• Title/Summary/Keyword: lytA

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Carbohydrate Concentration and Composition in Source and Sink Tissues of Two Tall Fescue Genotypes

  • Song, Beom-Heon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.43 no.4
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    • pp.273-278
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    • 1998
  • Carbohydrate metabolism and partitioning are dependent on relationships between sources and sinks which can be affected by rates of photosynthesis and respiration. Fructan, the major form of stored carbohydrate in tall fescue (festuca arundineacea Schreb.), changes in concentration during growth and in response to the environment. Objectives of this study were i) to examine the content and the composition of carbohydrates in five tissues (mature leaf blade, immature leaf blade, leaf elongation zone, terminal meristem, and root tips) of two tall fescue genotypes, one with high yield per tiller (HYT) and one with low yield per tiller (LYT), and ii) to compare the reserved and utilized carbohydrates among above five different tissues, particularly between the leaf elongation zone and root tips. The established vegetative tillers of the HYT and LYT genotypes were grown in a controlled-environment growth chamber. Water-soluble carbohydrate (WSC) in the leaf elongation zone was about 22% of dry weight in the HYT and about 19% in the LYT genotype. The root tip also had high WSC, about 12% of dry weight in the HYT and 6% in the LYT genotype. Hexoses and sucrose were the major components of total WSC in all tissues except the leaf elongation zone. The growing tissues (sinks), i.e., the leaf elongation zone and root tip, had a high proportion of low degree of polymerization fructan, i.e., 3 to 8 hexose units.

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Inhibition of Autolysis by Lipase LipA in Streptococcus pneumoniae Sepsis

  • Kim, Gyu-Lee;Luong, Truc Thanh;Park, Sang-Sang;Lee, Seungyeop;Ha, Jung Ah;Nguyen, Cuong Thach;Ahn, Ji Hye;Park, Ki-Tae;Paik, Man-Jeong;Pyo, Suhkneung;Briles, David E.;Rhee, Dong-Kwon
    • Molecules and Cells
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    • v.40 no.12
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    • pp.935-944
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    • 2017
  • More than 50% of sepsis cases are associated with pneumonia. Sepsis is caused by infiltration of bacteria into the blood via inflammation, which is triggered by the release of cell wall components following lysis. However, the regulatory mechanism of lysis during infection is not well defined. Mice were infected with Streptococcus pneumoniae D39 wild-type (WT) and lipase mutant (${\Delta}lipA$) intranasally (pneumonia model) or intraperitoneally (sepsis model), and survival rate and pneumococcal colonization were determined. LipA and autolysin (LytA) levels were determined by qPCR and western blotting. S. pneumoniae Spd_1447 in the D39 (type 2) strain was identified as a lipase (LipA). In the sepsis model, but not in the pneumonia model, mice infected with the ${\Delta}lipA$ displayed higher mortality rates than did the D39 WT-infected mice. Treatment of pneumococci with serum induced LipA expression at both the mRNA and protein levels. In the presence of serum, the ${\Delta}lipA$ displayed faster lysis rates and higher LytA expression than the WT, both in vitro and in vivo. These results indicate that a pneumococcal lipase (LipA) represses autolysis via inhibition of LytA in a sepsis model.

$pep^{27}$ and lytA in Vancomycin-Tolerant Pneumococci

  • Olivares, Alma;Trejo, Jose Olivares;Arellano-Galindo, Jose;Zuniga, Gerardo;Escalona, Gerardo;Vigueras, Juan Carlos;Marin, Paula;Xicohtencatl, Juan;Valencia, Pedro;Velazquez-Guadarrama, Norma
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1345-1351
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    • 2011
  • Vancomycin therapy failure due to the emergence of tolerance in pneumococci is increasing. The molecular mechanism of tolerance is not clear, but lytA and $pep^{27}$ are known to be involved. Our aim was to evaluate the expression of both genes in vancomycin-tolerant Streptococcus pneumoniae (VTSP) strains. Eleven VTSP strains from a total of 309 clinical isolates of S. pneumoniae from 1997 to 2006 were classified according to the criteria of Liu and Tomasz. All VTSP strains were evaluated for susceptibility according to CLSI criteria, serotype by the Quellung test, and clonality by PFGE. The expressions of lytA and $pep^{27}$ were analyzed in different growth phases by RT-PCR with and without vancomycin. Eighty-two percent of VTSP strains showed resistance to penicillin, and 100% were sensitive to vancomycin and cefotaxime. The most frequent serotypes of VTSP strains were 23F (4/11) and 6B (3/11). Clonal relationship was observed in only two strains. No significant changes were observed in $pep^{27}$ expression in the three phases of growth in VTSP strains with and without vancomycin. Interestingly, $pep^{27}$ expression in the stationary phase in the non-tolerant reference strain R6 was significantly higher. However, no significant differences in lytA expression were observed between VTSP and R6 strains during the phases of growth analyzed. The absence of changes in $pep^{27}$ expression in VTSP strains in the stationary phase may be related to their ability to tolerate high antibiotic concentrations, and thus, they survive and remain in the host under the antibiotic selective pressure reflected in therapeutic failure.

Changes of IgE production, splenic helper and suppressor T lymphocytes in mice infected with Paragonimus westermani (폐흡충(Paragonimus westermani) 감염이 흰쥐의 IgE 생성 및 비장림프구 아군분포에 미치는 영향)

  • Min, Deuk-Yeong;Ryu, Jae-Suk;Sin, Myeong-Heon
    • Parasites, Hosts and Diseases
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    • v.31 no.3
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    • pp.231-238
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    • 1993
  • Effects of Paragonims westermai infection were observed in mice on the change of serum IgE level, the number of peripheral eosinophils and the distribution of Thy $1.2^{+}{\;}(CD3),{\;}L3T4^{+}{\;}(CD4),{\;}and{\;}Lyt-2^{+}$ (CD8) splenic T Iymphocytes without mitogen serum IgE increased at 3 weeks after the infection and reached a peak on week 4 and maintained high levels of IgE until the 23r6 week. Peripheral eosinophil numbers Increased at the second week and attained peak level on week 9. The frequency of $L3T4^{+}$(CD4) and $Lyt-2^{+}$ (CD8) T Iymphocytes decreased slightly until 4 weeks after the infection, but not significantly. Absolute number of $L3T4^{+}{\;}and{\;}Lyt-2^{+}$ T Iymphocytes, and the ratio of L3T4/Lyt-2 were not markedly changed over the period of observation. The frequency of Thy $1.2^{+}$ (CD3) T lymphocytes in the infected group slightly decreased until 4 weeks after the infection and showed significant reductions at the 2nd and 4th week of the infection (p < 0.05).

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Mutation of a Transposed Amino Acid Triplet Repeat Enhances Coupling of m1 Muscarinic Receptor to Activation of Phospholipase C

  • Lee, Seok-Yong;Cho, Tai-Soon
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.206-206
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    • 1996
  • The C-terminus ends of the second putative transmembrane domains of both m1 and m2 muscarinic receptors contain a triplet of amino acid residues consisting of leucine (L), tyrosine (Y) and threonine (T), This triplet is repeated as LYT-LYT in m2 receptors at the interface between the second transmembrane domain and the first extracellular loop. Interestingly, however, it is repeated in a transposed fashion (LYT-TYL) in the sequence of m1 receptors. In this work we employed site-directed mutagenesis to investigate the possible significance of this unique sequence diversity for determining the distinct differential drug-receptor interaction and cellular function at m1 muscarinic receptor. Mutation of the LYTTYL sequence of m1 receptors to the corresponding m2 receptor LYTLYT sequence, however, did not result in a significant change in the binding affinity of the agonist carbachol or in the affinity of the majority of a series of receptor antagonists which are able to discriminate between wild-type m1 and m2 receptors. Surprisingly, the LYTLYT ml receptor mutant demonstrated markedly enhanced coupling to activation of phospholipase C without a change in its coupling to increased cyclic AMP formation. There was also an enhanced receptor sensitivity in transducing elevation of intracellular Ca$\^$2+/. These changes were not due to alterations in the rate of receptor. desensitization or sequestration, On the other hand, the reverse LYTLYT-LYTTYL mutation in the m2 receptor did not alter its coupling to inhibition of adenylate cyclase, but slightly enhanced its coupling to stimulation of PI hydrolysis, Our data suggest that the LYTTYL/LYTLYT sequence difference between ml and n12 muscarinic receptors is not involved in determining receptor pharmacology. On the other hand, while these differences might play a role in the modulation of muscarinic receptor coupling to PI hydrolysis, they are not important for specifying coupling of various subtypes of muscarinic receptors to different cellular signaling pathways.

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Evaluation of Urinary Antigen Test for Rapid Diagnosis of $Streptococcus$ $pneumoniae$ in Community-Acquired Pneumonia Patients

  • Yu, Mi-Young;Kim, In-Sik;Kang, Sang-Sun;Cha, Beong-Hun;Hyun, Sung-Hee
    • Biomedical Science Letters
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    • v.17 no.4
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    • pp.355-361
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    • 2011
  • We evaluated the performance of the NOW $Streptococcus$ $pneumoniae$ urinary antigen test, standard culture and polymerase chain reaction for detecting $S.$ $pneumoniae$. The urinary antigen test of pneumonia patients represented sensitivity at 72% and specificity at 79%. The results of PCR were targeting for autolysin ($lyt$A), pneumolysin ($ply$), and spn9828. The $lyt$A sensitivity and specificity stood at 56% and 87% respectively while $ply$ sensitivity reported 83% and specificity was 47%, sensitivity and specificity of spn9828 stood at 83% and 73% respectively. The results of urinary antigen test and three genes were all statistically meaningful within $P$ <0.05. When the urinary antigen test of $S.$ $pneumoniae$ was positive, the three kinds of genes were also likely to be positive. According to the result of urinary antigen test, the results of PCR presented a meaningful difference ($P$ <0.05). Especially, the urinary antigen test of $S.$ $pneumoniae$ was likely to be positive ($P$ <0.05) when more than two genes were positive in PCR results.

Enhanced Coupling of $M_1$ Muscarinic Receptors to Activation of Phospholipase C upon Mutation of a Transposed Amino Acid Triplet Repeat

  • Lee, Seok-Yong;Sung, Ki-Wug;Kim, Ok-Nyu;Lee, Sang-Bok
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.1
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    • pp.19-25
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    • 1997
  • The C-terminus ends of the second putative transmembrane domains of both $M_1$ and $M_2$ muscarinic receptors contain a triplet of amino acid residues consisting of leucine (L), tyrosine (Y) and threonine (T). This triplet is repeated as LYT-TYL in $M_1$ receptors at the interface between the second transmembrane domain and the first extracellular loop. Interestingly, however, it is repeated in a transposedfashion (LYT-LYT) in the sequence of $M_2$ receptors. In our previous work, we investigated the possible significance of this unique sequence diversity for determining the distinct differential receptor function at the two receptor subtypes. However, we found mutation of the LYTTYL sequence of $M_1$ receptors to the corresponding $M_2$ receptor LYTLYT sequence demonstrated markedly enhanced the stimulation of phosphoinositide (PI) hydrolysis by carbachol without a change in its coupling to increased cyclic AMP formation. In this work, thus, the enhanced stimulation of PI hydrolysis in the LYTLYT $M_1$ receptor mutant was further investigated. The stimulation of PI hydrolysis by carbachol was enhanced in the mutant $M_1$ receptor, and this change was not due to alterations in the rate of receptor desensitization or sequestration. The observed larger response to carbachol at mutant $M_1$ receptors was also not due to an artifact resulting from selection of CHO cells which express higher levels of G-proteins or phospholipase C. Our data suggest that although the LYTTYL sequence in $M_1$ muscarinic receptors is not involved in determining receptor pharmacology, mutation of the sequence enhanced the coupling of $M_1$ receptors to the stimulation of phospholipase C.

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Effects of Site-Mutagenesis of an Amino Acid Triplet Repeat at $M_1$ and $M_2$ Muscarinic Receptors on Receptor Function ($M_1$$M_2$ 무스카린성 수용체에서 아미노산 Triplet Repeat의 Site-Mutagenesis가 수용체기능에 미치는 영향)

  • Lee, Seok-Yong;Lee, Sang-Bok
    • The Korean Journal of Pharmacology
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    • v.32 no.3
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    • pp.311-321
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    • 1996
  • Both $M_1$ and $M_2$ muscarinic receptors contain a triplet of amino acid residues consisting of leucine (L), tyrosine (Y) and threonine (T) at C-terminus ends of the second putative transmembrane domains. This triplet is repeated as LYT-LYT in $M_2$ receptors at the interface between the second transmembrane domain and the first extracellular loop. Interestingly, however, it is repeated in a transposed fashion (LYT-TYL) in the sequence of $M_1$ receptors. In this work, we employed site-directed mutagenesis to investigate the possible significance of this unique sequence diversity for determining the distinct differential cellular function at the two receptor subtypes. Mutation of the LYTTYL sequence of $M_1$ receptors to the corresponding $M_2$ receptor LYTLYT sequence did not result in a significant change in the binding affinity of the agonist carbachol. The reverse mutation at the $M_2$ receptor also did not modify agonist affinity. Surprisingly, the LYTLYT $M_1$ receptor mutant demonstrated markedly enhanced coupling to activation of phospholipase C without a change in its coupling to increased cyclic AMP formation. There was also an enhanced receptor sensitivity in transducing elevation of intracellular $Ca^{2+}$. On the other hand, the reverse $LYTLYT{\rightarrow}LYTTYL$ mutation in the $M_2$ receptor did not alter its coupling to inhibition of adenylate cyclase, but slightly enhanced its coupling to stimulation of phosphoinositide (PI) hydrolysis. Our data suggest that the LYTTYL/LYTLYT sequence differences between $M_1$ and $M_2$ muscarinic receptors are not important for specifying ligand binding and coupling of various subtypes of muscarinic receptors to different cellular signaling pathways although they might play a role in the modulation of muscarinic reseptor coupling to PI hydrolysis.

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The Comparison of Shampoos for Skin Hydration by Measurement of Epidermal Capacitance in Normal Canine Skin (Epidermal Capacitance를 이용한 시판되는 치료용 샴푸의 정상적인 개 피부에 대한 가수효과)

  • Oh, Tae-Ho;Jae-Hoon heong;Jang, Kwang-Ho
    • Journal of Veterinary Clinics
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    • v.18 no.3
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    • pp.206-210
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    • 2001
  • Various commercial shampoos were frequently prescribed for dermatologic therapy in small animal practice. Skim hydration affected by the shampoos, however, was not evaluated routinely. In order to evaluate the skin hydration for the exact prescription of shampoos method to measure skin hydration of shampoos are needed. This study was undertaken to evaluate the skin hydration effect of shampoo on canine skin using Corneometer. Five healthy dogs were applied with 7 commercial shampoos: Humilac, Sebocalm, Sebolytics, Etiderm, Benzoyl peroxide, HyLyt and Zn-7 Derm. Skin hydrations were evaluated by measurement of electrical capacitance by Corneometer. A statistically significant increase in skim hydration was found 17(p<0.05) and 77 minutes(p<0.01) after application of Humilac indicating a humidifying effect of this product. A statistically significant decrease in skin hydration was found for the Benzoyl peroxide after 77 minutes(p<0.05). No statistically significant differences between the other shampoos were found. None of the products tested had any negative effect on the skin barrier function. The Corneometer was found useful for detecting skin hydration to shampoos and considered as a simple and useful tool for prescription of various shampoos routine practice.

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Molecular Diagnosis of Streptococcus pneumoniae in Middle Ear Fluids from Children with Otitis Media with Effusion (삼출성 중이염 소아의 중이액에서 폐구균의 분자적 진단)

  • Byun, Sung Wan;Kim, Han Wool;Yoon, Seo Hee;Park, In Ho;Kim, Kyung-Hyo
    • Pediatric Infection and Vaccine
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    • v.22 no.2
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    • pp.106-112
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    • 2015
  • Purpose: The long-term administration of antibiotics interferes with bacterial culture in the middle ear fluids (MEFs) of young children with otitis media with effusion (OME). The purpose of this study is to determine whether molecular diagnostics can be used for rapid and direct detection of the bacterial pathogen in culture-negative MEFs. Methods: The specificity and sensitivity of both polymerase chain reaction (PCR) and loop-mediated isothermal amplification (LAMP) to the lytA gene of Streptococcus pneumoniae were comparatively tested and then applied for pneumococcal detection in the clinical MEFs. Results: The detection limit of the PCR assay was approximately $10^4$ colony forming units (CFU), whereas that of LAMP was less than 10 CFU for the detection of S. pneumoniae. Both PCR and LAMP did not amplify nucleic acid at over $10^6$ CFU of H. influenzae or M. catarrhalis, both of which were irrelevant bacterial species. Of 22 culture-negative MEFs from children with OME, LAMP positivity was found in twelve MEFs (54.5%, 12/22), only three of which were PCR-positive (25%, 3/12). Our results showed that the ability of LAMP to detect pneumococcal DNA is over four times higher than that of PCR (P<0.01). Conclusions: As a high-resolution tool able to detect nucleic acid levels equivalent to <10 CFU of S. pneumoniae in MEFs without any cross-reaction with other pathogens, lytA -specific LAMP may be applied for diagnosing pneumococcus infection in OME as well as evaluating the impact of a pneumococcal conjugate vaccine against OME.