• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.8
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• pp.1051-1056
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• 2006

Response surface methodology was used to investigate the quality of clarified mixed apple and carrot juices using ultrafitration. Apple and carrot juices were blended at the ratio of 1:3, 1:1, and 3:1. A three-variable, three-level central composite design was employed where the independent variables were the blend ratio, temperature and average transmembrane pressure (ATP). With increasing temperature and pressure, flux linearly increased regardless of blending ratio. Blend juice with 75% apple showed the highest soluble sugar and total sugar content in apple and carrot blend juices. Soluble solid contents were more affected by blending ratio than temperature and ATP. Total sugar contents were greatly affected by temperature; increasing temperature led to higher total sugar content up to $25^{\circ}C$. Higher carrot ratio led to higher vitamin C content. In general, higher acidity was achieved by higher apple content and acidity was increased with increasing temperature. Turbidity increased for all samples as APT increased, with the blending ratio of 1:1 (apple:carrot) showing the highest turbidity. Viscosity was greatly changed in the blending ratio of 3:1 (apple:carrot) juice. The polynomial models developed by RSM were satisfactory to describe the relationships between the studied factors and the responses. Analytical optimization gave $flux=0.216\;L/m^2.h$, soluble $solids=10.39^{\circ}Brix$, total sugar=71.32 mg/mL, vitamin C=315.18 mg%, acidity=7.78 mL, turbidity=0.017, and viscosity=1.44 cp, when using a $temperature=44.97^{\circ}C$, ATP=113.57 kPa, and blend ratio=28.50%.

• Archives of Pharmacal Research
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• v.26 no.10
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• pp.846-854
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• 2003

The oligomerization of G-proteincoupled receptors (GPCRs) has been shown to occur by various mechanisms, such as via disulfide covalent linkages, non covalent (ionic, hydrophobic) interactions of the N-terminal, and/or transmembrane and/or intracellular domains. Interactions between GPCRs could involve an association between identical proteins (homomers) or non-identical proteins (heteromers), or between two monomers (to form dimers) or multiple monomers (to form oligomers). It is believed that muscarinic receptors may also be arranged into dimeric or oigomeric complexes, but no systematic experimental evidence exists concerning the direct physical interaction between receptor proteins as its mechanism. We undertook this study to determine whether muscarinic receptors form homomers or a heteromers by direct protein-protein interaction within the same or within different subtypes using a yeast two-hybrid system. Intracellular loops (i1, i2 and i3) and the C-terminal cytoplasmic tails (C) of human muscarinic (Hm) receptor subtypes, Hm1, Hm2 and Hm3, were cloned into the vectors (pB42AD and pLexA) of a two-hybrid system and examined for heteromeric or homodimeric interactions between the cytoplasmic domains. No physical interaction was observed between the intracellular domains of any of the Hm/Hm receptor sets tested. The results of our study suggest that the Hm1, Hm2 and Hm3 receptors do not form dimers or oligomers by interacting directly through either the hydrophilic intracellular domains or the C-terminal tail domains. To further investigate extracellular domain interactions, the N-terminus (N) and extracellular loops (o1 and o2) were also cloned into the two-hybrid vectors. Interactions of Hm2N with Hm2N, Hm2o1, Hm2o2, Hm3N, Hm3o1 or Hm3o2 were examined. The N-terminal domain of Hm2 was found to have no direct interaction with any extracellular domain. From our results, we excluded the possibility of a direct interaction between the muscarinic receptor subtypes (Hm1, Hm2 and Hm3) as a mechanism for homo- or hetero-meric dimerization/oligomerization. On the other hand, it remains a possibility that interaction may occur indirectly or require proper conformation or subunit formation or hydrophobic region involvement.

• IMMUNE NETWORK
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• v.11 no.2
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• pp.114-122
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• 2011

Background: The leukocyte common antigen (CD45) is a transmembrane-type protein tyrosine phosphatase that has five isoforms. Methods: We generated seven murine mAbs against human CD45 by injecting cells from different origins, such as human thymocytes, PBMCs, and leukemic cell lines. By using various immunological methods including flow cytometry, immunohistochemistry, and immunoprecipitation, we evaluated the reactivity of those mAbs to CD45 of thymus as well as tonsil lysates. Furthermore, we transiently transfected COS-7 cells with each of gene constructs that express five human CD45 isoforms respectively, and examined the specificities of the mAbs against the transfected isoforms. Results: In case of thymocytes, lymphocytes, and monocytes, all the seven mAbs demonstrated positive reactivities whereas none was reactive to erythrocytes and platelets. The majority of immune cells in formalin-fixed paraffin-embedded thymus and tonsil tissues displayed strong membranous immunoreactivity, and the main antigen was detected near 220 kDa in all cases. Among the mAbs, four mAbs (AP4, DN11, SHL-1, and P6) recognized a region commonly present in all the five isoforms. One mAb, YG27, recognized four isoforms (ABC, AB, BC, and O). Two mAbs, P1 and P14, recognized the isoforms that contain exon A encoded regions (ABC and AB). Conclusion: In this study, we confirmed that AP4, DN11, SHL-1, YG27 and P6, are mAbs reactive with the CD45 antigen whereas P1 and P14 are reactive with the CD45RA antigen.

• KSBB Journal
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• v.16 no.4
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• pp.403-408
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• 2001

Distillation condensate generated from downstream processing of microbial alcohol fermentation imposes a serious burden to biological wastewater treatment or anaerobic digestion due to its high contents of SS (suspended solids) and TN (total nitrogen), A pilot scale microfiltration of the stillage condensate with a stainless steel SCEPTER membrane of 0.1 ${\mu}$m pore size was carried out to remove SS which was mostly composed of microbial cell residue. A stable permeate flux was achieved when the decanter effluent containing 0.7% of SS was filtered under the conditions of X10 VCR (volume concentration ratio), 2.5 bar of TMP (transmembrane pressure), and 60$^{\circ}C$. When stillage condensate with 2.6% SS was treated directly with microfiltration, VCR below X3 was recommended for a long duration of filtration. The permeate and retentate obtained from microfiltration were recycled to make-up medium of fermentation. Adding permeate or retentate up to 30% of fermentation volume showed no distinguished undesirable influence during the course of alcohol fermentation. Although only slight improvements in the final amount of CO$_2$ evolution and alcohol content were observed, fermentation rate increased so that the required time to reach 450 L/ton of CO$_2$ evolution was shortened to 72% of that with normal media.

• Korean Chemical Engineering Research
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• v.43 no.2
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• pp.236-242
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• 2005

Membrane fouling by protein mixtures during microfiltration has been investigated for binary mixtures of bovine serum albumin (BSA), casein, lysozyme, pepsin, and ovalbumin. Filtration experiments were carried out using $0.2{\mu}m$ polycarbonate track-etched (PCTE) membrane in a stirred cell under constant transmembrane pressure (14 kPa) and concentration of hydrogen ion (pH=11) to study the effect of mixture composition on filtrate flux decline. Flux decline data were analyzed using a pore blockage-cake formation model developed recently. It was found that the model is in a good agreement with the experimental data. Fouling parameters such as the rate of pore blockage(${\alpha}$), the initial resistance of the protein deposit ($R_{po}$) and the increasing rate of the protein layer resistance(${\beta}$) were used to evaluate the rate of filtrate flow by membrane fouling in the binary mixture system. Generally, the trend of ${\alpha}$ is comparable with that of filtrate flux decline. It was also found that fast flux decreasing was observed over the binary mixture containing casein. The result is due to high value of the initial resistance of the protein deposit ($R_{po}$) over casein.

• BMB Reports
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• v.28 no.6
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• pp.527-532
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• 1995

Human taurine transporter has 12 transmembrane domains and its molecular weight is 69.6 kDa. The long cytoplasmic carboxy and amino termini might function as regulatory attachment sites for other proteins. Six potential protein kinase C phosphorylation sites have been reported in human taurine transporter. In this report, we studied the effects of phorbol 12-myristate 13-acetate (PMA) and glucocorticoid hormone on taurine transportation in the RAW 264.7, mouse macrophage cell line. When the cells were incubated with $[^{3}H]taurine$ in the presence or absence of $Na^+$ ion for 40 min at $37^{\circ}C$, the [$[^{3}H]taurine$ uptake rate was 780-times higher in the $Na^{+}-containing$ buffer than in the $Na^{+}-deficient$ buffer, indicating that this cell line expresses taurine transporter protein on the cell surface. THP1, a human promonocyte cell line, also showed a similar property. The $[^{3}H]taurine$ uptake rate was not influenced by the inflammatory inducing cytokines such as interleukin-1, gamma-interferon or interleukin-1+gamma-interferon, but was decreased by the PMA in the RAW 264.7 cell line. This suggests that activation of protein kinase C inhibits taurine transporter activity directly or indirectly. The inhibition of $[^{3}H]taurine$ uptake by PMA was time-dependent. Maximal inhibition occurred in one hr stimulation with PMA Increasing the treatment time beyond one h reduced the $[^{3}H]taurine$ uptake inhibition due to the depletion or inactivation of protein kinase C. The cell line also showed concentration-dependent $[^{3}H]taurine$ uptake under PMA stimulation. The phorbol-ester caused 23% inhibition at the concentration of 1 ${\mu}m$ PMA. The inhibition was significant even at a concentration as low as 10 nM PMA The reduced $[^{3}H]taurine$ uptake could be recovered by treatment with glucocorticosteroid hormone. Dexamethasone led to recover of the reduced taurine uptake induced by phorbol-ester, recovering maximally after one hr. This may suggest that macrophage cells require higher taurine concentration in a stressed state, for the secretion of glucocorticoid hormone is increased by hypothalamo-pituitary-adrenocortical (HPA) axis activation in the blood stream.

• Journal of Life Science
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• v.17 no.6 s.86
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• pp.783-790
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• 2007

G protein coupled receptors (GPCRs) transmit various extracellular signals into the cells. Upon binding of the ligands, conformational changes in the extracellular and/or transmembrane (TM) domains of CPCRs were propagated into the cytoplasmic (CP) domain of the molecule leading to the activation of their cognate heterotrimeric C proteins and kinases. Constitutively active GPCR mutants causing the activation of C Protein signaling even in the absence of ligand binding are of interest for the study of activation mechanism of GPCRs. Two classes of constitutively active mutations, categorized by their effects on the salt bridge between Ell3 and K296, were found in the TM domain of rhodopsin. Opsin mutants containing combinations of the mutations were constructed to study the conformational changes required for the activation of rhodopsin. Rhodopsin chromophore regenerated with 11-cis-retinal showed a thermal stability inversely correlated with its constitutive activity. In contrast, rhodopsin mutants exhibited a binding affinity to an agonist, all-trans-retinal, in a constitutive activity-dependent manner. In order to test whether the conformational changes responsible for the activation of trans-ducin (Gt) are the same as the conformation required for the recognition of rhodopsin kinase, analysis of the mutants were carried out with phosphorylation by rhodopsin kinase. Rhodopsin mutants containing combinations of different classes of the mutations showed a strong synergistic effect on the phosphorylation of the mutants in the dark as similar to that of Gt activation. The results suggest that at least two or three kinds of segmental and independent conformational changes are required for the activation of rhodopsin and the conformational changes responsible for activating rhodopsin kinase and Gt are similar to each other.

• Molecules and Cells
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• v.43 no.3
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• pp.228-235
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• 2020

The Drosophila transmembrane semaphorin Sema-1a mediates forward and reverse signaling that plays an essential role in motor and central nervous system (CNS) axon pathfinding during embryonic neural development. Previous immunohistochemical analysis revealed that Sema-1a is expressed on most commissural and longitudinal axons in the CNS and five motor nerve branches in the peripheral nervous system (PNS). However, Sema-1a-mediated axon guidance function contributes significantly to both intersegmental nerve b (ISNb) and segmental nerve a (SNa), and slightly to ISNd and SNc, but not to ISN motor axon pathfinding. Here, we uncover three cis-regulatory elements (CREs), R34A03, R32H10, and R33F06, that robustly drove reporter expression in a large subset of neurons in the CNS. In the transgenic lines R34A03 and R32H10 reporter expression was consistently observed on both ISNb and SNa nerve branches, whereas in the line R33F06 reporter expression was irregularly detected on ISNb or SNa nerve branches in small subsets of abdominal hemisegments. Through complementation test with a Sema-1a loss-of-function allele, we found that neuronal expression of Sema-1a driven by each of R34A03 and R32H10 restores robustly the CNS and PNS motor axon guidance defects observed in Sema-1a homozygous mutants. However, when wild-type Sema-1a is expressed by R33F06 in Sema-1a mutants, the Sema-1a PNS axon guidance phenotypes are partially rescued while the Sema-1a CNS axon guidance defects are completely rescued. These results suggest that in a redundant manner, the CREs, R34A03, R32H10, and R33F06 govern the Sema-1a expression required for the axon guidance function of Sema-1a during embryonic neural development.

• Membrane Journal
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• v.4 no.4
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• pp.221-231
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• 1994

The GR51PP(MWCO 50,000) and GR40PP(MWCO 100,000) membranes manufactured by DDS were used in ultrafiltration of dextran(Mw. : 500,000) solution in flat plate ultrafiltration cell filled with various types of turbulence promoters. The flux improvement by using turbulence promoter was higher in laminar flow region than in turbulent flow region. The maximum improvements of permeate flux were foud as 112% and 50% I laminar flow region and turbulent flow region, respectively. Also, the solute rejection of the ultrafiltration membrane was improved by turbulence promoters and its effect was significant in the high transmembrane pressure and laminar flow region. The smaller the spacer mesh size was used, the higher the flux improved, but the pressure drop in ultrafiltration cell also increased. In laminar flow region, pressure drop by the spacer was negligible, but in turbulent flow region it changed significantly depending upon the mesh size of the spacer and therefore, its mesh size must be baken into account in the design of the process. The predicted results of the modified mass transfer correlation had better agreement with experimental results than those of unmodified one, The modified mass transfer correlations for laminar and turbulent flow region are shown as follow. $N_{sh}=0.151(N_{Re})^{0.199}(N_{Sc})^{0.22}(N_{Scm})^{0.197}\;(625 $N_{sh}=0.0165(N_{Re})^{0.428}(N_{Sc})^{0.33}(N_{Scm})^{0.223}\;(5015

• Membrane Journal
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• v.21 no.4
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• pp.360-366
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• 2011

The effect of cross-flow velocity and transmembrane pressure (TMP) on membrane fouling was observed from pilot-scale operation of thermophilic anaerobic membrane bioreactor (AnMBR) treating food waste leachate. It was found that fouling rate was reduced significantly as cross-flow velocity increased at constant TMP mode of operation while this effectiveness was more pronounced at lower TMP. Higher TMP resulted in less permeable fouling layer possibly due to compressibility of foulant material on membrane surface. Particle sizes of membrane concentrate ranged from 10 to $100{\mu}m$, implying that shear-induced diffusion enhance back transport of these particle sizes away from the membrane effectively. From the continuous operation of AnMBR, it was confirmed that shear rate played an important role in the reduction of membrane fouling. Membrane autopsy works at the end of operation of AnMBR showed clearly that both organic and inorganic fouling were significant on membrane surface. Surface shear by cross-flow velocity was expected to be less effective to remove irreversible fouling which can be mainly caused by the adsorption of organic colloidal materials into membrane pores.