Hepatitis A virus (HAV) infection leads to acute liver failure and death through the intake of contaminated food. The polymerase chain reaction (PCR) has been used to detect HAV in food samples. HAV detection takes a long time, however, due to the virus concentration step required before PCR assay. In this study, a rapid method of detecting the HAVs present in lettuce using immunomagnetic separation combined with quantum dots (IMS-QDs) assay was developed. The detection limit of IMS-QDs for HAV was 10 $TCID_{50}/mL$, similar to the result that was obtained using RT-PCR combined with PEG or IMS. The application of IMS-QDs assay completed the viral detection within one hour, but this was not possible using PEG combined with RT-PCR. In conclusion, IMS-QDs assay is a rapid and efficient method for detecting HAV at a low concentration in agricultural products.
Sudden idiopathic sensorineural hearing loss is a disease that develops within several hours to several days. Its etiology has not yet been verified, but the disturbance of the circulation of blood in the inner ear, inner-ear hydrops, and viral infection are considered possible causes of the disease. This study was conducted to evaluate the effect of Panax ginseng extract, which is known to have a vasodilatory effect, on sudden sensorineural hearing loss. Sixty-nine patients suffering from sudden sensorineural hearing loss were admitted to Korea University Anam Hospital from March to December 2008. They were divided into the experimental (30 ears) and control (39 ears) groups. Ginseng extract (2700 mg/day, 4 weeks) was added to the therapeutic regimen in the experimental group. The effect of ginseng extract therapy was analyzed according to the factors relating to the prognosis. A considerable hearing improvement was documented in both groups (32.2 dB in the experimental group and 25.8 dB in the control group). However, there was little beneficial effect of ginseng extract on additional hearing improvement compared with control. The total recovery rate of the experimental group (80.0%) was better than that of the control group (58.9%), and the experimental group's high-tone hearing gain at 3 kHz (29.7 dB) was better than that of the control group (21.7 dB). The results of the study suggest that the effects of ginseng therapy tend to be superior to those of the conventional therapy, but the difference between the two is not statistically significant. The hearing gains tend to be in the higher frequencies and may be due to the promotion of cellular differentiation from the supporting cells.
Acute pancreatitis(AP) in children is not common but can be associated with severe morbidity rates and its diagnosis is often delayed. Thus, reported mortality rates range from 0 to 78%. We have treated 26 patients with AP from 5 to 17 years of age over the past 17 years. We are intended to assess the relevance of the prognostic criteria used to assess severity of adult AP and to review the etiology, clilical presentation, diagnosis, and management of AP in children. The authors retrospectively reviewed 26 children with AP managed in Kyung Hee University Hospital from 1978 to 1995. Among 26 patients with AP, male were 12, and female were 14. And the mean age of patients was 11.8 years. In 9(34.6%), no definitive cause was identified. Common causes of AP were trauma(23.1%) and biliary tract disease(23.1%). Other etiologies were viral infection(15.4%) and post ERCP(3.8%). The presenting features were abdominal pain(92.3%), vomiting(61.5%), fever(19.2%), submandibular pain(11.5%), and abdominal mass(7.6%). Back pain was rare(3.8%). Abdominal ultrasonographic findings were abnormal in 10 of 16 patients(62.5%) and abdominal CT findings were abnormal for 9 of 9 patients(100%). Seventeen patients(65.3%) were managed conservatively, and nine patients(34.6%) required surgical treatment. There was no mortality. To evaluate the severity of disease, we used the Imrie prognostic criteria used to assess the severity in adult AP. The number of positive criteria was correlated to the duration of hospitalization(r2=0.91) but statistically insignificant(p>0.05). But, the number of positive criteria was correlated to the operative incidence(r2=0.93) and statistically significant(p<0.05). The common causes of AP in children were unknown origin(34.6%), trauma(23.1%), and biliary tract disease(23.1%). Ultrasonography and computed tomography were useful imaging tools of AP in children. The Imrie criteria used to evaluate the severity in adult AP were suspected to be valuable to assess the severity of AP in children.
Kim, Seon-Ju;Kim, Hye-Kyung;Han, Young-Woo;Aleyas, Abi G.;George, Junu A.;Yoon, Hyun-A;Yoo, Dong-Jin;Kim, Koan-Hoi;Eo, Seong-Kug
Journal of Microbiology and Biotechnology
/
v.18
no.7
/
pp.1326-1334
/
2008
The prime-boost vaccination with DNA vaccine and recombinant viral vector has emerged as an effective prophylactic strategy to control infectious diseases. Here, we compared the protective immunities induced by multiple alternating immunizations with DNA vaccine (pCIgB) and replication-incompetent adenovirus (Ad-gB) expressing glycoprotein gB of pseudorabies virus (PrV). The platform of pCIgB-prime and Ad-gB-boost induced the most effective immune responses and provided protection against virulent PrV infection. However, priming with pCIgB prior to vaccinating animals by the DNA vaccine-prime and Ad-boost protocol provided neither effective immune responses nor protection against PrV. Similarly, boosting with Ad-gB following immunization with DNA vaccine-prime and Ad-boost showed no significant responses. Moreover, whereas the administration of Ad-gB for primary immunization induced Th2-type-biased immunity, priming with pCIgB induced Th1-type-biased immunity, as judged by the production of PrV-specific IgG isotypes and cytokine IFN-$\gamma$. These results indicate that the order and injection frequency of vaccine vehicles used for heterologous prime-boost vaccination affect the magnitude and nature of the immunity. Therefore, our demonstration implies that the prime-boost protocol should be carefully considered and selected to induce the desired immune responses.
Purpose: Aplasia Cutis Congenita (ACC) is a rare disease characterized by the focal defect of the skin at birth, frequently involving scalp, but it may affect any region of the body. There are no etiology known but some conditions such as intrauterine vascular ischemia, amniotic adherences and viral infections are associated. The ideal treatment for the ACC is not known. Superficial and relatively small sized defects (< $3{\times}5\;cm$) may heal spontaneously and large defects related with risks of infection and bleeding may require aggressive surgical treatment. Hyalomatrix$^{(R)}$ is a bilayer of an esterified hyaluronan scaffold beneath a silicone membrane. It has been used as a temporary dermal substitute to cover deep thickness skin defect and has physiological functions derive from the structural role in extracellular matrix and interaction with cell surface receptor. This material has been used for the wound bed pre-treatment for skin graft to follow and especially in uncooperative patient, like a newborn, this could be a efficient and aseptic way of promoting granulation without daily irritative wound care. For this reason, using Hyalomatrix$^{(R)}$ for the treatment of ACC was preferred in this paper. Methods: We report a case of a newborn with ACC of the vertex scalp and non-ossified partial skull defect. The large sized skin and skull defect ($6{\times}6\;cm$) was found with intact dura mater. No other complications such as bleeding or abnormal neurologic sign were accompanied. Escharectomy was performed and Hyalomatrix$^{(R)}$ was applied for the protection and the induction of acute wound healing for 3 months before the split-thickness skin graft. During the 3 months period, the dressing was renewed in aseptic technique for every 3 weeks. The skin graft was achieved on the healthy granulation bed. Results: The operative procedure was uneventful without necessity of blood transfusion. Postoperative physical examination revealed no additional abnormalities. Regular wound management was performed in out-patient clinic and the grafted skin was taken completely. No other problems developed during follow-up. Conclusion: Hyalomatrix$^{(R)}$ provides protective and favorable environment for wound healing. The combination of the use of Hyalomatrix$^{(R)}$ and the skin graft will be a good alternative for the ACC patients with relatively large defect on vertex.
While bacteriophage P2-P4 system is very useful experimental tool for the study of viral capsid assembly, there is no useful plasmid vector for the DNA manipulation in bacteriophage P2-P4 system. In this study, a new vector plasmid, P4 ash8 (sid71) kmr, was constructed by swapping the non-essential region of P4 DNA for kanamycin resistance(kmr) gene cassette of plasmid pUC4-K. P4 ash8 sid71 was starting material for the construction, since it tends to be maintained as a plasmid in the absence of the helper phage. The total size of this chimera was designed to be packaged into P4 or P2 size heads with induction by P2 infection. The conversion of plasmid P4 ash8 (sid71) kmr to bacteriophage was proved by burst size determination experiment and CsCl buoyant equilibrium density gradient experiment. Integrase destructed P4 derivative, P4 ash8 sid71 kmr intS, was able to be constructed easily by in vitro DNA manipulation of P4 ash8 sid71 kmr. The plasmid stability experiment with P4 ash8 sid71 kmr if/tS showed that the integrase of P4 affects the stable maintenance of plasmid P4 state.
Tobacco mosaic virus(TMV) coat protein cDNA was transformed to Nicotiana tabacum cv. NC82 and the transgenic tobacco plants resistant to TMV infection were isolated in the next generation. The expression of TMV coat protein cDNA and genetic stability of the fifth generation of TMV resistant transgenic tobacco plants at the higher temperature were investigated. The TMV coat protein cDNA was amplified by genomic PCR in all the TMV resistant transgenic tobacco plants. The TMV coat protein expressed in the transgenic tobacco plants was detected at very low level by immunoblot hybridization. Even in tansgenic plants that showed the viral symptom only on very late sucker growth (delay type plants), the coat protein expression in the suckers was much less than that of susceptible tobacco infected with TMV. The TMV coat protein expressed in the transgenic tobacco plants was below 0.01% of total protein. Transcription and expression of the coat protein cDNA in delay type plants were observbed at high temperature (38$^{\circ}C$), and TMV replication was suppressed at both 28$^{\circ}C$ and 38$^{\circ}C$. This indicates that unlike the resistance conferred by 'N' gene. TMV resistance of transgenic tobacco plant won't break down at high temperature.
Cucumber mosaic virus (CMV) leads to a cause of poor crop productivity and quality. To solve this problem, we attempted to develop a virus-resistance tobacco plants by using viral coat protein (CP) gene. Transgenic tobacco plants expressing CMV CP gene were analysed by the resistance upon CMV infection. The virus-resistance was measured in $\textrm{T}_{1}$, generation by the inhibition of plant growth and the expression of the mosaic symptoms infected with CMV. The transgenic lines were divided into four groups: highly resistant, resistant, moderate and susceptible based on their growth and symptom severity. Out of 39 transgenic lines, 16 lines showed significant virus-resistance. And of resistant lines, 2 lines were designated highly resistant based on the facts that they achieved similar plant height to that of non-infected tobacco plants and showed lower disease symptom than that of other lines. The steady state level of CP RNA and coat protein level were measured by northern blot and immunoblot analysis. The CP RNA was highly accumulated in most resistant and moderate lines but barely detected in susceptible lines. The coat protein was detected in most lines regardless of their resistance to CMV. from this result, virus-resistance appeared to correlate more with CP RNA level than the level of coat protein. However, in two highly resistant lines, CP RNA level was unexpectedly low. This unexpected phenomenon need to be further investigated.
Purpose : Yellow fever, a mosquito-borne viral hemorrhagic fever, is one of the most lethal diseases. Recently there have been an increasing number of Korean children who have travelled to yellow fever endemic zones and were administered yellow fever vaccine (YFV). Therefore, we carried out this study to provide child travelers with safety information of YFV. Methods : This study was conducted at the International Clinic of National Medical Center in Seoul between April 2007 and June 2008 for the evaluation of adverse events of YFV. One hundred twenty-five children received YFV (17-DD) and were prospectively monitored for adverse events through telephone interviews on day 3, 6, 9, 16, 23 and 30 after vaccination. Results : Adverse events were observed in 31 (24.8%) of 125 child travelers who received the YFV. The mean age was 12.5${\pm}$5.0 years. Sixty-six of the child travelers (52.8%) were males. The common adverse events were pain in 11 (8.8%), swelling in 8 (6.4%) and redness in 7 children (5.6%) at the injection site. The systemic adverse events included mild fever in 5 (4.0%), headache in 5 (4.0%), cough in 4 (3.2%), abdominal pain in 3 (2.4%), and vomiting in 2 children (1.6%). Most of the adverse events were detected within 7 days of administration and there were no differences in adverse events by gender or age. All travelers who had complained of symptoms improved spontaneously or following symptomatic treatment. Conclusion : This study showed that YFV is well-tolerated and there were no reports of severe adverse events. Studies are ongoing to clarify the cause and risk factors for rare adverse events.
Myeloid-derived suppressor cells (MDSCs) play an important role in impairing the function of T cells. We characterized MDSCs in two chronic hepatitis C (CHC) cohorts: a cross-sectional group that included 61 treatment-naive patients with CHC, 14 rapid virologic response (RVR) cases and 22 early virologic response (EVR) cases; and a longitudinal group of 13 cases of RVR and 10 cases of EVR after pegylated-interferon-${\alpha}$/ribavirin treatment for genotype 1b HCV infection. Liver samples from 32 CHC patients and six healthy controls were subjected to immunohistochemical analysis. MDSCs frequency in treatment-naive CHC was significantly higher than in RVR, EVR, or healthy subjects and was positively correlated with HCV RNA. Patients infected with HCV genotype 2a had a significantly higher frequency of MDSCs than those infected with genotype 1b. Decreased T cell receptor (TCR) ${\zeta}$ expression on $CD8^+$ T cells was significantly associated with an increased frequency of MDSCs in treatment-naive CHC patients and was restored by L-arginine treatment in vitro. Increased numbers of liver arginase-$1^+$ cells were closely associated with the histological activity index in CHC. The TCR ${\zeta}$ chain was significantly downregulated on hepatic $CD8^+$ T cells in CHC. During antiviral follow up, MDSCs frequency in peripheral blood mononuclear cells was directly correlated with the HCV RNA load in the plasma and inversely correlated with TCR ${\zeta}$ chain expression in $CD8^+$ T cells in both RVR and EVR cases. Notably, the RVR group had a higher frequency of MDSCs at baseline than the EVR group. Collectively, this study provides evidence that MDSCs might be associated with HCV persistence and downregulation of CD8 ${\zeta}$ chain expression.
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