Nakajo-Nishimura syndrome (NNS) is a disorder that segregates in an autosomal recessive fashion. Symptoms include periodic fever, skin rash, partial lipomuscular atrophy, and joint contracture. Here, we report a mutation in the human proteasome subunit beta type 8 gene (PSMB8) that encodes the immunoproteasome subunit β5i in patients with NNS. This G201V mutation disrupts the β-sheet structure, protrudes from the loop that interfaces with the β4 subunit, and is in close proximity to the catalytic threonine residue. The β5i mutant is not efficiently incorporated during immunoproteasome biogenesis, resulting in reduced proteasome activity and accumulation of ubiquitinated and oxidized proteins within cells expressing immunoproteasomes. As a result, the level of interleukin (IL)-6 and IFN-γ inducible protein (IP)-10 in patient sera is markedly increased. Nuclear phosphorylated p38 and the secretion of IL-6 are increased in patient cells both in vitro and in vivo, which may account for the inflammatory response and periodic fever observed in these patients. These results show that a mutation within a proteasome subunit is the direct cause of a human disease and suggest that decreased proteasome activity can cause inflammation.
Genetically engineered pigs with cell markers such as fluorescent proteins are highly useful in lines of research that include the tracking of transplanted cells or tissues. In this study, we produced transgenic-cloned pigs carrying a gene for the newly developed red fluorescent protein, humanized Kusabira-Orange (huKO), which was cloned from the coral stone Fungia concinna. The nuclear transfer embryos, reconstructed with fetal fibroblast cells that had been transduced with huKO cDNA using retroviral vector D Delta Nsap, developed efficiently in vitro into blastocysts (28.0%, 37/132). Nearly all (94.6%, 35/37) of the cloned blastocysts derived from the transduced cells exhibited clear huKO gene expression. A total of 429 nuclear transfer embryos were transferred to four recipients, all of which became pregnant and gave birth to 18 transgenic-cloned offspring in total. All of the pigs highly expressed huKO fluorescence in all of the 23 organs and tissues analyzed, including the brain, eyes, intestinal and reproductive organs, skeletal muscle, bone, skin, and hoof. Furthermore, such expression was also confirmed by histological analyses of various tissues such as pancreatic islets, renal corpuscles, neuronal and glial cells, the retina, chondrocytes, and hematopoietic cells. These data demonstrate that transgenic-cloned pigs exhibiting systemic red fluorescence expression can be efficiently produced by nuclear transfer of somatic cells retrovirally transduced with huKO gene.
The aim of this study is to establish a simple, objective blastocyst grading system using women's age and embryo developmental speed to predict clinical pregnancy following single vitrified-warmed blastocyst transfer (SVBT) by 6-year retrospective cohort study in a private infertility center.A total of 7,341 SVBT cycles divided into 2006-2011 (6,046 cycles) and 2012 cohort (1,295 cycles) were included. Clinical (CPR), ongoing pregnancy (OPR) and delivery rates (DR) were stratified by women's age (<35, 35-37, 38-39, 40-41, 42-45 years) and time to blastocyst expansion (<120, 120-129, 130-139, 140-149, >149 hours) as embryo developmental speed. In all the age groups, CPR, OPR, and DR decreased as the embryo developmental speed decreased (P < 0.0001). A simple 5-grade score based on women's age and embryo developmental speed was determined by actual clinical pregnancy rates observed in the 2006-2011 cohort. Subsequently the novel grading score was validated in the 2012 cohort (1295 cycles) finding an excellent association. In conclusion, in the present study we established a novel blastocyst grading system using women's age and embryo developmental speed as objective parameters.
Abstract. Miniature pigs have been recognized as valuable experimental animals in various fields such as medical and pharmaceutical research. However, the amount of information on somatic cell cloning in miniature pigs, as well as genetically modified miniature pigs, is much less than that available for common domestic pigs. The objective of the present study was to establish an efficient technique of cloning miniature pigs by somatic cell nuclear transfer. A high pregnancy rate was achieved following transfer of parthenogenetic (3/3) and cloned (5/6) embryos using female miniature pigs in the early pregnancy period as recipients after estrus synchronization with prostaglandin F2 alpha analog and gonadotrophins. The production efficiency of the cloned miniature pigs using male and female fetal fibroblasts as nucleus donors was 0.9% (2/215 and 3/331, respectively). Cloned miniature pigs were also produced efficiently (7.8%, 5/64) by transferring reconstructed embryos into the uteri of common domestic pigs. When donor cells transfected with the green fluorescent protein (GFP) gene were used in nuclear transfer, the production efficiency of the reconstructed embryos and rate of blastocyst development were comparable to those obtained by non-transfected cells. When transfected cell-derived reconstructed embryos were transferred to three common domestic pig recipients, all became pregnant, and a total of ten transgenic cloned miniature pigs were obtained (piglet production efficiency: 2.7%, 10/365). Hence, we were able to establish a practical system for producing cloned and transgenic-cloned miniature pigs with a syngeneic background. Key words: Embryo transfer, In vitro matured oocyte, Miniature pig, Nuclear transfer, Transgenic (J. Reprod. Dev. 54: [156][157][158][159][160][161][162][163] 2008) n recent years, techniques for producing cloned pigs by somatic cell nuclear transfer (SCNT) have been actively utilized to produce genetically modified pigs. A number of cloned pigs have been produced, including those with genes for GFP [1][2][3][4][5][6][7][8], as well as alpha1,3-galactosyltransferase knockout pigs [9][10][11][12][13][14][15][16]. In this manner, genetically modified pigs are being used in a wide variety of biomedical fields, ranging from basic research to organ transplantation.To date, more than ten breeds of miniature pigs have been established as experimental and companion animals [17]. As far as the use of pigs as experimental animals is concerned, miniature pigs are smaller and easier to handle than common domestic pigs, and they have been used in a variety of fields, such as medical and pharmacological research [10,[18][19][20][21][22][23][24][25]. However, compared with common domestic pigs, far less information regarding the production of cloned and genetically modified pigs is available for miniature pigs [10,13,14,[26][27][28][29][30][31].In the present study, we conducted a series of experiments including (i) validation of an estrus synchronization procedure for miniature pig recipients, (i...
Abstract. The aim of the present study was to investigate whether a combination of cytoplasmic lipid removal (delipation) and treatment by a microtubule stabilizer, paclitaxel, would lead to efficient cryopreservation of porcine in vitro matured (IVM) oocytes at the meiosis II (MII) stage. Vitrification and subsequent re-warming and culture of 109 untreated oocytes produced only 9 blastocysts (8.3%). On the other hand, the post-vitrification blastocyst rate was significantly improved (21/113, 18.6%, P<0.05) when oocytes were treated with 1 μM paclitaxel. Oocyte delipation also significantly increased the post-vitrification blastocyst rate compared with the untreated group (15/37, 40.5%, P<0.05). The delipation-and-paclitaxel group exhibited a significantly higher blastocyst rate (34/75, 45.3%, P<0.05) than the paclitaxel group, although it was not significantly higher than that for the delipation group. In transfer experiment, a total of 109 (18.6%) parthenogenetic blastocysts were obtained from 586 oocytes vitrified with the delipation-andpaclitaxel treatment. Transfer of 72 blastocysts to two recipients resulted in 14 (19.4%) somite stage fetuses. In conclusion, we demonstrated for the first time that by removing cytoplasmic lipid droplets from oocytes and performing a microtubule stabilization procedure, vitrified porcine IVM MII-stage oocytes could efficiently develop to the blastocyst stage while retaining the ability to develop into fetuses. Key words: Cryopreservation, Delipation, In vitro matured oocytes, Pig, Vitrification (J. Reprod. Dev. 56: [356][357][358][359][360][361] 2010) ryopreservation techniques for mammalian oocytes and embryos have been used for three main purposes: to preserve the genes of elite livestock animals and increase the efficiency of animal breeding, preserve valuable genetically modified animals and endangered species and use germ cells effectively in assisted reproductive technology. Thus far, embryo cryopreservation as a practical technique has been used in a variety of species, including experimental and livestock animals [1,2]. On the other hand, cryopreservation of oocytes remains impractical compared with embryos, and successful production of progeny from cryopreserved oocytes has been reported in limited species, including rabbits [3] [8] and rats [9]. As with porcine embryos, unfertilized porcine oocytes are highly sensitive to low temperature [10]. Consequently, cryopreserved porcine oocytes have yet to be used to successfully produce viable piglets.One of the reasons for the low-temperature sensitivity of porcine oocytes and embryos is their high intracellular lipid content. Nagashima and colleagues showed that the freezing tolerance of porcine oocytes [11] and early-stage embryos [12,13] can be dramatically increased by removing their cytoplasmic lipid droplets using a process called delipation.It is known that the meiotic spindle of the oocyte is extremely cryosensitive, and the impaired development of oocytes at meiosis II after cryopreservation has been attribut...
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.