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1.
Naomi JACK Tomoyuki MUTO Keigo IEMITSU Tamaki WATANABE Kazuhiro UMEYAMA Jun OHGANE Hiroshi NAGASHIMA 《The Journal of reproduction and development》2022,68(4):233
Recent developments in reproductive biology have enabled the generation of genetically engineered pigs as models for inherited human diseases. Although a variety of such models for monogenic diseases are currently available, reproduction of human diseases caused by haploinsufficiency remains a major challenge. The present study compares the phenotypes of mouse and pig models of Marfan syndrome (MFS), with a special focus on the expressivity and penetrance of associated symptoms. Furthermore, investigation of the gene regulation mechanisms associated with haploinsufficiency will be of immense utility in developing faithful MFS pig models. 相似文献
2.
Yuri ABE Tomokazu TAMURA Shiho TORII Shiho WAKAMORI Makoto NAGAI Kazuya MITSUHASHI Junki MINE Yuri FUJIMOTO Naofumi NAGASHIMA Fumi YOSHINO Yukihiko SUGITA Takushi NOMURA Masatoshi OKAMATSU Hiroshi KIDA Yoshihiro SAKODA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2016,78(1):61-70
In our previous study, we genetically analyzed bovine viral diarrhea viruses (BVDVs)
isolated from 2000 to 2006 in Japan and reported that subgenotype 1b viruses were
predominant. In the present study, 766 BVDVs isolated from 2006 to 2014 in Hokkaido,
Japan, were genetically analyzed to understand recent epidemics. Phylogenetic analysis
based on nucleotide sequences of the 5′-untranslated region of viral genome revealed that
766 isolates were classified as genotype 1 (BVDV-1; 544 isolates) and genotype 2 (BVDV-2;
222). BVDV-1 isolates were further divided into BVDV-1a (93), 1b (371) and 1c (80)
subgenotypes, and all BVDV-2 isolates were grouped into BVDV-2a subgenotype (222). Further
comparative analysis was performed with BVDV-1a, 1b and 2a viruses isolated from 2001 to
2014. Phylogenetic analysis based on nucleotide sequences of the viral glycoprotein E2
gene, a major target of neutralizing antibodies, revealed that BVDV-1a, 1b and 2a isolates
were further classified into several clusters. Cross-neutralization tests showed that
BVDV-1b isolates were antigenically different from BVDV-1a isolates, and almost BVDV-1a,
1b and 2a isolates were antigenically similar among each subgenotype and each E2 cluster.
Taken together, BVDV-1b viruses are still predominant, and BVDV-2a viruses have increased
recently in Hokkaido, Japan. Field isolates of BVDV-1a, 1b and 2a show genetic diversity
on the E2 gene with antigenic conservation among each subgenotype during the last 14
years. 相似文献
3.
Kazuhiro UMEYAMA Kasumi HONDA Hitomi MATSUNARI Kazuaki NAKANO Tatsuro HIDAKA Keito SEKIGUCHI Hironori MOCHIZUKI Yasuhiro TAKEUCHI Tsukasa FUJIWARA Masahito WATANABE Masaki NAGAYA Hiroshi NAGASHIMA 《The Journal of reproduction and development》2013,59(6):599-603
Somatic cell nuclear transfer (SCNT) is a useful technique for creating pig strains
that model human diseases. However, production of numerous cloned disease model pigs
by SCNT for large-scale experiments is impractical due to its complexity and
inefficiency. In the present study, we aimed to establish an efficient procedure for
proliferating the diabetes model pig carrying the mutant human hepatocyte nuclear
factor-1α gene. A founder diabetes transgenic cloned pig was generated by SCNT and
treated with insulin to allow for normal growth to maturity, at which point
epididymal sperm could be collected for cryopreservation. In vitro
fertilization and intrafallopian insemination using the cryopreserved epididymal
sperm resulted in diabetes model transgenic offspring. These results suggest that
artificial reproductive technology using cryopreserved epididymal sperm could be a
practical option for proliferation of genetically modified disease model pigs. 相似文献
4.
Wataru FUJII Asuka ONUMA Shin YOSHIOKA Keisuke NAGASHIMA Koji SUGIURA Kunihiko NAITO 《The Journal of reproduction and development》2015,61(6):589-593
The generation efficiencies of mutation-induced mice when using engineered zinc-finger nucleases (ZFNs) have
been generally 10 to 20% of obtained pups in previous studies. The discovery of high-affinity DNA-binding
modules can contribute to the generation of various kinds of novel artificial chromatin-targeting tools, such
as zinc-finger acetyltransferases, zinc-finger histone kinases and so on, as well as improvement of reported
zinc-finger recombinases and zinc-finger methyltransferases. Here, we report a novel ZFN pair that has a
highly efficient mutation-induction ability in murine zygotes. The ZFN pair induced mutations in all obtained
mice in the target locus, exon 17 of aminopeptidase Q gene, and almost all of the pups had biallelic
mutations. This high efficiency was also shown in the plasmid DNA transfected in a cultured human cell line.
The induced mutations were inherited normally in the next generation. The zinc-finger modules of this ZFN pair
are expected to contribute to the development of novel ZF-attached chromatin-targeting tools. 相似文献
5.
ABSTRACT: Arsenobetaine, the major arsenic compound in marine animals, is substantially non-toxic. It is, however, possible that arsenobetaine undergoes bacterial transformation during manufacturing of fermented fishery products. In the present study, therefore, three types of fish sauce were examined for arsenic concentrations and species compared with those in the raw materials (sardine, Japanese sandfish and Japanese common squid). Arsenic concentrations of the three types of fish sauce were almost equivalent to those in their raw materials, suggesting no accumulation of arsenic during fermentation. Arsenic speciation was performed by a combination of cation-exchange liquid chromatography (LC) and electrospray ionization (ESI)-single quadrupole mass spectrometry (MS). Optimal conditions of LC/ESI-MS were established to analyze seven arsenic compounds (arsenate, monomethylarsonic acid, dimethylarsinic acid, arsenobetaine, trimethylarsine oxide, arsenocholine and tetramethylarsonium ion) found in biological samples. When analyzed by LC/ESI-MS, the major arsenic compound in the raw materials was arsenobetaine as expected, while not arsenobetaine but dimethylarsinic acid was identified as the major arsenic compound in the three types of fish sauce. These results suggest that arsenobetaine in the raw materials is converted to dimethylarsinic acid but not to arsenate with high toxicity by bacterial actions during manufacturing of fish sauce. 相似文献
6.
Hitomi MATSUNARI Toshihiro KOBAYASHI Masahito WATANABE Kazuhiro UMEYAMA Kazuaki NAKANO Takahiro KANAI Taisuke MATSUDA Masaki NAGAYA Manami HARA Hiromitsu NAKAUCHI Hiroshi NAGASHIMA 《The Journal of reproduction and development》2014,60(3):230-237
The development and regeneration of the pancreas is of considerable interest because of the role of these processes in
pancreatic diseases, such as diabetes. Here, we sought to develop a large animal model in which the pancreatic cell lineage
could be tracked. The pancreatic and duodenal homeobox-1 (Pdx1) gene promoter was conjugated to Venus, a
green fluorescent protein, and introduced into 370 in vitro-matured porcine oocytes by intracytoplasmic
sperm injection-mediated gene transfer. These oocytes were transferred into four recipient gilts, all of which became
pregnant. Three gilts were sacrificed at 47–65 days of gestation, and the fourth was allowed to farrow. Seven of 16 fetuses
obtained were transgenic (Tg) and exhibited pancreas-specific green fluorescence. The fourth recipient gilt produced a litter
of six piglets, two of which were Tg. The founder Tg offspring matured normally and produced healthy first-generation (G1)
progeny. A postweaning autopsy of four 27-day-old G1 Tg piglets confirmed the pancreas-specific Venus expression.
Immunostaining of the pancreatic tissue indicated the transgene was expressed in β-cells. Pancreatic islets from Tg pigs were
transplanted under the renal capsules of NOD/SCID mice and expressed fluorescence up to one month after transplantation. Tg
G1 pigs developed normally and had blood glucose levels within the normal range. Insulin levels before and after sexual
maturity were within normal ranges, as were other blood biochemistry parameters, indicating that pancreatic function was
normal. We conclude that Pdx1-Venus Tg pigs represent a large animal model suitable for research on
pancreatic development/regeneration and diabetes. 相似文献
7.
ABSTRACT: Parvalbumin, a calcium-binding sarcoplasmic protein of approximately 12 kDa, represents the cross-reactive, major allergen in fish. In consideration of the fact that parvalbumin is contained at high levels not only in fish muscle but also in frog muscle, the present study was undertaken to clarify whether fish-allergic patients react to two parvalbumins (α- and β-parvalbumins) purified from the bullfrog Rana catesbeiana , which is sometimes consumed as a delicacy in Japan. In enzyme-linked immunosorbent assays (ELISA), sera from 12 of the 14 patients tested reacted equally to both parvalbumins purified from the Pacific mackerel Scomber japonicus and the bigeye tuna Thunnus obesus . Of the 12 sera positive to fish parvalbumins, eight sera also reacted to α- and β-parvalbumins of the bullfrog with different spectra: one serum reacted strongly to α-parvalbumin, six sera reacted strongly to β-parvalbumin and one serum reacted equally to both α- and β-parvalbumins. In addition, inhibition ELISA experiments revealed cross-reactivity between fish and bullfrog parvalbumins. Based on these results, it is proposed that fish-allergic patients should avoid the consumption of frog meat unless they are accurately diagnosed as lacking immunoglobulin E against frog. 相似文献
8.
9.
Masahito WATANABE Mirina KOBAYASHI Masaki NAGAYA Hitomi MATSUNARI Kazuaki NAKANO Miki MAEHARA Gota HAYASHIDA Shuko TAKAYANAGI Rieko SAKAI Kazuhiro UMEYAMA Nobuyuki WATANABE Masafumi ONODERA Hiroshi NAGASHIMA 《The Journal of reproduction and development》2015,61(3):169-177
Monomeric Plum (Plum), a far-red fluorescent protein with photostability and photopermeability, is potentially suitable for in vivo imaging and detection of fluorescence in body tissues. The aim of this study was to generate transgenic cloned pigs exhibiting systemic expression of Plum using somatic cell nuclear transfer (SCNT) technology. Nuclear donor cells for SCNT were obtained by introducing a Plum-expression vector driven by a combination of the cytomegalovirus early enhancer and chicken beta-actin promoter into porcine fetal fibroblasts (PFFs). The cleavage and blastocyst formation rates of reconstructed SCNT embryos were 81.0% (34/42) and 78.6% (33/42), respectively. At 36–37 days of gestation, three fetuses systemically expressing Plum were obtained from one recipient to which 103 SCNT embryos were transferred (3/103, 2.9%). For generation of offspring expressing Plum, rejuvenated PFFs were established from one cloned fetus and used as nuclear
donor cells. Four cloned offspring and one stillborn cloned offspring were produced from one recipient to which 117 SCNT embryos were transferred (5/117, 4.3%). All offspring exhibited high levels of Plum fluorescence in blood cells, such as lymphocytes, monocytes and granulocytes. In addition, the skin, heart, kidney, pancreas, liver and spleen also exhibited Plum expression. These observations demonstrated that transfer of the Plum gene did not interfere with the development of porcine SCNT embryos and resulted in the successful generation of transgenic cloned pigs that systemically expressed Plum. This is the first report of the generation and characterization of transgenic cloned pigs expressing the far-red fluorescent protein Plum. 相似文献
10.
在分析了日本珊瑚扇蟹铜铸熟若蟹甲壳组织和内脏组织中毒性含量的基础上,对毒素提取液进行分离纯化,并通过液相色谱法对其毒素种类进行分析。毒性分析结果显示,毒蟹个体的含毒量在37 500~116 000 MU范围,远远超过成人中毒致死量(5 000 MU),但个体间的毒含量有较大差异;毒蟹个体的甲壳组织的比毒性(909~1 610 MU/g)总体高于内脏组织(111~576 MU/g)。Bio-Gel-P2凝胶柱层析纯化结果显示,粗毒素提取液通过反复纯化可以达到纯化目的,毒素的比毒性从4.8 MU/mg上升到30 MU/mg左右;毒素进一步通过Bio-Rex 70离子交换柱层析的分离纯化结果显示,毒素中约有9%为膝沟藻毒素群(GTXs,麻痹性毒素中的一类)。高效液相色谱分析结果显示,膝沟藻毒素群的主要成分为GTX3和GTX2,并含有少量的dcGTX2和dcGTX3。 相似文献