• Title/Summary/Keyword: Key Infection

Search Result 386, Processing Time 0.027 seconds

Regulation of Tumor Neceosis Factor-${\alpha}$ Receptors and Signal Transduction Pathways

  • Han, Hyung-Mee
    • Toxicological Research
    • /
    • v.8 no.2
    • /
    • pp.343-357
    • /
    • 1992
  • Tumor necrosis factor-${\alpha}$(TNF), a polypeptide hormone secreted primarily by activated macrophages, was originally identified on the basis of its ability to cause hemorrhagic necrosis and tumor regression in vivo. Subsequently, TNF has been shown to be an important component of the host responses to infection and cancer and may mediate the wasting syndrome known as cachexia. These systemic actions of TNF are reflected in its diverse effects on target cells in vitro. TNF initiates its diverse cellular actions by binding to specific cell surface receptors. Although TNF receptors have been identified on most of animal cells, regulation of these receptors and the mechanisms which transduce TNF receptor binding into cellular responses are not well understood. Therefore, in the present study, the mechanisms how TNF receptors are being regulated and how TNF receptor binding is being transduced into cellular responses were investigated in rat liver plasma membranes (PM) and ME-180 human cervical carcinoma cell lines. $^{125}I$-TNF bound to high ($K_d=1.51{\pm}0.35nM$)affinity receptors in rat liver PM. Solubilization of PM with 1% Triton X-100 increased both high affinity (from $0.33{\pm}0.04\;to\;1.67{\pm}0.05$ pmoles/mg protein) and low affinity (from $1.92{\pm}0.16\;to\;7.57{\pm}0.50$ pmoles/mg protein) TNF binding without affecting the affinities for TNF, suggesting the presence of a large latent pool of TNF receptors. Affinity labeling of receptors whether from PM or solubilized PM resulted in cross-linking of $^{125}I$-TNF into $M_r$ 130 kDa, 90 kDa and 66kDa complexes. Thus, the properties of the latent TNF receptors were similar to those initially accessible to TNF. To determine if exposure of latent receptors is regulated by TNF, $^{125}I$-TNF binding to control and TNF-pretreated membranes were assayed. Specific binding was increased by pretreatment with TNF (P<0.05), demonstrating that hepatic PM contains latent TNF receptors whose exposure is promoted by TNF. Homologous up-regulation of TNF receptors may, in part, be responsible for sustained hepatic responsiveness during chronic exposure to TNF. As a next step, the post-receptor events induced by TNF were examined. Although the signal transduction pathways for TNF have not been delineated clearly, the actions of many other hormones are mediated by the reversible phosphorylation of specific enzymes or target proteins. The present study demonstrated that TNF induces phosphorylation of 28 kDa protein (p28). Two dimensional soidum dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) resolved the 28kDa phosphoprotein into two isoforms having pIs of 6.2 and 6.1. The pIs and relative molecular weight of p28 were consistent with those of a previously characterized mRNA cap binding protein. mRNA cap binding proteins are a class of translation initiation factors that recognize the 7-methylguanosine cap structure found on the 5' end of eukaryotic mRNAs. In vitro, these proteins are defined by their specific elution from affinity columns composed of 7-methylguanosine 5'-triphosphate($m^7$GTP)-Sepharose. Affinity purification of mRNA cap binding proteins from control and TNF treated ME-180 cells proved that TNF rapidly stimulates phosphorylation of an mRNA cap binding protein. Phosphorylation occurred in several cell types that are important in vitro models of TNF action. The mRNA cap binding protein phosphorylated in response to TNF treatment was purifice, sequenced, and identified as the proto-oncogene product eukaryotic initiation factor-4E(eIF-4E). These data show that phosphorylation of a key component of the cellular translational machinery is a common early event in the diverse cellular actions of TNF.

  • PDF

Expression, Purification and Antiserum Production of the Avian Influenza H9N2 Virus HA and NA Proteins (Avian Influenza H9N2 Virus의 HA와 NA 단백질 발현, 정제 및 항혈청 생산)

  • Lee, Hyun-Ji;Song, Byung-Hak;Kim, Jeong-Min;Yun, Sang-Im;Kim, Jin-Kyoung;Kang, Young-Sik;Koo, Yong-Bum;Jeon, Ik-Soo;Byun, Sung-June;Lee, Youn-Jeong;Kwon, Jun-Hun;Park, Jong-Hyeon;Joo, Yi-Seok;Lee, Young-Min
    • Korean Journal of Microbiology
    • /
    • v.44 no.3
    • /
    • pp.178-185
    • /
    • 2008
  • Avian influenza virus (AIV) is recognized as key to the emergence of pandemic influenza for humans; there are growing concerns that AIV H9N2 may become more efficient to transmit to humans in the near future, since the infection of poultry with AIV H9N2 has been common in recent years. In this study, we aimed to produce antisera recognizing the HA and NA proteins of AIV H9N2. Initially, coding sequences corresponding to the N-terminal regions of the HA and NA proteins of the Korean AIV H9N2 (A/Ck/Kr/MS96/96) isolated from a domestic chicken were amplified from the genomic RNA. Following cloning of the amplified cDNA fragments into pGEX4T-1 vector, two GST-fusion proteins (GST-HAln and GST-NAn) were expressed in E. coli BL21 and purified with glutathione sepharose columns; the recombinant GST-HAln and GST-NAn proteins were both used as immunogens in rabbits. The antigenicity of the rabbit antisera was analyzed by immunoblotting of the cell lysates prepared from AIV H9N2-infected MDCK cells. Overall, the recombinant HAln and NAn proteins fused to the C-terminus of GST and the rabbit antisera raised against the corresponding recombinant proteins would provide a valuable reagent for AIV diagnosis and basic research.

Role of Alveolar Macrophages in Productions of Prostaglandin D2 and E2 in the Inflamed Lung (프로스타글란딘 D2와 E2의 생성에 대한 허파 마크로파이지의 역할)

  • Joo, Myung-Soo
    • Journal of Life Science
    • /
    • v.20 no.6
    • /
    • pp.845-852
    • /
    • 2010
  • Our previous study showed that lungs infected by Pseudomonas, a gram-negative bacteria, produce prostaglandin $D_2$ ($PGD_2$) and prostaglandin $E_2$ ($PGE_2$), the two major prostanoids generated by cyclooxygenase-2 (COX-2), and that the ratio of $PGD_2$ and $PGE_2$ can affect the outcome of the bacterial lung infection. In this study, we sought to uncover the mechanism that determines the ratio of $PGD_2$ and $PGE_2$ produced in lung inflammation. When treated with lipopolysaccharide (LPS), primary alveolar macrophages, extracted from mouse lung, more $PGE_2$ was produced than $PGD_2$, whereas MH-S, a murine alveolar macrophage cell line, produced more $PGD_2$ than $PGE_2$ in a similar experiment. Western blot analyses showed that the kinetics of COX-2 expression in both cell types is similar and epigenetic silencing of COX-2 expression did not affect expressions of lipocalin-PGD synthase (L-PGDS) and PGE synthase (mPGES-1), major enzymes synthesizing $PGD_2$ and $PGE_2$ in inflammation, respectively, indicating no effect of COX-2 on expressions of the two enzymes. Expressions of L-PGDS and mPGES-1 were also similar in both cell types, suggesting no effect of the two key enzymes in determining the ratio of $PGD_2$ and $PGE_2$ in these cells. A single intraperitoneal injection of LPS to C57BL/6 mice induced COX-2 expression and, similar to alveolar macrophages, produced more $PGE_2$ than $PGD_2$ in the lung. These results suggest that the differential expressions of $PGD_2$ and $PGE_2$ in the lung reflect those in alveolar macrophages and may not be directly determined by the enzymes responsible for $PGD_2$ and $PGE_2$ synthesis.

Epidemics of Soybean Mosaic Virus and Varietal Resistance in Soybean (콩모자이크바이러스병(病)의 전염(傳染) 및 저항성(抵抗性) 검정(檢定)에 대하여)

  • Kim, Sang-Gyu;Lee, Key-Woon
    • Korean journal of applied entomology
    • /
    • v.25 no.2 s.67
    • /
    • pp.113-120
    • /
    • 1986
  • Soybeans collected from different locations in Korea were planted for tests of seed transmission of soybean mosaic virus (SMV). The percentage of seed transmission ranged from 20.5 to 29.5% in 14 seedlots including soybean cultivar Namhe and 12 to 19% in other 44 seedlots. However, no seed transmission of SMV was observed in soybean cultivar Younkiyongho. SMV was detected from embryo and cotyledon of soybean seeds. The infection of SMV was highly detected from premature seeds than from fully mature seeds, and higher from seeds harvested from plants infected before June 20 than plants infected around July 20 and August 20. No significant relationship was observed between seed transmission of SMV and mottling of seeds. The incidence of soybean mosaic disease followed by one month after peak of aphid population. The number of aphids was less on leaves of soybeans with short and dense trichomes whereas it was higher on leaves of soybean with long and sparse trichomes. Generally, the number of aphids was decreased on leaves with long and dense trichomes as the growth progressed. Soybean cultivar Columbus and 14 cultivars were susceptible, Chief and 14 cultivars were moderate, but Jangbaek and 17 other cultivars were resistant to SMV when inoculated with one isolate of SMV.

  • PDF

Criminal Law Issues in Epidemiological Investigations Under the INFECTIOUS DISEASE CONTROL AND PREVENTION ACT (감염병의 예방 및 관리에 관한 법률상 역학조사와 관련된 형사법적 쟁점)

  • Jang, Junhyuk
    • The Korean Society of Law and Medicine
    • /
    • v.23 no.3
    • /
    • pp.3-44
    • /
    • 2022
  • As a result of a close review focusing on the case of obstruction of epidemiological investigation by a religious group A in Daegu, which was a problem when the pandemic of Covid-19 infection began in Korea around February 2, 2020, when an epidemiological investigator requested a specific group to submit a list, While there have been cases where an act of not responding or submitting an edited omission list was sentenced to the effect that the act did not fall under an epidemiological investigation, in the case of non-submission of the visitor list for the B Center, even though a 'list of visitors' was requested. Regarding the fact of refusal without a justifiable reason, 'providing a list of persons entering the building is a key factual act that forms a link between epidemiological investigations accompanying an epidemiological investigation, and refusing to do so is also an act of refusal and obstruction of an epidemiological investigation. There are cases where it is possible to demand criminal punishment. Regardless of whether the request for submission of the membership list falls under the epidemiological investigation, there are cases in which the someones' actions correspond to the refusal or obstruction of the epidemiological investigation. A lower court ruling that if an epidemiological investigation is rejected or obstructed as a result of interfering with factual acts accompanying an epidemiological investigation, comprehensively considering whether or not the list has been diverted for purposes other than epidemiological investigation, the logic is persuasive. Epidemiological investigations such as surveys and human specimen collection and testing are conducted for each infectious disease patient or contact confirmed as a result of the epidemiological investigation, but epidemiological investigations conducted on individual individuals cannot exist independently of each other, and the This is because the process of identification and tracking is essential to an epidemiological investigation, and if someone intentionally interferes with or rejects the process of confirming this link, it will result in direct, realistic, and widespread interference with the epidemiological investigation. In this article, ① there are differences between an epidemiological investigation and a request for information provision under the Infectious Disease Control and Prevention Act, but there are areas that fall under the epidemiological investigation even in the case of a request for information, ② Considering the medical characteristics of COVID-19 and the continuity of the epidemiological investigation, the epidemiological investigator the fact that the act of requesting a list may fall under the epidemiological investigation, ③ that the offense of obstructing the epidemiological investigation in certain cases may constitute 'obstruction of Performance of Official Duties by Fraudulent Means', and ④ rejecting the request for information provision under the Infectious Disease Control and Prevention Act from September 29, 2020 In this case, it is intended to be helpful in the application of the Infectious Disease control and Prevention Act and the practical operation of epidemiological investigations in the future by pointing out the fact that a new punishment regulation of imprisonment or fine is being implemented.

Brief Introduction of Research Progresses in Control and Biocontrol of Clubroot Disease in China

  • He, Yueqiu;Wu, Yixin;He, Pengfei;Li, Xinyu
    • 한국균학회소식:학술대회논문집
    • /
    • 2015.05a
    • /
    • pp.45-46
    • /
    • 2015
  • Clubroot disease of crucifers has occurred since 1957. It has spread to the whole China, especially in the southwest and nourtheast where it causes 30-80% loss in some fields. The disease has being expanded in the recent years as seeds are imported and the floating seedling system practices. For its effective control, the Ministry of Agriculture of China set up a program in 2010 and a research team led by Dr. Yueqiu HE, Yunnan Agricultural University. The team includes 20 main reseachers of 11 universities and 5 institutions. After 5 years, the team has made a lot of progresses in disease occurrence regulation, resources collection, resistance identification and breeding, biological agent exploration, formulation, chemicals evaluation, and control strategy. About 1200 collections of local and commercial crucifers were identified in the field and by artificiall inoculation in the laboratories, 10 resistant cultivars were breeded including 7 Chinese cabbages and 3 cabbages. More than 800 antagostic strains were isolated including bacteria, stretomyces and fungi. Around 100 chemicals were evaluated in the field and greenhouse based on its control effect, among them, 6 showed high control effect, especially fluazinam and cyazofamid could control about 80% the disease. However, fluzinam has negative effect on soil microbes. Clubroot disease could not be controlled by bioagents and chemicals once when the pathogen Plasmodiophora brassicae infected its hosts and set up the parasitic relationship. We found the earlier the pathogent infected its host, the severer the disease was. Therefore, early control was the most effective. For Chinese cabbage, all controlling measures should be taken in the early 30 days because the new infection could not cause severe symptom after 30 days of seeding. For example, a biocontrol agent, Bacillus subtilis Strain XF-1 could control the disease 70%-85% averagely when it mixed with seedling substrate and was drenching 3 times after transplanting, i.e. immediately, 7 days, 14 days. XF-1 has been deeply researched in control mechanisms, its genome, and development and application of biocontrol formulate. It could produce antagonistic protein, enzyme, antibiotics and IAA, which promoted rhizogenesis and growth. Its The genome was sequenced by Illumina/Solexa Genome Analyzer to assembled into 20 scaffolds then the gaps between scaffolds were filled by long fragment PCR amplification to obtain complet genmone with 4,061,186 bp in size. The whole genome was found to have 43.8% GC, 108 tandem repeats with an average of 2.65 copies and 84 transposons. The CDSs were predicted as 3,853 in which 112 CDSs were predicted to secondary metabolite biosynthesis, transport and catabolism. Among those, five NRPS/PKS giant gene clusters being responsible for the biosynthesis of polyketide (pksABCDEFHJLMNRS in size 72.9 kb), surfactin(srfABCD, 26.148 kb, bacilysin(bacABCDE 5.903 kb), bacillibactin(dhbABCEF, 11.774 kb) and fengycin(ppsABCDE, 37.799 kb) have high homolgous to fuction confirmed biosynthesis gene in other strain. Moreover, there are many of key regulatory genes for secondary metabolites from XF-1, such as comABPQKX Z, degQ, sfp, yczE, degU, ycxABCD and ywfG. were also predicted. Therefore, XF-1 has potential of biosynthesis for secondary metabolites surfactin, fengycin, bacillibactin, bacilysin and Bacillaene. Thirty two compounds were detected from cell extracts of XF-1 by MALDI-TOF-MS, including one Macrolactin (m/z 441.06), two fusaricidin (m/z 850.493 and 968.515), one circulocin (m/z 852.509), nine surfactin (m/z 1044.656~1102.652), five iturin (m/z 1096.631~1150.57) and forty fengycin (m/z 1449.79~1543.805). The top three compositions types (contening 56.67% of total extract) are surfactin, iturin and fengycin, in which the most abundant is the surfactin type composition 30.37% of total extract and in second place is the fengycin with 23.28% content with rich diversity of chemical structure, and the smallest one is the iturin with 3.02% content. Moreover, the same main compositions were detected in Bacillus sp.355 which is also a good effects biocontol bacterial for controlling the clubroot of crucifer. Wherefore those compounds surfactin, iturin and fengycin maybe the main active compositions of XF-1 against P. brassicae. Twenty one fengycin type compounds were evaluate by LC-ESI-MS/MS with antifungal activities, including fengycin A $C_{16{\sim}C19}$, fengycin B $C_{14{\sim}C17}$, fengycin C $C_{15{\sim}C18}$, fengycin D $C_{15{\sim}C18}$ and fengycin S $C_{15{\sim}C18}$. Furthermore, one novel compound was identified as Dehydroxyfengycin $C_{17}$ according its MS, 1D and 2D NMR spectral data, which molecular weight is 1488.8480 Da and formula $C_{75}H_{116}N_{12}O_{19}$. The fengycin type compounds (FTCPs $250{\mu}g/mL$) were used to treat the resting spores of P. brassicae ($10^7/mL$) by detecting leakage of the cytoplasm components and cell destruction. After 12 h treatment, the absorbencies at 260 nm (A260) and at 280 nm (A280) increased gradually to approaching the maximum of absorbance, accompanying the collapse of P. brassicae resting spores, and nearly no complete cells were observed at 24 h treatment. The results suggested that the cells could be lyzed by the FTCPs of XF-1, and the diversity of FTCPs was mainly attributed to a mechanism of clubroot disease biocontrol. In the five selected medium MOLP, PSA, LB, Landy and LD, the most suitable for growth of strain medium is MOLP, and the least for strains longevity is the Landy sucrose medium. However, the lipopeptide highest yield is in Landy sucrose medium. The lipopeptides in five medium were analyzed with HPLC, and the results showed that lipopeptides component were same, while their contents from B. subtilis XF-1 fermented in five medium were different. We found that it is the lipopeptides content but ingredients of XF-1 could be impacted by medium and lacking of nutrition seems promoting lipopeptides secretion from XF-1. The volatile components with inhibition fungal Cylindrocarpon spp. activity which were collect in sealed vesel were detected with metheds of HS-SPME-GC-MS in eight biocontrol Bacillus species and four positive mutant strains of XF-1 mutagenized with chemical mutagens, respectively. They have same main volatile components including pyrazine, aldehydes, oxazolidinone and sulfide which are composed of 91.62% in XF-1, in which, the most abundant is the pyrazine type composition with 47.03%, and in second place is the aldehydes with 23.84%, and the third place is oxazolidinone with 15.68%, and the smallest ones is the sulfide with 5.07%.

  • PDF