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Molecular insights into the role of genetic determinants of congenital hypothyroidism

  • Kollati, Yedukondalu;Akella, Radha Rama Devi;Naushad, Shaik Mohammad;Patel, Rajesh K.;Reddy, G. Bhanuprakash;Dirisala, Vijaya R.
    • Genomics & Informatics
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    • v.19 no.3
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    • pp.29.1-29.10
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    • 2021
  • In our previous studies, we have demonstrated the association of certain variants of the thyroid-stimulating hormone receptor (TSHR), thyroid peroxidase (TPO), and thyroglobulin (TG) genes with congenital hypothyroidism. Herein, we explored the mechanistic basis for this association using different in silico tools. The mRNA 3'-untranslated region (3'-UTR) plays key roles in gene expression at the post-transcriptional level. In TSHR variants (rs2268477, rs7144481, and rs17630128), the binding affinity of microRNAs (miRs) (hsa-miR-154-5p, hsa-miR-376a-2-5p, hsa-miR-3935, hsa-miR-4280, and hsa-miR-6858-3p) to the 3'-UTR is disrupted, affecting post-transcriptional gene regulation. TPO and TG are the two key proteins necessary for the biosynthesis of thyroid hormones in the presence of iodide and H2O2. Reduced stability of these proteins leads to aberrant biosynthesis of thyroid hormones. Compared to the wild-type TPO protein, the p.S398T variant was found to exhibit less stability and significant rearrangements of intra-atomic bonds affecting the stoichiometry and substrate binding (binding energies, ΔG of wild-type vs. mutant: -15 vs. -13.8 kcal/mol; and dissociation constant, Kd of wild-type vs. mutant: 7.2E-12 vs. 7.0E-11 M). The missense mutations p.G653D and p.R1999W on the TG protein showed altered ΔG(0.24 kcal/mol and 0.79 kcal/mol, respectively). In conclusion, an in silico analysis of TSHR genetic variants in the 3'-UTR showed that they alter the binding affinities of different miRs. The TPO protein structure and mutant protein complex (p.S398T) are less stable, with potentially deleterious effects. A structural and energy analysis showed that TG mutations (p.G653D and p.R1999W) reduce the stability of the TG protein and affect its structure-functional relationship.

Overproduction of Bacterial Trypsin in Streptomyces - Optimization for Streptomyces griseus Trypsin Production by Recombinant Streptomyces (미생물을 이용한 트립신 과대 생산 연구 - Streptomyces용 숙주-벡터계를 이용한 트립신 유전자의 대량발현 최적화 -)

  • Kim, Jong-Hee;Hong, Soon-Kwang
    • Microbiology and Biotechnology Letters
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    • v.36 no.1
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    • pp.28-33
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    • 2008
  • The expression vector (pWHM3-TR1R2) for sprT gene encoding Streptomyces griseus trypsin (SGT) followed by two regulatory genes, sgtR1 and sgtR2, was introduced into Streptomyces lividans TK24 and Streptomyces griseus IFO 13350. Various media with different compositions were used to maximize the productivity of SGT in the recombinant trains. he SGT productivity was best when the transformant of S. lividans TK24 was cultivated in R2YE medium (0.74 unit/mL) at 5 days of cultivation. C5/L (0.66 unit/mL) medium also gave a good productivity, but Livid (0.08 unit/mL) and NDSK (0.06 unit/mL) yielded poor productivities. S. griseus IFO 13350/pWHM3-TR1R2 produced SGT by 1.518 unit/mL (C5/L), 1.284unit/mL (R2YE),0.932 unit/mL (NDSK), and 0.295 unit/mL (Livid) at 7 days of cultivation, which was much higher than those from S. lividans TK24/TR1R2. The SGT protein was purified from the culture broth of S. griseus IFO 13350/pWHM3-TR1R2 in C5/L to homogeneity via ammonium sulfate fractionation, and CM-sepharose and SP-sepharose column chromatographies. The specific activity of purified SGT was 69,252 unit/mg, and the final purification fold and recovery yield were 6.5 and 1.4%, respectively.

The Effect of miR-361-3p Targeting TRAF6 on Apoptosis of Multiple Myeloma Cells

  • Fan, Zhen;Wu, Zhiwei;Yang, Bo
    • Journal of Microbiology and Biotechnology
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    • v.31 no.2
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    • pp.197-206
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    • 2021
  • microRNA-361-3p (miR-361-3p) is involved in the carcinogenesis of oral cancer and pancreatic catheter adenocarcinoma, and has anti-carcinogenic effects on non-small cell lung cancer (NSCLC). However, its effect on multiple myeloma (MM) is less reported. Here, we found that upregulating the expression of miR-361-3p inhibited MM cell viability and promoted MM apoptosis. We measured expressions of tumor necrosis factor receptor-associated factor 6 (TRAF6) and miR-361-3p in MM cells and detected the viability, colony formation rate, and apoptosis of MM cells. In addition, we measured expressions of apoptosis-related genes Bcl-2, Bax, and Cleaved caspase-3 (C caspase-3). The binding site between miR-361-3p and TRAF6 was predicted by TargetScan. Our results showed that miR-361-3p was low expressed in the plasma of MM patients and cell lines, while its overexpression inhibited viability and colony formation of MM cells and increased the cell apoptosis. Furthermore, TRAF6, which was predicted to be a target gene of miR-361-3p, was high-expressed in the plasma of patients and cell lines with MM. Rescue experiments demonstrated that the effect of TRAF6 on MM cells was opposite to that of miR-361-3p. Upregulation of miR-361-3p induced apoptosis and inhibited the proliferation of MM cells through targeting TRAF6, suggesting that miR-361-3p might be a potential target for MM therapy.

Sensitivity of Gastric Cancer Cells to Chemotherapy Drugs in Elderly Patients and Its Correlation with Cyclooxygenase-2 Expression

  • Qiu, Zhen-Qin;Qiu, Zhen-Rong
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.8
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    • pp.3447-3450
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    • 2015
  • Objective: To explore the sensitivity of gastric cancer cells to chemotherapy drugs in elderly patients and its correlation with cyclooxygenase-2 (COX-2) expression in cancer tissue. Materials and Methods: Forty-three elderly patients with gastric cancer (observation group) and 31 young patients with gastrointestinal tumors (control group) who were all diagnosed by pathology and underwent surgery in the 89th Hospital of Chinese People's Liberation Army were selected. Drug sensitivity testing of tumor cells in primary culture was carried out in both groups using a methyl thiazolyl tetrazolium (MTT) method, and the expression of COX-2 and the factors related to multi-drug resistance (MDR) in cancer tissue were assessed by immunohistochemistry. Results: The inhibition rates (IR) of vincristine (VCR), 5-fluorouracil (5-FU), oxaliplatin (L-OHP), mitomycin (MMC) and epirubicin (eADM) on tumor cells in the observation group were dramatically lower than in the control group, with statistical significance (P<0.05 or P<0.01). The positive rates of COX-2, glutathione s-transferase-${\pi}$ (GST-${\pi}$) and P glycoprotein (P-gp) expression in cancer tissue in the observation group were all higher than in control group (P<0.05), while that of DNA topoisomerase $II{\alpha}$ ($TopoII{\alpha}$) expression lower than in the control group (P<0.01). In the observation group, COX-2 expression in cancer tissue had a significantly-positive correlation with GST-${\pi}$ and P-gp (r=0.855, P=0.000; r=0.240, P=0.026), but a negative correlation with $TopoII{\alpha}$ (r=-0.328, P=0.002). In the control group, COX-2 expression in cancer tissue was only correlated with P-gp positively (r=0.320, P=0.011). Bivariate correlation analysis displayed that COX-2 expression in cancer tissue in the observation group had a significantly-negative correlation with the IRs of 5-FU, L-OHP, paclitaxel (PTX) and eADM in tumor cells (r=-0.723, P=0.000; r=-0.570, P=0.000; r=-0.919, P=0.000; r=-0.781, P=0.000), but with hydroxycamptothecine (HCPT), VCR and 5-FU in the control group (r=-0.915, P=0.000; r=-0.890, P=0.000; r=-0.949, P=0.000). Conclusions: Gastric cancer cells in elderly patients feature stronger MDR, which may be related to high COX-2 expression.

Predictive Factors of Turnover Intention in New Nurses (신규간호사의 이직의도에 미치는 영향요인)

  • Lee, Jeong Hwa;Noh, Hye Ran;Park, Ju Young
    • Journal of muscle and joint health
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    • v.27 no.1
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    • pp.50-60
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    • 2020
  • Purpose: This study aimed to identify the effects of preceptor's leadership, preceptor-preceptee exchange relationship, and preceptee's self-leadership on newly hired nurses' turnover intention. Methods: Participants were 175 nurses working at general hospitals in D city. Data were collected from 164 nurses in May 2019 and were analyzed using multiple linear regression with SPSS/WIN 24.0 program. Results: Newly hired nurses' turnover intention was negatively correlated with preceptor's transactional leadership (r=-.23, p=.003), preceptor's emotional leadership (r=-.28, p<.001), preceptor-preceptee exchange relationship (r=-.34, p<.001), and preceptee's self-leadership (r=-.23, p=.004). The work unit (β=.35, p<.001), preceptor-preceptee exchange relationship (β=-.28, p=.002), present work experience (β=.27, p=.043), and marital status (β=-.19, p=.009) were significant predictors of the turnover intention (R2=.26, p<.001). Conclusion: Therefore, in order to reduce the degree of turnover of new nurses, improving the exchange relationship of preceptor-preceptee is required, and balancing between work and life should be considered especially for the married new nurses.

Effects of 60 Minutes Cardiopulmonary Resuscitation on Blood Lactic Acid Concentration, Heart Rate, and Rating of Perceived Exertion in Rescuers

  • Han, Seung-Eun;Ahn, Hee-Jeong;Shim, Gyu-Sik;Bang, Sung-Hwan;Song, Hyo-Suk
    • Journal of the Korea Society of Computer and Information
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    • v.27 no.8
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    • pp.195-202
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    • 2022
  • In this study, when cardiopulmonary resuscitation continued for a long time, the rescuer's blood lactic acid concentration, heart rate, and rating of perceived exertion were measured to identify the change in the rescuer's fatigue. Data collection was conducted from July 5 to July 9, 2021, with a total of 24 students, 12 students department of special warfare medical non-commissioned officer, and 12 students department of emergency medical technology at D University, undergoing a two-person alternative chest compression resuscitation for 60 minutes. As a result of the study, the rescuer's blood lactic acid concentration, heart rate, rating of perceived exertion, and chest compression speed were significantly changed according to the duration of CPR (p<.001, p<.001, p<.001, p<.001). blood lactic acid concentrations at every measurement cycle (30 minutes, 40 minutes, 50 minutes, 60 minutes) showed a significant positive correlation with heart rate (r=.696, p<.001, r=.672, p<.001, r=.709, p<.001, r=.782, p<.001), there was also a significant positive correlation with the rating of perceived exertion (r=.476, p<).05, r=.426, p<.05, r=.470, p<.05, r=.470, p<.05). Therefore, monitoring the fatigue of rescuers using heart rate and rating of perceived exertion will be useful for maintaining high-quality chest compression in situations where cardiopulmonary resuscitation is required for a long time.

Morphological Characteristics and Habitat Types of Rattus norvegicus and R. tanezumi Collected in Jeju Island (제주도에서 채집된 Rattus norvegicus와 R. tanezumi의 서식지 유형과 형태적 특성)

  • Kim, Tae-Wook;Joo, Sang-Min;Oh, Ah-Reum;Park, So-Jin;Han, Sang-Hyun;Oh, Hong-Shik
    • Korean Journal of Environment and Ecology
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    • v.27 no.5
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    • pp.550-560
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    • 2013
  • This study was investigated the morphological characters, secondary sexual dimorphism, and habitat patterns of Korean Rattus animals collected on Jeju Island from April 2005 to October 2012. Two wild rat species, namely R. norvegicus and R. tanezumi were identified on Jeju Island based on morphological characteristics and molecular data; however, R. rattus, which had recorded formerly, was not found in this study. Individuals of R. norvegicus were captured from urban, rural, and natural habitats, while those of R. tanezumi were specially found in animal farms and the surrounding areas. Comparing of morphological characters of two species, R. norvegicus had a shorter tail and ears than R. tanezumi (p<0.05), and the ratios of tail length and ear length to those of head-body length showed significantly differences between two species (p<0.05). The body weights (BW) of urban populations of R. norvegicus were significantly heavier than those of rural populations (p<0.05). No secondary sexual dimorphism was found in R. norvegicus, but females of R. tanezumi showed heavier BW than those of males (p<0.05). These findings suggest that it is necessary to revise the records for the existence of R. tanezumi and to confirm the animal fauna and elucidate the distribution and ecological characteristics from further studies using extensive sampling and detailed investigations on Jeju Island and also on the Korean Peninsula.

A Study on Family Caregivers' Burden of the Elderly with Dementia (가정내 치매노인간호자부담에 관한 조사연구)

  • Moon Hae Ree
    • Journal of Korean Public Health Nursing
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    • v.6 no.2
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    • pp.108-132
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    • 1992
  • The purpose of this study· is to identify sources of the caregiving burden, thereby suggesting social welfare alternetives for supporting family caregivers of the elderly with dementia. 23 family caregivers who participated in self-help group of family caregivers of dementia elderly at a welfare facility for the elderly in Seoul were seleted as the study sample. Zarit caregiving burden scale and Zarit functional impairment measurement for the elderly were adapted for this study and a number of additional variables were included in this study. Correlational analysis was utilized. The results of the study were summarized as follows : 1) The impairment of activities of daily living of the elderly was significantly related to the negative elderly. caregiver-family relationship(PADL:r=.6032 IADL : r=.5930 p<.05). 2) The impact on caregiver's health was very significantly related to the impact on the caregiver's task(r=.6233 p<.001). 3) The Impact on caregiver's health was very significantly related to the impact on the caregiver's social activity restriction(r=.6851 p<.001). 4) The impact on the caregiver's social activity was very significantly related to the impact on the caregiver's task(r=.6969 p<.001). 5) Caregiver's income was significantly related to the impact on the caregiver's task(r=.5252 p<.05). 6) Compensation(interpersonal relationship between the elderly and the caregivers, social praise and appreciation of the elderly) was important variables which affect to the feeling of the caregiving burden.

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A New Record of Prionospio depauperata (Annelida: Polychaeta: Spionidae) with DNA Barcoding Data of Four Prionospio Species in South Korea

  • Lee, Geon Hyeok;Yoon, Seong Myeong;Min, Gi-Sik
    • Animal Systematics, Evolution and Diversity
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    • v.36 no.4
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    • pp.382-386
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    • 2020
  • In this study, Prionospio depauperata Imajima, 1990 is newly reported in Korean fauna. Prionospio depauperata can be distinguished from other relatives by the four pairs of branchiae which are pinnate on chaetigers 2 and 5, and apinnate on chaetigers 3 and 4; caruncle extending to the end of chaetiger 2; and moderate dorsal crest present on chaetigers 7-13. The morphological diagnosis of P. depauperata are provided with the photographs of four Prionospio species. The mitochondrial cytochrome c oxidase subunit 1 (CO1), 16S ribosomal DNA (16S rDNA), and the nuclear 18S ribosomal DNA (18S rDNA) sequences of four Prionospio species from Korean waters, P. depauperata Imajima, 1990, P. japonica Okuda, 1935, P. krusadensis Fauvel, 1929, and P. membranacea Imajima, 1990, were determined for the first time. The inter-specific genetic distances among the congeners of four Prionospio species were 22.3-29.6% in CO1, 10.5-25.0% in 16S rDNA, and 0.3-3.6% in 18S rDNA.

A STUDY ON THE IDENTIFICATION OF Porphyromonas endodontalis BY PCR USING SPECIES SPECIFIC PRIMERS FOR THE 16S rDNA (16S rDNA sequence에 대한 종특이성 primer를 이용한 중합효소연쇄반응증폭에 의한 Porphyromonas endodontalis의 동정에 관한 연구)

  • Eom, Seung-Hee;Lim, Sung-Sam;Bae, Kwang-Shik
    • Restorative Dentistry and Endodontics
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    • v.24 no.1
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    • pp.13-25
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    • 1999
  • P. endodontalis which was known to be associated with the infected root canals and periapical lesions is very difficult to detect by culture methods or traditional methods. Detection of bacteria using polymerase chain reaction(PCR) for 16S ribosomal DNA(rDNA) is fast, simple, and accurate with relatively small amount of target cells. 16S rDNA consist of conserved regions those are same to all species, and variable regions which represent species specificity. The 16S rDNA sequences of P. endodontalis and P. gingivalis were aligned and two highly variable regions were selected as a pair of species specific oligonucleotide primers for P. endodontalis. And then the pair of primers for PCR amplification was synthesized to identify P. endodontalis. The sequences of the species specific primers for the 16S rDNA of P. endodontalis were as follows ; sense primer[endo1]: 5'-CTATATTCTTCTTTCTCCGCATGGAGGAGG-3' antisense primer[endo2]: 5'-GCATACCTTCGGTCTCCTCTAGCATAT-3' In this study, for the identification of P. endodontalis without culture from the mixed clinical samples, PCR was done with species specific primers for the 16S rDNA sequences of P. endodontalis. The results were as follows : 1. The species specificity of the primers for the 16S rDNA of P. endodntalis was determined by the PCR methods. About 490bp amplicon which was specific only for P. endodntalis was produced with P. endodontalis. No amplicon was produced by PCR with other strains similar to P. endodontalis. 2. The synthesized species specific primers reacted with conventionally identified P. endodontalis which we have in conservative dentistry laboratory. 3. The identification of P. endodontalis using PCR technique with samples collected from infected root canals or periapical lesions was more sensitive than that of culture methods. 4. Seven samples revealed including P. endodontalis by PCR technique. Five of them were related with pains, two of them with sinus tract, three of them with foul odor, and three of them with purulent drainage. P. endodontalis was shown to have great relation with pains.

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