• Title/Summary/Keyword: pH-sensitive

Search Result 914, Processing Time 0.028 seconds

Toxicological Effects of B(a)P on Preimplantation Mouse Embryos in Vitro (in vitro에서 B(a)P이 착상전 마우스 배자에 미치는 독성학적 영향에 관한 연구)

  • 박귀례;이유미;김판기;신재호;강태석;김주일;장성재
    • Journal of Environmental Health Sciences
    • /
    • v.24 no.2
    • /
    • pp.126-133
    • /
    • 1998
  • Effects of B(a)P on preimplantation mouse embryos in vitro were studied. Preimplantation mouse embryos were exposed to a concentration of 0.3, 1, 3 and 10 $\mu$M B(a)P for 72 hrs. The toxicological effects of B(a)P were evaluated by morphological observation of embryos up to the blastocyst stage, and by measuring DNA, RNA and protein synthesis by radioactive precursor incorporation. At 1 $\mu$M B(a)P did not affect preimplantation development but interfered with hatching and ICM formation. Suppressing effect of ICM formation was dose dependent. At the eight cell stage, the developmental rate was decreased at above 3 $\mu$M of B(a)P. At the blastocyst stage, attachment and trophoblast outgrowth were diminished at the 10 $\mu$M of B(a)P and ICM formation was decreased at 1 $\mu$M of B(a)P. Inner cell number of blastocyst was decreased dose dependently. So, number of ICM was one of the most sensitive and toxicological end point. The RNA incorporation rate of 0.1 $\mu ^3$H-uridine was dosedependent and the protein incroporation of 0.5 $\mu Ci ^{35}$S-methionine showed a significant decrease after 48 hrs. But the DNA incorporation rate of methyl-$^3$H thymidine was not affected. Our results suggested that B(a)P did not affect the DNA replication but transcription was inhibited by dose dependent manner. There delay of development during the blastocyst stage was mainly due to the inhibition of RNA synthesis followed by protein synthesis.

  • PDF

IDENTIFICATION OF PORPHYROMONAS ENDODONTALIS USING POLYMERASE CHAIN REACTION(RCR) (중합효소연쇄반응(Polymerase Chain Reaction)을 이용한 Porphyromonas endodontalis의 동정에 대한 연구)

  • Lee, Sang-Yup;Yoon, Soo-Han
    • Restorative Dentistry and Endodontics
    • /
    • v.23 no.1
    • /
    • pp.328-338
    • /
    • 1998
  • Porphyromonas endodontalis, an anaerobic Gram negative cocobacillus which was known to be associated with the infected root canals and periapical lesions, is very difficult to culture and to detect by the traditional method in that it requires much time to induce the specific black pigmentation, and it is very sensitive to oxygen and the antibiotics added in the culture medium. In this study, the nucleotide sequences of the 'probe h' (0.73kb), one of the specific DNA probes top. endodontalis (ATCC 35406) which had been developed by our department, was determined and then a pair of primers for PCR amplification was fabricated to identify P. endodontalis. The plasmids containing 'probe h' were purified by $Wizard^{TM}$ Midipreps DNA Purification System (Promega Corp.), and the nucleotide sequences of the 'probe h' were determined by the dideoxy chain termination method using TaqTrack Sequencing System (Promega Corp.) and detected by fluorescent labelling method. The sense/antisense PCR primers were designed with computer software (Lasergene, DNASTAR Ind. PCR was done with a programmable GeneAmp PCR System 2400 (Perkin Elmer-Cetus Co.). Each sample containing the whole genomic DNA of P. endodontalis and other black-pigmented Bacteroides was itailly denatured at $94^{\circ}C$ for 5 min and then subjected to 30 cycles, each of them consisting of 60s at $94^{\circ}C$, 60s at $60^{\circ}C$, and 90s. at $72^{\circ}C$. The amplified DNA was resolved electrophoretically in a 1.0 % agarose gel in 1X TAE buffer, stained with EtBr, and photographed on a UV transilluminator. The results were as follows : 1. The nucleotide sequences of 'probe h' (743 base pairs) were obtained by dideoxy chain termination method, and from that results the specific primers to P. endodontalis (ATCC 35406), 'Primer H1/ Primer H2', were designed. 2. It has been found that 'Primer H1/H2' could detect P. endodontalis (ATCC 35406) using PCR. 3. The PCR system with this primers may be a powerful technique to amplify the specific sequences of 'probe h' of P. endodontalis (ATCC 35406) that produce distinct identification of it from other black-pigmented Bacteroides, and this could help us to determine the nature of periapical disease.

  • PDF

Cultural Conditions of Lactobacillus sp. GM7311 for the Production of Bacteriocin (Lactobacillus sp. GM7311에 의한 박테리오신의 생산 조건)

  • LEE Myung Suk;CHANG Dong Suck;KANG Ji-Hee
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.30 no.5
    • /
    • pp.834-841
    • /
    • 1997
  • A lactic acid bacteria which showed antimicrobial activity was isolated from dairy products and identified as Lactobacillus sp. according to the morphological, physiological and biochemical properties, which was named Lactobacillus sp. SH 7311. The bacteriocin of Lactobacillus sp. GM 7311 showed a broad range of inhibitory spectrum against some gram positive and negative bacteria. Especially, Proteus mirabilis was highly sensitive to bacteriocin and used as indicator strain for further investigation. The optimal condition for the production of bacteriocin was showed on MRS broth at $37^{\circ}C$ and pM 6.0. Bacteriocin production of this strain cultured under optimal condition was increased late logarithmic phase to early stationary phase. This bacteriocin was fully active at the pH range $2.0\~5.0$, also was stable at $100^{\circ}C$ for 60 min. at pH 5.0, But about $40\%$ of bacteriocin activity was diminished by the treatment of acetone, ethanol, iso-butanol and ethyl ether during 2 hours at $4^{\circ}C$.

  • PDF

Reduced Graphene Oxide Field Effect Transistor for Detection of H+ Ions and Their Bio-sensing Application

  • Sohn, Il-Yung;Kim, Duck-Jin;Yoon, Ok-Ja;Tien, N.T.;Trung, T.Q.;Lee, N.E.
    • Proceedings of the Korean Vacuum Society Conference
    • /
    • 2012.02a
    • /
    • pp.195-195
    • /
    • 2012
  • Recently, graphene based solution-gated field-effect transistors (SGFETs) have been received a great attention in biochemical sensing applications. Graphene and reduced graphene oxide (RGO) possess various advantages such as high sensitivity, low detection limit, label-free electrical detection, and ease of fabrication due to their 2D nature and large sensing area compared to 1D nanomaterials- based nanobiosensors. Therefore, graphene or RGO -based SGFET is a good potential candidate for sensitive detection of protons (H+ ions) which can be applied as the transducer in various enzymatic or cell-based biosensing applications. However, reports on detection of H+ ions using graphene or RGO based SGFETs have been still limited. According to recent reports, clean graphene grown by CVD or exfoliation is electrochemically insensitive to changes of H+ concentration in solution because its surface does not have terminal functional groups that can sense the chemical potential change induced by varying surface charges of H+ on CVD graphene surface. In this work, we used RGO -SGFETs having oxygen-containing functional groups such as hydroxyl (OH) groups that effectively interact with H+ ions for expectation of increasing pH sensitivity. Additionally, we also investigate RGO based SGFETs for bio-sensing applications. Hydroloytic enzymes were introduced for sensing of biomolecular interaction on the surface of RGO -SGFET in which enzyme and substrate are acetylcholinesterase (AchE) and acetylcholine (Ach), respectively. The increase in H+ generated through enzymatic reaction of hydrolysis of Ach by AchE immobilized on RGO channel in SGFET could be monitored by the change in the drain-source current (Ids).

  • PDF

Prevalence of bacterial respiratory diseases in cattle (소 세균성 호흡기 질병 분포도 조사)

  • Jeong, Byeong-yeal;Jeon, Yong-soo
    • Korean Journal of Veterinary Research
    • /
    • v.44 no.3
    • /
    • pp.427-432
    • /
    • 2004
  • The objective of this study was to determine the prevalence of pneumonic bacteria in lungs and nasal swabs of cattle with respiratory diseases. From 95 pneumonic lungs of slaughtered cattle, 41 (43.2%) positive lungs were yielded with 54 pneumonic bacteria, which corresponded to P. multocida (n = 34), A. pyogenes (n = 14) and P. haemolytica (n = 6). One-hundred sixty seven pneumonic bacteria were isolated from 195 nasal swabs in calves, 64.7% (108 isolates) belonged to P. multocida, 16.2% to A. pyogenes, 13.8% to P. haemolytica and 5.4% to H. somnus. Fifty percents (n = 6) of isolates from pneumonic lungs of calves were identified as P. multocida. All isolates of P. multocida belonged to type A according to hyaluronidase test. Antimicrobial susceptibility tests showed that most isolates of P. multocida and P. haemolytica were sensitive to amoxicillin/clavulanic acid, cephalothin, ciprofloxacin, enrofloxacin, fluorophenicol and norfloxacin. The emergence of antimicrobial resistant Pasteurella spp. observed in this study, however, might limit such application. According to histopathological examination, pneumonia by mycoplasma or/and bacteria accounted for 92.8% among 69 pneumonic lungs of slaughtered cattle.

Characterization of Novel Salt-Tolerant Esterase Isolated from the Marine Bacterium Alteromonas sp. 39-G1

  • Won, Seok-Jae;Jeong, Han Byeol;Kim, Hyung-Kwoun
    • Journal of Microbiology and Biotechnology
    • /
    • v.30 no.2
    • /
    • pp.216-225
    • /
    • 2020
  • An esterase gene, estA1, was cloned from Alteromonas sp. 39-G1 isolated from the Beaufort Sea. The gene is composed of 1,140 nucleotides and codes for a 41,190 Da protein containing 379 amino acids. As a result of a BLAST search, the protein sequence of esterase EstA1 was found to be identical to Alteromonas sp. esterase (GenBank: PHS53692). As far as we know, no research on this enzyme has yet been conducted. Phylogenetic analysis showed that esterase EstA1 was a member of the bacterial lipolytic enzyme family IV (hormone sensitive lipases). Two deletion mutants (Δ20 and Δ54) of the esterase EstA1 were produced in Escherichia coli BL21 (DE3) cells with part of the N-terminal of the protein removed and His-tag attached to the C-terminal. These enzymes exhibited the highest activity toward p-nitrophenyl (pNP) acetate (C2) and had little or no activity towards pNP-esters with acyl chains longer than C6. Their optimum temperature and pH of the catalytic activity were 45℃ and pH 8.0, respectively. As the NaCl concentration increased, their enzyme activities continued to increase and the highest enzyme activities were measured in 5 M NaCl. These enzymes were found to be stable for up to 8 h in the concentration of 3-5 M NaCl. Moreover, they have been found to be stable for various metal ions, detergents and organic solvents. These salt-tolerant and chemical-resistant properties suggest that the enzyme esterase EstA1 is both academically and industrially useful.

Use of Anti-Phosphohistone H3 (PHH3) as a Mitosis Marker for Classifying Pulmonary Carcinoid Tumors

  • Seo, Bo-Ram;Hong, Young-Seob;Choi, Phil-Jo;Um, Soo-Jung;Seo, Jeong-Wook;Roh, Mee-Sook
    • Biomedical Science Letters
    • /
    • v.17 no.3
    • /
    • pp.197-202
    • /
    • 2011
  • Mitosis count is one of the most helpful morphologic features for distinguishing pulmonary typical carcinoid (TC) from atypical carcinoid (AC). However, identifying areas of highest mitotic activity is tedious and time-consuming, and mitosis count may vary substantially among pathologists. Anti-phosphohistone H3 (PHH3) is an antibody that specifically detects histone H3 only when phosphorylated at serine 10 or serine 28, an event that is concurrent with mitotic chromatin condensation and not observed during apoptosis. In this study, immunohistochemical staining for PHH3 was performed to determine whether PHH3 was a reliable and objective mitosis-specific marker for pulmonary carcinoid tumors. Seventeen cases of surgically resected pulmonary carcinoid tumors (12 TCs and 5 ACs) were obtained and classified according to the 2004 World Health Organization classification. Mitotic counts determined by PHH3 correlated to ones determined by hematoxylin and eosin (H&E) staining; however, PHH3 mitotic counts (mean mitotic counts: 1 in TCs and 3.2 in ACs) were slightly higher than H&E mitotic counts (mean mitotic counts: 0.25 in TCs and 1.8 in ACs). The mitotic counts determined by experienced observer were more correlated to those determined by inexperienced observer with the PHH3-based method (R=0.968, P<0.001) rather than H&E staining (R=0.658, P<0.001). These results suggest that the PHH3 mitotic counting method was more sensitive and simple for detecting mitoses compared to traditional H&E staining. Therefore, PHH3 immunohistochemistry may contribute to more accurate and reproducible diagnosis of pulmonary carcinoid tumors and may be a valuable aid for administrating appropriate clinical treatment.

Clinical Factors Related with Helicobacter Pylori Infection - Is there an Association with Gastric Cancer History in First-Degree Family Members?

  • Demirel, Busra B.;Akkas, Burcu Esen;Vural, Gulin Ucmak
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.3
    • /
    • pp.1797-1802
    • /
    • 2013
  • Background: The aim of this study was to assess clinical factors associated with Helicobacter pylori positivity and to evaluate the incidence of gastric carcinoma in first-degree family members of infected patients. A total of 580 patients (mean age:$38{\pm}17$) with gastrointestinal complaints underwent C-14 urea breath test (UBT). Patients were grouped as: Group-1, untreated patients (n:384); and Group-2, patients who previously treated with eradication triple therapy (n:196). C-14 UBT was performed 1-2 months after the completion of eradication therapy. Associations of H pylori positivity with age, gender, ABO and Rhesus groups, smoking, dietary habits, and history of gastric cancer in first-degree family members were evaluated. The frequency of H pylori positivity was significantly higher in group-1 (58%) compared to group-2 (20%), p=0.001. There were no correlations between H pylori positivity and age, gender, ABO groups, Rhesus subgroups, smoking and dietary habits in both patient groups. The frequency of gastric cancer in family members was significantly higher in patients with H pylori infection among group-1, compared to infected patients among group-2 (56% vs. 28.6% respectively, p=0.03). We observed a significant association between H pylori positivity and the presence of gastric cancer in first-degree relatives of group-1 patients. Our results provide some confirmation of the presence of a link between gastric cancer development and H pylori. C-14 UBT is a sensitive, reliable and a widely recommended test for the detection of H pylori infection and recurrence. We suggest that detection and eradication of H pylori may contribute to a reduced risk of gastric cancer in the family members of infected patients.

Characterization of Human ${\beta}-Carotene$ 15,15-dioxygenase Isolated from Recombinant Escherichia coli (유전자 재조합 기술에 의하여 제조된 인간 ${\beta}-carotene$ 15,15'-dioxygenase의 반응특성)

  • Shin, Won-Phil;Chang, Pahn-Shick
    • Korean Journal of Food Science and Technology
    • /
    • v.36 no.3
    • /
    • pp.440-447
    • /
    • 2004
  • Characteristics of human ${\beta}-carotene$ 15,15'-dioxygenase isolated by recombinant DNA technology was elucidated. Optimal pH and temperature were 9.0 and $40^{\circ}C$, respectively. Enzyme activity was temperature-sensitive. Enzyme was stable at pH 6.0-9.0 for 24 hr and under $5^{\circ}C$. Half-life of enzyme at $35^{\circ}C$ was 40 min. Crude preparations of enzyme were inhibited by ferrous ion-chelating agent and sulfhydryl-binding agent. GSH offsets inhibitory effect of PCMB. With increasing substrate concentrations, enzyme activity gave typical Michaelis-Menten curve, Based on Hanes-Woolf plot of data, $K_{m}\;and\;V_{max$ were $3.39{\times}10^{6}\;M\;and\;1.2\;pmol/mg$ protein/min, respectively.

Characterization of the Nature of Cd(II) Ion Binding Sites on Poly(acrylic acid) Using$^{113}Cd$NMR (카드뮴-113 NMR을 이용한 폴리아크릴산의 카드뮴(Ⅱ) 이온에 대한 결합자리의 특성연구)

  • Chung, Kun Ho;Koo, Ja-Eung;Shin, Hyun Sang;Moon, Hichung
    • Journal of the Korean Chemical Society
    • /
    • v.40 no.1
    • /
    • pp.44-49
    • /
    • 1996
  • The cadmium(Ⅱ)-carboxylate complexes of monocarboxylates and poly(acrylic acid) (PAA) in an aqueous solution were investigated using 113Cd NMR spectroscopy. From these spectral data the chemical shifts of the monocarboxylate (e.g. acetate, benzoate and propanoate) complexes of Cd(Ⅱ) were evaluated (CdL+: -22 to -24 ppm range; CdL2: -39 to -40 ppm range; L: carboxylate). The chemical shift of cadmium(Ⅱ) bound on PAA changed in value from -36.6 to -38.2 ppm when the [PAA]/[Cd] ratios were varied from 12 to 118 eq/mol at a constant pH of 6. The 113Cd chemical shift was sensitive to the change in solution pH: the chemical shift changed from 1.6 to -37.6 ppm when the pH was rasied from 1 to 6 at a constant [PAA]/[Cd] ratio of 39 eq/mol. These results show that under these conditions, the dominant species formed in solution is 1 : 2 complex (CdL2), and 1 : 1 complex (CdL+) is only formed at low pH, when polyanion concentration is low.

  • PDF