Cheongju, South Korea
Cheongju, South Korea

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PubMed | Centers for Disease Control and Prevention, Konkuk University and Optipharm Inc.
Type: Journal Article | Journal: Vaccine | Year: 2015

A natural recombinant nephropathogenic K40/09 strain of infectious bronchitis virus (IBV) was heat-adapted for possible future use as live attenuated vaccine. The K40/09 strain was selected during successive serial passages in specific-pathogen free (SPF) embryonated eggs at sub-optimal higher temperature (56C). Unlike the parental strain, the attenuated strain, designated K40/09 HP50, was found to be safe in 1-day-old SPF chicks, which showed neither mortality nor signs of morbidity, and rarely induced ciliostasis or histological changes in the trachea and kidney after intraocular and fine-spray administration. K40/09 HP50 provided almost complete protection against two distinct subgroups of a nephropathogenic strain (KM91-like and QX-like subgroup) and elicited the production of high titers of neutralizing antibody (neutralization index of 3.6). We conclude that the K40/09 HP50 vaccine virus is rapidly attenuated by heat adaptation and exhibits the desired level of attenuation, immunogenicity, and protective efficacy required for a live attenuated vaccine. These results indicate that the K40/09 vaccine could be helpful for the reduction of economic losses caused by recently emergent nephropathogenic IBV infection in many countries.


Oqani R.K.,Chungnam National University | Lin T.,Chungnam National University | Lee J.E.,Chungnam National University | Choi K.M.,Optipharm Inc. | And 2 more authors.
PLoS ONE | Year: 2016

Positive transcription elongation factor b (P-TEFb) is a RNA polymerase II carboxyl-terminal domain (Pol II CTD) kinase that phosphorylates Ser2 of the CTD and promotes the elongation phase of transcription. Despite the fact that P-TEFb has role in many cellular processes, the role of this kinase complex remains to be understood in mammalian early developmental events. In this study, using immunocytochemical analyses, we found that the P-TEFb components, CDK9, Cyclin T1 and Cyclin T2 were localized to nuclear speckles, as well as in nucleolar-like bodies in pig germinal vesicle oocytes. Using nascent RNA labeling and small molecule inhibitors, we showed that inhibition of CDK9 activity abolished the transcription of GV oocytes globally. Moreover, using fluorescence in situ hybridization, in absence of CDK9 kinase activity the production of ribosomal RNAs was impaired. We also presented the evidences indicating that P-TEFb kinase activity is essential for resumption of oocyte meiosis and embryo development. Treatment with CDK9 inhibitors resulted in germinal vesicle arrest in maturing oocytes in vitro. Inhibition of CDK9 kinase activity did not interfere with in vitro fertilization and pronuclear formation. However, when in vitro produced zygotes were treated with CDK9 inhibitors, their development beyond the 4-cell stage was impaired. In these embryos, inhibition of CDK9 abrogated global transcriptional activity and rRNA production. Collectively, our data suggested that P-TEFb kinase activity is crucial for oocyte maturation, embryo development and regulation of RNA transcription in pig. © 2016 Oqani et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


PubMed | Chungnam National University, Korea Research Institute of Bioscience and Biotechnology and Optipharm Inc.
Type: | Journal: Transgenic research | Year: 2016

Production of transgenic pigs for use as xenotransplant donors is a solution to the severe shortage of human organs for transplantation. The first barrier to successful xenotransplantation is hyperacute rejection, a rapid, massive humoral immune response directed against the pig carbohydrate GGTA1 epitope. Platelet activation, adherence, and clumping, all major features of thrombotic microangiopathy, are inevitable results of immune-mediated transplant rejection. Human CD39 rapidly hydrolyzes ATP and ADP to AMP; AMP is hydrolyzed by ecto-5-nucleotidase (CD73) to adenosine, an anti-thrombotic and cardiovascular protective mediator. In this study, we developed a vector-based strategy for ablation of GGTA1 function and concurrent expression of human CD39 (hCD39). An hCD39 expression cassette was constructed to target exon 4 of GGTA1. We established heterozygous GGTA1 knock-out cell lines expressing hCD39 from pig ear fibroblasts for somatic cell nuclear transfer (SCNT). We also described production of heterozygous GGTA1 knock-out piglets expressing hCD39 and analyzed expression and function of the transgene. Human CD39 was expressed in heart, kidney and aorta. Human CD39 knock-in heterozygous ear fibroblast from transgenic cloned pigs, but not in non-transgenic pigs cells. Expression of GGTA1 gene was lower in the knock-in heterozygous ear fibroblast from transgenic pigs compared to the non-transgenic pigs cell. The peripheral blood mononuclear cells (PBMC) from the transgenic pigs were more resistant to lysis by pooled complement-preserved normal human serum than that from wild type (WT) pig. Accordingly, GGTA1 mutated piglets expressing hCD39 will provide a new organ source for xenotransplantation research.


PubMed | Korea University, Inha University, National Institute of Animal Science, Seoul National University and 6 more.
Type: Journal Article | Journal: PloS one | Year: 2015

Since being domesticated about 10,000-12,000 years ago, domestic pigs (Sus scrofa domesticus) have been selected for traits of economic importance, in particular large body size. However, Yucatan miniature pigs have been selected for small body size to withstand high temperature environment and for laboratory use. This renders the Yucatan miniature pig a valuable model for understanding the evolution of body size. We investigate the genetic signature for selection of body size in the Yucatan miniature pig. Phylogenetic distance of Yucatan miniature pig was compared to other large swine breeds (Yorkshire, Landrace, Duroc and wild boar). By estimating the XP-EHH statistic using re-sequencing data derived from 70 pigs, we were able to unravel the signatures of selection of body size. We found that both selections at the level of organism, and at the cellular level have occurred. Selection at the higher levels include feed intake, regulation of body weight and increase in mass while selection at the molecular level includes cell cycle and cell proliferation. Positively selected genes probed by XP-EHH may provide insight into the docile character and innate immunity as well as body size of Yucatan miniature pig.


The present invention relates to a Korean-type porcine reproductive and respiratory syndrome virus (PRRSV) and a vaccine composition using the same, and a method for preventing porcine reproductive and respiratory syndrome. Korean-type porcine reproductive and respiratory syndrome virus with accession number KCTC 12096BP according to the present invention is Korean-type porcine reproductive and respiratory syndrome virus distinguished from European strains and North American strains, and a vaccine composition specific to Korean-type porcine reproductive and respiratory syndrome virus is produced, thereby preventing a Korean-type porcine reproductive and respiratory syndrome virus or specifically diagnosing the infection with Korean-type porcine reproductive and respiratory syndrome virus.


Patent
Optipharm. Co. and Yonsei University | Date: 2013-12-05

There are provided an improved cervical cancer diagnosing method and a diagnostic kit for same. According to the present invention, it is possible to more rapidly and accurately provide a patient group requiring a clinical treatment and a prevention treatment in terms of a technical aspect to predict that the limitation of the existing HPV DNA test method can be overcome, automate the RNA extraction from eliminated cells, and more rapidly provide more objective and accurate results because the result analysis can be performed by software by using the real-time RT-PCR.


The present invention relates to a Korean-type porcine reproductive and respiratory syndrome virus (PRRSV) and a vaccine composition using the same, and a method for preventing porcine reproductive and respiratory syndrome. Korean-type porcine reproductive and respiratory syndrome virus with accession number KCTC 12096BP according to the present invention is Korean-type porcine reproductive and respiratory syndrome virus distinguished from European strains and North American strains, and a vaccine composition specific to Korean-type porcine reproductive and respiratory syndrome virus is produced, thereby preventing a Korean-type porcine reproductive and respiratory syndrome virus or specifically diagnosing the infection with Korean-type porcine reproductive and respiratory syndrome virus.


There are provided a method for an early diagnosis and a screening test for a therapeutic agent of breast cancer by using tissue and blood, and a quantitative reverse transcription polymerase chain reaction kit for same. According to the present invention, it is possible to provide help in more effective treatment and diagnosis of breast cancer through expression rates of HER2 expressed in blood and a cancer-related marker in the blood in addition to a tissue specimen.


PubMed | Yonsei University and Optipharm Inc.
Type: | Journal: Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases | Year: 2016

Rapid and accurate identification of the causative pathogens of bloodstream infections (BSIs) is crucial for initiating an appropriate antimicrobial therapy which decreases the related morbidity and mortality rates. The aim of this study was to evaluate the usefulness of a newly developed multiplexed, bead-based bioassay system [i.e., the Quantamatrix Multiplexed Assay Platform (QMAP)], obtained directly from blood culture bottles, to simultaneously detect the presence of bacteria and identify the genes for antibiotic resistance.The QMAP system was used to evaluate 619 blood culture bottles from patients with BSIs and to compare the results of conventional culture methods.Using conventional bacterial cultures as the gold standard, the sensitivity, specificity, positive predictive value, and negative predictive value of the QMAP system for the detection of bacterial pathogens in positive blood culture (PBC) samples were 99.8% (n = 592, 95% CI 0.9852-1.000, p < 0.001), 100% (95% CI 0.983-1.000, p < 0.001), 100% (95% CI 0.9922-1.000, p < 0.001), and 99.5% (95% CI 0.9695-1.000, p < 0.001), respectively. In addition, sensitivity and specificity of the QMAP system for the identification of the genes for antibiotic resistance were 99.4% (n = 158, 95% CI 0.9617-0.9999, p < 0.009) and 99.6% (95% CI 0.9763-0.9999, p < 0.0001), respectively.Obtaining results using the QMAP system takes about 3 h, while culture methods can take up to 48-72 h. Therefore, analysis using the QMAP system is rapid and reliable for characterizing causative pathogens in BSIs.


PubMed | Ministry of Food & Drug Safety and Optipharm Inc.
Type: Journal Article | Journal: Clinical and experimental vaccine research | Year: 2016

Various new technologies have been applied for developing vaccines against various animal diseases. Virus-like particle (VLP) vaccine technology was used for manufacturing the porcine circovirus type 2 and RNA particle vaccines based on an alphavirus vector for porcine epidemic diarrhea (PED). Although VLP is classified as a killed-virus vaccine, because its structure is similar to the original virus, it can induce long-term and cell-mediated immunity. The RNA particle vaccine used a Venezuela equine encephalitis (VEE) virus gene as a vector. The VEE virus partial gene can be substituted with the PED virus spike gene. Recombinant vaccines can be produced by substitution of the target gene in the VEE vector. Both of these new vaccine technologies made it possible to control the infectious disease efficiently in a relatively short time.

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