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1.
Modulation of host immune responses by protozoal DNA 总被引:1,自引:0,他引:1
The pathology caused by acute Babesia bovis infection is similar to that seen in severe human malaria caused by Plasmodium falciparum infection, which is related to dysregulated production of inflammatory cytokines and nitric oxide (NO). We have observed induction of NO, inducible nitric oxide synthase (iNOS) and inflammatory cytokines in macrophages by B. bovis. Furthermore, proliferation of lymphocytes from individuals never exposed to certain protozoal pathogens can be induced by crude protozoal parasite extracts. We have repeatedly observed stimulation of naive PBMC from cattle to antigenic extracts of Babesia bovis. Based on recent studies demonstrating the mitogenicity of bacterial and other non-vertebrate DNAs for murine B cells and macrophages, the mitogenic properties of B. bovis DNA were examined. B. bovis and E. coli DNAs induced proliferation of PBMC and purified B cells from non-exposed cattle. Stimulatory activity was reduced by DNase treatment and methylation with CpG methylase, indicating the presence of stimulatory non-methylated CpG motifs in the B. bovis genome. B. bovis and E. coli DNAs enhanced IgG secretion by cultured B cells, stimulating IgG1 and more strongly, IgG2. Several hexameric CpG immunostimulatory sequences (ISS) active for murine B cells were identified in an 11 kb fragment of B. bovis DNA. An oligodeoxyribonucleotide containing one of these (AACGTT), located in the rhoptry associated protein-1 (rap-1) open reading frame, stimulated B cell proliferation. These studies identify a potential mechanism by which protozoal parasites may modulate host immune responses, leading to consequences such as hypergammaglobulinemia and splenomegaly. These results also support the use of ISS as vaccine adjuvants to enhance Type 1 immune responses in cattle. 相似文献
2.
Rhodococcus equi strains of intermediate virulence (IMV) for mice possess a 20kDa protein designated Virulence Associated Protein B (VapB) and a virulence plasmid of 79-100kb, and can be recovered from the submaxillary lymph nodes of pigs. The pathogenicity of such R. equi strains for foals is unknown. In this study, two foals, 42 and 43 days of age, were infected intratracheally with 10(6) and 10(9) cells of R. equi IMV strain A5, respectively. The foal infected with 10(9) cells of strain A5 became clinically ill, with the onset of illness (pyrexia and depression) occurring 21 days after inoculation. R. equi was isolated from the feces and tracheal washings of the foal from 14 to 28 days after inoculation. The foal infected with 10(6) cells of A5 showed no clinical signs, and no R. equi was isolated from any of the samples of feces or tracheal washings during the 28 days of observation. Two foals of 45 and 50 days of age were infected with 10(5) or 10(6) of virulent R. equi ATCC 33701 having 15-17kDa surface proteins designated VapA. Both exhibited severe clinical signs (pyrexia, depression and anorexia) at 12 and 13 days after inoculation. Histopathological examination revealed that strain A5 caused focal granulomatous pneumonia in the foals. R. equi IMV strain A5 was isolated from lung lesions of both foals and from the contents of the intestinal tracts of the foal infected with 10(9) bacteria. These results suggest that IMV R. equi having VapB is less virulent than virulent R. equi having VapA in foals. This finding supports our previous results on the pathogenicities of R. equi strains having these virulence-associated antigens assessed by mouse pathogenicity tests. 相似文献
3.
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
4.
Kraft pulp bleaching effluent supplemented with glucose was decolorized by a newly isolated fungusGeotrichum candidum Dec 1 (Dec 1) that showed a wide decolorizing spectrum to synthetic dyes. When the glucose concentration in the effluent was 30 g/l, the color removal and the reduction of absorbable organic halogens were 78% and 43% after 7 days culture, respectively. The average molecular weight of colored substances measured by gel filtration chromatography was lowered to less than 3000 from 5600 after 7 days culture. As the contribution of extracellular enzymes such as peroxidase (DyP), manganese peroxidase, and laccase to the decolorization of the kraft pulp bleaching effluent was small, Dec 1 appears to have a different mechanism of decolorizing kraft pulp bleaching effluent when compared with enzymes used to decolorize synthetic dyes. 相似文献
5.
Kazumi KISHIDA Mitsuhiro SAKASE Kenta MINAMI Miyuki M. ARAI Reiko SYOJI Namiko KOHAMA Takayuki AKIYAMA Akio OKA Hiroshi HARAYAMA Moriyuki FUKUSHIMA 《The Journal of reproduction and development》2015,61(6):519-524
The purposes of this study were to examine the relationship between male artificial insemination (AI)
fertility and sperm acrosomal conditions assessed by new and conventional staining techniques and to identify
possible reproductive dysfunctions causing low conception rates in AI using frozen-thawed spermatozoa with
poor acrosomal conditions in Japanese Black bulls. We investigated individual differences among bulls in the
results concerning (1) acrosomal conditions of frozen-thawed spermatozoa as assessed by not merely peanut
agglutinin-lectin staining (a conventional staining technique) but also immunostaining of acrosomal
tyrosine-phosphorylated proteins (a new staining technique), (2) routine AI using frozen-thawed spermatozoa as
assessed by pregnancy diagnosis, (3) in vivo fertilization of frozen-thawed spermatozoa and
early development of fertilized eggs as assessed by superovulation/AI-embryo collection tests and (4)
in vitro fertilization of frozen-thawed spermatozoa with oocytes. The percentages of
frozen-thawed spermatozoa with normal acrosomal conditions assessed by the abovementioned staining techniques
were significantly correlated with the conception rates of routine AI, rates of transferable embryos in
superovulation/AI-embryo collection tests and in vitro fertilization rates. These results are
consistent with new suggestions that the distribution of acrosomal tyrosine-phosphorylated proteins as well as
the acrosomal morphology of frozen-thawed spermatozoa are AI fertility-associated markers that are valid for
the prediction of AI results and that low conception rates in AI using frozen-thawed spermatozoa with poor
acrosomal conditions result from reproductive dysfunctions in the processes between sperm insemination into
females and early embryo development, probably failed fertilization of frozen-thawed spermatozoa with
oocytes. 相似文献
6.
Gene dropping analysis of founder contributions in a closed Japanese black cattle population 总被引:1,自引:0,他引:1
Takeshi HONDA Tetsuro NOMURA Moriyuki FUKUSHIMA Fumio MUKAI 《Animal Science Journal》2002,73(2):105-111
Gene dropping simulation was applied to Japanese Black cattle population in Hyogo prefecture, to examine the survivals of alleles originated from founder animals. In the analysis, unique alleles were assigned to founders, and the genotypes of all descendants along the actual pedigree were generated through Monte Carlo simulation following Mendelian segregation rules. By replicating this process 10 000 times, the distribution of frequencies of alleles from each founder was estimated. From the distribution, several quantities useful for the management of genetic diversity, such as the probability of allele extinction and the probability of alleles surviving at a critically low frequency were derived. The materials used were 68 781 animals born in 1955–1998 and their pedigree records traced back to the population in 1937 or before. The expected number of alleles retained in the population drastically decreased during the analyzed period, and reached to 57.9 in the population of 1998, which was only 3.3% of the total number of alleles assigned to founders. Detailed analysis of major founders with relatively high genetic contributions to the current population revealed that alleles from most of the major founders are now at high risk of future extinction. These results strongly suggest that for the management of genetic diversity, the genetic contributions of founders are not fully informative, and emphasize the importance of the detection of live animals having founder alleles with high extinction possibilities. 相似文献
7.
Yamato O Hayashi D Satoh H Shoda T Uchida K Nakayama H Sakai H Masegi T Murai A Iida T Hisada H Hisada A Yamasaki M Maede Y Arai T 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2008,70(8):813-818
GM2 gangliosidosis variant 0 (human Sandhoff disease) is a lysosomal storage disease caused by simultaneous deficiencies of acid beta-hexosaminidase (Hex) A and Hex B due to an abnormality of beta-subunit, a common component in these enzyme molecules, which is coded by the HEXB gene. In the present study, a retrospective diagnosis was performed in 2 previous suspected cases of feline Sandhoff-like disease using a DNA test to detect the causative mutation identified previously in 4 cats in 2 other families of Japanese domestic cats. Enzymic analysis was also performed using stored leukocytes and plasma collected from the subject families in order to investigate the usefulness of enzymic diagnosis and genotyping of carriers. The DNA test suggested that the 2 cases were homozygous recessive for the mutation. Consequently, 6 cats homozygous for the same mutation have been found in 4 separate locations of Japan, suggesting that this mutant allele may be spread widely in the Japanese domestic cat populations. In enzymic analysis, Hex A and Hex B activities in leukocytes and plasma measured using 4-methylumbelliferyl N-acetyl-beta-D-glucosaminide as a substrate were negligible in affected cats, compared with those in normal and carrier cats. However, there was a wide overlap in enzyme activity between normal and carrier cats. Therefore, it was concluded that enzymic analysis is useful for diagnosis of affected cats, but is not acceptable for genotyping of carriers. 相似文献
8.
Pool‐based genome‐wide association study identified novel candidate regions on BTA9 and 14 for oleic acid percentage in Japanese Black cattle
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Ayaka Nakajima Yuta Matsumoto Yoshinobu Uemoto Moriyuki Fukushima Emi Yoshida Eiji Iwamoto Takayuki Akiyama Namiko Kohama Eiji Kobayashi Takeshi Honda Kenji Oyama Hideyuki Mannen Shinji Sasazaki 《Animal Science Journal》2018,89(8):1060-1066
Fatty acid composition is an important indicator of beef quality. The objective of this study was to search the potential candidate region for fatty acid composition. We performed pool‐based genome‐wide association studies (GWAS) for oleic acid percentage (C18:1) in a Japanese Black cattle population from the Hyogo prefecture. GWAS analysis revealed two novel candidate regions on BTA9 and BTA14. The most significant single nucleotide polymorphisms (SNPs) in each region were genotyped in a population (n = 899) to verify their effect on C18:1. Statistical analysis revealed that both SNPs were significantly associated with C18:1 (p = .0080 and .0003), validating the quantitative trait loci (QTLs) detected in GWAS. We subsequently selected VNN1 and LYPLA1 genes as candidate genes from each region on BTA9 and BTA14, respectively. We sequenced full‐length coding sequence (CDS) of these genes in eight individuals and identified a nonsynonymous SNP T66M on VNN1 gene as a putative candidate polymorphism. The polymorphism was also significantly associated with C18:1, but the p value (p = .0162) was higher than the most significant SNP on BTA9, suggesting that it would not be responsible for the QTL. Although further investigation will be needed to determine the responsible gene and polymorphism, our findings would contribute to development of selective markers for fatty acid composition in the Japanese Black cattle of Hyogo. 相似文献
9.
Yamato O Masuoka Y Yonemura M Hatakeyama A Satoh H Kobayashi A Nakayama M Asano T Shoda T Yamasaki M Ochiai K Umemura T Maede Y 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(2):213-217
The present study was conducted to determine the clinical and clinico-pathologic characteristics of Shiba dogs with GM1 gangliosidosis, which is due to an autosomal recessively inherited deficiency of lysosomal acid beta-galactosidase activity. Clinical and clinico-pathological features were investigated in 10 homozygous Shiba dogs with GM1 gangliosidosis. The age at onset was 5 to 6 months and the dogs manifested progressive neurologic signs including loss of balance, intermittent lameness, ataxia, dysmetria and intention tremor of the head. The dogs were unable to stand by 10 months of age due to a progression of ataxia and spasticity in all limbs. Corneal clouding, a visual defect, generalized muscle rigospasticity, emotional disorder and a tendency to be lethargic were observed at 9 to 12 months. The dogs became lethargic from 13 months of age. The survival period seemed to be 14 to 15 months. As a clinico-pathologic feature, lymphocytes with abnormally large vacuoles were observed in peripheral blood (30 to 50% of total lymphocytes) through the lifetime of the dogs. The clinical and clinico-pathologic characteristics of this animal model are useful for not only the development and testing of potential methods of therapy, but also the diagnosis of affected homozygous Shiba dogs in veterinary clinics. 相似文献
10.
Osamu Yamato Asogi Kobayashi Hiroyuki Satoh Daiji Endoh Toru Shoda Yukiko Masuoka Ayano Hatakeyama Eun-Og Jo Tomoya Asano Madoka Yonemura Masahiro Yamasaki Yoshimitsu Maede 《Journal of veterinary diagnostic investigation》2004,16(4):299-304
In the present study, diagnostic methods for canine G(M1)-gangliosidosis were examined by comparing a DNA mutation assay with an enzyme assay. Sixty-two Shiba dogs of a pedigree with G(M1)-gangliosidosis were differentiated into 3 genotypes, i.e., normal, heterozygous, and homozygous affected dogs, using a DNA mutation assay, which consists of polymerase chain reaction amplification and the determination of restriction fragment length polymorphisms. The beta-galactosidase activity in leukocytes, umbilical cords, and plasma was measured using 4-methylumbelliferyl beta-D-galactoside and p-nitrophenyl beta-D-galactoside as artificial substrates and compared among the 3 genotypes. The results showed that it was possible to identify homozygous dogs with the enzyme assay using leukocytes and umbilical cords. When using leukocytes, heterozygous carriers could be differentiated from normal dogs in many cases. However, the use of the DNA mutation assay is essential for a complete determination of heterozygous carriers because of the overlap in the distribution of enzyme activity between these 2 groups. When umbilical cords were used, heterozygous carriers could not be differentiated from normal dogs because of no significant difference in enzyme activity between these 2 groups. The beta-galactosidase activity in plasma was not applicable to the diagnosis and genotyping of G(M1)-gangliosidosis in Shiba dogs. 相似文献