known to differ in central and peripheral areas of some regions. This study reports the essence of collagen fibril differences among different regions of the equine SDFT by transmission electron microscopic (TEM) and high-voltage electron microscopic observations and biochemical analysis. The distribution of large collagen fibrils increased but the density of collagen fibrils decreased from the proximal metacarpal region to the distal metacarpal region. Large collagen fibrils with an irregular cross-sectional profile were found more frequently in the middle metacarpal region than in other regions. Three-dimensional reconstruction of images of irregularly shaped collagen fibrils revealed that these fibrils are formed through fusion of small collagen fibrils with large ones. The amount of decorin, which reportedly inhibits the lateral fusion of collagen fibrils, decreased in the direction of the distal metacarpal region. On the other hand, the size of decorin gradually increased in the direction of the distal metacarpal region. These results suggest that regional differences in collagen fibril distribution and density of collagen fibrils in the SDFT are due, at least in part, to fusion of collagen fibrils and the concomitant regional differences in the amount and size of decorin.     

Counting absolute number of lymphocytes in quail whole blood by flow cytometry

In a previous study, we reported a new method for counting quail blood cells. After quail blood cells were stained with fluorescent lipophilic dye (DiOC6(3)), absolute counts of erythrocytes, granulocytes, and monocytes were obtained by means of flow cytometry (FC). The FC method has the potential for application to avian blood cells count; however, the method was unable to distinguish between lymphocytes and thrombocytes. In the present study, we improved the FC method to obtain separate counts of lymphocytes using DiOC5(3). After quail blood cells were stained with DiOC5(3), the cells were measured with FC. Each blood cell type was distinguished by means of their typical FL-1 (green fluorescence) and SSC (side scatter). Absolute numbers of erythrocytes, granulocytes, monocytes and lymphocytes in whole blood were obtained. The improved FC analysis worked equally well with chicken (Gallus gallus) and goose (Anser cygnoides) blood.     

Tetraploid cells of enhanced green fluorescent protein transgenic mice in tetraploid/diploid-chimeric embryos

We succeeded in noninvasively analyzing the distribution of tetraploid (4n) cells in tetraploid<-->diploid (4n<-->2n) chimeric embryos by using enhanced green fluorescent protein (EGFP) transgenic (Tg) mouse embryos. We also evaluated whether this technique of analyzing 4n-cells in EGFP Tg 4n<-->2n chimeric embryos could be used to determine which characteristics of 4n-cells cause the death of 4n-embryos and restricted distribution of 4n-cells in 4n<-->2n-chimeric embryos after implantation. In our experiments, the distribution of 4n-cells in 4n<-->2n-embryos was normal until an embryonic age of 3.5 days (E3.5). With respect to morphological development, there were no differences between 4n-, diploid (2n), 4n<-->2n-, and diploid/diploid (2n<-->2n) chimeric embryos, but the number of cells in the tetraploid (4n) blastocyst was smaller than expected. This decrease in the number of cells may have caused cell death or reduced the rate of cell division in 4n-cells, and may have restricted the distribution of 4n-cells in 4n<-->2n-chimeric embryos. This study demonstrated the utility of EGFP transgenic mouse embryos for relatively easy and noninvasive study of the sequential distribution of cells in chimeric embryos.     

Evaluation of effects of dietary carbohydrate on formation of struvite crystals in urine and macromineral balance in clinically normal cats

OBJECTIVE: To evaluate effects of dietary carbohydrate on urine volume; struvite crystal formation; and calcium, phosphorus, and magnesium balance in clinically normal cats. ANIMALS: 21 healthy adult cats (15 sexually intact males and 6 sexually intact females). PROCEDURE: Diets containing no carbohydrate source (control diet), control plus starch, or control plus fiber were given in a 3 X 3 Latin-square design. The diets were available ad libitum in study 1 (n = 12) and given under restrictions in study 2 (9) to equalize daily intakes of crude protein among the 3 groups. Formation of struvite crystals and balance of calcium, phosphorus, and magnesium were measured. RESULTS: Urine volume was lower in the starch group and fiber group in study 1, whereas no differences were detected among the groups in study 2. Urinary pH and struvite activity product were higher in the starch group in both studies, and the fiber group also had higher struvite activity product in study 2. In both studies, urinary concentrations of HCl-insoluble sediment were higher in the starch group and fiber group. In the fiber group, a net loss of body calcium, phosphorus, and magnesium was detected in study 2. CONCLUSIONS AND CLINICAL RELEVANCE: Starch and fiber in diets potentially stimulate formation of struvite crystals. Hence, reducing dietary carbohydrate is desirable to prevent struvite urolith formation. In addition, a net loss of body calcium, phosphorus, and magnesium during feeding of the fiber diet suggests that dietary inclusion of insoluble fiber could increase macromineral requirements of cats.     

Sequence comparison of the ORF 7 region of transmissible gastroenteritis viruses isolated in Japan

The 3' end region nucleotide sequence, including ORF7, of nine Japanese and two U.S.A. isolates of transmissible gastroenteritis virus (TGEV) were determined and compared. Nine Japanese TGEV strains have been isolated over the past 40 years (1956-1997). From the comparison of determined nucleotide sequences, we could divide the TGEV Japanese isolates into two groups and distinguish them from TGEV U.S.A. isolates.     

Anemia with chronic renal disorder and disrupted metabolism of erythropoietin in ICR-derived glomerulonephritis (ICGN) mice

The ICR-derived glomerulonephritis (ICGN) mouse, a new inbred mouse strain with a hereditary nephrotic syndrome, is considered to be a good model of human idiopathic nephrotic syndrome and notably exhibits proteinuria and hypoproteinemia from the neonatal stage. In chronic renal disorder (CRD), anemia is a major subsequent symptom (renal anemia). The precise cause of renal anemia remains unclear, primarily owing to the lack of appropriate spontaneous animal models for CRD. To establish adequate animal models for anemia with CRD, we examined the hematological-biochemical properties and histopathological characteristics. With the deterioration of renal function, ICGN mice developed a normochromic and normocytic anemia, and exhibited normochromic and microcytic at the terminal stage. The expression of erythropoietin (EPO) mRNA both in the kidneys and liver and the EPO leak into the urine were observed in ICGN mice, indicating a disrupted metabolism of EPO in ICGN mice. In addition, a lack of iron induced by the hemolysis in the spleen and the leak of transferrin into urine as proteinuria aggravated the anemic condition. In conclusion, the ICGN mouse is a good model for anemia with CRD.     

Variety in histochemical characteristics of the olfactory receptor cells in a flatfish, barfin flounder (Verasper moseri)

Variety in histochemical characteristics of the olfactory receptor cells (ORC) was examined by immunohistochemistry for protein gene product 9.5 (PGP9.5) and calretinin, and by lectin histochemistry with Phaseolus vulgaris leucoagglutinin (PHA-L) in the olfactory epithelium (OE) of the barfin flounder (Verasper moseri). PGP 9.5 immunoreactivity was observed in the ORC situated in the upper three fourths of the OE. Calretinin immunoreactivity was observed in the ORC which seemed to be immunonegative for PGP 9.5. These cells were located in the upper two thirds of the OE. PHA-L staining was observed in small subsets of the ORC. PGP 9.5 and calretinin immunoreactivities and PHA-L staining were also observed in the crypt cells unique to the fish OE. These findings suggest the different properties of olfactory perception among fish ORC.     

Morphogenesis of the olfactory pit in a flatfish, barfin flounder (Verasper moseri)

Morphogenesis of the olfactory pit (OP), olfactory lamella (OL) and olfactory epithelium (OE) was examined by scanning electron and light microscopy in the barfin flounder (Verasper moseri). At day 0 after hatch, the OP was already formed. At day 14, the cellular differentiation of the OE was prominent. At day 42, the OP became a cavity by the formation of its roof. At day 56, the first OL extended remarkably and was lined with the OE on both sides. The OL increased in number with development. These findings suggest that the OE is functionally active at day 14. The formation of the OL in the OP may be initiated by the stimulus when the barfin flounder touched at the bottom of the sea.