• Title/Summary/Keyword: Ligation

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The New LM-PCR/Shifter Method for the Genotyping of Microorganisms Based on the Use of a Class IIS Restriction Enzyme and Ligation-Mediated PCR

  • Krawczyk, Beata;Leibner-Ciszak, Justyna;Stojowska, Karolina;Kur, Jozef
    • Journal of Microbiology and Biotechnology
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    • v.21 no.12
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    • pp.1336-1344
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    • 2011
  • This study details and examines a novel ligation-mediated polymerase chain reaction (LM-PCR) method. Named the LM-PCR/Shifter, it relies on the use of a Class IIS restriction enzyme giving restriction fragments with different 4-base, 5' overhangs, this being the Shifter, and the ligation of appropriate oligonucleotide adapters. A sequence of 4-base, 5' overhangs of the adapter and a 4-base sequence of the 3' end of the primer(s) determine a subset of the genomic restriction fragments, which are amplified by PCR. The method permits the differentiation of bacterial species strains on the basis of the different DNA band patterns obtained after electrophoresis in polyacrylamide gels stained with ethidium bromide and visualized in UV light. The usefulness of the LM-PCR/Shifter method for genotyping is analyzed by a comparison with the restriction endonuclease analysis of chromosomal DNA by the pulsed-field gel electrophoresis (REA-PFGE) and PCR melting profile (PCR MP) methods for isolates of clinical origin. The clustering of the LM-PCR/Shifter fingerprinting data matched those of the REA-PFGE and PCR MP methods. We found that the LM-PCR/Shifter is rapid, and offers good discriminatory power and excellent reproducibility, making it a method that may be effectively applied in epidemiological studies.

Effects of Ethyl Pyruvate on Allodynia, TNF-${\alpha}$ Expression, and Apoptosis in the Dorsal Root Ganglion after Spinal Nerve Ligation Injury

  • Choi, Dae-Kee;Leem, Jeong-Gill;Shin, Jin-Woo;Suh, Jeong-Hun
    • The Korean Journal of Pain
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    • v.25 no.4
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    • pp.213-220
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    • 2012
  • Background: It has been demonstrated that the expression of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) and apoptotic cell death in the dorsal root ganglion (DRG) following spinal nerve constriction injury play a role in the initiation and continuation of hyperalgesia and allodynia. The present study was designed to investigate the effects of ethyl pyruvate (EP) on mechanical and cold allodynia, TNF-${\alpha}$ expression, and apoptosis in DRG after spinal nerve ligation injury. Methods: Rats were divided into 3 groups: control, pre-EP, and post-EP. EP (50 mg/kg) was intraperitoneally injected 30 minutes before (pre-EP) or after (post-EP) surgery. Behavioral tests to determine mechanical and cold allodynia were conducted before surgery and 4 and 7 days after surgery. Seven days after surgery, TNF-${\alpha}$ protein levels in DRG were evaluated by enzyme-linked immunosorbent assay, and DRG apoptosis was determined by immunohistochemical detection of activated caspase-3. Results: Treatment with EP significantly reduced mechanical and cold allodynia following spinal nerve ligation injury. TNF-${\alpha}$ protein levels in the pre-EP ($4.7{\pm}1.2$ pg/200 ${\mu}g$; P < 0.001) and post-EP ($6.4{\pm}1.8$ pg/200 ${\mu}g$; P < 0.001) groups were 2-3 times lower than the control group ($14.4{\pm}1.2$ pg/200 ${\mu}g$). The percentages of neurons and satellite cells that co-localized with caspase-3 were also significantly lower in the pre-EP and post-EP groups than the control group. Conclusions: These results demonstrate that EP has a strong anti-allodynic effect that acts through the inhibition of TNF-${\alpha}$ expression and apoptosis in DRG after spinal nerve ligation injury.

미숙아와 신생아의 동맥과 개존증에 대한 수술요법(소개흉 결찰술과 흉강경 보조하의 clipping과의 비교)

  • 장지원;한재진;원용순;원태희;안재호;박영식;최수승
    • Journal of Chest Surgery
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    • v.33 no.1
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    • pp.26-31
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    • 2000
  • Background: This study was aimed at analyzing the preoperative conditions post-operative results indication and methods of surgical closure of patent ducturs arterio년 in prematures low birth weight infants and neonates. Patients and Methods: We retro-spectively studied two groups of patients (prematures group and neonates group) who underwent surgical closure of the patent ductus arteriosus between March 1995 and June 1998. Results: The premature group consisted of 9 patients(3 males and 6 females) Their mean gestational period was 30.7 weeks(ranging from 26 weeks to 33 weeks) mean age 27.8 days(11 days to 55 days) and mean body weight 1.56 kg. Prominent preoperative symptoms were dependency on mechanical ventilation generalized edema and hepatomegaly. We performed PDA ligation via thoracotomy in all premature patients. The neonate group consisted of 16 patients and their mean body weight was 3.75 kg. Major symptoms of this group was tachypnea and intercostal retraction resistant to medical treatment. We performed video-assisted PDA clipping to them all. There were no postoperative complications or operation-related mortality in both groups. Comparing the ratio of size of PDA(mm)/body weight(kg) the ratio of premature group (ligation through thoracotomy) was higher than that of neonate group ( video-assisted clipping) that is 3,89:1.21(p=0.03) Conclusion : We conclude that the surgical closure of PDA can be a safe method of treatment for prematures low birth weight infants and neonates with compromised general conditions. Choice of surgical technique depends on the surgeon's preference but there was a tend-ency to choose the ligation method through thoracotomy for patients with small body weight and large PDA.

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The Effects of Transcutaneous Electrical Nerve Stimulation (TENS) on the Neuropathic Pain in Peripheral Nerve Injury (말초신경 손상에 의한 신경병증성 통증에 TENS가 미치는 효과)

  • Lee, Soon-Hyun;Song, Chang-Ho
    • Journal of the Korean Society of Physical Medicine
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    • v.8 no.1
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    • pp.79-89
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    • 2013
  • PURPOSE: To identify the effects of single trial transcutaneous electrical nerve stimulation (TENS) application on chronic neuropathic pain and the repeated TENS application to development of neuropathic pain following peripheral nerve injury. METHODS: First, 20 rats were given the median nerve ligation to induce chronic neuropathic pain. After the ligation, neuropathic pain was assessed by measuring the forepaws withdrawal threshold to von Frey filaments for 3 weeks. Afterward, rats were randomly divided into TENS group and placebo-TENS group. TENS (frequency 100Hz, pulse width $200{\mu}s$) was applied to the forearm for 20 minutes. Second, 34 rats were randomly allocated into two group after median nerve ligation: TENS group and placebo-TENS group. Both interventions were applied to the forearm for 20 minutes from 1 day to 3 weeks after injury. Neuropathic pain to mechanical was measured on each rat for 3 weeks. RESULTS: Exeprimental rats showed a clear neuropathic pain-like behaviors, such as reduced forepaw withdrawal threshold to mechanical stimulation for 3 weeks, after median nerve ligation. And, TENS decreased effectively the chronic neuropathic pain originated from median nerve injury. TENS also diminished the development of neuropathic pain after nerve injury. CONCLUSION: Our animal model studying for neuropathic pain following median nerve injury may be useful to investigate peripheral neuropathic pain in human. Also, TENS may be used to mediate chronic neuropathic pain and to prevent the development of neuropathic pain following median nerve injury.

Up-Regulation of Interleukin-4 Receptor Expression by Interleukin-4 and CD40 Ligation via Tyrosine Kinase-Dependent Pathway

  • Kim, Hyun-Il;So, Eui-Young;Yoon, Suk-Ran;Han, Mi-Young;Lee, Choong-Eun
    • BMB Reports
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    • v.31 no.1
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    • pp.83-88
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    • 1998
  • Recently a B cell surface molecule, CD40, has emerged as a receptor mediating a co-stimulatory signal for B cell proliferation and differentiation. To investigate the mechanism of synergy between interleukin-4 (IL-4) and CD40 ligation in B cell activation, we have examined the effect of CE40 cross-linking on the IL-4 receptor expression in human B cells using anti-CE40 antibody. We observed that IL-4 and anti-CD40 both induce IL-4 receptor gene expression with a rapid kinetics resulting in a noticeable accumulation of IL-4 receptor mRNA within 4 h. While IL-4 caused a dose-dependent induction of surface IL-4 receptor expression, the inclusion of anti-CD40 in the IL-4-treated culture, further up-regulated the IL-4-induced IL-4 receptor expression as analyzed by flow cytometry. Pretreatment of B cells with inhibitors of protein tyrosine kinase (PTK) resulted in a significant inhibition of both the IL-4- and anti-CD40-induced IL-4 receptor mRNA levels, while protein kinase C (PKC) inhibitors had no effects. These results suggest that IL-4 and CD40 ligation generate B cell signals, which via PTK-dependent pathways, lead to the synergistic induction of IL-4 receptor gene expression. The rapid induction of IL-4 receptor gene expression through the tyrosine kinase-mediated signal transduction by B cell activating stimuli, would provide cells capacity for an efficient response to IL-4 in the early phase of IL-4 action, and may in part constitute the molecular basis of the reported anti-CD40 co-stimulatory effect on the IL-4-induced response.

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A Study on the Utilization of Dietary [15N]urea in Cecal Ligated Chickens (맹장 결찰계(Cecal-ligated Chicken)를 이용한 [15N]urea의 이용성에 관한 연구)

  • Son, Jang-Ho
    • Korean Journal of Poultry Science
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    • v.38 no.1
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    • pp.37-43
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    • 2011
  • The effect of cecal ligation on the utilization of dietary [15N]urea in chickens fed 5 % protein diet plus urea were investigated. Nitrogen balance and utilization tended (P<0.01) to increase by cecal ligation. Total uric acid excretion was significantly decrease by (P<0.01) cecal ligation in chickens from origin of diet and urea (P<0.01). Excretion of ammonia was increased in chickens from origin of diet, where as it decreased in chickens an urea diet (P<0.01). Amount of urea nitrogen excretion from origin of urea was significantly decrease (P<0.01) by cecal ligation, but cecal ligated chicken fed 5% protein diet with urea showed 51.6% urea utilization. Result obtained in present study indicates that ceca is having beneficial role for urea utilization in chicken fed protein deficient diets, but ceca do not always positive role for nitrogen utilization.

Antifibrotic Effect of Stephania tetrandra on Experimental Liver Fibrosis Induced by Bile Duct Ligation and Scission in Rats

  • Nan, Ji-Xing;Park, Eun-Jeon;Lee, Sung-Hee;Park, Pil-hoon;Kim, Ji-Young;Ko, Geon-il;Sohn, Dong-Hwan
    • Archives of Pharmacal Research
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    • v.23 no.5
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    • pp.501-506
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    • 2000
  • We examined the antifibrotic effect of a methanol extract from Stephania tetrandra (ST) on experimental liver fibrosis. liver fibrosis was induced by bile duct ligation and scission (BDL/S) in rats. In BDL/S rats, activity levels of aspartate transaminase (AST), alanine transaminse (ALT), alkaline phosphatase (ALP), concentration of total bilirubin in serum, and hydroxyproline content of the liver were significantly increased. The ST treatment (either 100 ${m}g/kg/day$ or 200 ${m}g/kg/day$, p.o. for 4 weeks) in BDL/S rats reduced the serum AST, ALT and ALP activity levels significantly (p<0.01). Similarly, when compared to the control group, the concentration of hydroxyproline in the livers of the BDL/S rats treated with 100${m}g$ or 200${m}g$ ST treated rats decreased by 40% and 33% respectively, when compared to the BDL/S control group (p<0.01). The morphological characteristics of fibrotic liver that were observed in the BDL/S control group, improved in the ST treated BDL/S group. In the fibrotic liver of BDL/S rats treated with ST, a marked reduction in the numbers of alpha smooth muscle cell actin positive stellate cells was observed. These results indicate that doses of either 100 or 200 ${m}g/kg/day$ of methanol extract from S. tetrandra, had an antifibrotic effect in rats with liver fibrosis induced by bile duct ligation and scission.

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The Change of Pulmonary Arterial Pressures after Left Lung Transplantation and Ligation of Right Pulmonary Artery in Dogs (황견에서 좌측 폐이식수술 및 폐동맥결찰 수술후 폐동맥압 변하에 관한 연구)

  • 이두연
    • Journal of Chest Surgery
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    • v.27 no.5
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    • pp.345-352
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    • 1994
  • We have performed left lung transplantation followed by ligation of right pulmonary artery in 14 dogs at the Chest Disease Research Institute, Yonsei University College of Medicine from May 1992 to February 1994. Excised left lung was perfused with 1500cc of 4$^{\circ}$C cold Euro-Collin`s[E-C] solution at a pressure of 30cmH2O through main pulmonary artery and preserved in 4$^{\circ}$C cold E-C solution for one hour. Left lung transplantation were proceeded in order of left atrium, left main bronchus, left pulmonary artery and right pulmonary artery ligation as usual method. The femoral artery and pulmonary artery pressures were monitored for more than 5 hours after the transplantations in 14 dogs. Six recipient dogs had elevated mean pulmonary artery pressure to greater than 30mmHg after the left lung transplantation and ligation of right pulmonary artery. The cause of elevated mean pulmonary artery pressure was due to inadequate preservation resulting in ischemic damage to donor lungs in 3 cases, and inadequate surgical techniques in 3 cases. Two recipient dogs without surgical complications died immediate post-operatively due to hemorrhagic shock. The bleeding focuses were LA anastomotic site in one case and femoral artery puncture site in another case. The remaining 6 recipient dogs showed mean pulmonary arterial pressure less than 30mmHg. However, one dog had spontaneous pneumothorax in post-operative 4 days, and another dog had rejection phenomenon in post-operative 5 days which was confirmed by pathologic findings of extracted transplanted lung. One dog succumbed of severe hemoptysis which was due to lung abscess with pin point stenosis of bronchial anastomosis in post-operative 38 days. In conclusion, elevated mean pulmonary arterial pressure greater than 30mmHg in immediate postoperative period can be due to inadequate preservation of extracted lung or poor surgical techniques. And the two dogs succumbed of hemorrhagic shock even though the mean pulmonary arterial pressure was less than 30mmHg. It is thought that careful preservation of the extracted donor lung in 4oC E-C solution and complete surgical techniques are the most important factors early and late complications.

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Temporal changes of periodontal tissue pathology in a periodontitis animal model

  • Hyunpil Yoon;Bo Hyun Jung;Ki-Yeon Yoo;Jong-Bin Lee;Heung-Sik Um;Beom-Seok Chang;Jae-Kwan Lee
    • Journal of Periodontal and Implant Science
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    • v.53 no.4
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    • pp.248-258
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    • 2023
  • Purpose: This study aimed to characterize the early stages of periodontal disease and determine the optimal period for its evaluation in a mouse model. The association between the duration of ligation and its effect on the dentogingival area in mice was evaluated using micro-computed tomography (CT) and histological analysis. Methods: Ninety mice were allocated to an untreated control group or a ligation group in which periodontitis was induced by a 6-0 silk ligation around the left second maxillary molar. Mice were sacrificed at 1, 2, 3, 4, 5, 8, 11, and 14 days after ligature placement. Alveolar bone destruction was evaluated using micro-CT. Histological analysis was performed to assess the immune-inflammatory processes in the periodontal tissue. Results: No significant difference in alveolar bone loss was found compared to the control group until day 3 after ligature placement, and a gradual increase in alveolar bone loss was observed from 4 to 8 days following ligature placement. No significant between-group differences were observed after 8 days. The histological analysis demonstrated that the inflammatory response was evident from day 4. Conclusions: Our findings in a mouse model provide experimental evidence that ligature-induced periodontitis models offer a consistent progression of disease with marginal attachment down-growth, inflammatory infiltration, and alveolar bone loss.

THE EFFECT OF LIGATION METHOD ON THE FRICTIONAL FORGE BETWEEN ORTHODONTIC BRACKET AND ARCHWIRE (결찰양식이 교정용 브라켓과 교정선 사이의 마찰력에 미치는 영향)

  • Shin, Hyun-Jeong;Kwon, Oh-Won;Kim, Kyo-Han
    • The korean journal of orthodontics
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    • v.28 no.5 s.70
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    • pp.813-823
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    • 1998
  • The frictional force has been considered as an harmful factor in an active unit where tooth movement occurs, but as an advantageous factor in anchor unit that resist tooth movement. That is, efficient tooth movement is planned by using ligation methods that have low levels of bracket-wire frictional force and the anchorage control can be achieved by using ligation methods that have high levels of bracket-wire frictional force that result in binding of the bracket accompanied by little or no tooth movement. The purpose of this study was to evaluate the frictional force generated between bracket and wire in accordance with the methods of ligation, the material of ligation and the passage of time under artificial saliva. Tested were 0.017x0.022 inch stainless steel wires in standard edgewise twin brackets for upper central incisors in a 0.018-inch slot. The wires were ligated into the brackets with elastomeric modules and stainless steel ligatures. Whole tie, half tie, twisting tie and double overlay tie were done with elastomeric modules. With 0.009-inch stainless steel ligature whole tie and half tie were done by needle holder and whole tie by ligature tying plier. With 0.012-inch stainless steel ligature whole ties were done by needle holder. Whole tie groups of elastomeric module were kept in artificial saliva bath at $37^{\circ}C$ for 28 days. The frictional force was recorded by means of an Instron universial testing instrument (4202 INSTRON, Instron Co., U.S.A.) at initial, 7, 14, 21, and 28 days. The results for ligated samples in a simulated oral environment revealed the fellowing : ${\cdot}$In elastomeric module whole tie, 28 days group was significantly greater mean static frictional force than any other group but there were no significant differences among any other group (p>0.05). ${\cdot}$Elastomeric module twisting ties were significantly greater mean static frictional forces than any other ligation method but there were no significant differences between twisting tie and double overlay tie (p>0.05). Twisting tie, double overlay tie, whole tie, half tie showed differences in decreasing order. ${\cdot}$Stainless steel half tie produced lower mean static frictional force than whole tie, ligation by ligature tying plier produced greater mean static frictional force than by needle holder and ligation with 0.012-inch stainless steel ligature produced greater mean static frictional force than with 0.009-inch stainless steel ligature (p<0.05). ${\cdot}$There were no significant differences between the mean static frictional forces of elastomeric whole tie and stainless steel whole tie (p>0.05).

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