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Monitoring of Chicken RNA Integrity as a Function of Prolonged Postmortem Duration

  • Malila, Yuwares;Srimarut, Yanee;U-chupaj, Juthawut;Strasburg, Gale;Visessanguan, Wonnop
    • Asian-Australasian Journal of Animal Sciences
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    • 제28권11호
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    • pp.1649-1656
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    • 2015
  • Gene expression profiling has offered new insights into postmortem molecular changes associated with meat quality. To acquire reliable transcript quantification, high quality RNA is required. The objective of this study was to analyze integrity of RNA isolated from chicken skeletal muscle (pectoralis major) and its capability of serving as the template in quantitative real-time polymerase chain reaction (qPCR) as a function of postmortem intervals representing the end-points of evisceration, carcass chilling and aging stages in chicken abattoirs. Chicken breast muscle was dissected from the carcasses (n = 6) immediately after evisceration, and one-third of each sample was instantly snap-frozen and labeled as 20 min postmortem. The remaining muscle was stored on ice until the next rounds of sample collection (1.5 h and 6 h postmortem). The delayed postmortem duration did not significantly affect $A_{260}/A_{280}$ and $A_{260}/A_{230}$ ($p{\geq}0.05$), suggesting no altered purity of total RNA. Apart from a slight decrease in the 28s:18s ribosomal RNA ratio in 1.5 h samples (p<0.05), the value was not statistically different between 20 min and 6 h samples ($p{\geq}0.05$), indicating intact total RNA up to 6 h. Abundance of reference genes encoding beta-actin (ACTB), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), hypoxanthine-guanine phosphoribosyltransferase (HPRT), peptidylprolylisomerase A (PPIA) and TATA box-binding protein (TBP) as well as meat-quality associated genes (insulin-like growth factor 1 (IGF1), pyruvate dehydrogenase kinase isozyme 4 (PDK4), and peroxisome proliferator-activated receptor delta (PPARD) were investigated using qPCR. Transcript abundances of ACTB, GAPDH, HPRT, and PPIA were significantly different among all postmortem time points (p<0.05). Transcript levels of PDK4 and PPARD were significantly reduced in the 6 h samples (p<0.05). The findings suggest an adverse effect of a prolonged postmortem duration on reliability of transcript quantification in chicken skeletal muscle. For the best RNA quality, chicken skeletal muscle should be immediately collected after evisceration or within 20 min postmortem, and rapidly preserved by deep freezing.

Korean Red Ginseng alleviates dehydroepiandrosterone-induced polycystic ovarian syndrome in rats via its antiinflammatory and antioxidant activities

  • Choi, Jong Hee;Jang, Minhee;Kim, Eun-Jeong;Lee, Min Jung;Park, Kyoung Sun;Kim, Seung-Hyun;In, Jun-Gyo;Kwak, Yi-Seong;Park, Dae-Hun;Cho, Seung-Sik;Nah, Seung-Yeol;Cho, Ik-Hyun;Bae, Chun-Sik
    • Journal of Ginseng Research
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    • 제44권6호
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    • pp.790-798
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    • 2020
  • Background: Beneficial effects of Korean Red Ginseng (KRG) on polycystic ovarian syndrome (PCOS) remains unclear. Methods: We examined whether pretreatment (daily from 2 hours before PCOS induction) with KRG extract in water (KRGE; 75 and 150 mg/kg/day, p.o.) could exert a favorable effect in a dehydroepian-drosterone (DHEA)-induced PCOS rat model. Results: Pretreatment with KRGE significantly inhibited the elevation of body and ovary weights, the increase in number and size of ovarian cysts, and the elevation of serum testosterone and estradiol levels induced by DHEA. Pretreatment with KRGE also inhibited macrophage infiltration and enhanced mRNA expression levels of chemokines [interleukin (IL)-8, monocyte chemoattractant protein-1), proinflammatory cytokines (IL-1β, IL-6), and inducible nitric oxide synthase in ovaries induced by DHEA. It also prevented the reduction in mRNA expression of growth factors (epidermal growth factor, transforming growth factor-beta (EGF, TGF-β)) related to inhibition of the nuclear factor kappa-light-chain-enhancer of activated B cell pathway and stimulation of the nuclear factor erythroid-derived 2-related factor 2 pathway. Interestingly, KRGE or representative ginsenosides (Rb1, Rg1, and Rg3(s)) inhibited the activity of inflammatory enzymes cyclooxygenase-2 and iNOS, cytosolic p-IκB, and nuclear p-nuclear factor kappa-light-chain-enhancer of activated B in lipopolysaccharide-induced RAW264.7 cells, whereas they increased nuclear factor erythroid-derived 2-related factor 2 nuclear translocation. Conclusion: These results provide that KRGE could prevent DHEA-induced PCOS via antiinflammatory and antioxidant activities. Thus, KRGE may be used in preventive and therapeutic strategies for PCOS-like symptoms.

키토산이 배양중인 치주인대세포에 미치는 영향 (Effects of Chitosan on Human Periodontal Ligament Cells in Vitro)

  • 김옥수;정현주
    • Journal of Periodontal and Implant Science
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    • 제31권1호
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    • pp.163-180
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    • 2001
  • The aim of this study was to evaluate the effects of chitosan coating on the attachment, proliferation, functional and morphological change of periodontal ligament cells. Primary human periodontal ligament cells were cultured in dulbecco's modified Eagle's medium with 10% fetal bovine serum and 1% antibiotics. In experimental group, cells of 4th to 7th passage were inoculated in the multiwell plates coated with chitosan in concentration of 0.22, 0.2, and $2mg/m{\ell}$. Cell counting and MTT assay were done after 0.5, 1.5, 3, 6 and 24 hours of incubation to evaluate the cell attachment, and then after 2 and 7 days of culture to evaluate the cell proliferation. The alkaline phosphatase activity was measured after 4 and 7 days of culture and the ability to produce mineralized modules was evaluated after 21 days of culture. The results were as follows : 1. The morphology of periodontal ligament cells on the chitosan coating was round or spheric. Round cells were aggregated after 6 hours of culture. Aggregated cells on the chitosan coated surface showed nodule-like appearance after 24 hours of culture and not achieved confluency at 7 days. 2. During early period of culture, the attachment of periodontal ligament cells were inhibited by chitosan coating. Inhibition of cell attachment tended to increase with the concentration of chitosan. 3. At the chitosan concentration of 0.02 and $0.2mg/m{\ell}$, periodontal ligament cells were more rapidly proliferated at 7 days, compared to the control group. At the concentration of $2mg/m{\ell}$, the proliferation of periodontal ligament cells was inhibitied(p<0.01). 4. Alkaline phosphatase activity of periodontal ligament cells was increased in chitosan coated group, especially at the concentration of $0.02mg/m{\ell}$after 4 days of culture.5. Periodontal ligament cells produced mineralized nodules on chitosan coated wells without the addition of mineralized nodule forming materials (ascorbic acid, ${\beta}-glycerophosphat$, dexamethasone). With the addition of mineralized nodule forming materials, periodontal ligament cells produced more mineralized nodules at the concentration of $0.02mg/m{\ell}$, compared to the control. In summary, the attachment, proliferation, cell activity, and alkaline phosphatase activity of periodontal ligament cells depended on the concentration of coated chitosan. Chitosan stimulated mineralized nodule formation by periodontal ligament cells. At the appropriate concentration($0.02mg/m{\ell}$), chitosan could increase alkaline phosphatase activity and stimulate the formation of mineralized nodule by periodontal ligament cells. These results suggest that chitosan can be used as an adjunct for bone graft material, and the matrix of tissue engineering for periodontal regeneration, especially bone regeneration.

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한반도 남서부, 황해 대륙붕에서 자생하는 해록석의 지구화학적 변화 (Geochemical Variation of Authigenic Glauconite from Continental Shelf of the Yellow Sea, off the SW Korea)

  • 이찬희;이성록;이치원;최석원
    • 자원환경지질
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    • 제30권4호
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    • pp.303-312
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    • 1997
  • 황해 대륙붕의 준고화된 표층퇴적물에서 자생하는 해록석들을 산출상태와 외부조직에 따라 괴상형, 균열형 및 다공질형으로 분류하였다. 이 해록석은 현세의 해침에 의하여 퇴적된 표층의 사질퇴적물내에 산포상으로 산출되며, 보통 0.1~1 mm 크기의 직경을 갖는다. x-선 회절분석 결과로 계산된, 해록석의 단위포와 크기는 $a=5.242{\AA}$, $b=9.059{\AA}$, $c=10.163{\AA}$, ${\beta}=100.5^{\circ}$, $V=474.53{\AA}^3$ 이고, 화학조성과 단위포의 크기로 계산된 밀도는 $2.60{\pm}0.45gm/cc$ 이다. 이 광물은 가열실험시 $10{\AA}$의 회절선이 증가하는 것으로 볼때 일정한 팽윤층을 갖는 것으로 보인다. $O_{10}(OH)_2$를 기준으로 평균조성을 구하면, 팔면체 자리의 Fe 함량은 1.19~2.06 이고, Al 함량은 0.18~0.76 이다. Fe와 Al은 서로 치환관계를 보이며 다공질형에서 괴상형으로 갈수록 Fe의 함량은 증가하고 Al은 감소한다. Mg의 함량은 0.35~0.54로서 Al이 높을수록 Mg의 양도 증가한다. K의 함량은 0.34~0.71의 범위를 보이며 다공질형에서 괴상형으로 갈수록 증가한다. 괴상형 또는 균열형 해록석은 질서/무질서의 혼합층 운모이며, 다공질형의 해록석은 혼합층 일라이트/스멕타이트로서 7~27%의 팽윤층을 포함한다. 이 광물은 표층 퇴적물이 퇴적된후에 극미립의 퇴적입자를 핵으로, 환원환경하에서 발생하는 퇴적물의 분해 및 생물체 파편과의 반응에 따라 자생한 것으로 추정된다.

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NF-κB 신호경로에서 CLK3의 새로운 음성 조절자로서의 기능 (CLK3 is a Novel Negative Regulator of NF-κB Signaling)

  • 전별은;권찬성;이지은;우예린;김상우
    • 생명과학회지
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    • 제32권11호
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    • pp.833-840
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    • 2022
  • 만성 염증은 종양의 발생 및 진행과 밀접하게 연관되어 있다. 핵인자 kappa B (NF-κB)는 5개의 전사인자로 구성되며 염증 반응에 필수적인 역할을 한다. 다양한 암에서 NF-κB의 조절장애가 보고되고 있으며 NF-κB 조절이 암 치료에 있어 핵심 표적이 된다. 본 연구에서는 CDC Like Kinase 3 (CLK3)를 NF-κB 신호전달 경로를 조절하는 새로운 키네이스임을 확인하였다. 우리는 CLK3가 정규 및 비정규 NF-κB 신호전달경로를 억제하는 것을 밝혔다. CLK3 과발현 또는 knock-down 세포주를 이용한 루시퍼레이즈 분석 결과, 이 키네이스는 TNFα와 PMA가 유도하는 정규 NF-κB 신호전달경로 활성을 억제하였다. 또한 CLK3 과발현은 잘 알려진 비정규 NF-κB 신호경로 유도제인 NF-κB-inducing kinase (NIK) 또는 CD40에 의한 NF-κB 활성을 저해하였다. 추가적으로 CLK3의 NF-κB 신호전달 저해기전을 설명하고자 TNFα 처리 후 웨스턴 블롯 분석으로 이 키네이스 영향권 내에 있는 NF-κB 신호경로 분자들을 식별하였다. 그 결과 CLK3가 TAK1, IKKα/α, p65, IκBα 및 ERK1/2-MAPK의 인산화/활성화를 저해하여 TNFα 처리로 유도된 NF-κB 및 MAPK 신호경로를 모두 억제함을 밝혔다. 앞으로의 연구는 CLK3가 정규 및 비정규 NF-κB 경로를 억제하는 기작을 밝히는데 초점을 맞출 것이다. 위 연구 결과들을 토대로 CLK3가 NF-κB 신호전달경로의 새로운 음성 조절자로써 기능함을 제시하였다.

대봉감의 항산화, 항염증 및 면역증강 효과 (Beneficial Effects of Daebong Persimmon against Oxidative Stress, Inflammation, and Immunity in vivo)

  • 이희재;임소영;강민경;박정진;정현정;양수진
    • 한국식품영양과학회지
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    • 제44권4호
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    • pp.491-496
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    • 2015
  • 본 연구는 대표적인 떫은 감 품종인 대봉감과 반시의 항산화, 항염증 및 면역증강 효과를 탐색, 비교하였다. 쥐 모델을 대상으로 4주간 대봉감과 반시의 섭취는 식이섭취량과 체중 변화에 영향을 미치지 않았다. B지역의 대봉감 섭취는 고지방식이 대조군 대비 유의적으로 카탈레이즈 활성 수준을 증가시켰으며, 간 조직의 지질과산화물인 말론디알데하이드 수준은 고지방식이 대조군 대비 A지역 대봉감을 섭취한 군에서 유의적으로 감소하였다. 또한 A지역 대봉감의 섭취는 염증인자인 IL-1 beta의 수준을 고지방식이군 대비 유의적으로 감소시켰다. 또 다른 대표적인 염증인자인 IL-6는 고지방식이 대조군과 비교하여 A지역과 B지역의 대봉감을 섭취한 군에서 유의적으로 감소시켰다. TNF-alpha 수준은 A지역의 대봉감을 섭취한 군에서 고지방식이대조군 대비 유의적으로 감소되었다. A지역 대봉감 섭취는 IgG의 수준을 고지방식이 대조군 대비 유의적으로 증가시켰다. IGF-1의 수준은 고지방식이 대조군과 비교하여 A, B, C 지역 대봉감 섭취군에서 증가하였다. 이상의 결과로부터 대봉감의 섭취는 우수한 항염증 효과와 면역증강 효과가 있을 것으로 사료된다.

우라늄화합물로 오염된 금속폐기물의 전해제염 (Electrochemical Decontamination of Metallic Wastes Contaminated with Uranium Compounds)

  • 양영미;최왕규;오원진;유승곤
    • 방사성폐기물학회지
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    • 제1권1호
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    • pp.11-23
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    • 2003
  • 국내의 가동 중지된 우라늄 변환시설의 해체 시 우라늄 화합물로 오염되어 대량으로 발생될 금속폐기물의 재활용 또는 자체처분을 위한 제염기술로 전해제염 공정의 적용성을 평가하였다. 이를 위하여 우라늄 변환시설 내부설비의 주 구성 재료인 SUS-304 와 Inconel-600 금속시편을 대상으로 전해용해 실험을 수행하였다. SUS-304 와 Inconel-600 금속시편에 대한 전해용해 성능에 있어서 중성염 전해용액으로 $Na_2$SO$_4$가 가장 효과적이었으나, 우라늄변환시설의 가동 시 질산 매질과 주로 접촉했던 설비 표면의 이력과 시설 가동 중 발생한 우라늄 폐액의 성상을 고려하여 $Na_2$SO$_4$ 전해용액 내에서의 SUS-304 시편에 대한 전해용해와 비교해서 약 30%, 그리고 Inconel-600 시편에 대해서는 거의 동등한 성능을 보인 NaNO$_3$ 중성염 용액을 금속성폐기물의 전해제염 용액으로 선정하였다. 본 연구에서는 NaNO$_3$ 중성염 전해용액에서 전류밀도, 전해시간 및 전해 용액의 농도가 SUS-304 및 Inconel-600 금속시편의 전해용해 성능에 미치는 영향을 조사하였다. 이 실험결과를 바탕으로 실제 우라늄 변환시설로부터 인출하여 $UO_2$, AUC 및 ADU 등의 우라늄 화합물로 오염된 시편에 대해 전류밀도 100mA/$\textrm{cm}^2$, IM NaNO$_3$ 전해용액 내에서 전해 제염 실증시험을 수행하였으며, 오염물의 종류 및 오염준위의 대소와는 관계없이 모든 시편에 대하여 10분 이내의 짧은 시간 내에 자체처분 기준치 이하로 $\alpha$$\beta$ 방사능 준위를 감소시킴으로써 본 중성염 전해제염이 매우 성공적임을 확인하였다.nely regimented hierarchical language. I try, in this paper, to develop the idea that hierarchical regimentation of Korean language uses is not humane. 1 of for the main argument for the thesis as what follows: How could one justify the hierarchical regimentation of a language like Korean\ulcorner Only if there is an essential structure in which the fine grades of differences of social positions of all the people are distinct; The essentialism here involved is not plausible. And I may add that language is to be used fur the purposes of communication, rationalization and expression. If true, language use is a genuine art of liberation or humanization. Any overt hierarchical language tends to damage those purposes and more to enforce those oppressive elements already existing in the community. Then, a hierarchical language is to defeat its own purpose.중 행정부가 북한에 대해 실시한 포용정책이 어떠한 성과를 거두고 어떠한 문제점을 간과하고 있는가에 대해 논의하고, 대북 정책의 새로운 지평을 논의하는 것을 목적으로 하고 있다. 1) 포용 정책은

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삶기에 의한 국수호박의 영양성분 함량 변화 (Quantitative Changes of Nutritional Composition of Spaghetti Squash by Boiling)

  • 한혜경;강민승;나종민;윤현녀;김수연;김세나;김정봉;박홍주;조영숙;김소영
    • 한국식품조리과학회지
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    • 제27권6호
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    • pp.815-823
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    • 2011
  • This study was carried out to analyze the change of major nutrient components of spaghetti squash by boiling. The moisture, crude protein, fat, ash and carbohydrate contents in fresh squash were 94.2%, 0.6%, 0.1%, 0.7% and 4.4% respectively as against 95.1%, 0.5%, 0.1%, 0.5% and 3.8% in boiled squash. The decrease in protein and ash contents of the boiled sample were found to be significant. Major component of the minerals were potassium and the fresh and boiled squash had the contents of 330 mg and 256 mg, respectively. There were no differences of dietary fiber between the fresh and boiled squash. Beta-carotene contents of the fresh and boiled spaghetti squash were $0.69{\mu}g$ and $2.22{\mu}g$, respectively. The contents of tocopherol were decreased as like 4.3 mg and 2.0 mg. A total of 17 kinds of amino acids were isolated from squash and they were decreased by boiling and the high content of amino acids in order were aspartic acid, glutamic acid, arginine, lysine, and leucine in raw squash. Particularly, total amino acid of fresh squash were 6,739.5 mg per 100 g edible portion and higher than that of boiled squash(4,820.3 mg). Total polyphenolic compound of the fresh squash from $297.3{\mu}g/mg$ was slightly decreased to $253.3{\mu}g/mg$ by boiling.

Isolation and Genetic Transformation of Primordial Germ Cell (PGC)-Derived Cells from Cattle, Goats, Rabbits and Rats

  • Lee, C.K.;Moore, K.;Scales, N.;Westhusin, M.;Newton, G.;Im, K.S.;Piedrahita, J.A.
    • Asian-Australasian Journal of Animal Sciences
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    • 제13권5호
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    • pp.587-594
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    • 2000
  • At present embryonic stem (ES) cells with confirmed pluripotential properties are only available in the mouse. Recently, we were able to isolate, culture and genetically transform primordial germ cell (PGC)-derived cells from pig embryos and demonstrate their ability to contribute to chimera development in the pig. In order to determine whether the system we developed could be used to isolate embryonic germ (EG) cells from other mammalian species, we placed isolated PGCs from cattle, goats, rabbits and rats in culture. Briefly, PGCs were isolated from fetuses of cow (day 30-50), goat (day 25), rabbit (day 15-18) and rat (day 11-12), and plated on STO feeder cells in Dulbecco's modified Eagle's medium (DMEM): Ham's F10 medium (1:1) supplemented with 0.01 mM nonessential amino acids, 2 mM L-glutamine, 0.1 mM $\beta$ - mercaptoethnol, soluble recombinant human stem cell factor (SCF; 40ng/ml), human basic fibroblast growth factor (bFGF; 20ng/ml) and human leukemia inhibitory factor (LIF; 20ng/ml). For maintenance of the cells, colonies were passed to fresh feeders every 7-10 days. In all species tested, we were able to obtain and maintain colonies with ES-like morphology. Their developmental potential was tested by alkaline phosphatase (AP) staining and in vitro differentiation assay. For genetic transformation, cells were electroporated with a construct containing the green fluorescent protein (GFP) under the control of the cytomegalovirus (CMV) promoter. GFP-expressing colonies were detected in cattle, rabbits and rats. These results suggest that PGC-derived cells from cattle, goats, rabbits and rats can be isolated, cultured, and genetically transformed, and provide the basis for analyzing their developmental potential and their possible use for the precise genetic modification of these species.

Porphyromonas Gingivalis Invasion of Human Aortic Smooth Muscle Cells

  • Lee, Seoung-Man;Lee, Hyeon-Woo;Lee, Jin-Yong
    • International Journal of Oral Biology
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    • 제33권4호
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    • pp.163-177
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    • 2008
  • Periodontal disease, a form of chronic inflammatory bacterial infectious disease, is known to be a risk factor for cardiovascular disease (CVD). Porphyromonas gingivalis has been implicated in periodontal disease and widely studied for its role in the pathogenesis of CVD. A previous study demonstrating that periodontopathic P. gingivalis is involved in CVD showed that invasion of endothelial cells by the bacterium is accompanied by an increase in cytokine production, which may result in vascular atherosclerotic changes. The present study was performed in order to further elucidate the role of P. gingivalis in the process of atherosclerosis and CVD. For this purpose, invasion of human aortic smooth muscle cells (HASMC) by P. gingivalis 381 and its isogenic mutants of KDP150 ($fimA^-$), CW120 ($ppk^-$) and KS7 ($relA^-$) was assessed using a metronidazole protection assay. Wild type P. gingivalis invaded HASMCs with an efficiency of 0.12%. In contrast, KDP150 failed to demonstrate any invasive ability. CW120 and KS7 showed relatively higher invasion efficiencies, but results for these variants were still negligible when compared to the wild type invasiveness. These results suggest that fimbriae are required for invasion and that energy metabolism in association with regulatory genes involved in stress and stringent response may also be important for this process. ELISA assays revealed that the invasive P. gingivalis 381 increased production of the proinflammatory cytokine interleukin (IL)-$1{\beta}$ and the chemotactic cytokines (chemokine) IL (interleukin)-8 and monocyte chemotactic (MCP) protein-1 during the 30-90 min incubation periods (P<0.05). Expression of RANTES (regulation upon activation, normal T cell expressed and secreted) and Toll-like receptor (TLR)-4, a pattern recognition receptor (PRR), was increased in HASMCs infected with P. gingivalis 381 by RT-PCR analysis. P. gingivalis infection did not alter interferon-$\gamma$-inducible protein-10 expression in HASMCs. HASMC nonspecific necrosis and apoptotic cell death were measured by lactate dehydrogenase (LDH) and caspase activity assays, respectively. LDH release from HASMCs and HAMC caspase activity were significantly higher after a 90 min incubation with P. gingivalis 381. Taken together, P. gingivalis invasion of HASMCs induces inflammatory cytokine production, apoptotic cell death, and expression of TLR-4, a PRR which may react with the bacterial molecules and induce the expression of the chemokines IL-8, MCP-1 and RANTES. Overall, these results suggest that invasive P. gingivalis may participate in the pathogenesis of atherosclerosis, leading to CVD.