The neuropeptide substance P(SP) has been implicated in the mediation of inflammation and immune-mediated disease such as arthritis. Recently, it was reported that SP was markedly increased around the blood vessels in inflamed gingiva as well as in close association with the inflammatory cell infiltrate. These results support that SP may contribute to the pathophysiology of neuronal inflammation in human periodontal tissues. SP may regulate inflammatory/immune responses by stimulating the proliferation of human T cells, differentiation and antibody-secreting potential of B cells, macrophage respiratory burst, connective tissue proliferation, and the secretion of cytokines from monocytes and T cells. Here, I studied potential role of SP as a costimulatory chemical signal in inflammatory/immune responses, by determining the released proinflammatory cytokines such as $MIP-1{\alpha}$, $IL-1{\beta}$, and IL-6 from culture supernatants of homogeneous immune cell lines. Serum free cell supernatants were concentrated with TCA precipitation, fractionated with SDS-PAGE, and subjected into western blot analysis. Among 15 cell lines tested, macrophage/monocyte cell line RAW264.7 and WRl9m.1 showed the highest level of induction of $MIP-1{\alpha}$ when stimulated with LPS. Discrete IL-6 bands with multiple forms of molecular mass were detected from supernatants of B cell lines A20(32kDa), Daudi(32, 35kDa), and SKW6.4(29kDa), which were expressed constitutively. $IL-1{\beta}$ could not be detected by the method of western blot analysis from supernatants of all cell lines tested except RAW264.7, WRl9m.1, and erythroid cell line K562 which showed the least amount of $IL-{\beta}$ secretion. SP $10^{-9}M$ with suboptimal dose of LPS treatment showed synergistic induction of $MIP-1{\alpha}$ release from RAW264.7 or WR19m.1, and also IL-6 release from A20, but this synergism is not the case in costimulation of RAW264.7 or WRl9m.1 with SP $10^{-9}M$ and TPA. Although treatment of T cell line CTLL-R8 with SP $10^{-7}M$ or PHA+TPA induced modest level of $MIP-1{\alpha}$ secretion, synergism was not observed when they are applied together. These findings all together suggest the possibility of a regulatory role of SP in inflammatory/immune reaction through differential modulation of bioactivities of other chemical cosignals.
Journal of the Korea Institute of Information and Communication Engineering
/
v.8
no.4
/
pp.926-932
/
2004
This study deals with the statistical analyses on the relationship between the red tide formation and the meteorological factors in the Korean coastal waters. From 1995 to 2002, the red tide was observed every year and the number of occurrences increased as well. The red tide mostly occurred in July, August, September and October. from multiple linear regression, the meteorological factors governing the mechanisms of the increase in the number of red tide occurrences are found to be a water temperature, rainfall, sunshine duration and wind velocity. But water temperature as the limited factor controlling the growth of phytoplankton (Cochlodinium polykrikoids) in 15∼$30^{\circ}c$. NO = 8.089 - 0.319WT + 0.019RF + 0.141SD + 0.l19WV (R = 0.897) in August NO = 7.531 - 0.327WT + 0.027RF + 0.208SD + 0.208WV (R = 0.894) in September Here, NO is the number of occurrence for red tide, WT is water temperature, RF is rainfall, SD is sunshine duration and WV is wind velocity, respectively. The necessary times till the day of red tide occurrence verse the day when water temperature reaches $15^{\circ}c$ are 78∼104 days, then it should be divided the coastal waters into 4 areas by the comparison among the accumulated sunshine duration, water temperature and rainfall as follows; the South West Coast (SW), South Middle Coast (SM), South East Coast(SE) and East South Coast (ES). The coastal areas that red tide occurs were complicated and various by change of marine environments. Usually red tide with a high concentrations (individual number, cells/ml) appeared in SM and SE. It was found that the general situations for the frequencies of red tide formation are mainly concentrated to 24.5∼$25^{\circ}c$ (high water temperature) and eve. 1000 cells/ml (high individual number) such as the category of red tide warning.
In order to observe the antigenic localization in the tissues of the adult Clonorchis sinensis, immunogold labeling method was applied using serum immunoglobulins (IgG) of either worm·infected rabbits (group I) or antigen-immunized rabbits (group II) (by the body quid obtained from the adult worms). The electron micrographs of the sectioned worm tissue antigens, embedded in Lowicryl HM 20 medium and stained with protein A-gold complex (particle sixte: 12 nm), were compared between the group I and group II. The gold particles were observed in the interstitial matrix of the worm parenchyma, the epithelial lamellae of the cecum, and the cecal lumen both in group I and II. But the particles were in general more concentrated in group II. The gold particles were not observed on the basal lamina of the tegument or on vitelline glands in group I, while they were highly concentrated on those areas in group II. There were also differences in the antigenicity of interstitial matrix(reacted with group I IgG) and head part(reacted with group II IgG) of the sperm cells in the seminal receptacle. Conclusively, it is suggested that the substances comprising the basal lamina of the tegument or vitelline glands act as specific antigens reacting with antigen(body quid) immunized rabbit IgG. On the other hand, the substances in the cecal lumen and cecal epithelial lamellae are thought to be the specific antigen that react with the worm-infected rabbit IgG.
The poxvirus group is considered to be a typical cytoplasmic inclusion forming virus. Every poxvirus has been reported to produce only one kind of inclusion in the infected tissues. A vague concept that inclusions of poxviruses are eosinophilic or acidophilic has prevailed. Although many papers and theories about the nature of the inclusion have been presented, most of them are not quite convincing on the point of the relations with virus multiplication, and an analysis of papers published showed that there seem to be many discrepancies in the descriptions of the nature of the poxvirus inclusions. Comparative studies on host-virus interaction with cowpox, orf, swinepox and fowlpox viruses which selected from each Group (I-IV) of poxviruses were performed from the morphological and virological standpoints. At first, in cowpox virus-FL cell system, as a comparative model, cytoplasmic inclusion, nucleic acid metabolism by autoradiography and detection of viral antigen by immunofluorescence were studied and obtained the results as follows: 1. The focus-like cytopathic effect (CPE) at early stage developed to entire culture at terminal stage of infection, and also the developing status of CPE was correlated to viral doses for inoculation. Two kinds of cytoplasmic inclusions which named A and B type were easily observed by Giemsa, hematoxylin-eosin (H & E) and May-Greenwald Giemsa (MGG) stainings in the infected cells. The B type inclusions were formed at early stage of infection and the A type inclusions were produced subsequently the B type formation. The B type which common type inclusion in poxviruses was a small compact or aggregate at early stage and developed to a large diffuse body at terminal stage of infection. On the other hand, the A type inclusion which depend upon the kind of virus was appeared as round and discrete shape, and its size and number was increased gradually during the culture period. It was characteristic to form distinct halos around the both types of inclusions in acid fixed, H & E stained preparations of infected cultures. The B type inclusion was always positive in Feulgen reaction and showed as DNA containing body but the A type inclusion was not. 2. In the relationship between inclusion and DNA metabolism of infected cells by the qualitative autoradiography using 3H-thymidine, the appearance of silver grains was coincided with B type inclusion but not with A type inclusion. This showed that the DNA synthesis was proceeded in all B type inclusions except those in the terminal stage with a diffuse form. This suggested that the B type inclusions are only sites of DNA synthesis and this was proceeded after the cell infection independently. The activity of DNA synthesis of the inclusions was nearly the same as that of the nucleic of normal cells and non-inclusion bearing cells. and non-inclusion bearing cells. Regardless of the size of the degree of DNA synthesis of the B type inclusion, inclusion bearing cells all showed remarkable suppression of nuclear DNA synthesis. 3. By the direct fluorescent antibody technique viral antigen in infected cells was detected. The B type inclusions have been proved to contain a great deal of viral antigen, whereas the basic substance of A type inclusion did not show antigenicity except the round edge. It was suggested that the round edge fluorescence might be caused by the glare of cytoplasmic viral antigen which pushed out and concentrated by the A type inclusion development. 4. Hemorrhagic red pock formations on chorioallantoic membrane of embryonated chicken egg had proved the characteristic of used viral strain. 5. By the above studies on the nature of two types of inclusions and the role they play in virus multiplication, it was concluded that the B type inclusion must be the site of the synthesis of viral DNA and protein as well as the site of the virus.
Cyanobacterial resting cells, such as akinetes, are important seed cells for cyanobacteria's early development and bloom. Due to their importance, various methods have been attempted to isolate resting cells present in the sediment. Ludox is a solution mainly used for cell separation in marine sediments, but finding an accurate method for use in freshwater is difficult. This study compared the two most commonly used Ludox methods (direct sediment treatment and sediment distilled water suspension treatment). Furthermore, we proposed a highly efficient method for isolating cyanobacterial resting cells and eDNA amplification from freshwater sediments. Most of the resting cells found in the sediment were akinete to the Nostocale and were similar to those of Dolichospermum, Cylindrospermum, and Aphanizomenon. Twenty times more akinetes were found in the conical tube column using the sediment that had no treatment than in the sample treated by suspending the sediment in distilled water. Akinete separated through Ludox were mainly spread over the upper and lower layers in the column rather than concentrated at a specific depth in the column layer. The mibC, Geo, and 16S rDNA genes were successfully amplified using the sediment directly in the sample. However, the amplification products of all genes were not found in the sample in which the sediment was suspended in distilled water. Therefore, 5 g to 10 g of sediment is used without pretreatment when isolating cyanobacterial resting cells from freshwater sediment. Cell isolation and gene amplification efficiency are high when four times the volume of Ludox is added. The Ludox treatment method presented in this study isolates cyanobacterial resting cells in freshwater sediment, and the same efficiency may not appear in other biotas. Therefore, to apply Ludox to the separation of other biotas, it is necessary to conduct a pre-experiment to determine the sediment pretreatment method and the water layer where the target organism exists.
The purpose of this study was to evaluate and compare different elution and concentration methods for optimization of human rotavirus (HRV) detection method using real-time RT-PCR and cell culture techniques. The leafy vegetable samples (lettuce, Chinese cabbage) were artificially inoculated with HRV. Viruses were extracted from the vegetables by two different elution buffers, buffer A (100 mM Tris-HCl, 50 mM glycine, 3% beef extract, pH 9.5) and buffer B (250 mM Threonine, 300 mM NaCl, pH 9.5), and the extracted viruses were concentrated by filtration and PEG precipitation sequentially. To determine infectivity of the viruses, the viruses recovered from the samples were infected to the MA-104 cells, and integrated cell culture real-time RT-PCR was performed at 1, 48, 72, 96, 120, 144, 168 h post-infection (p.i.). The elution buffer A was more efficient in extracting the virus from the produce samples tested than the buffer B, 29.54% and 18.32% of recoveries, respectively. The sensitivity of real-time RT-PCR method was markedly improved when the virus was concentrated by the filtration method. When the viruses were eluted and concentrated by buffer A and filtration, respectively, the average recovery rate was approximately 51.89%. When the viruses recovered from samples were infected to MA-104 cell, infectious HRV was detected within 48 h p.i. by ICC/real-time RT-PCR, whereas cytopathic effects were not observed until 72 h p.i. The optimized detection method evaluated in this study could be useful for rapid and reliable detection of HRV in fresh produce products and applied for detection of other food-borne viruses.
The $LaCrO_3$-dispersed Cr alloys for metallic interconnect of solid oxide fuel cell were prepared as a function of $LaCrO_3$ content in the range of 5 to 25 vol.% and were sintered at 1500$^{\circ}C$ under an Ar atmosphere with 5 vol.% $H_2$. The sintering and oxidation behaviors of these alloys were examined. The alloys indicated a good sinterability above 95% relative density at a given sintering condition, and their sintering densities is independent on $LaCrO_3$ content. The $LaCrO_3$ particles of the sintered alloys were concentrated on interfaces of Cr particles, and the size of the Cr particles increased with decreasing $LaCrO_3$ content, which is caused by inhibited grain growth of Cr particle by $LaCrO_3$ particle. The oxidation test showed all $LaCrO_3$-dispersed Cr alloys have good oxidation resistance as compared with pure Cr, which is attributed to presence of $LaCrO_3$ at the interface at which the oxidation reaction occurs rapidly. The Cr alloys with about 15 vol.% $LaCrO_3$ are very resistant to oxidation.
Kim, Jae-Won;Kim, Bong-Seok;Choi, Cheol-Young;Lee, Jung-Sick
Applied Microscopy
/
v.33
no.3
/
pp.243-250
/
2003
Ultrastructural changes of the male germ cells and structure of spermatozoa in Paralichthys olivaceus were examined by means of the light and transmission electron microscopes. The spermatogonium has a large nucleus with a single nucleus with a single nucleolus in the interphase. Primary spermatocytes are identified by the formation of the synaptonemal complex in the karyoplasm. The secondary spermatocytes are more concentrated and contains numerous cell organelle in the cytoplasm. The nucleus of spermatid in spermiogenesis is more condensed in the karyoplasm, and show spherical structure in shape. Mitochondria of the spermatids are observed in the lower portion of the nucleus. The spermatozoon consists of the head, mid piece and tail. The acrosome is not observed in the head. Axial filaments of the flagellum consists of nine pairs of the peripheral microtubules and one pair of the central microtubules.
CFD is used to analyze gas flow characteristics, power absorption, electron temperature, electron density and chemical species profile of an internal antenna type inductively coupled plasma system. An optimized grid generation technology is used for a complex real-scale models for industry. A bare metal antenna shows concentrated power absorption around rf a feeding line. Skin depth of power absorption for a system is modeled to 50 mm, which is reported 53 mm by experiments. For an application of bipolar plates for hydrogen fuel cells, multi-sheet loading ICP nitriding system is proposed using an internal ICP antenna. It shows higher atomic nitrogen density than reported simple pulsed dc nitriding systems. Minimum gap between sheets for uniform nitriding is modeled to be 39 mm.
A total of 220 tadpoles was captured in 6 areas and total 117 frogs, Rana nigromacuzata, were collected in 11 areas in Korea. They were examined for their infection status by the metacercariae of Fibricola seoulensis by peptic digestion technique and by histological observation with hematoBylineosin staining. This study was carried out from August, 1983 to September, 1984. Followings are the results. 1. The tadpoles of R. nigromaculata were positive for the metacercariae from 3.3% to 100% by area. The number of metacercariae per infected tadpole ranged from 1 to 584, and the mean number Per tadpole ranged from 7.6 to 221 by area. 2. The metacercariae from 16 tadpoles were counted by the body portion. A great majority of the metacercariae was collected from abdominal cavity, 98.3% of 484 counted larvae. And 6(1.2%) larvae were from proximal tail and 2(0.4%) from trunk. 3. Histological sections of tadpoles showed many metacercariae in abdominal cavity but none in other parts. The larvae were free in the spaces among intestinal loops or around primitive liver. A few larvae were in duct-like tissues near trunk wall. There was little infiltration of inaammatory cells. 4. The metacercarial infection rates of frogs ranged from 0% to 100% by area. The larval burden was 1 to 470 by infected frogs, and mean number ranged from 1 to 175.6 by area. By above results, it is suggested that the cercariae of F. seoulensis may infect R. nigromaculata already in the stage of tadpole. Almost all of the metacercariae were concentrated in abdominal cavity of tadpoles. According to the infection status of frogs, this nuke is prevalent almost nationwidely in rice paddies in Korea.
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