• Title/Summary/Keyword: Cellular capacity

Search Result 505, Processing Time 0.064 seconds

Analysis of calmodulin binding property of IQ motifs of IQGAP1 (IQGAP1내에 존재하는 IQ 부위들의 CaM 결합 특성 분석)

  • Jang, Deok-Jin
    • Analytical Science and Technology
    • /
    • v.24 no.6
    • /
    • pp.527-532
    • /
    • 2011
  • IQ motif-containing GTPase-activating protein 1 (IQGAP1), which is a well-known $Ca^{2+}$-independent calmodulin (CaM) binding protein, is involved in various cellular functions such as cell proliferation and cell migration. IQGAP1 has four repeated IQ motifs, which are crucial for CaM binding. It has been shown that all four IQ motifs of IQGAP1 can bind to $Ca^{2+}$/CaM, while the third and fourth IQ motifs of IQGAP1 can bind to apoCaM. However, it has not been clear whether the CaM binding of IQ motifs of IQGAP1 was mediated directly or indirectly. In this study, we examined whether the binding between CaM and each IQ motif of IQGAP1 was direct in vitro. As a result, we found that IQ1 motif has a weak $Ca^{2+}$-dependent CaM binding. In contrast, IQ3 has a $Ca^{2+}$-dependent CaM binding. All other motifs have no significant CaM binding. We also found that IQ(2.7-3) and IQ(3.5-4.4) have CaM binding capacity. This finding indicates that IQ motifs of IQGAP1 plays a dynamic role via different motif interactions with $Ca^{2+}$/CaM or proCaM.

Desmin Binding Property of Nebulin Isoforms

  • Jeon Eun-Hee;Lee Yeong-Mi;Lee Min-A;Kim Ji-Hee;Choi Jae-Kyong;Park Eun-Ran;Kim Hyun-Suk;Ahn Seung-Ju;Min Byung-In;Joo Young-Mi;Kim Chong-Rak
    • Biomedical Science Letters
    • /
    • v.12 no.2
    • /
    • pp.73-79
    • /
    • 2006
  • Nebulin is a giant ($600{\sim}900$ kDa), modular sarcomeric protein proposed to regulate the assembly, and to specify the precise lengths of actin filamints in vertebrate skeletal muscles. Recently, There is an evidence that the nebulin also expressed in non muscle tissue, brain and liver. We identified a new isoform of nebulin from adult brain library by PCR screening. It contains two simple-repeats exon 165, 166 and linker-repeats exon $154{\sim}161$ except exon 159. The nebulin modules M160 to M170 (exon 150 to exon 161) has been shown to bind desmin. In mature striated muscle, desmin intermediate filaments surround Z-discs and link individual myofibrils laterally at their Z-discs and to other intracellular structures, including the costameres and the intercalated discs of the sarcolemma, sarcoplasmic reticulum, mitochondria, T-tubules, and nuclei. Therefore, it is an interesting possibility that the differential splice pathways within the linker region of nebulin modify the affinity of nebulin's interaction with desmin. The specific interactions of nebulin and desmin were confirmed in vivo by yeast two hybrid experiments. To verify in the cellular level the interaction between nebulin isoform and desmin, we transfected COS-7 cell with EGFP-tagged nebulin and DsRed-tagged desmin. Based on evidence showing that despite exon 159 was deleted, the new isoform of nebulin was interact with desmin. This suggest that nebulin in brain may interact with another intermediate filament. The conservation of these ligand-binding capacity in brain and skeletal nebulins suggest that nebulins may have conserved roles in brain and skeletal muscle.

  • PDF

Adventitial Fibroblast Abormality in Thoracic Aortic Aneurysms and Aortic Dissections

  • Suh, Jong-Hui;Yoon, Jeong-Seob;Kim, Hwan-Wook;Jo, Keon-Hyon
    • Journal of Chest Surgery
    • /
    • v.44 no.6
    • /
    • pp.406-412
    • /
    • 2011
  • Background: Development of thoracic aortic aneurysms and aortic dissections (TAAD) is attributed to unbearable wall tension superimposed on defective aortic wall integrity and impaired aortic repair mechanisms. Central to this repair mechanisms are well-balanced and adequately functional cellular components of the aortic wall, including endothelial cells, smooth muscle cells (SMCs), inflammatory cells, and adventitial fibroblasts. Adventitial fibroblasts naturally produce aortic extracellular matrix (ECM), and, when aortic wall is injured, they can be transformed into SMCs, which in turn are involved in aortic remodeling. We postulated the hypothesis that adventitial fibroblasts in patients with TAAD may have defects in ECM production and SMC transformation. Materials and Methods: Adventitial fibroblasts were procured from the adventitial layer of fresh aortic tissues of patients with TAAD (Group I) and of multi-organ donors (Group II), and 4-passage cell culture was performed prior to the experiment. To assess ECM production, cells were treated with TNF-${\alpha}$ (50 pM) and the expression of MMP-2/MMP-3 was analyzed using western blot technique. To assess SMC transformation capacity, cells were treated with TGF-${\beta}1$ and expression of SM ${\alpha}$-actin, SM-MHC, Ki-67 and SM calponin was evaluated using western blot technique. Fibroblasts were then treated with TGF-${\beta}1$ (10 pM) for up to 10 days with TGF-${\beta}1$ supplementation every 2 days, and the proportion of transformed SMC in the cell line was measured using immunofluorescence assay for fibroblast surface antigen every 2 days. Results: MMP-3 expression was significantly lower in group I than in group II. TGF-${\beta}1$-stimulated adventitial fibroblasts in group I expressed less SM ${\alpha}$-actin, SM-MHC, and Ki-67 than in group II. SM-calponin expression was not different between the two groups. Presence of fibroblast was observed on immunofluorescence assay after more than 6 days of TGF-${\beta}1$ treatment in group I, while most fibroblasts were transformed to SMC within 4 days in group II. Conclusion: ECM production and SMC transformation are compromised in adventitial fibroblasts from patients with TAAD. This result suggests that functional restoration of adventitial fibroblasts could well be a novel approach for the prevention and treatment of TAAD.

Exploring the Metabolomic Responses of Bacillus licheniformis to Temperature Stress by Gas Chromatography/Mass Spectrometry

  • Dong, Zixing;Chen, Xiaoling;Cai, Ke;Chen, Zhixin;Wang, Hongbin;Jin, Peng;Liu, Xiaoguang;Permaul, Kugenthiren;Singh, Suren;Wang, Zhengxiang
    • Journal of Microbiology and Biotechnology
    • /
    • v.28 no.3
    • /
    • pp.473-481
    • /
    • 2018
  • Owing to its high protein secretion capacity, simple nutritional requirements, and GRAS (generally regarded as safe) status, Bacillus licheniformis is widely used as a host for the industrial production of enzymes, antibiotics, and peptides. However, as compared with its close relative Bacillus subtilis, little is known about the physiology and stress responses of B. licheniformis. To explore its temperature-stress metabolome, B. licheniformis strains ATCC 14580 and B186, with respective optimal growth temperatures of $42^{\circ}C$ and $50^{\circ}C$, were cultured at $42^{\circ}C$, $50^{\circ}C$, and $60^{\circ}C$ and their corresponding metabolic profiles were determined by gas chromatography/mass spectrometry and multivariate statistical analyses. It was found that with increased growth temperatures, the two B. licheniformis strains displayed elevated cellular levels of proline, glutamate, lysine, pentadecanoic acid, hexadecanoic acid, heptadecanoic acid, and octadecanoic acid, and decreased levels of glutamine and octadecenoic acid. Regulation of amino acid and fatty acid metabolism is likely to be associated with the evolution of protective biochemical mechanisms of B. licheniformis. Our results will help to optimize the industrial use of B. licheniformis and other important Bacillus species.

Establishment of Functional Cells for Vascular Defect Disease from Human Embryonic Stem Cell via Region Sorting Depending on Cell Volume (세포 크기 차이를 이용한 유세포 분석을 통한 인간배아줄기세포 유래 기능성 혈관세포의 확립)

  • Lee, Ji-Hye;Kim, Ju-Mi;Chung, Hyung-Min;Chae, Jung-Il
    • Microbiology and Biotechnology Letters
    • /
    • v.39 no.4
    • /
    • pp.364-373
    • /
    • 2011
  • Human embryonic stem cells have been highlighted as a valuable cellular source in the regenerative medicine field, due to their pluripotency. However, there is the challenge of the establishment of specific functional cell type forms of undifferentiated human embryonic stem cells (hESC). To establish and purify functional cell types from hESCs, we differentiated undifferentiated hESCs into vascular lineage cells and sorted the specific cell population from the whole cell population, depending on their cell volume, and compared them with the non-sorted cell population. We observed that about 10% of the PECAM positive population existed in the VEGF induced differentiating human embryoid body (hEB), and differentiated hEBs were made into single cells for cell transplantation. After making single cells, we performed cell sorting using a fluorescence-activated cell sorter (FACs), according to their cell volume on the basis of FSC region gating, and compared their therapeutic capacity with the non-sorted cell population through cell transplantation into hindlimb ischemic disease model mice. 4 Weeks after cell transplantation, the recovery rate of blood perfusion reached 54% and 17% in the FSC regions of sorted cells- and non-sorted cells, respectively. This result suggests that derivation of a functional cell population from hESCs can be performed through cell sorting on the basis of cell volume after preliminary differentiation induction. This approach may then greatly contribute to overcoming the limitations of marker sorting.

Investigation of Nanofiber and Thermosensitive Scaffold for Intervertebral Disc through Organ Culture (기관배양을 통한 추간판 재생용 나노파이버 및 온도 감응성 지지체에 대한 검증)

  • Lee, Yong-Jae;Shin, Ji-Won;Shin, Ho-Jun;Kim, Chan-Hwan;Park, Ki-Dong;Bae, Jin-Woo;Seo, Hyoung-Yeon;Kim, Young-Jick;Shin, Jung-Woog
    • Journal of Biomedical Engineering Research
    • /
    • v.28 no.4
    • /
    • pp.512-519
    • /
    • 2007
  • The purpose of this study is to investigate the potential of a novel tissue engineering approach to regenerate intervertebral disc. In this study, thermosensitive scaffold (chitosan-Pluronic hydrogel) and nanofiber were used to replace the nucleus pulposus (NP) and annulus fibrosus of a degenerated intervertebral disc, leading to an eventual regeneration of the disc using the minimally invasive surgical procedure and organ culture. In preliminary study, disc cells were seeded into the scaffolds and cellular responses were assessed by MTT assay and scanning electron microscopy (SEM). Based on these results, we could know that tissue engineered scaffolds might provide favorable environments for the regeneration of tissues. Organ culture was performed in fresh porcine spinal motion segments with endplates on both sides. These spinal motion segments were classified into three groups: control (Intact), injured NP (Defect), and inserting tissue engineered scaffolds (Insert). The specimens were cultivated for 7 days, subsequently structural stability, cell proliferation and morphological changes were evaluated by the relaxation time, quantity of DNA, GAG and histological examination. In these results, inserting group showed higher relaxation time, reduced decrement of DNA contents, and accumulated GAG amount. Consequently, the tissue engineered scaffolds used in this study seen to be a promising base scaffolds for regenerative intervertebral disc due to its capacity to absorb external dynamic loading and the possible ideal environment provided for disc cell growing.

No short-term effects of calorie-controlled Mediterranean or fast food dietary interventions on established biomarkers of vascular or metabolic risk in healthy individuals

  • Parcina, Marijo;Brune, Maik;Kaese, Vareska;Zorn, Markus;Spiegel, Rainer;Vojvoda, Valerija;Fleming, Thomas;Rudofsky, Gottfried;Nawroth, Peter Paul
    • Nutrition Research and Practice
    • /
    • v.9 no.2
    • /
    • pp.165-173
    • /
    • 2015
  • BACKGROUND/OBJECTIVES: This study addressed the question whether the composition of supposedly 'healthy' or 'unhealthy' dietary regimes has a calorie-independent short-term effect on biomarkers of metabolic stress and vascular risk in healthy individuals. SUBJECTS/METHODS: Healthy male volunteers (age $29.5{\pm}5.9years$, n = 39) were given a standardized baseline diet for two weeks before randomization into three groups of different dietary regimes: fast food, Mediterranean and German cooking style. Importantly, the amount of calories consumed per day was identical in all three groups. Blood samples were analyzed for biomarkers of cardiovascular risk and metabolic stress after two weeks of the baseline diet and after two weeks of the assigned dietary regime. RESULTS: No dietary intervention affected the metabolic or cardiovascular risk profile when compared in-between groups or compared to baseline. Subjects applied to the Mediterranean diet showed a statistically significant increase of uric acid compared to baseline and compared to the German diet group. Plasma concentrations of urea were significantly higher in both the fast food group and the Mediterranean group, when compared to baseline and compared to the German diet group. No significant differences were detected for the levels of vitamins, trace elements or metabolic stress markers (8-hydroxy-2-deoxyguanosine, malondialdehyde and methylglyoxal, a potent glycating agent). Established parameters of vascular risk (e.g. LDL-cholesterol, lipoprotein(a), homocysteine) were not significantly changed in-between groups or compared to baseline during the intervention period. CONCLUSIONS: The calorie-controlled dietary intervention caused neither protective nor harmful short-term effects regarding established biomarkers of vascular or metabolic risk. When avoiding the noxious effects of overfeeding, healthy individuals can possess the metabolic capacity to compensate for a potentially disadvantageous composition of a certain diet.

Performance Analysis of Population-Based Bandwidth Reservation Scheme with Various Request Reservation Ratios (요청 예약 비율에 따른 Population-Based Bandwidth Reservation 구조의 성능 분석)

  • Kwon, Se-Dong;Han, Man-Yoo;Park, Hyun-Min
    • The KIPS Transactions:PartC
    • /
    • v.9C no.3
    • /
    • pp.385-398
    • /
    • 2002
  • To accommodate the increasing number of mobile terminals in the limited radio spectrum, wireless systems have been designed as micro/picocellular architectures for a higher capacity. This reduced coverage area of a cell has caused a higher rate of hand-off events, and the hand-off technology for efficient process becomes a necessity to provide a stable service. Population-based Bandwidth Reservation(PBR) Scheme is proposed to provide prioritized handling for hand-off calls by dynamically adjusting the amount of reserved bandwidth of a cell according to the amount of cellular traffic in its neighboring cells. We analyze the performance of the PBR scheme according to the changes of a fractional parameter, f, which is the ratio of request reservation to the total amount of bandwidth units required for hand-off calls that will occur for the next period. The vague of this parameter, f should be determined based on QoS(Quality of Service) requirement. To meet the requirement the value of Parameter(f) must be able to be adjusted dynamically according to the changing traffic conditions. The best value of f can be determined by a function of the average speed of mobile stations, average call duration, cell size, and so on. This paper considers the average call duration and the cell size according to the speed of mobile stations. Although some difference exists as per speed, in the range of 0.4 $\leq$ f $\leq$ 0.6, Blocking Probability, Dropping Probability and Utilization show the best values.

System Throughput of Cognitive Radio Multi-hop Relay Networks (무선인지 멀티홉 릴레이 네트워크의 시스템 스루풋)

  • Hassan, I.;Rho, Chang-Bae;Song, Ju-Bin
    • Journal of the Institute of Electronics Engineers of Korea TC
    • /
    • v.46 no.4
    • /
    • pp.29-39
    • /
    • 2009
  • The need for radio spectrum is recently considered as a huge hurdle towards the rapid development of wireless networks. Large parts of the spectrum are allocated to licensed radio services in proprietary way. However, enormous success of the wireless services and technologies in the unlicensed bands has brought new ideas and innovations. In recent years cognitive radio has gained much attention for solving the spectrum scarcity problem. It changes the way spectrum is regulated so that more efficient spectrum utilization is possible. Multi-hop relay technology on the other hand has intensively been studied in the area of ad hoc and peer-to-peer networks. But in cellular network, only recently the integration of multi-hop capability is considered to enhance the performance significantly. Multi-hop relaying can extend the coverage of the cell to provide high data rate service to a greater distance and in the shadowed regions. Very few papers still exist that combine these methods to maximize the spectrum utilization. Thus we propose a network architecture combining these two technologies in a way to maximize the system throughput. We present the throughput capacity equations for the proposed system model considering various system parameters like utilization factor by the primary users and primary users' transmission radius and through extensive numerical simulations we analyze the significance of work.

Comparative Analysis of Host Insect Immunodepression Induced by Two Entomopathogenic Bacteria, Xenorhabdus nematophilus and Staphylococcus gallinarum, with Differential Pathogenicities (병원력 차이를 보이는 두 곤충병원세균(Xenorhabdus nematophilus와 Staphylococcus gallinarum)의 면역저하 능력 비교 분석)

  • 박영진;김길호;김용균
    • Korean journal of applied entomology
    • /
    • v.42 no.4
    • /
    • pp.353-360
    • /
    • 2003
  • Immunodepression can be required for entomopathogenic bacteria to induce their potent pathogenicities to the target insects. Here, we raise a hypothesis that the capacity of a pathogenic bacterium to induce the target insect immunodepression has positive relationship with the degree of pathogenicity. X. nematophilus had 1,200 times as potent as another entomopathogenic bacterium, Staphylococcus gallinarum against the fifth instar larvae of silkworm, Bombyx mori, when they were Injected into the hemocoel. Although both bacteria had significant cytotokic effect on the hemocytes of B. mori, X. nematophilus gave faster and greater cytotoxicity than did S. gallinarum. In cellular immune reactions, B. mori could form 20 hemocyte nodules against the bacterial injection with 5${\times}$10$\^$5/ cells. The number of the hemocyte nodules was significantly depressed when live X. nematophilus was inject-ed, but not in S. gallinarum. Activation of prophenoloxidase (proPO) was depressed in the bacterial injection. The depression of PO activation was significantly greater in X. nematophilus infection than in S. gallinarum injection. Lysozyme activity was induced by the injection of S. gallinarum at 4 h after the treatment, but not induced in X. nematophilus at all the time. These results showed that X. nemato-philus induced greater immunodepression against B. mori and resulted in higher pathogenicity than did S. gallinarum. Therefore, this study suggests that the immunodepression induced by entomopathogenic bacteria has positive relationship with their pathogenicity.