The use of nanoparticles as a delivery system for a specific antigen could solve many limitations of mucosal vaccine applications, such as low immunogenicity, or antigen protection and stabilization. In this study, we tested the ability of nasally administered chitosan nanoparticles loaded with glycoprotein B of murine cytomegalovirus to induce an immune response in an animal model. The choice of chitosan nanoparticle type was made by in vitro evaluation of sorption efficiency and antigen release. Three types of chitosan nanoparticles were prepared: crosslinked with tripolyphosphate, coated with hyaluronic acid, and in complex with polycaprolactone. The hydrodynamic size of the nanoparticles by dynamic light scattering, zeta potential, Fourier transform infrared spectroscopy, scanning electron microscopy, stability, loading efficiency, and release kinetics with ovalbumin were evaluated. Balb/c mice were immunized intranasally using the three-dose protocol with nanoparticles, gB, and adjuvants Poly(I:C) and CpG ODN. Subsequently, the humoral and cell-mediated antigen-specific immune response was determined. On the basis of the properties of the tested nanoparticles, the cross-linked nanoparticles were considered optimal for further investigation. The results show that nanoparticles with Poly(I:C) and with gB alone raised IgG antibody levels above the negative control. In the case of mucosal IgA, only gB alone weakly induced the production of IgA antibodies compared to saline-immunized mice. The number of activated cells increased slightly in mice immunized with nanoparticles and gB compared to those immunized with gB alone or to negative control. The results demonstrated that chitosan nanoparticles could have potential in the development of mucosal vaccines.
Background: The opitmal ventilator setting during partial liquid ventilation(PLV) is controversial. This study investigated the effects of various gas exchange parameters during PLV in normal rabbit lungs in order to aid in the development of an optimal ventilator setting during PLV. Methods: Seven New-Zealand white rabbits were ventilated in pressure-controlled mode with the following settings; tidal volume($V_T$) 8 mL/kg, positive end-expiratory pressure(PEEP) 4 $cmH_2O$, inspiratory-to-expiratory ratio(I:E ratio) 1:2, fraction of inspired oxygen($F_TO_2$) 1.0. The respiration rate(RR) was adjusted to keep $PaCO_2$ between 35~45 mmHg. The ventilator settings were changed every 30 min in the following sequence : (1) Baseline, as the basal ventilator setting, (2) Inverse ratio, I:E ratio 2:1, (3) high PEEP, adjust PEEP to achieve the same mean inspiratory pressure (MIP) as in the inverse ratio, (4) High $V_T$, $V_T$ 15 mL/kg, (5) high RR, the same minute ventilation (MV) as in the High $V_T$. Subsequently, the same protocol was repeated after instilling 18 mL/kg of perfluorodecalin for PLV. The parameters of gas exchange, lung mechanics, and hemodynamics were examined. Results: (1) The gas ventilation(GV) group showed no significant changes in the $PaO_2$ at all phases. The $PaCO_2$ was lower and the pH was higher at the high $V_T$ and high RR phases(p<0.05). No significant changes in the lung mechanics and hemodynamics parameters were observed. (2) The baseline $PaO_2$ for the PLV was $312{\pm}$ mmHg. This was significantly lower when decreased compared to the baseline $PaO_2$ for GV which was $504{\pm}81$ mmHg(p=0.001). During PLV, the $PaO_2$, was significantly higher at the high PEEP($452{\pm}38$ mmHg) and high $V_T$ ($461{\pm}53$ mmHg) phases compared with the baseline phase. However, it did not change significantly during the inverse I:E ratio or the high RR phases. (3) The $PaCO_2$ was significantly lower at high $V_T$ and RR phases for both the GV and PLV. During the PLV, $PaCO_2$ were significantly higher compared to the GV (p<0.05). (4) There were no important or significant changes in of baseline and high RR phases lung mechanics and hemodynamics parameters during the PLV. Conclusion: During PLV in the normal lung, adequate $V_T$ and PEEP are important for optimal oxygenation.
Currently, thanks to the major stride made in developing wired and wireless communication technology, a variety of IT services are available on land. This trend is leading to an increasing demand for IT services to vessels on the water as well. And it is expected that the request for various IT services such as two-way digital data transmission, Web, APP, etc. is on the rise to the extent that they are available on land. However, while a high-speed information communication network is easily accessible on land because it is based upon a fixed infrastructure like an AP and a base station, it is not the case on the water. As a result, a radio communication network-based voice communication service is usually used at sea. To solve this problem, an additional frequency for digital data exchange was allocated, and a ship ad-hoc network (SANET) was proposed that can be utilized by using this frequency. Instead of satellite communication that costs a lot in installation and usage, SANET was developed to provide various IT services to ships based on IP in the sea. Connectivity between land base stations and ships is important in the SANET. To have this connection, a ship must be a member of the network with its IP address assigned. This paper proposes a SANET-CC protocol that allows ships to be assigned their own IP address. SANET-CC propagates several non-overlapping IP addresses through the entire network from land base stations to ships in the form of the tree. Ships allocate their own IP addresses through the exchange of simple requests and response messages with land base stations or M-ships that can allocate IP addresses. Therefore, SANET-CC can eliminate the IP collision prevention (Duplicate Address Detection) process and the process of network separation or integration caused by the movement of the ship. Various simulations were performed to verify the applicability of this protocol to SANET. The outcome of such simulations shows us the following. First, using SANET-CC, about 91% of the ships in the network were able to receive IP addresses under any circumstances. It is 6% higher than the existing studies. And it suggests that if variables are adjusted to each port's environment, it may show further improved results. Second, this work shows us that it takes all vessels an average of 10 seconds to receive IP addresses regardless of conditions. It represents a 50% decrease in time compared to the average of 20 seconds in the previous study. Also Besides, taking it into account that when existing studies were on 50 to 200 vessels, this study on 100 to 400 vessels, the efficiency can be much higher. Third, existing studies have not been able to derive optimal values according to variables. This is because it does not have a consistent pattern depending on the variable. This means that optimal variables values cannot be set for each port under diverse environments. This paper, however, shows us that the result values from the variables exhibit a consistent pattern. This is significant in that it can be applied to each port by adjusting the variable values. It was also confirmed that regardless of the number of ships, the IP allocation ratio was the most efficient at about 96 percent if the waiting time after the IP request was 75ms, and that the tree structure could maintain a stable network configuration when the number of IPs was over 30000. Fourth, this study can be used to design a network for supporting intelligent maritime control systems and services offshore, instead of satellite communication. And if LTE-M is set up, it is possible to use it for various intelligent services.
Currently, an arterial spin labeling (ASL) magnetic resonance imaging (MRI) technique does not routinely used in clinical studies to measure perfusion in brain because optimization of imaging protocol is required to obtain optimal perfusion signals. Therefore, the objective of this study was to investigate changes of perfusion-weighed signal intensities with varying several parameters on a pulsed arterial spin labeling MRI technique obtained from a 3T MRI system. We especially evaluated alternations of ASL-MRI signal intensities on special brain areas, including in brain tissues and lobes. The signal targeting with alternating radiofrequency (STAR) pulsed ASL method was scanned on five normal subjects (mean age: 36 years, range: 29~41 years) on a 3T MRI system. Four parameters were evaluated with varying: 1) the labeling gap, 2) the labeling delay time, 3) the labeling thickness, and 4) the slice scan order. Signal intensities were obtained from the perfusion-weighted imaging on the gray and white matters and brain lobes of the frontal, parietal, temporal, and occipital areas. The results of this study were summarized: 1) Perfusion-weighted signal intensities were decreased with increasing the labeling gap in the bilateral gray matter areas and were least affected on the parietal lobe, but most affected on the occipital lobe. 2) Perfusion-weighted signal intensities were decreased with increasing the labeling delay time until 400 ms, but increased up to 1,000 ms in the bilateral gray matter areas. 3) Perfusion-weighted signal intensities were increased with increasing the labeling thickness until 120 mm in both the gray and white matter. 4) Perfusion-weighted signal intensities were higher descending scans than asending scans in both the gray and white matter. We investigated changes of perfusion-weighted signal intensities with varying several parameters in the STAR ASL method. It should require having protocol optimization processing before applying in patients. It has limitations to apply the ASL method in the white matter on a 3T MRI system.
A fibrinolytic protease (PoFE) was purified from the cultured mycelia of the edible oyster mushroom Pleurotus ostreatus, using a combination of various chromatographies. The purification protocol resulted in an 876-fold purification of the enzyme, with a final yield of 6.5%. The apparent molecular mass of the purified enzyme was estimated to be 32 kDa by SDS-PAGE, fibrin-zymography, and size exclusion using FPLC. The optimal reaction pH value and temperature were pH 6.5 and $35^{\circ}C$, respectively. PoFE effectively hydrolyzed fibrinogen, preferentially digesting the $A{\alpha}$-chain and the $B{\beta}$-chain over the ${\gamma}$-chain. Enzyme activity was enhanced by the addition of $Ca^{2+},\;Zn^{2+},\;and\;Mg^{2+}$ ions. Furthermore, PoFE activity was potently inhibited by EDTA, and it was found to exhibit a higher specificity for the chromogenic substrate S-2586 for chymotrypsin, indicating that the enzyme is a chymotrypsin-like metalloprotease. The first 19 amino acid residues of the N-terminal sequence were ALRKGGAAALNIYSVGFTS, which is extremely similar to the metalloprotease purified from the fruiting body of P. ostreatus. In addition, we cloned the PoFE protein, encoding gene, and its nucleotide sequence was determined. The cDNA of cloned PoFE is 867 nucleotides long and consists of an open reading frame encoding 288 amino acid residues. Its cDNA showed a high degree of homology with PoMEP from P. ostreatus fruiting body. The mycelia of P. ostreatus may thus represent a potential source of new therapeutic agents to treat thrombosis.
In this study we purified a fibrinolytic enzyme from Cordyceps militaris using a combination of ion-exchange chromatography on a DEAE Sephadex A-50 column, gel filtration chromatography on a Sephadex G-75 column, and FPLC on a HiLoad 16/60 Superdex 75 column. This purification protocol resulted in a 191.8-fold purification of the enzyme and a final yield of 12.9 %. The molecular mass of the purified enzyme was estimated to be 52 kDa by SDS-PAGE, fibrin-zymography, and gel filtration chromatography. The first 19 amino acid residues of the N-terminal sequence were ALTTQSNV THGLATISLRQ, which is similar to the subtilisin-like serine protease PR1J from Metarhizium anisopliae var. anisopliase. This enzyme is a neutral protease with an optimal reaction pH and temperature of 7.4 and $37^{\circ}C$, respectively. Results for the fibrinolysis pattern showed that the enzyme rapidly hydrolyzed the fibrin $\alpha$-chain followed by the $\gamma$-$\gamma$ chains. It also hydrolyzed the $\beta$-chain, but more slowly. The A$\alpha$, B$\beta$, and $\gamma$ chains of fibrinogen were also cleaved very rapidly. We found that enzyme activity was inhibited by $Cu^{2+}$ and $Co^{2+}$, but enhanced by the additions of $Ca^{2+}$ and $Mg^{2+}$ ions. Furthermore, fibrinolytic enzyme activity was potently inhibited by PMSF and APMSF. This enzyme exhibited a high specificity for the chymotrypsin substrate S-2586 indicating it's a chymotrypsin-like serine protease. The data we present suggest that the fibrinolytic enzyme derived from the edible and medicinal mushroom Cordyceps militaris has fibrin binding activity, which allows for the local activation of the fibrin degradation pathway.
Kim, Hyoung-Mo;Kim, Jin-Kyong;Shin, Kwan-Suk;Kim, Jun-Hyung;Jung, Jae-Chil;Kim, Tai-Jin
KSBB Journal
/
v.23
no.4
/
pp.317-322
/
2008
The Biochemical Oxygen Demand(BOD) is one of important parameters for the most widely used method of organic pollution in wastewater and wastewater treatment effluent. As the conventional BOD test needs 5-day long incubation period, it is thus incompatible with real time control of wastewater treatment plant. To resolve this problem, in the present study an on-line Dissolved Oxygen(DO) monitoring system was used to observe the transient change of dissolved oxygen concentration in livestock wastewater. The system was composed of BOD sensor, amplifier and computer. It was observed that the concentration of the microorganism in the livestock wastewater was relatively constant during the growth period of initial one hour, which allowed the assumption of the constant Oxygen Uptake Rate(OUR) within one hour of measurement. It was thus concluded that the present scheme provided a protocol for automatic measurement of BOD in livestock wastewater, which can be applicable to optimal control of livestock wastewater treatment plant.
The Journal of Korean Institute of Communications and Information Sciences
/
v.37C
no.9
/
pp.829-840
/
2012
Recently, in network communication environments, it is changing very fast from wired to wireless. The open shortest path firtst (OSPF), one of link state routing protocols, mainly used in wired networks, is the routing method to select optimal traffic path as identifying the link state of neighbor routers. The traditional OSPF cost functions performs with first fixed cost permanently, unless the router link is changed. However, in wireless networks, the performance of links show big difference by other environment factors. The bit error rate (BER), a parameter which can quite affect link state in wireless networks, is not considered in the traditional OSPF cost functions. Only a link bandwidth is considered in the traditional OSPF cost functions. In this paper, we verify the various parameters which can affect link performance, whether it is permissible to use as the parameter of proposed cost functions. To propose new cost functions, we use the effective bandwidth. This bandwidth is calculated by proposed formula using the BER of the network link and link bandwidth. As applied by the proposed triggering condition, the calculated effective bandwidth decrease the unstable of network by generating less link state update messages in wireless networks that frequently changes the link state. Simulation results show that the proposed cost functions significantly outperforms the traditional cost functions in wireless networks in terms of the services of VoIP and data transmission.
Park, Seong-Ok;Yoon, Hyun-A;Aleyas, Abi George;Lee, John-Hwa;Chae, Joon-Seok;Eo, Seong-Kug
IMMUNE NETWORK
/
v.5
no.2
/
pp.89-98
/
2005
Background: DNA vaccination represents an anticipated approach for the control of numerous infectious diseases. Used alone, however, DNA vaccine is weak immunogen inferior to viral vectors. In recent, heterologous prime-boost vaccination leads DNA vaccines to practical reality. Methods: We assessed prime-boost immunization strategies with a DNA vaccine (minigene, $gB_{498-505}$ DNA) and recombinant vaccinia virus $(vvgB_{498-505})$ expressing epitope $gB_{498-505}$ (SSIEF ARL) of CD8+ T cells specific for glycoprotein B (gB) of herpes simplex virus (HSV). Animals were immunized primarily with $gB_{498-505}$ epitope-expressing DNA vaccine/recombinant vaccinia virus and boosted with alternative vaccine type expressing entire Ag. Results: In prime-boost protocols using vvgBw (recombinant vaccinia virus expressing entire Ag) and $vvgB_{498-505}$, CD8+ T cell-mediated immunity was induced maximally at both acute and memory stages if primed with vvgBw and boosted with $vvgB_{498-505}$ as evaluated by CTL activity, intracellular IFN-staining, and MHC class I tetramer staining. Similarly $gB_{498-505}$ DNA prime-gBw DNA (DNA vaccine expressing entire Ag) boost immunization elicited the strongest CD8+ T cell responses in protocols based on DNA vaccine. However, the level of CD8+ T cell-mediated immunity induced with prime-boost vaccination using DNA vaccine expressing epitope or entire Ag was inferior to those based on vvgBw and $vvgB_{498-505}$. Of particular interest CD8+ T cell-mediated immunity was optimally induced when $vvgB_{498-505}$ was used to prime and gB DNA was used as alternative boost. Especially CD7+ T cell responses induced by such protocol was longer lasted than other protocols. Conclusion: These facts direct to search for the effective strategy to induce optimal CD8+ T cell-mediated immunity against cancer and viral infection.
Lee Young-Soo;Shin Dong-Bae;Park Soo-Jin;Kim Jin-Yong;Kim Hee-Sang;Ha Du-Hae
Clinics in Shoulder and Elbow
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v.3
no.2
/
pp.95-101
/
2000
Purpose : The purpose of this study was to evaluate the efficacy of oblique coronal MR images, oblique axial images of neutral, internal rotation and external rotation positions in the diagnosis of SLAP Ⅱ lesion. Materials and Methods: MR arthrography of the glenohumeral joint was evaluated retrospectively in 16 patients(16 shoulders) who underwent arthroscopic surgery(mean age; 38 years old, Male; 13, Female; 3). Oblique coronal fat-suppressed Tl-weighted spin echo images were performed with each shoulder in the neutral position of the arm and oblique axial images were performed in neutral, internal and external rotations of the arm respectively. The preoperative findings of MR were classified as definite tear, possible tear and no tear. Arthroscopic findings were correlated with MR findings of several different position of the arm. Results: Arthroscopic surgery revealed 8 SLAP Ⅱ lesion, 2 SLAP I lesion, and 6 normal superior labrum respectively. The accuracy of diagnosis in the 8 SLAP Ⅱ lesion were high on oblique axial image in external rotation which were interpreted as 8 definite tear, to compare with oblique axial images in neutral position which were interpreted as 4 definite tear, 3 possible tear, 1 no tear. The 6 normal superior labrum lesion were interpreted as no tear in all three position. The 2 SLAP I lesion were interpreted as 1 definite tear, 1 no tear on oblique axial image in neutral position and 1 definite tear, 1 possible tear on oblique axial image in external rotation. Conclusion: This study showed that axial MR images in external rotation of the arm combined with oblique coronal images have proved to be effective to detect SLAP Ⅱ lesion, and should be considered in imaging protocol for MR arthrography of the SLAP Ⅱ lesion.
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