Laboratory diagnosis of canine GM2-gangliosidosis using blood and cerebrospinal fluid.     

Osamu Yamato  Hiroyuki Satoh  Naoaki Matsuki  Kenichiro Ono  Masahiro Yamasaki  Yoshimitsu Maede 《Journal of veterinary diagnostic investigation》2004,16(1):39-44

In the present study, laboratory techniques were used to diagnose canine GM2-gangliosidosis using blood and cerebrospinal fluid (CSF) that can be collected noninvasively from living individuals. Lysosomal acid beta-hexosaminidase (Hex) was measured spectrofluorometrically using 4-methylumbelliferyl N-acetyl-beta-D-glucosaminide and 4-methylumbelliferyl 7-(6-sulfo-2-acetamido-2-deoxy-beta-D-glucopyranoside) as substrates. Main isoenzymes A and B of Hex in leukocytes were also analyzed using cellulose acetate membrane electrophoresis. GM2-ganglioside in CSF was detected and determined quantitatively by using thin-layer chromatography/enzyme-immunostaining method with anti-GM2-ganglioside antibody. In normal dogs, Hex activities could be determined in leukocytes, serum, and CSF and the total activities were markedly reduced in all the enzyme sources in a dog with Sandhoff disease. Electrophoresis of a leukocyte lysate from a normal dog showed that the Hex A and Hex B were not separated distinctively with formation of a broad band, whereas there were no bands in electrophoresis of a lysate from a dog with Sandhoff disease, showing a deficiency in the total enzyme activity. GM2-ganglioside could be detected and determined quantitatively in as little as 100 microl of canine CSE GM2-ganglioside in CSF in a dog with Sandhoff disease increased to 46 times the normal level. In conclusion, the methods in the present study are useful for diagnosis of canine GM2-gangliosidosis. These techniques enable definitive and early diagnosis of canine GM2-gangliosidosis even if tissues and organs cannot be obtained.  相似文献   

Familial Congenital Methemoglobinemia in Pomeranian Dogs Caused by a Missense Variant in the NADH‐Cytochrome B5 Reductase Gene          下载免费PDF全文

H. Shino  Y. Otsuka‐Yamasaki  T. Sato  K. Ooi  O. Inanami  R. Sato  M. Yamasaki 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2018,32(1):165-171

Background

In veterinary medicine, congenital methemoglobinemia associated with nicotinamide adenine dinucleotide (NADH)‐cytochrome b5 reductase (b5R) deficiency is rare. It has been reported in several breeds of dogs, but little information is available about its etiology.

Objectives

To analyze the NADH‐cytochrome b5 reductase gene, CYB5R3, in a Pomeranian dog family with methemoglobinemia suspected to be caused by congenital b5R deficiency.

Animals

Three Pomeranian dogs from a family with methemoglobinemia were analyzed. Five healthy beagles and 5 nonrelated Pomeranian dogs without methemoglobinemia were used as controls.

Methods

Methemoglobin concentration, b5R activity, and reduced glutathione (GSH) concentration were measured, and a turbidity index was used to evaluate Heinz body formation. The CYB5R3 genes of the affected dog and healthy dogs were analyzed by direct sequencing.

Results

Methemoglobin concentrations in erythrocytes of the affected dogs were remarkably higher than those of the control dogs. The b5R activity of the affected dogs was notably lower than that of the control dogs. DNA sequencing indicated that this Pomeranian family carried a CYB5R3 gene missense variant (ATC→CTC at codon 194) that resulted in the replacement of isoleucine (Ile) by leucine (Leu).

Conclusions and Clinical Importance

This dog family had familial congenital methemoglobinemia caused by b5R deficiency, which resulted from a nonsynonymous variant in the CYB5R3 gene. This variation (c.580A>C) led to an amino acid substitution (p.Ile194Leu), and Ile194 was located in the proximal region of the NADH‐binding motif. Our data suggested that this variant in the canine CYB5R3 gene would affect function of the b5R in erythrocytes.  相似文献   

Glycine max and Glycine soja are capable of cold acclimation          下载免费PDF全文

J. Robison  N. Arora  Y. Yamasaki  M. Saito  J. Boone  B. Blacklock  S. Randall 《Journal of Agronomy and Crop Science》2017,203(6):553-561

Soybean has been considered a cold intolerant species; based largely upon seed germination and soil emergent evaluations. This study reports a distinct acquisition of cold tolerance, in seedlings, following short acclimation periods. Diversity in cold responses was assessed in eight cultivars of Glycine max and six accessions of G. soja. All varieties of soybean significantly increased in freezing tolerance following acclimation. This study indicates soybean seedlings are indeed capable of sensing cold and acquiring cold tolerance. Germination rates after cold imbibition were negatively correlated with maturity group, but positively correlated with cold acclimation potential in G. soja. Seed fatty acid composition was varied between the species, with Glycine soja accessions containing about 2‐times more linolenic acid (18:3) than G. max. Furthermore, high levels of linoleic acid (18:2) in seeds were positively correlated with germination rates following cold imbibition in G. soja only. We suggest that domestication has not impacted the overall ability of soybean to cold acclimate at the seedling stage and that there is little variation within the domesticated species for ability to cold acclimate. Thus, this brief comparative study reduces the enthusiasm for the “wild” species as an additional source of genetic diversity for cold tolerance.  相似文献   

Repeatable and objective method for evaluating angiogenesis using real‐time RT‐PCR of endoglin expression in canine tumours     

Mayu Kimura  Masahiro Yamasaki  Hiroshi Satoh  Naohiro Uchida 《Veterinary and comparative oncology》2021,19(1):34-43

  相似文献   

Genetic relationships between the dominant genotypes of Phytophthora infestans in Hokkaido, Japan     

Seishi Akino  Masayasu Kato  Kiyotaka Gotoh  Shigeo Naito  Akira Ogoshi 《Journal of General Plant Pathology》2005,71(3):200-203

The genetic characteristics of the dominant genotypes of Phytophthora infestans in Japan (US-1, JP-1, Japanese A1-A, A1-B) were compared. Differences were evident in the peptidase genotype, amplified fragment length polymorphism, and RG57 DNA fingerprints. Almost all of the fingerprint bands for the Japanese genotype A1-B were also present in JP-1 and Japanese A1-A, and few bands were unique to Japanese A1-B. These results suggest that the Japanese A1-B genotype was generated from sexual reproduction involving Japanese A1-A and JP-1 or related genotypes.  相似文献   

A Generalized Two-Dimensional Gaussian Model of Disease Foci of Head Blight of Wheat Caused by Gibberella zeae     

Paulitz TC  Dutilleul P  Yamasaki SH  Fernando WG  Seaman WL 《Phytopathology》1999,89(1):74-83

ABSTRACT A generalized two-dimensional Gaussian model is proposed to describe disease foci of head blight of wheat in plots (100 to 2,500 m(2)) originating from small areas (1 to 16 m(2)) inoculated with Gibberella zeae-colonized corn kernels. These anisotropic, asymmetrical foci arose from ascospores produced in perithecia. The model is Z = exp[-(AX(2) + BY(2) + CXY + DX + EY + F)], in which Z = the incidence of seed or spikelet infection at point (X,Y) located in the plot, exp = the exponential function, X = the abscissa or spatial coordinate of the point along a given axis (approximately parallel to the average wind vector during the period of spore release in these experiments), Y = the ordinate or spatial coordinate of the point along the axis perpendicular to the X axis (approximately perpendicular to the wind direction in these experiments), A and B = the quadratic coefficients of the second-order polynomial AX(2) + BY(2) + CXY + DX + EY + F, C = the bilinear coefficient, D and E = the linear coefficients, and exp(-F) = the incidence of seed or spikelet infection at the focus peak in which X = 0 and Y = 0. The generalized two-dimensional Gaussian model was tested on data from a circular or isotropic focus, an elliptical or anisotropic focus with two axes of symmetry, and two anisotropic foci with one and zero axis of symmetry. Its goodness-of-fit (r(2) and adjusted r(2)) was compared with the inverse power, modified inverse power, exponential, and classical Gaussian models. Submodels using only the linear terms, only the quadratic terms, or combinations selected from stepwise regression procedures using various probabilities to enter and to stay and a procedure maximizing the adjusted r (2) were also considered. Spatial analysis of the residuals was performed using Geary's c coefficient at the first distance class. For the circular and elliptical foci, our model provided a fit similar to the modified inverse power and exponential models. However, for anisotropic foci with one or zero axis of symmetry arising from ascospores influenced by wind direction, the generalized two-dimensional Gaussian model provided a better fit. For these anisotropic foci, the linear term X but not the quadratic term X(2) was generally retained in the model, indicating an exponential gradient in the direction parallel to the wind. In all models, the quadratic term Y(2) was retained, along with Y in some cases, indicating that the gradient in the direction roughly perpendicular to the wind was Gaussian or Gaussian-exponential in shape. The bilinear term XY provided an indication of the orientation of the focus in relation to the axes of the sampling grid. This model has the versatility and parameters (quadratic, bilinear, and linear) to better describe the anisotropy of foci from wind-dispersed spores.  相似文献   

Age-related changes of genital systems in the female Crj:CD (SD) IGS rats during sexual maturation     

Noda S  Sawaki M  Shiraishi K  Yamasaki K  Yamaguchi R 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2002,64(4):315-319

The age-related changes of vaginal opening, body weight, the weights of the uterus and ovary, together with histological examination, serum 17beta-estradiol (E2) and progesterone levels were examined in intact female Crj:CD (SD) IGS rats between 21 and 36 days of age to understand the basic biological profile of changes of the female genital system during sexual maturation in the rat for female pubertal assays. With the beginning of the elevation of serum E2 level from 28 days of age, all parameters except body weight started to show drastic change until 31 days of age. The highest incidence of vaginal opening was recorded at 34 days of age. On macroscopic examinations, a number of rats showed uterine imbibition but vaginal opening. Immediately after the confirmation of the vaginal opening, the genital systems of three rats were observed microscopically. Both ovaries already had multiple corpora lutea, and degeneration of endometrial epithelial cells was observed. In conclusion, we obtained essential data on genital tract development of female Crj:CD (SD) IGS rats for in vivo screening assays that will contribute to detect potential endocrine active chemicals. In addition, it is assumed that the first ovulation precedes or occurs simultaneously with vaginal opening.  相似文献   

Use of canine red blood cell with high concentrations of potassium, reduced glutathione, and free amino acid as host cells for in vitro cultivation of Babesia gibsoni     

Yamasaki M  Asano H  Otsuka Y  Yamato O  Tajima M  Maede Y 《American journal of veterinary research》2000,61(12):1520-1524

OBJECTIVE: To determine the usefulness of canine RBC with high concentrations of potassium, reduced glutathione (GSH), and amino acid(i.e., HK cells) for in vitro cultivation of Babesia gibsoni. ANIMALS: RBC were obtained from 3 dogs that had inherited HK cells and from 3 genetically unaffected dogs that, therefore, had RBC with lower potassium (LK) concentrations (i.e., LK cells). PROCEDURES: First, B. gibsoni were cultivated using HK or LK cells in alpha-modification of Eagle medium, consisting of Earle salts with glutamine and without ribosides, deoxyribosides, and sodium bicarbonate under a humidified atmosphere containing 5% CO2 at 37 C. Second, parasites were cultivated with LK- or HK-cell lysates. Finally, HK cells were separated into 3 fractions (bottom, middle, top layers) by density gradient centrifugation, and B. gibsoni were cultivated with each of the HK-cell fractions. In addition, the concentrations of free amino acids and reduced glutathione (GSH) in each HK-cell fraction were measured. RESULTS: B. gibsoni preferentially multiplied in HK-cell cultures rather than in LK-cell cultures. Furthermore, the addition of HK-cell lysate to the culture medium resulted in enhanced multiplication of the parasites. Higher multiplication of the parasites was observed in HK cells from the top layer, compared with HK cells from the middle and bottom layers. The HK cells from the top layer had higher concentrations of glutamate, aspartate, and GSH, compared with HK cells from the middle and bottom layer. CONCLUSIONS: Canine HK cells are useful host cells for in vitro cultivation of B. gibsoni, and the high concentrations of glutamate, aspartate, and GSH may result in enhancement of multiplication of the parasites in HK cells.  相似文献   

Effects of carbon and nitrogen amendment on soil carbon and nitrogen mineralization in volcanic immature soil in southern Kyushu,Japan     

Ayaka Yamasaki  Ryunosuke Tateno  Hideaki Shibata 《Journal of Forest Research》2011,16(5):414-423

Soil N mineralization is affected by microbial biomass and respiration, which are limited by available C and N. To examine the relationship between C and N for soil microbial dynamics and N dynamics, we conducted long-term laboratory incubation (150 days) after C and N amendment and measured changes in C and N mineralization, microbial biomass C, and dissolved C and N throughout the incubation period. The study soil was volcanic immature soil from the southern part of Japan, which contains lower C and N compared with other Japanese forest soils. Despite this, the area is covered by well-developed natural and plantation forests. Carbon amendment resulted in an increase in both microbial biomass and respiration, and net N mineralization decreased, probably due to increasing microbial immobilization. In contrast, N amendment resulted in a decrease in microbial respiration and an increase in net N mineralization, possibly due to decreased immobilization by microbes. Amendment of both C and N simultaneously did not affect microbial biomass and respiration, although net N mineralization was slightly increased. The results suggested that inhibitory effect on microbial respiration by N amendment should be reduced if carbon availability is higher. Thus, soil available C may limit microbial biomass and respiration in this volcanic immature soil. Even in immature soil where C and N substrate is low, soil C, such as plant root exudates and materials from above- and belowground dead organisms, might help to maintain microbial activity and N mineralization in this study site.  相似文献   

Rapid and simple mutation screening of G(M1) gangliosidosis in Shiba dogs by direct amplification of deoxyribonucleic acid from various forms of canine whole-blood specimens.     

Osamu Yamato  Eun-Og Jo  Toru Shoda  Masahiro Yamasaki  Yoshimitsu Maede 《Journal of veterinary diagnostic investigation》2004,16(5):469-472

This report describes a rapid and simple method for mutation screening of G(M1) gangliosidosis in Shiba dogs by direct amplification of DNA from canine whole-blood specimens using a novel polymerase chain reaction (PCR) reagent cocktail, which can eliminate the DNA extraction process and amplify the genomic DNA directly from human or murine whole blood. The strategy of this mutation screening is based on the identification of a nucleotide deletion by restriction enzyme analysis, coupled with the direct PCR amplification. The target sequence of the canine beta-galactosidase gene could be amplified directly from various forms of canine whole-blood specimens, including anticoagulated blood, blood stored frozen for 1 year, dried blood held in filter paper for 1 year at room temperature, and dry powder of blood stripped from Giemsa-stained blood films, which had been prepared 10 years earlier, resulting in the determination of genotypes in all the specimens. This method simplified the molecular diagnosis and carrier screening of G(M1) gangliosidosis in Shiba dogs, making it simple to examine specimens from the large, widely distributed population of these dogs.  相似文献