• IMMUNE NETWORK
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• v.3 no.4
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• pp.268-275
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• 2003

Background: Hemopoietic cells require the constant presence of growth factors for survival in vitro and in vivo. Caspases have been known as central executors of apoptotic cell death. We have, therefore, investigated the pathways that regulate caspase activity and apoptosis using the $CD34^+$ cell line, TF-1 which requires GM-CSF for survival. Methods: Apoptosis was measured by annexin V staining and mitochondrial membrane potential was measured by DiOC6 labelling. Intracellular pH was measured using pH sensitive fluorochrome, BCECF or SNARF-1, followed by flow cytometry analysis. Caspase activation was analyzed by PARP cleavage using anti-PARP antibody. Results: Removal of GM-CSF induceed PARP cleavage, a hallmark of caspase activity, concomitant with pHi acidification and a drop in mitochondrial potential. Treatment with ZVAD, a competitive inhibitor of caspases, partially rescued cell death without affecting pHi acidification and the reduction of mitochondrial potential, suggesting that both these events act upstream of caspases. Overexpression of Bcl-2 prevented cell death induced by GM-CSF deprivation as well as pHi acidification and the reduction in mitochondrial membrane potential. In parental cells maintained with GM-CSF, EIPA, a competitive inhibitor of $Na^+/H^+$ antiporter induced apoptosis, accompanied by a drastic reduction in mitochondrial potential. In contrast, EIPA induced apoptosis in Bcl-2 transfectants without causing mitochondrial membrane depolarization. Conclusion: Taken together, our results suggest that the regulation of $H^+$fluxes, either through a mitochondriondependent or independent pathway, is central to caspase activation and apoptosis.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.3
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• pp.975-978
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• 2012

P63 is a gene product required in cell cycle regulation which plays vital roles in tumor differentiation. Aims of the present study were to assess the frequency, pattern, sensitivity and specificity of two p63 protein clones P63 4A4 and P63 4A4+Y4A3 in squamous cell carcinomas (SCCs). Thirty cases of head and neck region SCC diagnosed on the basis of H&E staining were examined along with 60 cases of head and neck region biopsies other than squamous cell carcinoma, negative on H&E staining, were taken as control. Immunostaining was performed on slides according to the Thermo Scientific UltraVision LP detection System. P63 4A4+Y4A3 clone is more sensitive 96.6% in comparison to 86% in P63 4A4 with having greater NPV of 98.3%. The results signify the importance of P63 4A4+Y4A3 marker over the old markers and may be used as a confirmatory marker of squamous cell carcinoma.

• Journal of Sensor Science and Technology
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• v.9 no.2
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• pp.106-112
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• 2000

A FET-type reference electrode(REFET) is an effective method to eliminate typical problems with ISFET(ion sensitive field-effect transistor) such as drift, temperature, light-dependence and miniaturization of reference electrode. However, it is difficult to make the highly reliable REFET with excellent long-term stability and reproducibility. In this paper, an ion-blocking membrane was applied to the REFET for the PET-type electrolyte sensors(pH, pNa-ISFET). The fabricated REFET indicated the stable sensitivity (55.4 mV/pH, 53.5 mV/decade) and good linearity in the pH and pNa measurement. In the measurement, ISFET/Pt/REFET configuration showed excellent stability and reproducibility.

• Journal of Korean Society of Environmental Engineers
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• v.22 no.11
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• pp.1985-1994
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• 2000

This research was designed to investigate the effect of initial adsorbed amount of phenol, temperature, and pH on the desorption reaction of phenol from spent activated carbon loaded with phenol. Methanol, acetone, and N,N-dimethylformamide( DMF) were used as test organic solvents. The initial adsorbed quantities of phenol investigated here were 166.1mg/g, 180.7mg/g, and 197.9mg/g. The effect of temperature was evaluated from 15 to $55^{\circ}C$ with an interval of $10^{\circ}C$, while that of pH was investigated under acidic. neutral. and alkaline conditions. The extent of phenol desorption was proportional to the strength of dipole moment such as methanol < acetone < DMF. Over 90% desorption of phenol was achieved by acetone and DMF. The quantity of des orbed phenol by the organic solvents decreases with increasing the initial adsorbed amount of phenol. DMF is affected least by the initially adsorbed amount of phenol. An increase in reaction temperature leads to higher desorption of phenol. Desorption reaction by methanol is most sensitive to the temperature. As the pH of solvents increases. the desorption rate is also increasing. At pH=12. the desorption rate of phenol by methanol increases sharply by 10%. Although methanol demonstrated the weakest desorption power. the desorption capacity of methanol would approach that of acetone and DMF by adjusting temperature and pH. Methanol may emerge as a promising solvent for removing phenol from activated carbon because of acceptable regeneration efficiency as well as relatively cheap price.

• The Korean Journal of Physiology and Pharmacology
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• v.12 no.4
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• pp.177-183
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• 2008

The layers of keratinocytes form an acid mantle on the surface of the skin. Herein, we investigated the effects of acidic pH on the membrane current and $[Ca^{2+}]_c$ of human primary keratinocytes from foreskins and human keratinocyte cell line (HaCaT). Acidic extracellular pH ($pH_e{\leq}5.5$) activated outwardly rectifying $Cl^-$ current ($I_{Cl,pH}$) with slow kinetics of voltage-dependent activation. $I_{Cl,pH}$ was potently inhibited by an anion channel blocker 4,4'-diisothiocyanostilbene-2,2'-disulphonic acid (DIDS, 73.5% inhibition at 1${\mu}$M). $I_{Cl,pH}$ became more sensitive to $pH_e$ by raising temperature from $24^{circ}C$ to $37^{circ}C$. HaCaT cells also expressed $Ca^{2+}$-activated $Cl^-$ current ($I_{Cl,Ca}$), and the amplitude of $I_{Cl,Ca}$ was increased by relatively weak acidic $pH_e$ (7.0 and 6.8). Interestingly, the acidic $pH_e$ (5.0) also induced a sharp increase in the intracellular [$Ca^{2+}$] (${\triangle}[Ca^{2+}]_{acid}$) of HaCaT cells. The ${\triangle}[Ca^{2+}]_{acid}$ was independent of extracellular $Ca^{2+}$, and was abolished by the pretreatment with PLC inhibitor, U73122. In primary human keratinocytes, 5 out of 28 tested cells showed ${\triangle}[Ca^{2+}]_{acid}$. In summary, we found $I_{Cl,pH}$ and ${\triangle}[Ca^{2+}]_{acid}$ in human keratinocytes, and these ionic signals might have implication in pathophysiological responses and differentiation of epidermal keratinocytes.

• Journal of fish pathology
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• v.2 no.2
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• pp.99-108
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• 1989

Anaethetics are needed for handling fish, especially for transportation, tagging and grading. Among them, MS-222 has been popular in aquaculture since it has an excellent anaesthetic effect. However, MS-222 is more expensive than other chemicals. Benzocaine (Ethyl-p-aminobenzoate) has a similar molecular formula and equivalent anaesthetic effect to MS-222, and is cheaper. The purpose of this study was first to compare anaesthetic effects (Benzocaine) under various conditions : temperature, concentration, pH and body weight. Second purpose was to compare actual anaesthetic effects at 50 ppm benzocaine at ambient temperature and pH for the grading of tilapia. The results of this study are as follows : 1. The effect of anaesthesia at $24^{\circ}C$ was better with low pH, that is 5.6 than high pH 6.6 and 7.6. 2. The anaesthetic effect was not different at different body weight form 11g to 1,350g. 3. The fish were anaesthetized in 4~10 minutes at 50 ppm benzocaine at temperature. $20{\sim}24^{\circ}C$ and pH 6.8~7.3 and recovered in 4~6 minutes when they were put back in the fresh water after 30 minutes anaesthesia. 4. Benzocaine was more sensitive at pH fluctuation than temperature. 5. Twenty four hour-TLm of Benzocaine was 50 ppm at $24^{\circ}C$, pH 6.8 when the fish were put back in the fresh water after 120 minutes.

• The Plant Pathology Journal
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• v.36 no.3
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• pp.218-230
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• 2020

Net blotch of barley caused by Pyrenophora teres (Died.) Drechsler, is one of the most destructive diseases on barley in Algeria. It occurs in two forms: P. teres f. teres and P. teres f. maculata. A total of 212 isolates, obtained from 58 fields sampled in several barley growing areas, were assessed for fungicide sensitivity by target gene analysis. F129L and G137R mitochondrial cytochrome b substitution associated with quinone outside inhibitors (QoIs) resistance, and succinate dehydrogenase inhibitors (SDHIs) related mutations (B-H277, C-N75S, C-G79R, C-H134R, and C-S135R), were analyzed by pyrosequencing. In vitro sensitivity of 45 isolates, towards six fungicides belonging to three chemical groups (QoI, demethylase inhibitor, and SDHI) was tested by microtiter technique. Additionally, sensitivity towards three fungicides (azoxystrobin, fluxapyroxad, and epoxiconazole) was assessed in planta under glasshouse conditions. All tested isolates were QoI-sensitive and SDHI-sensitive, no mutation that confers resistance was identified. EC₅₀ values showed that pyraclostrobin and azoxystrobin are the most efficient fungicides in vitro, whereas fluxapyroxad displayed the best disease inhibition in planta (81% inhibition at 1/9 of the full dose). The EC₅₀ values recorded for each form of net blotch showed no significant difference in efficiency of QoI treatments and propiconazole on each form. However, in the case of fluxapyroxad, epoxiconazole and tebuconazole treatments, analysis showed significant differences in their efficiency. To our knowledge, this study is the first investigation related to mutations associated to QoI and SDHI fungicide resistance in Algerian P. teres population, as well as it is the first evaluation of the sensitivity of P. teres population towards these six fungicides.

• Journal of Microbiology and Biotechnology
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• v.34 no.6
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• pp.1229-1238
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• 2024

This study aimed to develop and assess a chitosan biomedical antibacterial gel ZincOxide-GrapheneOxide/Chitosan/β-Glycerophosphate (ZnO-GO/CS/β-GP) loaded with nano-zinc oxide (ZnO) and graphene oxide (GO), known for its potent antibacterial properties, biocompatibility, and sustained drug release. ZnO nanoparticles (ZnO-NPs) were modified and integrated with GO sheets to create 1% and 3% ZnO-GO/CS/β-GP thermo-sensitive hydrogels based on ZnO-GO to Chitosan (CS) mass ratio. Gelation time, pH, structural changes, and microscopic morphology were evaluated. The hydrogel's antibacterial efficacy against Porphyromonas gingivalis, biofilm biomass, and metabolic activity was examined alongside its impact (MC3T3-e1). The findings of this study revealed that both hydrogel formulations exhibited temperature sensitivity, maintaining a neutral pH. The ZnO-GO/CS/β-GP formulation effectively inhibited P. gingivalis bacterial activity and biofilm formation, with a 3% ZnO-GO/CS/β-GP antibacterial rate approaching 100%. MC3T3-e1 cells displayed good biocompatibility when cultured in the hydrogel extract.The ZnO-GO/CS/β-GP thermo-sensitive hydrogel demonstrates favorable physical and chemical properties, effectively preventing P. gingivalis biofilm formation. It exhibits promising biocompatibility, suggesting its potential as an adjuvant therapy for managing and preventing peri-implantitis, subject to further clinical investigations.

• Korean Journal of Microbiology
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• v.52 no.2
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• pp.230-235
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• 2016

This study was carried out to analyze the differences in enzyme activity and stability between the native Fe superoxide dismutase (FeSOD) and the 6xHis-tagged superoxide dismutase (6xHis-FeSOD) of Streptomyces subrutilus P5. The optimum pHs for both native FeSOD and 6xHis-FeSOD were 7, while the pH range of the activity was narrower for the 6xHis-FeSOD. The native FeSOD was stable at pH 4-9, but the 6xHis-FeSOD lost its stability at pH > 9. The temperatures of the optimum activities were same for both types of enzymes. However, the heat stability of the 6xHis-FeSOD was clearly decreased; even at $20^{\circ}C$ the enzyme lost the activity after 360 min. In contrast, the native FeSOD was stable after 720 min at below $40^{\circ}C$. $H_2O_2$ inhibition was occurred already at 0.5 mM for the 6xHis-tagged enzyme. Therefore, from the results that the 6xHis-FeSOD retained the enzyme activity at pH 6-7 and $20-40^{\circ}C$, it can be assumed that the protein structure became destabilized under different storage conditions and sensitive to the enzyme inhibitor.

• Korean Journal of Poultry Science
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• v.43 no.1
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• pp.31-38
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• 2016

Consumers demand fresh and high-quality eggs. Egg quality may be represented by shell color, shell weight, egg weight, shell thickness, shell density, albumen height, yolk color, albumen pH and viscosity. Various factors such as strain, age of hen, storage temperature, humidity, the presence of $CO_2$ and storage time affect egg quality. Therefore, we investigated the effects of storage time and temperature on egg quality to define the freshness of Korean market eggs. A total of 1,800 eggs were used for this experiment and were separated into 45 treatments with 40 eggs in each. The treatments were consisted of 15 storage periods (2 d to 30 d) and 3 storage temperatures ($2^{\circ}C$, $12^{\circ}C$, $25^{\circ}C$). Each egg was weighed and broken, and the height of the thick albumen, Haugh units (HU), egg shell color and yolk color were measured by a QCM+system. We also observed the physiochemical properties of eggs such as yolk pH, albumen pH and albumen viscosity. The egg weight, shell weight, albumen height, HU and albumen viscosity significantly decreased with increasing storage time and temperature. However, the albumen and yolk pH significantly increased with increasing storage period and temperature. The interaction effects between the storage period and temperature were significant for shell weight, shell density, egg weight, albumen height, HU, yolk color, yolk pH, albumen pH and albumen viscosity. In the analysis of the correlation with egg quality, the storage temperature exhibited a higher correlation coefficient than the storage period. In conclusion, storage time and temperature are the major factors affecting egg quality, but the storage temperature is a more sensitive determinant of egg quality deterioration compared with the storage period.