Association of Genetic Variants in Complement Factor H and Factor H-Related Genes with Systemic Lupus Erythematosus Susceptibility

Zhao, Jian,Wu, Hui,Khosravi, Melanie,Cui, Huijuan,Qian, Xiaoxia,Kelly, Jennifer A.,Kaufman, Kenneth M.,Langefeld, Carl D.,Williams, Adrienne H.,Comeau, Mary E.,Ziegler, Julie T.,Marion, Miranda C.,Adl Public Library of Science 2011 PLoS genetics Vol.7 No.5

<▼1><P>Systemic lupus erythematosus (SLE), a complex polygenic autoimmune disease, is associated with increased complement activation. Variants of genes encoding complement regulator factor H (CFH) and five CFH-related proteins (CFHR1-CFHR5) within the chromosome 1q32 locus linked to SLE, have been associated with multiple human diseases and may contribute to dysregulated complement activation predisposing to SLE. We assessed 60 SNPs covering the <I>CFH</I>-<I>CFHRs</I> region for association with SLE in 15,864 case-control subjects derived from four ethnic groups. Significant allelic associations with SLE were detected in European Americans (EA) and African Americans (AA), which could be attributed to an intronic <I>CFH</I> SNP (rs6677604, in intron 11, <I>P</I><SUB>meta</SUB> = 6.6×10<SUP>−8</SUP>, OR = 1.18) and an intergenic SNP between <I>CFHR1</I> and <I>CFHR4</I> (rs16840639, <I>P</I><SUB>meta</SUB> = 2.9×10<SUP>−7</SUP>, OR = 1.17) rather than to previously identified disease-associated <I>CFH</I> exonic SNPs, including I62V, Y402H, A474A, and D936E. In addition, allelic association of rs6677604 with SLE was subsequently confirmed in Asians (AS). Haplotype analysis revealed that the underlying causal variant, tagged by rs6677604 and rs16840639, was localized to a ∼146 kb block extending from intron 9 of <I>CFH</I> to downstream of <I>CFHR1</I>. Within this block, the deletion of <I>CFHR3</I> and <I>CFHR1</I> (<I>CFHR3-1</I>Δ), a likely causal variant measured using multiplex ligation-dependent probe amplification, was tagged by rs6677604 in EA and AS and rs16840639 in AA, respectively. Deduced from genotypic associations of tag SNPs in EA, AA, and AS, homozygous deletion of <I>CFHR3-1</I>Δ (<I>P</I><SUB>meta</SUB> = 3.2×10<SUP>−7</SUP>, OR = 1.47) conferred a higher risk of SLE than heterozygous deletion (<I>P</I><SUB>meta</SUB> = 3.5×10<SUP>−4</SUP>, OR = 1.14). These results suggested that the <I>CFHR3-1</I>Δ deletion within the SLE-associated block, but not the previously described exonic SNPs of <I>CFH</I>, might contribute to the development of SLE in EA, AA, and AS, providing new insights into the role of complement regulators in the pathogenesis of SLE.</P></▼1><▼2><P><B>Author Summary</B></P><P>Systemic lupus erythematosus (SLE) is a complex autoimmune disease, associated with increased complement activation. Previous studies have provided evidence for the presence of SLE susceptibility gene(s) in the chromosome 1q31-32 locus. Within 1q32, genes encoding complement regulator factor H (CFH) and five CFH-related proteins (CFHR1-CFHR5) may contribute to the development of SLE, because genetic variants of these genes impair complement regulation and predispose to various human diseases. In this study, we tested association of genetic variants in the region containing <I>CFH</I> and <I>CFHRs</I> with SLE. We identified genetic variants predisposing to SLE in European American, African American, and Asian populations, which might be attributed to the deletion of <I>CFHR3</I> and <I>CFHR1</I> genes but not previously identified disease-associated exonic variants of <I>CFH</I>. This study provides the first evidence for consistent association between <I>CFH/CFHRs</I> and SLE across multi-ancestral SLE datasets, providing new insights into the role of complement regulators in the pathogenesis of SLE.</P></▼2>

HT-2 toxin affects development of porcine parthenotes by altering DNA and histone methylation in oocytes matured in vitro

Zhang, Y.,Jia, R.X.,Pan, M.H.,Lu, Y.,Cui, X.S.,Kim, N.H.,Sun, S.C. Butterworths, etc 2017 Theriogenology Vol. No.

T-2 toxin is a type A mycotoxin produced by various Fusarium species, while HT-2 toxin is a major metabolite of T-2 toxin. Both T-2 toxin and HT-2 toxin are known to have deleterious effects on animals. Our previous work showed that HT-2 treatment caused the failure of porcine oocyte maturation. In this study, we reported that HT-2 also affected porcine embryo development. In HT-2 toxin treated group, all the percentages of embryos in 2-cell, 4-cell and blastocyst stage were significantly lower compared with those in control groups. We then explored the causes from the epigenetic modification aspect of the oocytes. The analysis of fluorescence intensity showed that 5-methyl cytosine (5 mC) level was increased after exposure to HT-2 toxin in porcine oocytes, indicating that the general DNA methylation level increased in the treated porcine oocytes. In addition, histone modifications were also affected, since our results showed that H3K4me2 and H3K9me2 levels were increased in the oocytes from HT-2-treated group. Therefore, our results indicated that HT-2 toxin decreased porcine embryo developmental competence through altering the epigenetic modifications of oocytes.

Deoxynivalenol exposure induces autophagy/apoptosis and epigenetic modification changes during porcine oocyte maturation

Han, J.,Wang, Q.C.,Zhu, C.C.,Liu, J.,Zhang, Y.,Cui, X.S.,Kim, N.H.,Sun, S.C. Academic Press 2016 Toxicology and applied pharmacology Vol.300 No.-

Deoxynivalenol (DON) is a widespread trichothecene mycotoxin which contaminates agricultural staples and elicits a complex spectrum of toxic effects on humans and animals. It has been shown that DON impairs oocyte maturation, reproductive function and causes abnormal fetal development in mammals; however, the mechanisms remain unclear. In the present study, we investigate the possible reasons of the toxic effects of DON on porcine oocytes. Our results showed that DON significantly inhibited porcine oocyte maturation and disrupted meiotic spindle by reducing p-MAPK protein level, which caused retardation of cell cycle progression. In addition, up-regulated LC3 protein expression and aberrant Lamp2, LC3 and mTOR mRNA levels were observed with DON exposure, together with Annexin V-FITC staining assay analysis, these results indicated that DON treatment induced autophagy/apoptosis in porcine oocytes. We also showed that DON exposure increased DNA methylation level in porcine oocytes through altering DNMT3A mRNA levels. Histone methylation levels were also changed showing with increased H3K27me3 and H3K4me2 protein levels, and mRNA levels of their relative methyltransferase genes, indicating that epigenetic modifications were affected. Taken together, our results suggested that DON exposure reduced porcine oocytes maturation capability through affecting cytoskeletal dynamics, cell cycle, autophagy/apoptosis and epigenetic modifications.

An efficient two-photon fluorescent probe for human NAD(P)H:quinone oxidoreductase (hNQO1) detection and imaging in tumor cells

Kwon, N.,Cho, M.,Park, S.,Kim, D.,Nam, S. J.,Cui, L.,Kim, H.,Yoon, J. unknown 2017 Chemical communications Vol. No.

<P>A new quinone propionic acid locked TP fluorophore which can be used for human NAD(P)H:quinone oxidoreductase (hNQO1) detection was developed. The probe, TPQ, which displays high selectivity and anti-interference ability, was successfully applied to endogenous hNQO1 imaging and for the identification of different cancer cells.</P>

Formulation, release characteristics and bioavailability of novel monolithic hydroxypropylmethylcellulose matrix tablets containing acetaminophen

Cao, Q.R.,Choi, Y.W.,Cui, J.H.,Lee, B.J. Elsevier Science Publishers 2005 Journal of controlled release Vol.108 No.2

Effect of incorporating pharmaceutical excipients on the in vitro release profiles and the release mechanism of monolithic hydroxypropylmethylcellulose (4000 cps) matrix tablets (m-HPMC tablets) in terms of mimicking the dual drug release character of bi-layered Tylenol® ER tablets was studied. We also compared the in vitro release profiles of optimized m-HPMC matrix tablet and Tylenol® ER tablet in water, pH 1.2 gastric fluid, and pH 6.8 intestinal fluid, and in vivo drug bioavailabilities in healthy human volunteers. Acetaminophen was used as the model drug. The m-HPMC tablets were prepared using a wet granulation method followed by direct compression. Release profiles and swelling rates of m-HPMC tablets were found to be highly influenced by the types and amounts of pharmaceutical excipients incorporated. Starch 1500 (Prejel®) and sodium lauryl sulfate (SLS) played a key role in determining the dissolution rate of m-HPMC tablets. Additional excipients, i.e., microcrystalline cellulose (Avicel® PH101) and NaH<SUB>2</SUB>PO<SUB>4</SUB> were used to tune the release profiles of m-HPMC tablets. The effect of pharmaceutical excipients on drug release from HPMC-based matrix tablets was found to be mainly due to a change in hydrophilic gel expansion and on physical interactions between the drug and HPMC. The optimized m-HPMC tablet with a balanced ratio of Prejel®, SLS, Avicel® PH101, and NaH<SUB>2</SUB>PO<SUB>4</SUB> in the formulation showed dual release profiles in water, pH 1.2 gastric fluid, and pH 6.8 intestinal fluid in vitro. Dual release was defined as immediate drug release within few minutes followed by extended release over 8 h. The similarity factors of m-HPMC tablets and bi-layered Tylenol® ER tablets were 79.8, 66.1, and 82.7 in water, gastric fluid and intestinal fluid, respectively, indicating the equivalence of the two release profiles. No significant in vivo bioavailability differences were observed in healthy human volunteers. The developed m-HPMC tablet with dual release characteristics can be easily prepared using a conventional high-speed tablet machine and could provide an alternative to commercially available bilayered Tylenol® ER tablets.

Low temperature plasma induces angiogenic growth factor via up-regulating hypoxia-inducible factor 1α in human dermal fibroblasts

Cui, H.S.,Joo, S.Y.,Lee, D.H.,Yu, J.H.,Jeong, J.H.,Kim, J.B.,Seo, C.H. Academic Press 2017 Archives of biochemistry and biophysics Vol.630 No.-

Numerous studies on the application of low temperature plasma (LTP) have produced impressive results, including antimicrobial, antitumor, and wound healing effects. Although LTP research has branched out to include medical applications, the detailed effects and working mechanisms of LTP on wound healing have not been fully investigated. Here, we investigated the potential effect of inducing growth factor after exposure to LTP and demonstrated the increased expression of angiogenic growth factor mediated by LTP-induced HIF1α expression in primary cultured human dermal fibroblasts. In cell viability assays, fibroblast viability was reduced 6 h and 24 h after LTP treatment for only 5 min, and pre-treating with NAC, a ROS scavenger, prevented cell loss. Fibroblast migration significantly increased at 6 h and 24 h in scratch wound healing assays, the expression of cytokines significantly changed, and regulatory growth factors were induced at 6 h and 24 h after exposure to LTP in RT-PCR or ELISAs. Specifically, LTP treatment significantly induced the expression of HIF1α, an upstream regulator of angiogenesis. Pre-treatment with the inhibitor CAY10585 abolished HIF1α expression and prevented LTP-induced angiogenic growth factor production according to immunoblotting, immunocytochemistry, and ELISA results. Taken together, our results provide information on the molecular mechanism by which LTP application may promote angiogenesis and will aid in developing methods to improve wound healing.

탄수화물의 종류가 IN VITRO 반추위박테리아에 의한 OLEIC ACID 의 HYDROGENATION 및 INCORPORATION 에 미치는 효과

송만강,왕제휘,최향순 한국영양사료학회 1999 韓國營養飼料學會誌 Vol.23 No.6

본 시험은 첨가하는 탄수화물의 종류가 반추위 박테리아에 의한 발효특성, 박테리아 성장 및 oleic acid(C_(18:1))의 hydrogenation과 박테리아 lipid로의 incorporation을 조사하고자 in vitro 방법으로 실시 되었다. 200㎖ non-selective basal broth medium에 dextrose, starch 또는 cellulose powder를 0.20%로(w/v) 첨가하였으며, 탄수화물이 첨가되지 않은 control을 포함한 4처리 각각을 위해 nylon 천(2×3㎝)을 이용하여 평균 87.4㎎의 C_(18:1) 과 I^-(14)C_(18:1) 2.09uCi를 흡착시켰다. Oleic acid와 각각의 탄수화물이 첨가된 broth medium에 8겹의 cheese cloth로 여과시킨 반추위액 3㎖를 첨가하여 혐기적인 방법으로 39℃의 진탕배양기에서 24시간 배양하였다. 이와는 별도로 반추위액만이 포함된 broth medium(blank)을 동일한 조건에서 배양하였다. 탄수화물의 첨가는 blank와 control에 비하여 배양 12시간 이후 배양액(broth medium)의 pH를 낮추었으며 배양 6시간에서는 dextrose 첨가구에서 가장 낮았다(p<0.0001). 배양액 암모니아 농도의 경우 12시간의 배양 이후 탄수화물 첨가구의 pH보다 blank와 control에서 더 증가되었으나(p<0.0111) 첨가한 탄수화물 종류에 따른 뚜렷한 차이는 없었다. 배양 종료시 배양액의 총 VFA 농도는 blank 및 control에 비하여 3종류의 탄수화물 첨가구 모두에서 현저히(p<0.0002) 증가되었다. 또한 배양시간이 경과함에 따라 acetate(C₂)의 조성 비율이 점차 감소되었던 반면 propionate(C₃)의 조성 비율은 점차 증가되었다. 배양 12시간에서는 starch와 cellulose 첨가구에서 C₂의 조성비율이 낮았으나(p<0.0002), 종료시점인 24시간에서는 blank와 control의 경우에 비하여 모든 탄수화물 첨가구에서 더 낮은 것으로 나타났다 (p<0.0001). Propionate의 경우 C₂와는 반대의 경향을 보였다. Butyrate 조성 비율은 24시간에 dextrose와 staph 첨가구에서(p<0.0001) 더 높았다. 한편, 24시간의 배양 후 박테리아 수는 dextrose 첨가구에서 가장 높았으나(p<0.0001) 다른 처리구 간에는 차이가 없었다. Nylon 천에서 배양액으로 유출된 C_(18:1)의 bydrogenation 율(%)은 배양 6시간 및 12시간에서 상대적으로 pH가 낮은 dextrose나 starch 첨가구에서 높았던 반면 cellulose 첨가구에서 낮은 경향을 보였다. 또한 C_(18:1)이나 stearic acid의 형태로 박테리아 lipid로의 incorporation된 율은 탄수화물 첨가구 중 pH가 가장 높았던 cellulose 첨가구에서 11.1%로 가장 증가된 경향이었다. 그러나 hydrogenation 및 incorporation 모두 탄수화물 첨가여부 또는 탄수화물의 종류에 따른 현저한 차이는 없었다. An in vitro study was conducted to examine the effect of carbohydrate sources(dextrose, starch or cellulose) added to the broth medium on fermentation characteristics, bacterial growth, hydrogenation of oleic acid(C_(18:1)) and direct incorporation of C_(18) fatty acids by mined ruminal bacteria in vitro. Carbohydrates were added to 200㎖ non-selective basal broth medium at the level of 0.2%(w/v). Oleic acid(87.4㎎) and 1-^(14)C_(18:1)(2.09uCi) were absorbed into the nylon cloth(2 × 3㎝), and the two pieces of nylon cloth were placed to 200㎖ broth medium for each treatment. Three mls of rumen fluid strained through 8 layers of cheese cloth were added to each broth medium, then was incubated anaerobically in the shaking incubator of 39℃ for 24 hour. The broth media of control which do not contain the carbohydrates and blank which contain rumen fluid only were also incubated. Addition of carbohydrates to the broth medium tended to decrease the pH of broth media after 12h and 24h incubations compared to those of blank and control, and the lowest(p<0.0001) pH was observed from the dextrose addition at the 6h incubation. Ammonia concentrations in the broth media of blank and control after 12h incubation slightly increased(p<0.0111) compared to those in the carbohydrates added media, but there were no differences in pH among carbohydrate sources. Volatile fatty acid concentration in the carbohydrates added broth media increased(p<0.0002) compared to those in blank and control after 24h incubation. As incubation time passed molar proportion of acetate(C₂) decreased gradually but propionate(C₃) proportion increased. Proportions of C₂ were lower (p<0.0002) for the starch and cellulose added media at 12h incubation while increased C₂ proportions were observed from all the carbohydrate added media compared to those from blank and control when the incubation was terminated(24h). Opposite results to the C₂ were observed from C₃. Increased(p<0.0001) molar proportions of butyrate were found from the dextrose and starch added media after 24h incubation. Highest number of viable bacteria was observed from the dextrose added medium after 24h incubation. Percent hydrogenation of C_(18:10) tended to increase for the dextrose and starch added media which were relatively low in pH after 6h and 12h incubation while incorporation(%) of C_(18:1) or C_(18:0) into the bacterial lipids tended to increase for the celluiose addod medium which was high in pH.

Onset of Pronuclear Formation and DNA Synthesis in Porcine Oocytes following Intracytoplasmic Injection of Porcine or Murine Spematozoa

Cui, X.S.,Kim, B.K.,Jun, S.H.,Jin, D.I.,Lee, S.H.,Park, C.S.,Kim, N.H. 충남대학교 형질전환복제돼지연구센터 2004 논문집 Vol. No.8

The onset of pronucleus formation and DNA synthesis in porcine oocytes following the injection of porcine or murine sperm was determined in order to obtain insights into species-specific paternal factors that contribute to fertilization. After 44h in vitro maturation, spermatozoa was injected into the cytoplasm of oocytes. After injection, all oocytes were transferred to NCSU23 medium and cultured at 39℃ under 5% CO₂ in air. Similar frequencies of oocytes with female pronuclei were observed after injection with porcine sperm or with murine sperm. In contrast, male pronuclei formed 8 to 9 h following the injection of porcine sperm, and 6 to 8 h following the injection of murine sperm. After pronucleus formation maternally derived microtubules were assembled and appeared to move both male and female pronuclei to the oocyte center. A few porcine oocytes entered metaphase 22 h after the infection of murine sperm, but normal cell division was not observed. The mean time of onset of S-phase in male pronuclei was 9.7 h following porcine sperm injection and 7.4 h following mouse sperm injection. These results suggested that DNA synthesis was delayed in both pronuclei until the sperm chromatin fully decondensed, and the sperm nuclear decondensing activity and microtubule nucleation abilities of the male centrosome are celt cycle dependent.

Single poly-EEPROM with stacked MIM and n-well capacitor

Cui, Z.-Y.,Choi, M.-H.,Kim, Y.-S.,Lee, H.-G.,Kim, K.-W.,Kim, N.-S. IET 2009 Electronics letters Vol.45 No.3

<P>The new structure of electrically erasable programmable read-only memory (EEPROM), using a capacitor of stacked metal-insulator-metal (MIM) and <I>n</I>-well, is proposed. The oxide capacitance in the <I>n</I>-well region is effectively applied without sacrificing the cell area and the control gate coupling ratio. Therefore, for the same program-voltage rating, the proposed cell allows the EEPROM to have a higher speed handling capability even with a quite small cell size. Measured results show that the programming speed of the proposed cell is almost the same as that of the conventional MIM control gate cell. In an endurance test of 10 000 program/erase cycles, the shift of program threshold voltage is found to be 1.4 V without degradation of read currents.</P>

Apoptosis and Apoptosis Related Gene Expression in Preimplantation Porcine Diploid Parthenotes Developing In Vitro

Cui, X.S.,Kim, I.H.,Kim, N.H. 충남대학교 형질전환복제돼지연구센터 2004 논문집 Vol. No.8

This study was conducted to determine effects of polyvinyl alcohol (PVA), fetal bovine serum (FBS) bovine serum albumin (BSA) and epidermal growth factor (EGF) on blastocoel formation, total cell number, apoptosis and apoptosis-related gene expression of porcine diploid parthenotes developing in vitro. The addition of 0.4% BSA to the culture medium enhanced the development of 2-cell stage parthenotes to the blastocysts stage (P<0.01). FBS reduced cell numbers of blastocysts (P<0.01) and increased percentage of apoptosis in the blastocysts (P<0.001). However, while BSA increased cell numbers, it did so only when EGF was present. Either agent on its own had no effect. Similarly, apoptosis in the blastocysts was not influenced by either agent on its own but was reduced when both BSA and EGF were present. Furthermore, semi-quantitative reverse-transcriptase polymerase chain reaction revealed that EGF enhanced the mRNA expression of Bcl-xL, in the presence of 0.4% BSA but BSA and EGF alone had no effect, and EGF and/or BSA did not influence Bak gene expression in the blastocyst stage parthenotes. However FBS reduced Bcl-xL mRNA expression (P<0.05) and enhanced Bak expression. This result suggests that apoptosis related genes expression is significantly affected by supplements, which may result in alteration of apoptosis and embryo viability of porcine embryos developing in vitro.