• Restorative Dentistry and Endodontics
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• 제24권1호
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• pp.1-12
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• 1999

Bacterial infection of the pulp results in the development of a periapical lesion with the concomitant resorption of periapical bone. The cytokines are believed to play an important role in this matter. The purpose of this study was to find the relationship among the presence of black pigmented bacteria, the levels of cytokines(TNF-${\alpha}$, -${\beta}$, IL-$1{\beta}$, and TGF-${\beta}1$), and the amount of bone resorption in periapical and pulpal diseases. For the purpose, the patients were grouped into chronic apical pathosis, acute apical pathosis, acute pulpitis, and a healthy control group. Root canal samples were taken from periapical tissue exudates during routine endodontic treatment, and the venous blood was taken from each patients. The samples were processed to measure local and systemic levels of the cytokines using enzyme linked immunosorbent assay(ELISA). Bacterial content of Porphyromonas endodontalis, Porphyromonas gingivalis, and Prevotella nigrescens were measured by indirect immunofluorescence method and the size of the periapical lesions were measured from the radiographs. The following results were obtained: 1. The levels of bone resorptive cytokines(TNF-${\alpha}$, TNF-${\beta}$, and IL-$1{\beta}$) in exudates from acute and chronic apical pathoses were significantly higher than those from acute pulpitis and the normal pulps(p<0.05). 2. IL-$1{\beta}$ were the highest among the bone resorptive cytokines in apical pathoses. However, no statistical difference between acute and chronic lesions were found(p>0.05). 3. The levels of TGF-${\beta}1$ in exudates from acute pulpitis and chronic apical pathoses were significantly higher than those from acute apical pathoses and the normal pulps(p<0.05). However, there were no significant correlations among the levels of bone resorptive cytokines. 4. The levels of TNF-${\beta}$ in serum were significantly higher than those from the exudates while serum TGF-${\beta}1$ concentrations were significantly lower(p<0.05). 5. Exudates from the canals in which the P. nigrescens were detected showed significantly higher levels of IL-$1{\beta}$ than those from the canals without the microorganism(p<0.05). 6. There were no significant correlations among the levels of the cytokines, the amount of bone destruction, and the presence of acute and chronic symptoms(p>0.05).

• 대한한의학회지
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• 제26권2호
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• pp.217-230
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• 2005

Objectives: Chungpaesagan-tang (CPSGT), which is frequently used for treating patients of cerebrovascular disease, has not been reported by clinical doctors concerning the effect of neuronal aptosis caused by brain ischemia. To study the effect of CPSGT on focal cerebral ischemia in normal and diabetic rats and SHR, focal cerebral ischemia was induced by transient MCAO, and after onset CPSGT was administrated. Methods: Rats (Sprague-Dawley) were divided into four groups: sham-operated group, MCA-occluded group, CPSGT­administrated group after MCA occlusion, and normal group. The MCA was occluded by intraluminal method. CPSGT was administrated orally twice (l and 4 hours) after middle cerebral artery occlusion. All groups were sacrificed at 24 hours after the surgery. The brain tissue Was stained with $2\%$ triphenyl tetrazolium chloride (TTC) or $1\%$ cresyl violet solution, to examine effect of CPSGT on ischemic brain tissue. The blood samples were obtained from the heart.~. Tumor necrosis $factor-\alpha$ level and interleukin-6 level of serum was measured from sera using enzyme-linked immunoabsorbent assay (ELISA). Then changes of immunohistochemical expression of $TNF-\alpha$ in ischemic damaged areas were observed. Results: In NC+MCAO+CP and DM+MCAO+CP, CPSGT significantly (p<0.01) decreased the number of neuron cells compared to the control group. CPSGT markedly reduced (p<0.01) the infarct size of the forebrain in distance from the interaural line on cerebral ischemia in diabetic rats. CPSGT significantly reduced the $TNF-\alpha$ expression in penumbra region of damaged hemisphere in diabetic rats. Conclusions: CPSGT had a protective effect on cerebral ischemia in SD rats, especially in diabetic rats compared with normal SD rats.

• 대한한의학회지
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• 제27권4호
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• pp.1-11
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• 2006

Objectives : We investigated the effect of Dohongsamul-tang (DHSMT) on the production of various cytokines in lipopolysaccaride (LPS) stimulated peripheral mononuclear cells (PBMCs) from CI patients. Methods: Cell viability was determined using MTT assay. ELISA was carried out for investigating $TNF-{\alpha}$, $IL-1{\beta}$, IL-6, IL-8, IL-4, and $TGF-{\beta}$ 1 Results : The amount of tumor necrosis factor (TNF)-${\alpha}$, interleukin (IL)-$1{\beta}$, IL-6, IL-8, IL-4, and transforming growth factor(TGF)-${\beta}$ 1 in PBMC culture supernatant significantly increased in the LPS treated cells compared to unstimulated cells. We show that DHSMT inhibited the production of TNF-${\alpha}$, IL-1${\beta}$, IL-6, IL-8, and IL-4 induced by LPS in a dose dependent manner. The maximal inhibition rate of the TNF-${\alpha}$, IL-1${\beta}$, IL-6, IL-8, and IL-4 production by pretreatment of DHSMT (1.0mg/ml) was 38.52 ${\pm}$ 2.5% (P < 0.01), 44.02 ${\pm}$ 3.5% (P < 0.05), 45.32 ${\pm}$ 2.3% (P < 0.01), 42.30 ${\pm}$ 3.1% (P < 0.05), and 49.70　${\pm}$　3.1%(P ＆lt; 0.05), respectively. On the other hand, DMSMT significantly increased the LPS-induced TGF-${\beta}$ 1 production(P<0.05). Conclusions : Taken together. these results suggest that DHSMT might have regulatory effects on cytokine production, which might explain its beneficial effect in the treatment of CI.

• 한국잠사곤충학회지
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• 제53권1호
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• pp.55-60
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• 2015

본 연구의 목적은 누에형질전환 기술을 이용하여 누에체액에서 melittin 항균펩타이드를 생산하는 것으로서, 본 실험에서는 누에유래의 액틴3 프로모터를 이용하여 melittin 항균펩타이드를 발현시켰다. 누에형질전환체 선발을 위해서는 3xP3 promoter와 EGFP 유전자를 이용하여 선발하였고, 300개의 누에알에 microinjection 하여 F1 세대에서 11 bloods의 누에형질전환체를 선발하였다. 선발된 누에형질전환체는 초기배 단계의 눈과 신경조직, 유충과 번데기 그리고 성충의 눈에서 EGFP 형광단백질이 발현되는 것을 확인할 수 있었다. 또한 G2 세대 누에형질전환체를 5령 5일 유충까지 사육 후, 체액을 채취한 후 전처리 하였다. 이 시료를 항균활성검정을 하였고, 총 10마리의 누에를 선발할 수 있었다. 이렇게 선발 된 누에는 서로 교배를 통해서 계대사육을 하였다. 이러한 과정으로 선발된 G3세대 누에형질전환체를 이용하여 앞의 과정과 동일한 방법으로 항균할성을 검정하였다. 그 결과 대조군으로 사용된 시그마사의 melittin(0.016 mg/ml)과 거의 동일한 항균활성을 나타내었다. 이상의 결과에서 melittin 항균펩타이드를 생산하는 누에형질전환체가 성공적으로 제작되었음을 확인할 수 있었다.

• 한국수정란이식학회지
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• 제28권2호
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• pp.157-162
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• 2013

Methoxychlor (MXC) was developed to be a replacement for the banned pesticide DDT. HPTE [2,2-bis (p-hydroxyphenyl)-1,1,1-trichloroethane], which is an in vivo metabolite of MXC, has strong oestrogenic and anti-androgenic effects. MXC and HPTE are thought to produce potentially adverse effects by acting through oestrogen and androgen receptors. Of the two, HPTE binds to sex-steroid receptors with greater affinity, and it inhibits testosterone biosynthesis in Leydig cells by inhibiting cholesterol side-chain cleavage enzyme activity and cholesterol utilisation. In a previous study, MXC was shown to induce Leydig cell apoptosis by decreasing testosterone concentrations. I focused on the effects of MXC on male mice that resulted from interactions with sex-steroid hormone receptors. Sex-steroid hormones affect other organs including the kidney and liver. Accordingly, I hypothesised that MXC can act through sex-steroid receptors to produce adverse effects on the testis, kidney and liver, and I designed our experiments to confirm the different effects of MXC exposure on the male reproductive system, kidney and liver. In these experiments, I used pre-pubescent ICR mice; the puberty period in ICR mice is from postnatal day (PND) 45 to PND60. I treated the experimental group with 0, 100, 200, 400 mg MXC/kg b.w. delivered by an intra-peritoneal injection with sesame oil used as vehicle for 4 weeks. At the end of the experiment, the mice were sacrificed under anaesthesia. The testes and accessory reproductive organs were collected, weighed and prepared for histological investigation. I performed a chemiluminescence immune assay to observe the serum levels of testosterone, LH and FSH. Blood biochemical determination was also performed to check for other effects. There were no significant differences in our histological observations or relative organ weights. Serum testosterone levels were decreased in a dose-dependent manner; a greater dose resulted in the production of less testosterone. Compared to the control group, testosterone concentrations differed in the 200 and 400 mg/kg dosage groups. In conclusion, I observed markedly negative effects of MXC exposure on testosterone concentrations in pre-pubescent male mice. From our biochemical determinations, I observed some changes that indicate renal and hepatic failure. Together, these data suggest that MXC produces adverse effects on the reproductive system, kidney and liver.

• 대한임상검사과학회지
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• 제43권4호
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• pp.171-178
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• 2011

Glycated hemoglobin ($HbA_{1c}$) is a most preferably used baseline of diabetes, implicating average blood glucose levels over a 2-3 month period of time. Recently the American Diabetes Association has recommended the $HbA_{1c}$ assay as one of the criteria for diabetes. Although some studies provide data with "estimated average glucose", by converting the $HbA_{1c}$ results from simple linear regression, the results are not applicable to whole diabetes. We compared the relationship between $HbA_{1c}$ and estimated average glucose by anemia degree of diabetic patients in Korea. The data from the 2008~2009 Korean National Health and Nutrition Examination Survey were used. Analysis was done for 1,257 diabetes subjects with $HbA_{1c}$ results. The distribution of subjects was 34.1% in 60's, 25.9% in 70's, 21,6% in 50's, showing that there was more than 80% in over 50's. To take a close look of the differences depending on the anemic degree, we applied WHO criteria (hemoglobin<13.0 in men and hemoglobin<12.0 in women) and divided anemia degree. The regression equation for A1c and estimated average glucose was $eAG_{mg/dL}=24.3{\times}A1c-32.0$ ($R^2=0.54$, p<0.001) in all subjects, $eAG_{mg/dL}=33.1{\times}A1c-96.1$ ($R^2=0.52$, p<0.001) in slight anemia ($11.0{\leq}$Hb<13.0 in men, $10.0{\leq}$Hb<12.0 in women), and $eAG_{mg/dL}=13.5{\times}A1c+34.9$ ($R^2=0.12$, p =0.075) in moderate anemia (Hb<11.0 in men, Hb<10.0 in women). The regression was not significant in moderate anemia. The relationship between HbA1c and eAG was lower correlation than ADAG study, and eAG showed lower value in all ranges among $HbA_{1c}$ 5~13%. Such as a korea where, there are many diabetic patients among the old aged and higher prevalence rate of anemia, we should be extra careful when we reflect eAG using $HbA_{1c}$ and need to establish criteria which can be applicable to koreans.

• Journal of Microbiology and Biotechnology
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• 제17권10호
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• pp.1670-1674
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• 2007

Duffy binding protein (DBP) plays a critical role in Plasmodium vivax invasion of human red blood cells. We previously reported a single-chain antibody fragment (scFv) that was specific to P. vivax DBP (PvDBP). However, the stabilization and the half-life of scFvs have not been studied. Here, we investigated the effect of PEGylated scFvs on their biological activity and stability in vitro. SDS-PAGE analysis showed that three clones (SFDBII-12, -58, and -92) were formed as monomers (about 70 kDa) with PEGylation. Clone SFDBII-58 gave the highest yield of PEGylated scFv. Binding analysis using BIAcore between DBP and scFv showed that both SFDBII-12 and -58 were decreased approximately by two folds at the level of binding affinity to DBP after PEGylation. However, the SFDBII-92 clone still showed a relatively high level of binding affinity ($K_D=1.02{\times}10^{-7}\;M$). Binding inhibition assay showed that PEGylated scFv was still able to competitively bind the PvDBP and playa critical role in inhibiting the interactions between PvDBP protein expressed on the surface of Cos-7 cells and Duffy receptor on the surface of erythrocytes. When both scFvs and their PEGylated counterparts were exposed to trypsin, scFv was completely degraded only after 24 h, whereas 35% of PEGylated scFvs remained intact, maintaining their stability against the proteolytic attack of trypsin until 72 h. Taken together, these results suggest that the PEGylated scFvs retain their stability against proteolytic enzymes in vivo, with no significant loss in their binding affinity to target antigen, DBP.

• Journal of Microbiology and Biotechnology
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• 제23권5호
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• pp.637-643
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• 2013

We have demonstrated that 1-deoxynojirimycin (DNJ) isolated from Bacillus subtilis MORI could enhance the levels of adiponectin and its receptors in differentiated 3T3-L1 adipocytes, which has been shown to be effective in lowering blood glucose levels and enhancing insulin sensitivity. DNJ was not toxic to differentiated 3T3-L1 adipocytes for up to a concentration of $5{\mu}M$. In terms of expression levels of adiponectin and its receptors (AdipoR1 and AdipoR2), DNJ in concentrations as low as $0.5{\mu}M$ elevated both mRNA and protein levels of adiponectin and transcript levels of AdipoR1 and AdipoR2. In addition, DNJ increased phosphorylation of 5' adenosine monophosphate-activated protein kinase (AMPK) in a statistically significant manner. Finally, treatment with DNJ resulted in increased mRNA expression of glucose transporter 4 (GLUT4), which encodes for a glucose transporter, along with a significant increase in glucose uptake into the adipocytes based on results of a 2-deoxy-D-[$^3H$] glucose uptake assay. Our findings indicate that DNJ may greatly facilitate glucose uptake into adipose tissues by increasing the action of adiponectin via its up-regulated expression as well as its receptor genes. In addition, the glucose-lowering effects of DNJ may be achieved by an increased abundance of GLUT4 protein in the plasma membrane, as a consequence of the increased transcript levels of the GLUT4 gene and the activation of AMPK.

• Clinical and Experimental Reproductive Medicine
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• 제29권3호
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• pp.215-222
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• 2002

Objective : Previous studies have suggested that hyperhomocysteinemia and methylenetetrahydrofolate reductase (MTHFR C677T) mutations are associated with increased risk of recurrent spontaneous abortion (RSA). Recently, a second site polymorphism in MTHFR, 1298A-->C, which changes a glutamic acid into an alanine residue, was shown to be associated with a decreased enzyme activity. We tested whether the variant alleles of MTHFR C677T and A1298C are risk factor (biomarker) for RSA. Materials and Methods: We analyzed DNA from a case-control study in the Korean DNA was extracted from blood samples of 118 patients with RSA and 123 healthy fertile patients as the controls. MTHFR variant alleles were determined by a PCR-restriction fragment length polymorphism assay. Results: We found no evidence for an association between 677TT genotype and risk of RSA (OR=1.95, 95% CI=$0.84{\sim}4.50$, p=0.12). However, the MTHFR 1298AC (OR=0.36, 95% CI=$0.20{\sim}0.63$, p=0.0004) and 1298AC+CC (OR=0.35, 95% CI=$0.20{\sim}0.61$, p=0.0002) genotypes were lower among 118 RSA cases compared with 123 controls, conferring a 2.8-fold decrease in risk of RSA, respectively. Moreover, the combined genotypes of MTHFR 677CC/1298AC (OR=0.30, 95% CI=$0.10{\sim}0.88$, p=0.029) and 677CT/1298AC (OR=0.77, 95% CI=$0.60{\sim}0.99$, p=0.043) also showed significantly lower risk than those with MTHFR 677CC/1298AA type. Conclusion: MTHFR 1298AC, MTHFR 677CC/1298AC and 677CT/1298AC genotypes may represent genetic markers for the protection of RSA at least in Korean women.

• Asian-Australasian Journal of Animal Sciences
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• 제12권3호
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• pp.395-399
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• 1999

Three experiments were conducted to determine the effects of conditioning a complex (20% whey, 10% lactose, 4% plasma protein, 4% wheat gluten and 2% blood meal) diet on growth performance of weanling pigs. In Exp. 1,180 pigs (average initial BW of 6.4 kg) were fed the experimental diet (1.7% lysine) during a 7-d growth assay. Treatments were a meal control (M), standard (ST), and expander (EX) conditioned mash or pellets. Rate and efficiency of gain were decreased by 39% and 21% (p<0.005) respectively, for pigs fed EX diets compared to those fed the ST diet. In Exp. 2,196 pigs (average initial BW of 6.5 kg) were used to determine the effects of EX operating conditions on nutritional value of a pelleted complex diet. When steam conditioning temperature (prior to expanding) was $54^{\circ}C$, increasing cone pressure of the EX from 0 to 7 to 14 kg/cm2 resulted in linear decreases in rate of gain of weaned pigs (p<0.006), suggesting heat damage of the diet. Increasing conditioning temperature (i.e., adding steam) of the diets from 46 to 54 to $63^{\circ}C$ (cone pressure at $12kg/cm^2$) resulted in improved rate of gain (p<0.04) of the pigs. However, none of the pigs fed expanded diets compared favorably to the pigs fed the conditioned $(54^{\circ}C)$ pellets processed with no cone pressure. In Exp. 3,168 pigs (average initial BW of 6.6 kg) were used to determine the effects of expanding the various components of the diet. Treatments were M and ST pellets as controls, EX-corn, EX-corn soybean-meal, EX corn-soybean meal-oil, and EX-complete diet. Efficiency of gain was increased by 13% with EX portions of the diet compared to the mash control, but there was a marked decrease in performance when the complete diet was expanded (p<0.001). Expanded corn-soybean meal-oil supported the greatest ADG with a 19% increase compared to the average of the EX corn and EX corn-soybean meal treatments (p<0.005). In conclusion, our results suggest no benefit from expanding complete phase-I diets.