• Title/Summary/Keyword: Forensic Medicine

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Sustained release of alginate hydrogel containing antimicrobial peptide Chol-37(F34-R) in vitro and its effect on wound healing in murine model of Pseudomonas aeruginosa infection

  • Shuaibing Shi;Hefan Dong;Xiaoyou Chen;Siqi Xu;Yue Song;Meiting Li;Zhiling Yan ;Xiaoli Wang ;Mingfu Niu ;Min Zhang;Chengshui Liao
    • Journal of Veterinary Science
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    • v.24 no.3
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    • pp.44.1-44.17
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    • 2023
  • Background: Antibiotic resistance is a significant public health concern around the globe. Antimicrobial peptides exhibit broad-spectrum and efficient antibacterial activity with an added advantage of low drug resistance. The higher water content and 3D network structure of the hydrogels are beneficial for maintaining antimicrobial peptide activity and help to prevent degradation. The antimicrobial peptide released from hydrogels also hasten the local wound healing by promoting epithelial tissue regeneration and granulation tissue formation. Objective: This study aimed at developing sodium alginate based hydrogel loaded with a novel antimicrobial peptide Chol-37(F34-R) and to investigate the characteristics in vitro and in vivo as an alternative antibacterial wound dressing to treat infectious wounds. Methods: Hydrogels were developed and optimized by varying the concentrations of crosslinkers and subjected to various characterization tests like cross-sectional morphology, swelling index, percent water contents, water retention ratio, drug release and antibacterial activity in vitro, and Pseudomonas aeruginosa infected wound mice model in vivo. Results: The results indicated that the hydrogel C proved superior in terms of cross-sectional morphology having uniformly sized interconnected pores, a good swelling index, with the capacity to retain a higher quantity of water. Furthermore, the optimized hydrogel has been found to exert a significant antimicrobial activity against bacteria and was also found to prevent bacterial infiltration into the wound site due to forming an impermeable barrier between the wound bed and external environment. The optimized hydrogel was found to significantly hasten skin regeneration in animal models when compared to other treatments in addition to strong inhibitory effect on the release of pro-inflammatory cytokines (interleukin-1β and tumor necrosis factor-α). Conclusions: Our results suggest that sodium alginate -based hydrogels loaded with Chol-37(F34-R) hold the potential to be used as an alternative to conventional antibiotics in treating infectious skin wounds.

Microsatellite Alterations of Plasma DNA in Non Small Cell Lung Cancer (비소세포폐암 환자의 혈장 DNA를 이용한 Microsatellite 분석)

  • Kim, Kyu-Sik;Kim, Eun-Jung;Kim, Soo-Ock;Oh, In-Jae;Park, Chang-Min;Jeong, Ju-Yeon;Kim, Yu-Il;Lim, Sung-Chul;Park, Jong-Tae;Kim, Young-Chul
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.4
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    • pp.352-358
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    • 2005
  • Microsatellites are short tandem repeated nucleotide sequences that are present throughout the human genome. Variations in the repeat number or a loss of heterozygosity around the microsatellites have been termed a microsatellite alteration (MA). A MA reflects the genetic instability caused by an impairment in the DNA mismatch repair system and is suggested to be a novel tumorigenic mechanism. A number of studies have reported that MA in the DNA extracted from the plasma occurs at varying frequencies among patients with a non-small cell lung carcinoma (NSCLC). The genomic DNA from 9 subjects with a non-small cell lung cancer (squamous cell cancer 6, adenocarcinoma 2, non-small cell lung cancer1) and 9 age matched non-cancer control subjects (AMC: tuberculosis 3, other inflammatory lung disease 6) and 12 normal control subjects (NC) were extracted from the peripheral blood leukocytes and plasma. Three microsatellite loci were amplified with the primers targeting the Gene Bank sequence D21S1245, D3S1300, and D3S1234. MA in the form of an allelic loss or a band shift was examined with 6% polyacrylamide gel electrophoresis and silver staining. None (0/12) of the NC subjects less than 40 years of age showed a MA in any of the three markers, while 88.9%(8/9) of the AMC above 40 showed a MA in at least one of the three markers (p<0.05). Sixty percent(6/10) of the control subjects with a smoking history showed a MA in one of the three markers, while 9.1%(1/11) of the control subjects without smoking history showed a MA (p<0.05). However, not only did 66.7%(6/9) of lung cancer patients show a MA in at least one of the three markers but so did 88.9%(8/21) of the AMC patients (p>0.05). In conclusion, a MA in the D21S1245, D3S1300, and D3S1234 loci using DNA extracted from the plasma was detected in 66.7% of lung cancer while no MA was found in the young non-smoking control subjects. However, many of the non-cancer control subjects (aged smokers) also showed a MA, which compromised the specificity of the MA analysis as a screening test. Therefore, a further study with a larger sample size will be needed.

Measurement of facial soft tissues thickness using 3D computed tomographic images (3차원 전산화단층찰영 영상을 이용한 얼굴 연조직 두께 계측)

  • Jeong Ho-Gul;Kim Kee-Deog;Han Seung-Ho;Shin Dong-Won;Hu Kyung-Seok;Lee Jae-Bum;Park Hyok;Park Chang-Seo
    • Imaging Science in Dentistry
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    • v.36 no.1
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    • pp.49-54
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    • 2006
  • Purpose : To evaluate accuracy and reliability of program to measure facial soft tissue thickness using 3D computed tomographic images by comparing with direct measurement. Materials and Methods : One cadaver was scanned with a Helical CT with 3 mm slice thickness and 3 mm/sec table speed. The acquired data was reconstructed with 1.5 mm reconstruction interval and the images were transferred to a personal computer. The facial soft tissue thickness were measured using a program developed newly in 3D image. For direct measurement, the cadaver was cut with a bone cutter and then a ruler was placed above the cut side. The procedure was followed by taking pictures of the facial soft tissues with a high-resolution digital camera. Then the measurements were done in the photographic images and repeated for ten times. A repeated measure analysis of variance was adopted to compare and analyze the measurements resulting from the two different methods. Comparison according to the areas was analyzed by Mann-Whitney test. Results : There were no statistically significant differences between the direct measurements and those using the 3D images (p>0.05). There were statistical differences in the measurements on 17 points but all the points except 2 points showed a mean difference of 0.5 mm or less. Conclusion : The developed software program to measure the facial soft tissue thickness using 3D images was so accurate that it allows to measure facial soft tissues thickness more easily in forensic science and anthropology.

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Effects of the Temperature and Light Intensity on the Growth and Microcystin Production of Three Species of Microcystis (M. aeruginosa, M. ichthyoblabe, M. viridis) (Microcystis 3종(M. aeruginosa, M. ichthyoblabe, M. viridis)의 성장과 microcystins 생성에 대한 온도 및 조도의 영향)

  • Lee, Kyung-Lak;Jheong, Weon-Hwa;Kim, Jin-Hee;Kim, Han-Soon
    • Korean Journal of Ecology and Environment
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    • v.43 no.3
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    • pp.400-408
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    • 2010
  • The growth and microcystins production characteristics of three species of Microcystis (M. aeruginosa, M. ichthyoblabe, M. viridis) isolated from Yeongchun dam and Ankei dam in Kyungpook Province, South Korea were investigated at temperatures of $15{\sim}35^{\circ}C$ and light intensities of $35{\sim}180\;{\mu}mol\;m^{-2}\;s^{-1}$. All of the three species exhibited the highest growth rates (${\mu}_{max}$) over the $30^{\circ}C$. The maximum growth rates of M. aeruginosa and M. ichthyoblabe was observed at $70\;{\mu}mol\;m^{-2}\;s^{-1}$, while M. viridis showed maximum growth rate at $35\;{\mu}mol\;m^{-2}\;s^{-1}$. The maximum production of total microcystins was observed at $20^{\circ}C$, and the production of microcystins decreased according as temperature increase. The highest microcystins production of M. aeruginosa, M. ichthyoblabe and M. viridis observed at light intensities of $120\;{\mu}mol\;m^{-2}\;s^{-1}$, $70\;{\mu}mol\;m^{-2}\;s^{-1}$ and $35\;{\mu}mol\;m^{-2}\;s^{-1}$, respectively. The concentration of microcyst in production and microcystin types of three species according to temperatures and light intensities showed clear difference between the species.

A Mouse Model of Photochemically Induced Spinal Cord Injury

  • Piao, Min Sheng;Lee, Jung-Kil;Jang, Jae-Won;Kim, Soo-Han;Kim, Hyung-Seok
    • Journal of Korean Neurosurgical Society
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    • v.46 no.5
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    • pp.479-483
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    • 2009
  • Objective : A mouse model of spinal cord injury (SCI) could further increase our basic understanding of the mechanisms involved in injury and repair of the nervous system. The purpose of this study was to investigate whether methods used to produce and evaluate photochemical graded ischemic SCI in rats, could be successfully adapted to mice, in a reliable and reproducible manner. Methods : Thirty female imprinting control region mice (weighting 25-30 g, 8 weeks of age) were used in this study. Following intraperitoneal injection of Rose bengal, the translucent dorsal surface of the T8-T9 vertebral laminae of the mice were illuminated with a fiber optic bundle of a cold light source. The mice were divided into three groups; Group 1 (20 mg/kg Rose bengal, 5 minutes illumination), Group 2 (20 mg/kg Rose bengal, 10 minutes illumination), and Group 3 (40 mg/kg Rose bengal, 10 minutes illumination). The locomotor function, according to the Basso-Beattie-Bresnahan scale, was assessed at three days after the injury and then once per week for four weeks. The animals were sacrificed at 28 days after the injury, and the histopathology of the lesions was assessed. Results : The mice in group 1 had no hindlimb movement until seven days after the injury. Most mice had later recovery with movement in more than two joints at 28 days after injury. There was limited recovery of one joint, with only slight movement, for the mice in groups 2 and 3. The histopathology showed that the mice in group 1 had a cystic cavity involving the dorsal and partial involvement of the dorsolateral funiculi. A larger cavity, involving the dorsal, dorsolateral funiculi and the gray matter of the dorsal and ventral horns was found in group 2. In group 3, most of the spinal cord was destroyed and only a thin rim of tissue remained. Conclusion : The results of this study show that the photochemical graded ischemic SCI model. described in rats, can be successfully adapted to mice, in a reliable and reproducible manner. The functional deficits are correlated an increase in the irradiation time and, therefore, to the severity of the injury. The photothrombotic model of SCI, in mice with 20 mg/kg Rose bengal for 5 minutes illumination, provides an effective model that could be used in future research. This photochemical model can be used for investigating secondary responses associated with traumatic SCI.

Vasa Vasorum Densities in Human Carotid Atherosclerosis Is Associated with Plaque Development and Vulnerability

  • Joo, Sung-Pil;Lee, Seung-Won;Cho, Yong-Hwan;Kim, You-Sub;Seo, Bo-Ra;Kim, Hyung-Seok;Kim, Tae-Sun
    • Journal of Korean Neurosurgical Society
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    • v.63 no.2
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    • pp.178-187
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    • 2020
  • Objective : The extensive vasa vasorum network functions as a conduit for the entry of inflammatory cells or factors that promote the progression of angiogenesis and plaque formation. Therefore, we investigated the correlation between the carotid vasa vasorum activities and carotid plaque vulnerability using indocyanine green video angiography (ICG-VA) during carotid endarterectomy (CEA). Methods : Sixty-nine patients who underwent CEA were enrolled prospectively from September 2015 to December 2017. During CEA, a bolus of ICG was injected intravenously before and after resecting the atheroma. Additionally, we performed immunohistochemistry using CD68 (a surface marker of macrophages), CD117 (a surface marker of mast cells), and CD4 and CD8 (surface markers of T-cells) antibodies to analyze the resected plaque specimens. Results : The density of active vasa vasorum was observed in all patients using ICG-VA. The vasa vasorum externa (VVE) and interna (VVI) were seen in 11 (16%) and 57 patients (82.6%), respectively. Macroscopically, the VVE-type patterns were strongly associated with preoperative angiographic instability (81.8%, p=0.005) and carotid plaque vulnerability (90.9%, p=0.017). In contrast, the VVI-type patterns were weakly associated with angiographic instability (31.6%) and plaque vulnerability (49.1%). CD68-stained macrophages and CD117-stained mast cells were observed more frequently in unstable plaques than in stable plaques (p<0.0001, p=0.002, respectively). Conclusion : The early appearance of VVE, along with the presence of many microvessel channels that provided nutrients to the developing and expanding atheroma during ICG-VA, was strongly associated with unstable carotid plaques. The degree of infiltration of macrophages and mast cells is possibly related to the formation of unstable plaques.

Molecular Genetic Characteristics of Methicillin-Resistant Staphylococcus aureus (MRSA) Isolated From Nasal Cavity of University Students (대학생들의 비강으로부터 분리된 메티실린 내성 황색포도알균의 분자유전학적 특성)

  • Lee Eun gwang;Oh Dae Hwan;Sunjin Jung;Sohyun Park;Yeonim Choi
    • Journal of the Health Care and Life Science
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    • v.9 no.2
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    • pp.303-308
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    • 2021
  • Staphylococcus aureus and methicillin resistant S. aureus (MRSA) is a major cause of nosocomial infections and is one of the most commonly isolated bacterial species in the hospital and continues to be an important pathogens in both community and hospital-acquires infection. The purpose of this study is to investigate the carrier rate of S. aureus and MRSA in the community and molecular genetic characteristics of these organisms. The identification of S. aureus and MRSA were done by the procedures in Murray's manual of Clinical Microbiology and antibiotic susceptibility patterns by the Clinical and Laboratory Standards Institute(CLSI). MRSA strains were confirms by oxacillin disk diffusion method. forty-six strains (71.9%) of S. aureus were isolated from the nasal specimens of 64 students in health science university. twenty-two strains (22%) of 46 S. aureus were resistant to penicillin and oxacillin. twenty-two strains of the 46 S. aureus isolates were methicillin-resistant Staphylococcus aureus (MRSA). The mecA genes in MRSA were detected by polymerase chain reaction (PCR). Community and nosocomial infections caused by methicillin-resistant Staphylococcus aureus are a significant problem worldwide. There continuous epidemiological study is to investigate the prevalence of MRSA in community acquired infections.

The Effects of Storage of Human Saliva on DNA Isolation and Stability (인체타액의 보관이 DNA 분리와 안정도에 미치는 영향)

  • Kim, Yong-Woo;Kim, Young-Ku
    • Journal of Oral Medicine and Pain
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    • v.31 no.1
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    • pp.1-16
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    • 2006
  • The most important progress in diagnostic sciences is the increased sensitivity and specificity in diagnostic procedures due to the development of micromethodologies and increasing availability of immunological and molecular biological reagents. The technological advances led to consider the diagnostic use of saliva for an array of analytes and DNA source. The purpose of the present study was to compare DNA from saliva with those from blood and buccal swab, to evaluate diagnostic and forensic application of saliva, to investigate the changes of genomic DNA in saliva according to the storage temperature and period of saliva samples, and to evaluate the integrity of the DNA from saliva stored under various storage conditions by PCR analysis. Peripheral venous blood, unstimulated whole saliva, stimulated whole saliva, and buccal swab were obtained from healthy 10 subjects (mean age: $29.9{\pm}9.8$ years) and genomic DNA was extracted using commercial kit. For the study of effects of various storage conditions on genomic DNA from saliva, stimulated whole saliva were obtained from healthy 20 subjects (mean age: $32.3{\pm}6.6$ years). After making aliquots from fresh saliva, they were stored at room temperature, $4^{\circ}C$, $-20^{\circ}C$, and $-70^{\circ}C$. Saliva samples after lyophilization and dry-out procedure were stored at room temperature. After 1, 3, and 5 months, the same experiment was performed to investigate the changes in genomic DNA in saliva samples. In case of saliva aliquots stored at room temperature and dry-out samples, the results in 2 weeks were also included. Integrity of DNA from saliva stored under various storage conditions was also evaluated by PCR amplification analysis of $\beta$-globin gene fragments (989-bp). The results were as follows: 1. Concentration of genomic DNA extracted from saliva was lower than that from blood (p<0.05), but there were no significant differences among various types of saliva samples. Purities of genomic DNA extracted from stimulated whole saliva and lyophilized one were significantly higher than that from blood (p<0.05). Purity of genomic DNA extracted from buccal swab was lower than those from various types of saliva samples (p<0.05). 2. Concentration of genomic DNA from saliva stored at room temperature showed gradual reduction after 1 month, and decreased significantly in 3 and 5 months (p<0.05, p<0.01, respectively). Purities of DNA from saliva stored for 3 and 5 months showed significant differences with those of fresh saliva and stored saliva for 1 month (p<0.05). 3. In the case of saliva stored at $4^{\circ}C$ and $-20^{\circ}C$, there were no significant changes of concentration of genomic DNA in 3 months. Concentration of DNA decreased significantly in 5 months (p<0.05). 4. There were no significant differences of concentration of genomic DNA from saliva stored at $-70^{\circ}C$ and from lyophilized one according to storage period. Concentration of DNA showed decreasing tendency in 5 months. 5. Concentration of genomic DNA immediately extracted from saliva dried on Petri dish were 60% compared with that of fresh saliva. Concentration of DNA from saliva stored at room temperature after dry-out showed rapid reduction within 2 weeks (p<0.05). 6. Amplification of $\beta$-globin gene using PCR was successful in all lyophilized saliva stored for 5 months. At the time of 1 month, $\beta$-globin gene was successfully amplified in all saliva samples stored at $-20^{\circ}C$ and $-70^{\circ}C$, and in some saliva samples stored at $4^{\circ}C$. $\beta$-globin gene was failed to amplify in saliva stored at room temperature and dry-out saliva.