• Title/Summary/Keyword: mechanisms

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Adhesion-induced generation of oxygen free radical from human alveolar macrophages and its mechanisms (폐포대식세포의 부착에 의한 산소유리기 분비능 활성화 및 그 기전)

  • Chung, Man-Pyo;Yoo, Chul-Gyu;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.2
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    • pp.210-220
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    • 1996
  • Background : Neutrophils or monocytes separated in vitro by the adherence to plastic surface are known to be activated by surface adherence itself and subsequent experimental data might be altered by surface adherence. In the process of surface adherence, adhesion molecules have a clear role in intracellular signal pathway of cellular activation. Human alveolar macrophages(HAM) are frequently purified by the adherence procedure after bronchoalveolar lavage. But the experimental data of many reports about alveolar macrophages have ignored the possibility of adhesion-induced cellular activation. Method : Bronchoalveolar lavage was performed in the person whose lung of either side was confirmed to be normal by chest CT. With the measurement of hydrogen peroxide release from adherent HAM to plastic surface and non-adherent HAM with or without additional stimulation of phorbol myristate acetate(PMA) or N-formyl-methionyl-leucyl-phenylalanine (fMLP), we observed the effect of the adherence to plastic surface. We also evaluated the effect of various biological surfaces on adhesion-induced activation of HAM. Then, to define the intracellular pathway of signal transduction, pretreatment with cycloheximide, pertussis toxin and anti-CD11/CD18 monoclonal antibody was done and we measured hydrogen peroxide in the culture supernatant of HAM. Results : 1) The adherence itself to plastic surface directly stimulated hydrogen peroxide release from human alveolar macrophages and chemical stimuli such as phorbol myristate acetate(PMA) or N-formyl-methionyl-leucyl-phenylalanine(fMLP) colud not increase hydrogen peroxide release in these adherent macrophages which is already activated. 2) PMA activated human alveolar macrophages irrespective of the state of adhesion. However, fMLP stimulated the release of hydrogen peroxide from the adherent macrophages, but not from the non-adherent macrophages. 3) HAM adherent to A549 cell(type II alveolar epithelium-like human cell line) monolayer released more hydrogen peroxide in response to both PMA and fMLP. This adherence-dependent effect of fMLP was blocked by pretreatment of macrophages with cycloheximide, pertussis toxin and anti-CD18 monoclonal antibody, Conclusion : These results suggest that the stimulatory effect of PMA and fMLP can not be found in adherent macrophage because of the activation of human alveolar macrophage by the adherence to plastic surface and the cells adhered to biologic surface such as alveolar epithelial cells are appropriately responsive to these stimuli. It is also likely that the effect of fMLP on the adherent macrophage requires new protein synthesis via G protein pathway and is dependent on the adhesion between alveolar macrophages and alveolar epithelial cells by virtue of CD11/CD18 adhesion molecules.

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EARLY DETECTION OF INITIAL DENTAL CARIES USING A $DIFOTI^{TM}$ (Digital Imaging Fiber-Optic Trans-illumination을 이용한 초기 법랑질 우식병소의 조기 진단)

  • Yeom, Hae-Woong;Yoo, Seung-Hoon;Kim, Jong-Soo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.31 no.4
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    • pp.587-597
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    • 2004
  • Over the past 20 years, great strides have been made in research regarding the mechanisms involved in the progression of carious lesions, but new equipment and research tools need to be developed to continue these advancements in caries research. Various methods have been applied to reduce the incidence of carious lesions, which have led to a significant decrease in the number of occlusal caries, but a concurrent increase in the proportion of proximal carious lesions. New diagnostic equipment has been developed to detect early stage carious lesions, and these have demonstrated excellent laboratory results and show promise in clinical applications. The research presented here examines the efficacy of the newly developed $DIFOTI^{TM}$ system in detecting proximal carious lesions compared to traditional intraoral exam and bitewing radiography, possible problems or deficiencies of using the system in clinic, possible improvements that can be made to the system, and the efficacy of detecting early, reversible carious lesions that can be remineralized by preventative fluoride applications. The subject pool consisted of 23 grammer school age patients just prior to entering the mixed dentition phase. Each patient was given a thorough oral examination, radiographic examination consisting of bitewing radiographs of the posterior teeth, and $DIFOTI^{TM}$ examination of the anterior and posterior teeth. Each examination was carried out two times by two examiners, and the data were statistically analyzed. The results are as follows: 1. The mean alpha value of reliability test of the visual oral examination was as follows; occlusal surface was 0.8470. mesial surface was 0.6430, distal surface was 0.5727. lingual surface was 0.2807 and distal surface was 0.2339. When the examination was limited to posterior teeth, the mean alpha value was as follows; occlusal surface was 0.8577, distal surface was 0.8211, lingual surface was 0.7728, buccal surface was 0.7152 and mesial surface was 0.6782. 2. The alpha value of reliability test of the radiographic analysis of carious lesions of the occlusal, mesial, and distal surfaces was 0.8500. 3. The alpha value of reliability test of the $DIFOTI^{TM}$ diagnostic analysis of carious lesions of the occlusal, buccal, lingual, mesial, and distal surfaces was determined to be 0.7917. 4. The $DIFOTI^{TM}$ diagnostic system was found to be the most accurate means of detecting occlusal, buccal, and lingual surface carious lesions (p<0.05), while mesial and distal proximal carious lesions were most accurately assessed using bitewing radiography (p<0.05).

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Seismic study of the Ulleung Basin crust and its implications for the opening of the East Sea (탄성파 탐사를 통해 본 울릉분지의 지각특성과 동해형성에 있어서의 의미)

  • Kim, Han Jun
    • Journal of the Korean Geophysical Society
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    • v.2 no.1
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    • pp.9-26
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    • 1999
  • The Ulleung Basin (Tsushima Basin) in the southwestern East Sea (Japan Sea) is floored by a crust whose affinity is not known whether oceanic or thinned continental. This ambiguity resulted in unconstrained mechanisms of basin evolution. The present work attempts to define the nature of the crust of the Ulleung Basin and its tectonic evolution using seismic wide-angle reflection and refraction data recorded on ocean bottom seismometers (OBSs). Although the thickness of (10 km) of the crust is greater than typical oceanic crust, tau-p analysis of OBS data and forward modeling by 2-D ray tracing suggest that it is oceanic in character: (1) the crust consists of laterally consistent upper and lower layers that are typical of oceanic layers 2 and 3 in seismic velocity and gradient distribution and (2) layer 2C, the transition between layer 2 and layer 3 in oceanic crust, is manifested by a continuous velocity increase from 5.7 to 6.3 km/s over the thickness interval of about 1 km between the upper and lower layers. Therefore it is not likely that the Ulleung Basin was formed by the crustal extension of the southwestern Japan Arc where crustal structure is typically continental. Instead, the thickness of the crust and its velocity structure suggest that the Ulleung Basin was formed by seafloor spreading in a region of hotter than normal mantle surrounding a distant mantle plume, not directly above the core of the plume. It seems that the mantle plume was located in northeast China. This suggestion is consistent with geochemical data that indicate the influence of a mantle plume on the production of volcanic rocks in and around the Ulleung Basin. Thus we propose that the opening models of the southwestern East Sea should incorporate seafloor spreading and the influence of a mantle plume rather than the extension of the crust of the Japan Arc.

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Purification of Human HtrA1 Expressed in E. coli and Characterization of Its Serine Protease Activity (E. coli에서 발현된 human HtrA1 단백질의 정제와 HtrA1의 serine protease 활성 조건에 관한 연구)

  • Kim, Kyung-Hee;Kim, Sang-Soo;Kim, Goo-Young;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.16 no.7 s.80
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    • pp.1133-1140
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    • 2006
  • Human HtrA1 (High temperature requirement protein A1) is a homologue of the E. coli periplasmic serine protease HtrA. A recent study has demonstrated that HtrA1 is a serine protease involved in processing of insulin like growth factor binding protein (ICFBP), indicating that it serves as an important regulator of IGF activity. Additionally, several lines of evidence suggest a striking correlation between proteolytic activity of HtrA1 serine protease and the pathogenesis of several diseases; however, physiological roles of HtrA1 remain to be elucidated. We used the pGEX bacterial expression system to develop a simple and rapid method for purifying HtrA1, and the recombinant HtrA1 protein was utilized to investigate the optimal conditions in executing its proteolytic activity. The proteolytically active HtrA1 was purified to approximately 85% purity, although the yield of the recombinant HtrA1 protein was slightly low $460{\mu}g$ for 1 liter E. coli culture). Using in vitro endoproteolytic cleavage assay, we identified that the HtrA1 serine protease activity was dependent on the enzyme concentration and the incubation time and that the best reaction temperature was $42^{\circ}C$ instead of $37^{\circ}C$. We arbitrary defined one unit of proteolytic activity of the HtrA1 serine protease as 200nM of HtrA1 that cleaves half of $5{\mu}M\;of\;{\beta}-casein$ during 3 hr incubation at $37^{\circ}C$. Our study provides a method for generating useful reagents to investigate the molecular mechanisms by which HtrA1 serine protease activity contributes in regulating its physiological function and to identify natural substrates of HtrA1.

A Study on the Gene Expression of Adipogenic Regulators by an Herbal Composition (생약복합물에 의한 지방세포형성 조절자의 유전자 발현 연구)

  • Lee, Hae-Yong;Kang, Ryun-Hwa;Bae, Sung-Min;Chae, Soo-Ahn;Lee, Jung-Ju;Oh, Dong-Jin;Park, Suk-Won;Cho, Soo-Hyun;Shim, Yae-Jie;Yoon, Yoo-Sik
    • Journal of Life Science
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    • v.20 no.5
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    • pp.729-735
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    • 2010
  • In our previous study, it was reported that an herbal mixture, SH21B, inhibits fat accumulation and adipogenesis both in vitro and in vivo models of obesity. SH21B is a mixture composed of seven herbs: Scutellaria baicalensis Georgi, Prunus armeniaca Maxim, Ephedra sinica Stapf, Acorus gramineus Soland, Typha orientalis Presl, Polygala tenuifolia Willd, and Nelumbo nucifera Gaertner (Ratio 3:3:3:3:3:2:2). The aim of this study was to investigate the detailed molecular mechanisms of the effects of SH21B on various regulators of the adipogenesis pathway. During the adipogenesis of 3T3-L1 cells, SH21B significantly decreased the expression levels of central transcription factors of adipogenesis, such as peroxisome proliferator-activated receptor (PPAR)$\gamma$ and CCAAT/enhancer binding protein (C/EBP)$\alpha$. To elucidate the detailed molecular mechanism of the anti-adipogenic effects of SH21B, we examined the expression levels of the various pro-adipogenic or anti-adipogenic regulators of adipogenesis upstream of $PPAR{\gamma}$ and C/$EBP{\alpha}$. The mRNA levels of Krox20 and Kruppel-like factor (KLF) 15, which are pro-adipogenic regulators, were significantly down-regulated by SH21B treatment, whereas the mRNA levels of C/$EBP{\gamma}$ and KLF5 were not changed. KLF2 and C/EBP homologous protein (CHOP), which are anti-adipogenic regulators, were significantly up-regulated by SH21B treatment. These results suggest that the molecular mechanism of the anti-adipogenic effect of SH21B involves both the down-regulations of pro-adipogenic regulators, such as Krox20 and KLF15, and the up-regulations of anti-adipogenic regulators, such as KLF2 and CHOP, which results in the suppression of central transcription factors of adipogenesis including $PPAR{\gamma}$ and C/$EBP{\alpha}$.

Transcriptome Analyses for the Anti-Adipogenic Mechanism of an Herbal Composition (생약복합물의 지방세포형성억제 기전규명을 위한 전사체 분석)

  • Lee, Hae-Yong;Kang, Ryun-Hwa;Bae, Sung-Min;Chae, Soo-Ahn;Lee, Jung-Ju;Oh, Dong-Jin;Park, Suk-Won;Cho, Soo-Hyun;Shim, Yae-Jie;Yoon, Yoo-Sik
    • Journal of Life Science
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    • v.20 no.7
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    • pp.1054-1065
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    • 2010
  • SH21B is a natural composition composed of seven herbs: Scutellaria baicalensis Georgi, Prunus armeniaca Maxim, Ephedra sinica Stapf, Acorus gramineus Soland, Typha orientalis Presl, Polygala tenuifolia Willd and Nelumbo nucifera Gaertner (Ratio 3:3:3:3:3:2:2). In our previous study, we reported that SH21B inhibited adipogenesis and fat accumulation in 3T3-L1 cells through modulation of various regulators in the adipogenesis pathway. The aim of this study was to analyze the transcriptome profiles for the anti-adipogenic effects of SH21B in 3T3-L1 cells. Total RNAs from SH21B-treated 3T3-L1 cells were reverse-transcribed into cDNAs and hybridized to Affymetrix Mouse Gene 1.0 ST array. From microarray analyses, we identified 2,568 genes of which expressions were changed more than two-fold by SH21B, and the clustering analyses of these genes resulted in 9 clusters. Three clusters among the 9 showed down-regulation by SH21B (cluster 4, cluster 6 and cluster 9), and two clusters showed up-regulation by SH21B (cluster 7 and cluster 8) during the adipogenesis of 3T3-L1 cells. It was found that many genes related to cell proliferation and adipogenesis were included in these clusters. Clusters 4, 6 and 9 included genes which were related with adipogenesis induction and cell cycle arrest. Clusters 7 and 8 included genes related to cell proliferation as well as adipogenesis inhibition. These results suggest that the mechanisms of the anti-adipogenic effects of SH21B may be the modulation of genes involved in cell proliferation and adipogenesis.

Anti-proliferative Activities of Solvent Fractions of Lees Extracts in Human Colorectal HCT116 Cells (대장암 세포주에서 주박 추출물의 유기용매 분획물의 항성장 활성)

  • Kang, Hyung-Taek;Lee, Seung Hoon;Kim, Soon Young;Kim, Mi-Sun;Shin, Woo-Chang;Sohn, Ho-Yong;Kim, Jong-Sik
    • Journal of Life Science
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    • v.24 no.9
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    • pp.967-972
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    • 2014
  • In the present study, we prepared eighty-five different kinds of lees extracts and their solvent fractions and investigated their anti-proliferative activities against human colorectal cancer HCT116 cells. HCT116 cells were treated with eighty-five solvent fractions of lees extracts and then cell viability was measured using MTS assay. Among the treated solvent fractions, three solvent fractions (KSD-E1-3, KSD-E2-3, and KSD-E4-3) were selected based on cell viability assay. In addition, we performed an oligo DNA microarray analysis to analyze the gene expression changes by treatment of KSD-E1-3 in HCT116 cells. Among the upregulated genes, we selected 4 genes (NAG-1, ATF3, p21, and DDIT3) and performed RT-PCR using gene-specific primers. Among the treated solvent fractions, KSD-E1-3 dramatically induced the expressions of the four selected genes. In addition, we investigated whether the upregulations of those genes were dependent on the transcription factor p53's presence using p53 null HCT116 cells. The results indicate that the upregulations of NAG-1, ATF3, and DDIT3 are not dependent on the p53 presence, whereas p21 is dependent on the p53 presence. These findings may help to understand the molecular mechanisms of the anti-proliferative activity mediated by rice wine lees in human colorectal cancer cells.

Cytoprotective Effects of Schisandrin A against Hydrogen Peroxide-induced Oxidative Stress in SW1353 Human Chondrocytes (SW1353 인간 연골세포에서 산화적 스트레스에 대한 schisandrin A의 세포 보호 효과)

  • Jeong, Jin-Woo;Choi, Eun Ok;Kwon, Da Hye;Kim, Bum Hoi;Park, Dong Il;Hwang, Hye Jin;Kim, Byung Woo;Choi, Yung Hyun
    • Journal of Life Science
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    • v.27 no.9
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    • pp.1070-1077
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    • 2017
  • Chondrocyte apoptosis induced by reactive oxygen species (ROS) plays an important role in the pathogenesis of osteoarthritis. Schisandrin A, a bioactive compound found in fruits of the Schisandra genus, has been reported to possess multiple pharmacological and therapeutic properties. Although several studies have described the antioxidant effects of analogues of schisandrin A, the underlying molecular mechanisms of this bioactive compound remain largely unresolved. The present study investigated the cytoprotective effect of schisandrin A against oxidative stress (hydrogen peroxide [$H_2O_2$]) in SW1353 human chondrocyte cells. The results showed that schisandrin A preconditioning significantly inhibited $H_2O_2-induced$ growth inhibition and apoptotic cell death by blocking the degradation of poly (ADP-ribose) polymerase proteins and down-regulating pro-caspase-3. These antiapoptotic effects of schisandrin A were associated with attenuation of mitochondrial dysfunction and normalization of expression changes of proapoptotic Bax and antiapoptotic Bcl-2 in $H_2O_2-stimulated$ SW1353 chondrocytes. Furthermore, schisandrin A effectively abrogated $H_2O_2-induced$ intracellular ROS accumulation and phosphorylation of histone H2AX at serine 139, a widely used marker of DNA damage. Thus, the present study demonstrates that schisandrin A provides protection against $H_2O_2-induced$ apoptosis and DNA damage in SW1353 chondrocytes, possibly by prevention of ROS generation. Collectively, our data indicate that schisandrin A has therapeutic potential in the treatment of oxidative disorders caused by overproduction of ROS.

Intracerebral Regional and Vasculature-Specific Distributions of Ischemic Cerebrovascular Diseases: Using MRI and MRA (MRI와 MRA를 이용한 허혈성 뇌혈관 질환의 뇌혈관별 분포에 대한 연구)

  • Kim, Ham-Gyum
    • Journal of radiological science and technology
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    • v.33 no.3
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    • pp.223-230
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    • 2010
  • The purpose of this study was to utilize Magnetic Resonance Imaging (MRI) and Magnetic Resonance Angiography (MRA) to analyze intracerebral regional distributions (hot spot) of ischemic cerebrovascular diseases which were characterized by stenosis and occlusion cerebral vasculature, except for cerebrovascular diseases induced by rupture of cerebral vasculature in terms of Korean people's cerebrovascular diseases, so that it could apply the findings of analysis to clinical practices. This study focused only on analyzing intracerebral regional distributions of ischemic cerebrovascular diseases that are characterized by stenosis and occlusion cerebral vasculature, because there are different etiologic mechanisms of ischemic cerebrovascular diseases like hemorrhagic cerebrovascular diseases (caused by rupture of cerebral vasculature) and cerebral infarction (induced by atheromatous arteriosclerosis). As a result, this study could come to the following findings of analysis: 1. According to sex ratio analysis, it was found that male group comprised larger portion of total 626 subjects in this study than female one (55.0% > 45.0%). 2. According to analysis on actual intracerebral regional distributions of ischemic cerebrovascular diseases, it was found that most subjects (37.5 %) were attacked by such diseases on the right side of cerebral vasculature, which was followed by left side of cerebral vasculature (35.1%) and bilateral cerebral vasculature (27.3%) respectively. 3. According to analysis on actual intracerebral regional distributions of ischemic cerebrovascular diseases, it was found that internal carotid artery (ICA) comprised the largest portion (38.9%) of those distributions, which was followed by middle cerebral artery (MCA, 35.7%), posterior cerebral artery (PCA, 13.4%), anterior cerebral artery (ACA, 6.0%) and vertebral artery (VA, 3.3%) respectively. 4. It was found that there was no subject attacked by any disease on A-com region, and there was only one male subject attacked by cerebrovascular diseases on P-com region. 5. It was found that female group was more susceptible to the attack of cerebrovascular diseases on MCA region than male one (54.6% > 42.2%), which means significant differences depending upon sex on statistical basis ($x^2$ = 9.64, p < .01). 6. It was found that male group was more susceptible to the attack of cerebrovascular diseases on ICA region (56.4% > 46.8%), which means significant differences depending upon sex on statistical basis ($x^2$ = 5.71, p < .05). 7. Moreover, it was also found that male group was more susceptible to the attack of cerebrovascular diseases on BA region (2.3% > 0.4%), which means significant differences depending upon sex on statistical basis ($x^2$ = 4.25, p < .05). 8. However, it was found that there was not any significant difference in intracerebral vasculature-specific distributions of cerebrovascular diseases depending on age of subjects, and stenosis comprised larger portion of cerebrovascular diseases than occlusion.

Behavior of NOM Fouling in Submerged Photocatalytic Membrane Reactor Combined with $TiO_2$ Nanoparticles ($TiO_2$ 나노입자/UV 결합 침지형 중공사막 시스템에서 자연유기물의 파울링거동)

  • Park, Seung-Soo;Seo, Hyung-Jun;Kim, Jeong-Hwan
    • Membrane Journal
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    • v.21 no.1
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    • pp.46-54
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    • 2011
  • In this study, combined effect of airflow rate, $TiO_2$ concentration, solution pH and $Ca^{+2}$ addition on HA (humic acid) fouling in submerged, photocatalytic hollow-fiber microfiltraiton was investigated systematically. Results showed that UV irradiation alone without $TiO_2$ nanoparticles could reduce HA fouling by 40% higher than the fouling obtained without UV irradiation. Compared to the HA fouling without UV irradiation and $TiO_2$ nanoparticles, the HA fouling reduction was about 25% higher only after the addition of $TiO_2$ nanoparticles. Both adsorptive and hydrophilic properties of $TiO_2$ nanoparticles for the HA can be involved in mitigating membrane fouling. It was also found that the aeration itself had lowest effect on fouling mitigation while the HA fouling was affected significantly by solution pH. Transient behavior of zeta potential at different solution pHs suggested that electrostatic interactions between HA and $TiO_2$ nanoparticles should improve photocatalytic efficiency on HA fouling. $TiO_2$ concentration was observed to be more important factor than airflow rate to reduce HA fouling, implying that surface reactivity on $TiO_2$ naoparticles should be important fouling mitigation mechanisms in submerged, photocatalyic microfiltraiton. This was further supported by investigating the effect of $Ca^{+2}$ addition on fouling mitigation. At higher pH (= 10), addition of $Ca^{+2}$ can play an important role in bridging between HA and $TiO_2$ nanoparticles and increasing surface reactivity on nanoparticles, thereby reducing membrane fouling.