• Title/Summary/Keyword: Molecular techniques

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Ultrashort Echo Time MRI (UTE-MRI) Quantifications of Cortical Bone Varied Significantly at Body Temperature Compared with Room Temperature

  • Jerban, Saeed;Szeverenyi, Nikolaus;Ma, Yajun;Guo, Tan;Namiranian, Behnam;To, Sarah;Jang, Hyungseok;Chang, Eric Y.;Du, Jiang
    • Investigative Magnetic Resonance Imaging
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    • v.23 no.3
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    • pp.202-209
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    • 2019
  • Purpose: To investigate the temperature-based differences of cortical bone ultrashort echo time MRI (UTE-MRI) biomarkers between body and room temperatures. Investigations of ex vivo UTE-MRI techniques were performed mostly at room temperature however, it is noted that the MRI properties of cortical bone may differ in vivo due to the higher temperature which exists as a condition in the live body. Materials and Methods: Cortical bone specimens from fourteen donors ($63{\pm}21$ years old, 6 females and 8 males) were scanned on a 3T clinical scanner at body and room temperatures to perform T1, $T2^*$, inversion recovery UTE (IR-UTE) $T2^*$ measurements, and two-pool magnetization transfer (MT) modeling. Results: Single-component $T2^*$, $IR-T2^*$, short and long component $T2^*s$ from bi-component analysis, and T1 showed significantly higher values while the noted macromolecular fraction (MMF) from MT modeling showed significantly lower values at body temperature, as compared with room temperature. However, it is noted that the short component fraction (Frac1) showed higher values at body temperature. Conclusion: This study highlights the need for careful consideration of the temperature effects on MRI measurements, before extending a conclusion from ex vivo studies on cortical bone specimens to clinical in vivo studies. It is noted that the increased relaxation times at higher temperature was most likely due to an increased molecular motion. The T1 increase for the studied human bone specimens was noted as being significantly higher than the previously reported values for bovine cortical bone. The prevailing discipline notes that the increased relaxation times of the bound water likely resulted in a lower signal loss during data acquisition, which led to the incidence of a higher Frac1 at body temperature.

Review on Graphene Oxide-based Nanofiltration Membrane (산화그래핀 기반 나노여과막의 최신 연구동향)

  • Kim, Dae Woo
    • Membrane Journal
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    • v.29 no.3
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    • pp.130-139
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    • 2019
  • Various two-dimensional nano materials such as graphene, zeolite, and metal-organic framework have been utilized to develop an ultra-thin high-performance membrane for water purification, gas separation, and so on. Particularly, in the case of graphene oxide, synthesis methods and thin film coating techniques have been accumulated and established since early 2000s, therefore graphene oxide has been rapidly applied to membrane field. The multi-layered graphene oxide thin film can filter molecules separately by the molecular sieving of interlayer spacing between adjacent layers, and it is also possible to separate various materials depending on the surface functional groups or the degree of interaction to intercalated materials. This review mainly focuses on the nanofiltration application of graphene oxide. The major factors affecting the separation performance of graphene oxide membrane in solvent are summarized and other technical elements required for the commercialization of graphene oxide membranes will be discussed including stability issue and fabrication method.

Effect of Ghrelin on Memory Impairment in a Rat Model of Vascular Dementia (그렐린이 혈관성 치매 쥐의 기억 손상에 미치는 효과)

  • Park, Jong-Min;Kim, Youn-Jung
    • Journal of Korean Academy of Nursing
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    • v.49 no.3
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    • pp.317-328
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    • 2019
  • Purpose: The purpose of this study was to identify the effect of ghrelin on memory impairment in a rat model of vascular dementia induced by chronic cerebral hypoperfusion. Methods: Randomized controlled groups and the posttest design were used. We established the representative animal model of vascular dementia caused by bilateral common carotid artery occlusion and administered $80{\mu}g/kg$ ghrelin intraperitoneally for 4 weeks. First, behavioral studies were performed to evaluate spatial memory. Second, we used molecular biology techniques to determine whether ghrelin ameliorates the damage to the structure and function of the white matter and hippocampus, which are crucial to learning and memory. Results: Ghrelin improved the spatial memory impairment in the Y-maze and Morris water maze test. In the white matter, demyelination and atrophy of the corpus callosum were significantly decreased in the ghrelin-treated group. In the hippocampus, ghrelin increased the length of hippocampal microvessels and reduced the microvessels pathology. Further, we confirmed angiogenesis enhancement through the fact that ghrelin treatment increased vascular endothelial growth factor (VEGF)-related protein levels, which are the most powerful mediators of angiogenesis in the hippocampus. Conclusion: We found that ghrelin affected the damaged myelin sheaths and microvessels by increasing angiogenesis, which then led to neuroprotection and improved memory function. We suggest that further studies continue to accumulate evidence of the effect of ghrelin. Further, we believe that the development of therapeutic interventions that increase ghrelin may contribute to memory improvement in patients with vascular dementia.

Complete Genome Sequencing and Infectious cDNA Clone Construction of Soybean Mosaic Virus Isolated from Shanxi

  • Wang, Defu;Cui, Liyan;Zhang, Li;Ma, Zhennan;Niu, Yanbing
    • The Plant Pathology Journal
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    • v.37 no.2
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    • pp.162-172
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    • 2021
  • Soybean mosaic virus (SMV) is the predominant viral pathogen that affects the yield and quality of soybean. The natural host range for SMV is very narrow, and generally limited to Leguminosae. However, we found that SMV can naturally infect Pinellia ternata and Atractylodes macrocephala. In order to clarify the molecular mechanisms underlying the cross-family infection of SMV, we used double-stranded RNA extraction, rapid amplification of cDNA ends polymerase chain reaction and Gibson assembly techniques to carry out SMV full-length genome amplification from susceptible soybeans and constructed an infectious cDNA clone for SMV. The genome of the SMV Shanxi isolate (SMV-SX) consists of 9,587 nt and encodes a polyprotein consisting of 3,067 aa. SMV-SX and SMV-XFQ008 had the highest nucleotide and amino acid sequence identities of 97.03% and 98.50%, respectively. A phylogenetic tree indicated that SMV-SX and SMV-XFQ018 were clustered together, sharing the closest relationship. We then constructed a pSMV-SX infectious cDNA clone by Gibson assembly technology and used this clone to inoculate soybean and Ailanthus altissima; the symptoms of these hosts were similar to those caused by the virus isolated from natural infected plant tissue. This method of construction not only makes up for the time-consuming and laborious defect of traditional methods used to construct infectious cDNA clones, but also avoids the toxicity of the Potyvirus special sequence to Escherichia coli, thus providing a useful cloning strategy for the construction of infectious cDNA clones for other viruses and laying down a foundation for the further investigation of SMV cross-family infection mechanisms.

Ginsenoside compound K ameliorates palmitate-induced atrophy in C2C12 myotubes via promyogenic effects and AMPK/autophagy-mediated suppression of endoplasmic reticulum stress

  • Kim, Tae Jin;Pyun, Do Hyeon;Kim, Myeong Jun;Jeong, Ji Hoon;Abd El-Aty, A.M.;Jung, Tae Woo
    • Journal of Ginseng Research
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    • v.46 no.3
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    • pp.444-453
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    • 2022
  • Background: Compound K (CK) is among the protopanaxadiol (PPD)-type ginsenoside group, which produces multiple pharmacological effects. Herein, we examined the effects of CK on muscle atrophy under hyperlipidemic conditions along with its pro-myogenic effects. Further, the molecular pathways underlying the effects of CK on skeletal muscle have been justified. Methods: C2C12 myotubes were treated with palmitate and CK. C2C12 myoblasts were differentiated using CK for 4-5 days. For the in vivo experiments, CK was administered to mice fed on a high-fat diet for 8 weeks. The protein expression levels were analyzed using western blotting analysis. Target protein suppression was performed using small interfering (si) RNA transfection. Histological examination was performed using Jenner-Giemsa and H&E staining techniques. Results: CK treatment attenuated ER stress markers, such as eIF2a phosphorylation and CHOP expression and impaired myotube formation in palmitate-treated C2C12 myotubes and skeletal muscle of mice fed on HFD. CK treatment augmented AMPK along with autophagy markers in skeletal muscle cells in vitro and in vivo experiments. AMPK siRNA or 3-MA, an autophagy inhibitor, abrogated the impacts of CK in C2C12 myotubes. CK treatment augmented p38 and Akt phosphorylation, leading to an enhancement of C2C12 myogenesis. However, AMPK siRNA abolished the effects of CK in C2C12 myoblasts. Conclusion: These findings denote that CK prevents lipid-induced skeletal muscle apoptosis via AMPK/autophagy-mediated attenuation of ER stress and induction of myoblast differentiation. Therefore, we may suggest the use of CK as a potential therapeutic approach for treating muscle-wasting conditions associated with obesity.

ESTIMATION OF NITROGEN-TO-IRON ABUNDANCE RATIOS FROM LOW-RESOLUTION SPECTRA

  • Kim, Changmin;Lee, Young Sun;Beers, Timothy C.;Masseron, Thomas
    • Journal of The Korean Astronomical Society
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    • v.55 no.2
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    • pp.23-36
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    • 2022
  • We present a method to determine nitrogen abundance ratios with respect to iron ([N/Fe]) from molecular CN-band features observed in low-resolution (R ~ 2000) stellar spectra obtained by the Sloan Digital Sky Survey (SDSS) and the Large Sky Area Multi-Object Fiber Spectroscopic Telescope (LAMOST). Various tests are carried out to check the systematic and random errors of our technique, and the impact of signal-to-noise (S/N) ratios of stellar spectra on the determined [N/Fe]. We find that the uncertainty of our derived [N/Fe] is less than 0.3 dex for S/N ratios larger than 10 in the ranges Teff = [4000, 6000] K, log g = [0.0, 3.5], [Fe/H] = [-3.0, 0.0], [C/Fe] = [-1.0, +4.5], and [N/Fe] = [-1.0, +4.5], the parameter space that we are interested in to identify N-enhanced stars in the Galactic halo. A star-by-star comparison with a sample of stars with [N/Fe] estimates available from the Apache Point Observatory Galactic Evolution Experiment (APOGEE) also suggests a similar level of uncertainty in our measured [N/Fe], after removing its systematic error. Based on these results, we conclude that our method is able to reproduce [N/Fe] from low-resolution spectroscopic data, with an uncertainty sufficiently small to discover N-rich stars that presumably originated from disrupted Galactic globular clusters.

Head-to-Head Comparison between Xpert MTB/RIF Assay and Real-Time Polymerase Chain Reaction Assay Using Bronchial Washing Specimens for Tuberculosis Diagnosis

  • Son, Eunjeong;Jang, Jinook;Kim, Taehwa;Jang, Jin Ho;Chung, Jae Heun;Seol, Hee Yun;Yeo, Hye Ju;Yoon, Seong Hoon;Lee, Seung Eun;Cho, Woo Hyun;Kim, Yun Seong;Jeon, Doosoo
    • Tuberculosis and Respiratory Diseases
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    • v.85 no.1
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    • pp.89-95
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    • 2022
  • Background: With the introduction of Xpert MTB/RIF assay (Xpert), its incorporation into tuberculosis (TB) diagnostic algorithm has become an important issue. The aim of this study was to evaluate the performance of the Xpert assay in comparison with a commercial polymerase chain reaction (PCR) assay. Methods: Medical records of patients having results of both Xpert and AdvanSure TB/NTM real-time PCR (AdvanSure) assays using the same bronchial washing specimens were retrospectively reviewed. Results: Of the 1,297 patients included in this study, 205 (15.8%) were diagnosed with pulmonary TB. Using mycobacterial culture as the reference method, sensitivity of the Xpert assay using smear-positive specimens was 97.5%, which was comparable to that of the AdvanSure assay (96.3%, p=0.193). However, the sensitivity of the Xpert assay using smear-negative specimens was 70.6%, which was significantly higher than that of the AdvanSure assay (52.9%, p=0.018). Usng phenotypic drug susceptibility testing as the reference method, sensitivity and specificity for detecting rifampicin resistance were 100% and 99.1%, respectively. Moreover, a median turnaround time of the Xpert assay was 1 day, which was significantly shorter than 3 days of the AdvanSure assay (p<0.001). Conclusion: In comparison with the AdvanSure assay, the Xpert assay had a higher sensitivity using smear-negative specimens, a shorter turnaround time, and could reliably predict rifampin resistance. Therefore, the Xpert assay might be preferentially recommended over TB-PCR in Korean TB diagnostic algorithm.

High efficiency of homemade culture medium supplemented with GDF9-β in human oocytes for rescue in vitro maturation

  • Mohsenzadeh, Mehdi;Khalili, Mohammad Ali;Anbari, Fatemeh;Vatanparast, Mahboubeh
    • Clinical and Experimental Reproductive Medicine
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    • v.49 no.2
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    • pp.149-158
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    • 2022
  • Objective: Optimizing culture media for the incubation of immature oocytes is a vital strategy to increase the oocyte maturation rate during in vitro maturation (IVM) programs. This study evaluated the IVM and fertilization rates of human germinal vesicle (GV) and metaphase I (MI) oocytes using two different maturation media (commercial and homemade) with or without growth differentiation factor 9-β (GDF9-β). supplementation. Methods: Immature oocytes from intracytoplasmic sperm injection (ICSI) cycles were collected and assigned to one of two IVM culture media (commercial or homemade; cleavage-stage base). After maturation, MII oocytes were examined under an inverted microscope for the presence of the polar body, zona pellucida (ZP) birefringence, and meiotic spindle (MS) visualization after maturation in four conditions (commercial or homemade medium, with or without GDF9-β. ICSI was done for matured oocytes, and fertilization was confirmed by the visualization of two distinct pronuclei and two polar bodies. Results: No significant differences were found between the two culture media in terms of the time and rate of oocyte maturation or the rate of fertilization (p>0.05). Growth factor supplementation increased the 24-hour maturation rate for both GV and MI oocytes only in homemade medium. The maturation rate after 24 hours was higher for MI oocytes (p<0.05). Similar results were observed for MS visualization and ZP structure in both types of media (p>0.05). Conclusion: Higher rates of oocyte maturation and fertilization were observed after application of homemade medium supplemented with GDF9-β. Therefore, this combination may be recommended as an alternative for clinical IVM programs.

Evaluation of the cost-effectiveness of ASF detection with or without the use of on-field tests in different scenarios, in Sardinia

  • Cappai, Stefano;Loi, Federica;Rolesu, Sandro;Coccollone, Annamaria;Laddomada, Alberto;Sgarangella, Francesco;Masala, Sergio;Bitti, Giuseppe;Floris, Vincenzo;Desini, Pietro
    • Journal of Veterinary Science
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    • v.21 no.2
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    • pp.14.1-14.10
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    • 2020
  • African swine fever (ASF) is a highly contagious disease of domestic pigs and wild boars (WBs). Without a vaccine, early antibody and antigen detection and rapid diagnosis are crucial for the effective prevention of the disease and the employment of control measures. In Sardinia, where 3 different suid populations coexisted closely for a long time, the disease persists since 1978. The recent ASF eradication plan involves more stringent measures to combat free-ranging pigs and any kind of illegality in the pig industry. However, critical issues such as the low level of hunter cooperation with veterinary services and the time required for ASF detection in the WBs killed during the hunting season still remain. Considering the need to deliver true ASF negative carcasses as early as possible, this study focuses on the evaluation and validation of a duplex pen-side test that simultaneously detects antibodies and antigens specific to ASF virus, to improve molecular diagnosis under field conditions. The main goal was to establish the specificity of the two pen-side tests performed simultaneously and to determine their ability to detect the true ASF negative carcasses among the hunted WBs. Blood and organ samples of the WBs hunted during the 2018/2019 hunting seasons were obtained. A total of 160 animals were tested using the pen-side kit test; samples were collected for virological and serological analyses. A specificity of 98% was observed considering the official laboratory tests as gold standards. The new diagnostic techniques could facilitate faster and cost-effective control of the disease.

Exon Capture - Principle and Applications to Phylogenomics and Population Genomics of Fishes (엑손 포획 - 원리와 어류의 계통유전체학 및 집단유전체학으로의 응용)

  • Li, Chenhong
    • Korean Journal of Ichthyology
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    • v.33 no.4
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    • pp.205-216
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    • 2021
  • Phylogenetic reconstruction based on one locus or a few loci can be misleading due to gene-tree/species-tree discordance. Species delimitation and intraspecific studies also often suffered from low resolution because of insufficient statistic power when few loci were used. Exon capture method is one of the most efficient way to collect genome-scale data, which can significantly augment studies that aimed to investigate patterns and histories of organisms at both intraspecific and high level. Here, I showed the advancement of shifting from single-gene method to genomic approach and the benefit of applying exon capture method comparing to alternative genomic techniques. Then, I explained the principle of exon capture method as well as providing detailed recommendations for applying this method. Finally, I demonstrated exon capture method using two applications and discussed future perspectives of this technology.