• Title/Summary/Keyword: Biomarker Detection

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1D Proton NMR Spectroscopic Determination of Ethanol and Ethyl Glucuronide in Human Urine

  • Kim, Siwon;Lee, Minji;Yoon, Dahye;Lee, Dong-Kye;Choi, Hye-Jin;Kim, Suhkmann
    • Bulletin of the Korean Chemical Society
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    • v.34 no.8
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    • pp.2413-2418
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    • 2013
  • Forensic and legal medicine require reliable data to indicate excessive alcohol consumption. Ethanol is oxidatively metabolized to acetate by alcohol dehydrogenase and non-oxidatively metabolized to ethyl glucuronide (EtG), ethyl sulfate (EtS), phosphatidylethanol, or fatty acid ethyl esters (FAEE). Oxidative metabolism is too rapid to provide biomarkers for the detection of ethanol ingestion. However, the non-oxidative metabolite EtG is a useful biomarker because it is stable, non-volatile, water soluble, highly sensitive, and is detected in body fluid, hair, and tissues. EtG analysis methods such as mass spectroscopy, chromatography, or enzyme-linked immunosorbent assay techniques are currently in use. We suggest that nuclear magnetic resonance (NMR) spectroscopy could be used to monitor ethanol intake. As with current conventional methods, NMR spectroscopy doesn't require complicated pretreatments or sample separation. This method has the advantages of short acquisition time, simple sample preparation, reproducibility, and accuracy. In addition, all proton-containing compounds can be detected. In this study, we performed $^1H$ NMR analyses of urine to monitor the ethanol and EtG. Urinary samples were collected over time from 5 male volunteers. We confirmed that ethanol and EtG signals could be detected with NMR spectroscopy. Ethanol signals increased immediately upon alcohol intake, but decreased sharply over time. In contrast, EtG signal increased and reached a maximum about 9 h later, after which the EtG signal decreased gradually and remained detectable after 20-25 h. Based on these results, we suggest that $^1H$ NMR spectroscopy may be used to identify ethanol non-oxidative metabolites without the need for sample pretreatment.

Determination of Monoglycoceramides in Biological Samples using Enzymatic Deacylation and Reverse-phase HPLC (역상HPLC컬럼을 이용한 생체 내 단당세라마이드 분석)

  • Choi, Mi-Hwa;Choi, Kyeong-Mi;Ji, So-Young;Lee, Youn-Sun;Cho, Ju-Hyun;Lee, Yong-Moon;Yun, Yeo-Pyo;Yoo, Hwan-Soo
    • YAKHAK HOEJI
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    • v.54 no.5
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    • pp.354-361
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    • 2010
  • Glycosphingolipids are structural components of mammalian cell membranes and are involved in essential cellular physiology such as cell-cell interaction, recognition, transmembrane signaling, proliferation and cell death. In this study, the simple quantitative method of monoglycoceramides-containing glucosylceramide and galactosylceramide was developed. The glycosylceramides extracted from culture cells and rat plasma were resolved by TLC, deacylated by SCDase and analyzed by HPLC-fluorescence detector at an excitation wavelength of 340 nm and an emission wavelength of 455 nm. Limit of detection was approximately 0.1 pmol and limit of quantification was about 1 pmol for both monoglycoceramide standards. The recoveries of standard glucosylceramides from intra- and inter-day assays were 113.8 and 88.8% and those of galactosylceramides were 110.7 and 123.9%, respectively. The monoglycoceramide contents of SW-620 cells and rat plasma were $141.5{\pm}5$ pmol/$1{\times}10^6$ cells and $3.9{\pm}0.3{\mu}M$, respectively. The present analytical method provides a reproducible quantification and total content of monoglycoceramide which may be as a potential biomarker for lipid imbalance-related human diseases.

Comparison Between Two Detection Methods for HPV16, HPV18 and P16Ink4a Biomarkers in Diagnosis of Abnormal Cervical Cytology

  • Khazaei, Sedigheh;Izadi, Babak;Mirbahari, Seyed Ghasem;Madani, Seyed Hamid;Khosravi, Shohre Malek;Alagha, Mohsen Emami;Sajadimajd, Soraya
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.12
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    • pp.5223-5227
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    • 2016
  • Background: Cervical cancer is the second most common cancer among women in many populations. While the Pap smear is a well established screening test it suffers from both false-positive and false-negative results in diagnosis of cancers and precancerous states. In this study, immunocytochemistry of the P16 biomarker and HPV-PCR were compared for their diagnostic potential. Materials and methods: In the study, we obtained pairs of specimens from 45 women with cervical dysplasia. One sample was placed in a liquid-based solution, and processed for staining of sections with antibodies to P16. HPV-PCR was performed on the other and the results obtained were analyzed by T-test using SPSS v. 15. Results: Using HPV-PCR 71% of the samples were found to be infected with either HPV 16 or HPV 18, and the rate of infection did not have a statistically significant relationship with higher grades of dysplasia (p= 0.253). In contrast, with immunocytochemistry evaluation of P16, 64% of the specimens were positive, but the percentage of positive results significantly increased with higher grades of dysplasia (p= 0.0001). Conclusion: Employment of the P16 marker as an optional test might be preferable over HPV-PCR for cervical dysplasia in our geographical region.

Peripapillary Retinal Nerve Fiber Layer Thicknesses Did Not Change in Long-term Hydroxychloroquine Users

  • Lee, Eun Jung;Kim, Sang Jin;Han, Jong Chul;Eo, Doo Ri;Lee, Min Gyu;Ham, Don-Il;Kang, Se Woong;Kee, Changwon;Lee, Jaejoon;Cha, Hoon-Suk;Koh, Eun-Mi
    • Korean Journal of Ophthalmology
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    • v.32 no.6
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    • pp.459-469
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    • 2018
  • Purpose: To evaluate changes in the peripapillary retinal nerve fiber layer (RNFL) thicknesses using spectral-domain optical coherence tomography (SD-OCT) in hydroxychloroquine (HCQ) users. Methods: The medical records of HCQ users were retrospectively reviewed. In these HCQ users, an automated perimetry, fundus autofluorescence photography, and SD-OCT with peripapillary RNFL thickness measurements were performed. The peripapillary RNFL thicknesses were compared between the HCQ users and the control groups. The relationships between the RNFL thicknesses and the duration or cumulative dosage of HCQ use were analyzed. Results: This study included 77 HCQ users and 20 normal controls. The mean duration of HCQ usage was $63.6{\pm}38.4$ months, and the cumulative dose of HCQ was $528.1{\pm}3.44g$. Six patients developed HCQ retinopathy. Global and six sectoral RNFL thicknesses of the HCQ users did not significantly decrease compared to those of the normal controls. No significant correlation was found between the RNFL thickness and the duration of use or cumulative dose. The eyes of those with HCQ retinopathy had temporal peripapillary RNFL thicknesses significantly greater than that of normal controls. Conclusions: The peripapillary RNFL thicknesses did not change in the HCQ users and did not correlate with the duration of HCQ use or cumulative doses of HCQ. RNFL thickness is not a useful biomarker for the early detection of HCQ retinal toxicity.

Real-time Highly Sensitive Measurement of Myocardial Infarction Biomarkers Using Silicon-based Ellipsometric Biosensors (실리콘 기반 타원편광계식 바이오센서를 이용한 심근경색 생체표지자의 실시간 초고감도 진단 농도 측정)

  • Min, Yoon Gi;Cho, Hyun Mo;Jo, Jae Heung
    • Korean Journal of Optics and Photonics
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    • v.30 no.2
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    • pp.59-66
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    • 2019
  • We report highly sensitive detection of myocardial infarction biomarkers, such as myoglobin and cTnI, within several hundred seconds using a rotating-analyzer ellipsometer and a biosensor with biochips fabricated on a $SiO_2$-coated tilted silicon substrate. We choose the running buffer to be pure phosphate-buffered saline (PBS) or 10% mixed human serum. When we choose the running buffer to be pure PBS, we obtain diagnostic densities of pure myocardial infarction biomarkers of up to 1 ng/ml and 5 pg/ml respectively. Meanwhile, when we use PBS with 10% human serum, the measured densities of myoglobin and cTnI were up to 1 ng/mL and 1 pg/mL respectively. The measured diagnostic densities are less than 1/15 and 1/80 (in cases of myoglobin and cTnI respectively) of those referenced by the World Health Organization.

A study of the effectiveness of using the serum procalcitonin level as a predictive test for bacteremia in acute pyelonephritis

  • Lee, Ga Hee;Lee, Yoo Jin;Kim, Yang Wook;Park, Sihyung;Park, Jinhan;Park, Kang Min;Jin, Kyubok;Park, Bong Soo
    • Kosin Medical Journal
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    • v.33 no.3
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    • pp.337-346
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    • 2018
  • Objectives: Serum procalcitonin (PCT) is a specific biomarker that rises after bacterial infection, and levels of PCT are known to correlate with the severity and mortality of patients with pneumonia and sepsis. However, the usefulness of PCT levels in acute pyelonephritis is unknown. This study aimed to evaluate the effectiveness of using the PCT level as a predictive test for bacteremia in acute pyelonephritis. Methods: Between January 2012 and June 2013, 140 patients diagnosed with acute pyelonephritis were admitted to Haeundae Paik Hospital. Serum PCT, C-reactive protein (CRP), and white blood cell (WBC) levels at pre- and post- treatment were measured. Blood and urine cultures were obtained from all patients. The levels of PCT, CRP, and WBCs were each compared between the blood culture-positive and blood culture-negative groups to assess their effectiveness in predicting bacteremia. Results: Pre-treatment PCT level was 0.77 ng/mL (95% CI: 0.42-1.60 ng/mL) in the blood culture-negative group and 4.89 ng/mL (95% CI: 2.88-9.04 ng/mL) in the blood culture-positive group, and the increase between the two groups was statistically significant. The area under the receiver operating characteristic curve of PCT level for prediction of bacteremia was 0.728. A cut-off value of 1.23 ng/mL indicated a sensitivity of 79.0 % and specificity of 60.0 % for PCT level. Conclusions: Serum PCT level is a useful predictive test for bacteremia in acute pyelonephritis. Through the early detection of bacteremia, serum PCT level can help estimate the prognosis and predict complications such as sepsis.

Helicobacter pylori GroEL Seropositivity Is Associated with an Increased Risk of Opisthorchis viverrini-Associated Hepatobiliary Abnormalities and Cholangiocarcinoma

  • Jala, Isabelle;Almanfaluthi, Muhammad Luthfi;Laha, Thewarach;Kanthawong, Sakawrat;Tangkawattana, Sirikachorn;Saichua, Prasert;Suttiprapa, Sutas;Sripa, Banchob
    • Parasites, Hosts and Diseases
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    • v.59 no.4
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    • pp.363-368
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    • 2021
  • Despite the synergistic effect of Opisthorchis viverrini and Helicobacter pylori co-infection on pathogenesis of severe hepatobiliary abnormalities (HBA) including advanced periductal fibrosis and replace with cholangiocarcinoma (CCA) have been established, the immune response to H. pylori in O. viverrini infected population has never been explored. Hence, this study aimed to investigate the antibody responses to 2 immunogenic H. pylori proteins in O. viverrini-infected patients with HBA and CCA. The risk analysis by multinomial logistic regression revealed that GroEL seropositivity was associated with higher risks of hepatobiliary abnormalities and CCA with adjusted odds ratios (95% confidence intervals) of 2.11 (95% CI=1.20-3.71, P=0.008) and 2.13 (95% CI=1.21-3.75, P=0.009), respectively. These findings indicate that GroEL seropositivity might be a biomarker for early detection of O. viverrini associated HBA and CCA.

Identification of plasma miRNA biomarkers for pregnancy detection in dairy cattle

  • Lim, Hyun-Joo;Kim, Hyun Jong;Lee, Ji Hwan;Lim, Dong Hyun;Son, Jun Kyu;Kim, Eun-Tae;Jang, Gulwon;Kim, Dong-Hyeon
    • Journal of Animal Reproduction and Biotechnology
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    • v.36 no.1
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    • pp.35-44
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    • 2021
  • A pregnancy diagnosis is an important standard for control of livestock's reproduction in paricular dairy cattle. High reproductive performance in dairy animals is a essential condition to realize of high life-time production. Pregnancy diagnosis is crucial to shortening the calving interval by enabling the farmer to identify open animals so as to treat or re-breed them at the earliest opportunity. MicroRNAs are short RNA molecules which are critically involved in regulating gene expression during both health and disease. This study is sought to establish the feasible of circulating miRNAs as biomarkers of early pregnancy in cattle. We applied Illumina small-RNA sequencing to profile miRNAs in plasma samples collected from 12 non-pregnant cows ("open" cows: samples were collected before insemination (non-pregnant state) and after pregnancy check at the indicated time points) on weeks 0, 4, 8, 12 and 16. Using small RNA sequencing we identified a total of 115 miRNAs that were differentially expressed weeks 16 relative to non-pregnancy ("open" cows). Weeks 8, 12 and 16 of pregnancy commonly showed a distinct increase in circulating levels of miR-221 and miR-320a. Through genome-wide analyses we have successfully profiled plasma miRNA populations associated with pregnancy in cattle. Their application in the field of reproductive biology has opened up opportunities for research communities to look for pregnancy biomarker molecules in dairy cattle.

Comparative co-expression analysis of RNA-Seq transcriptome revealing key genes, miRNA and transcription factor in distinct metabolic pathways in diabetic nerve, eye, and kidney disease

  • Asmy, Veerankutty Subaida Shafna;Natarajan, Jeyakumar
    • Genomics & Informatics
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    • v.20 no.3
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    • pp.26.1-26.19
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    • 2022
  • Diabetes and its related complications are associated with long term damage and failure of various organ systems. The microvascular complications of diabetes considered in this study are diabetic retinopathy, diabetic neuropathy, and diabetic nephropathy. The aim is to identify the weighted co-expressed and differentially expressed genes (DEGs), major pathways, and their miRNA, transcription factors (TFs) and drugs interacting in all the three conditions. The primary goal is to identify vital DEGs in all the three conditions. The overlapped five genes (AKT1, NFKB1, MAPK3, PDPK1, and TNF) from the DEGs and the co-expressed genes were defined as key genes, which differentially expressed in all the three cases. Then the protein-protein interaction network and gene set linkage analysis (GSLA) of key genes was performed. GSLA, gene ontology, and pathway enrichment analysis of the key genes elucidates nine major pathways in diabetes. Subsequently, we constructed the miRNA-gene and transcription factor-gene regulatory network of the five gene of interest in the nine major pathways were studied. hsa-mir-34a-5p, a major miRNA that interacted with all the five genes. RELA, FOXO3, PDX1, and SREBF1 were the TFs interacting with the major five gene of interest. Finally, drug-gene interaction network elucidates five potential drugs to treat the genes of interest. This research reveals biomarker genes, miRNA, TFs, and therapeutic drugs in the key signaling pathways, which may help us, understand the processes of all three secondary microvascular problems and aid in disease detection and management.

Development and Validation of a Machine Learning-based Differential Diagnosis Model for Patients with Mild Cognitive Impairment using Resting-State Quantitative EEG (안정 상태에서의 정량 뇌파를 이용한 기계학습 기반의 경도인지장애 환자의 감별 진단 모델 개발 및 검증)

  • Moon, Kiwook;Lim, Seungeui;Kim, Jinuk;Ha, Sang-Won;Lee, Kiwon
    • Journal of Biomedical Engineering Research
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    • v.43 no.4
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    • pp.185-192
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    • 2022
  • Early detection of mild cognitive impairment can help prevent the progression of dementia. The purpose of this study was to design and validate a machine learning model that automatically differential diagnosed patients with mild cognitive impairment and identified cognitive decline characteristics compared to a control group with normal cognition using resting-state quantitative electroencephalogram (qEEG) with eyes closed. In the first step, a rectified signal was obtained through a preprocessing process that receives a quantitative EEG signal as an input and removes noise through a filter and independent component analysis (ICA). Frequency analysis and non-linear features were extracted from the rectified signal, and the 3067 extracted features were used as input of a linear support vector machine (SVM), a representative algorithm among machine learning algorithms, and classified into mild cognitive impairment patients and normal cognitive adults. As a result of classification analysis of 58 normal cognitive group and 80 patients in mild cognitive impairment, the accuracy of SVM was 86.2%. In patients with mild cognitive impairment, alpha band power was decreased in the frontal lobe, and high beta band power was increased in the frontal lobe compared to the normal cognitive group. Also, the gamma band power of the occipital-parietal lobe was decreased in mild cognitive impairment. These results represented that quantitative EEG can be used as a meaningful biomarker to discriminate cognitive decline.