A microscope is the fundamental research and diagnostic apparatus for clinical investigation of signaling transduction, morphological changes and physiological tracking of cells and intact tissues from patients in the biomedical laboratory science. Proper use, care and maintenance of microscope with comprehensive understanding in mechanism are fully requested for reliable image data and accurate interpretation for diagnosis in the clinical laboratory. The standard operating procedure (SOP) for light microscopes includes performance procedure, brief information of all mechanical parts of microscopes with systematic troubleshooting mechanism depending on the laboratory capacity. Maintenance program encompasses cleaning objective, ocular lenses and inner optics; replacement and calibration of light source; XY sample stage management; point spread function (PSF) measurement for confocal laser scanning microscope (CLSM); quality control (QC) program in fluorescent microscopy; and systematic troubleshooting. Laser safety is one of the concern for medical technologists engaged in CLSM laboratory. Laser safety guideline based on the laser classification and risk level, and advisory lab wear for CLSM users are also expatiated in this overview. Since acquired image data presents a wide range of information at the moment of acquisition, well-maintained microscopes with proper microscopic maintenance program are impulsive for its interpretation and diagnosis in the clinical laboratory.
Natural products have always been an attractive source in terms of novel anti-metastatic compounds which can hinder MMP expression and activity. Corydalis heterocarpa is a salt marsh plant found in the seashores throughout Korea. Its yellow flowers and spikes have been an ingredient in folk medicine to treat spasm and contractions. The present study assessed the potential of different solvent-based fractions from the crude extract of Corydalis heterocarpa (CHE), a halophyte with reported bioactivities, to suppress the PMA-induced MMP expression in human fibrosarcoma HT-1080 cells. The solvent fractions which were named after the solvent used for fractionation (n-hexane, 85% aqueous (aq.) methanol (MeOH), n-butanol (BuOH), and H2O were shown to inhibit the both elevated mRNA and protein expression levels of MMP-2 and MMP-9 and simultaneously relieved the suppression on the expression of the endogenous MMP inhibitors TIMP-1 and TIMP-2. Results indicated that the CHE fractions might intervene with the PMA-induced activation of the MAPK signaling which is the upstream activator of MMP overexpression. Among tested samples, 85% aq. MeOH and n-hexane fractions of CHE was determined to be the most active and future studies to isolate the bioactive substances responsible for the regulation of the MMP expression are, therefore, urged. In conclusion, C. heterocarpa was shown to be a potential source of anti-metastatic compounds and n-Hexane and MeOH fractions might yield lead molecules to develop novel MMP inhibitors.
Like vertebrates, insects synthesize various eicosanoids after the committed catalytic step of phospholipase A2 (PLA2). However, the subsequent biosynthetic steps exhibit some deviation from those of vertebrates. Due to little composition of arachidonic acid in insect phospholipids, PLA2 releases linoleic acid, which is another polyunsaturated fatty acid and relatively rich in insect phospholipids, to synthesize arachidonic acid via chain extension and desaturation. Resulting arachidonic acid is then oxygenated into a prostaglandin (PG), PGH2, by a specific peroxidase called peroxynectin, but not by cyclooxygenase. PGH2 is then isomerized to various PGs such as PGA2, PGD2, PGE2, PGI2, and a thromboxane (TXB2). All four epoxyeicosatrienoic acids such as 5,6-EET, 8,9-EET, 11,12-EET, and 14,15-EET are also synthesized from arachidonic acid by oxygenation of vertebrate types of monooxygenases. However, the other type of eicosanoids called leukotrienes are found in insect tissues but their synthetic pathway is unclear. Eicosanoids mediate various insect physiological processes such as metabolism, excretion, immunity, and reproduction. Thus, identification of novel compounds interrupting eicosanoid biosynthesis would be a novel approach to develop insecticides. This review focuses on PGs and their immune mediation.
Despite several advances in identification of cardiac transcription factors, there are still needs to find new bioactive molecules that promote cardiomyogenesis from stem cells to highly efficient myocardial differentiation. We analyzed Illumina expression microarray data of mouse embryonic stem cells (mESCs)-derived cardiomyocytes. 276 genes were upregulated (≥ 4fold) in mESCs-derived cardiomyocytes compared undifferentiated ESCs. Secreted phosphoprotein 2 (Spp2) is one of candidates and is known to inhibit bone morphogenetic protein 2 (BMP2) signal transduction as a pseudoreceptor for BMP2. However, its function in cardiomyogenesis is unknown. We confirmed that Spp2 expression increased during the differentiation into functional cardiomyocytes using mESCs, TC-1/Kh2 and E14. Interestingly, Spp2 secretion transiently increased 3 days after formation of embryoid bodies (EBs), indicating that the extracellular secretion of Spp2 is involved in the differentiation of ESCs into cardiomyocytes. To characterize Spp2, we performed experiments using the C2C12 mouse myoblast cell line, which has the property of shifting the differentiation pathway from myoblastic to osteoblastic by treatment with BMP2. Similar to the differentiation of ESCs, transcription of Spp2 increased as C2C12 myoblasts differentiated into myotubes. In particular, Spp2 secretion increased dramatically in the early stage of differentiation. Furthermore, treatment with Spp2-Flag recombinant protein promoted the differentiation of C2C12 myoblasts into myotubes. Taken together, we suggest a novel bioactive protein Spp2 that differentiates ESCs into cardiomyocytes. This may be useful for understanding the molecular pathways of cardiomyogenesis and for experimental or clinical promotion of stem cell therapy for ischemic heart diseases.
Asia-pacific Journal of Multimedia Services Convergent with Art, Humanities, and Sociology
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v.7
no.1
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pp.395-405
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2017
To confirm potential anti-cancer activities of ethylacetate (EtOAc) fraction from Orostachys japonicus on the A549 human lung cancer cells, this study examined. As a result of conducting MTS assay for measuring cell viability, the EtOAc fraction inhibited the proliferation of A549 cells in a dose-dependent manner. To investigate whether the inhibiting A549 cell viability was caused by apoptosis, this study analyzed chromatin condensation in A549 cells using DAPI staining. The morphological changes such as the formation of nuclear condensation were formed in a dose-dependent manner. Also, this study performed Annexin V-FITC staining for detecting phosphatidylinositol (PS). As a result of Annexin V-FITC staining to investigate level of early and late apoptosis, the apoptosis level treated with EtOAc fraction was higher than that of control. RT-PCR was performed to study the correlation between G2/M cell cycle arrest and cell cycle control genes. The anti-cancer activity of EtOAc fraction was accompanied by inhibition of CDK1, 4, cyclin B1 and D1 mRNA. This study also examined the expression of various marker proteins: p53, Bax, Bcl-2 and pro-caspase 3. Western blotting revealed that p53 and Bax proteins were up-regulated, and Bcl-2 and pro-caspase 3 proteins down-regulated in a time and dose-dependent manner.
Jeong Woong Park;Marc Ndimukaga;Jaeyoung Heo;Ki-Duk Song
Korean Journal of Poultry Science
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v.50
no.4
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pp.193-202
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2023
Influenza IAVs are encapsulated negative-strand RNA viruses that infect many bird species' respiratory systems and can spread to other animals, including humans. This work reanalyzed previous microarray datasets to identify common and specific differentially expressed genes (DEGs) in chickens, as well as their biological activities. There were 760 and 405 DEGs detected in HPAIV and LPAIV-infected chicken cells, respectively. HPAIV and LPAIV have 670 and 315 DEGs, respectively, with both viruses sharing 90 DEGs. Because of HPAIV infection, numerous genes were implicated in a fundamental biological function of the cell cycle, according to the functional annotation of DEGs. Of the targeted genes, expressions of CDC Like Kinase 3 (CLK3), Nucleic Acid Binding Protein 1 (NABP1), Interferon-Inducible Protein 6 (IFI6), PIN2 (TERF1) Interacting Telomerase Inhibitor 1 (PINX1), and Cellular Communication Network Factor 4 (WISP1) were altered in DF-1 cells treated with polyinosinic:polycytidylic acid (PIC), a toll-like receptor 3 (TLR3) ligand, suggesting that transcription of these genes be controlled by TLR3 signaling. To gain a better understanding of the pathophysiology of AIVs in chickens, it is crucial to focus more research on unraveling the mechanisms through which AIV infections may manipulate host responses during the infection process. Insights into these mechanisms could facilitate the development of novel therapeutic strategies.
Vinclozolin (VCZ) is a systemic fungicide commonly used in fruits, vegetables and the wine industry. VCZ and its metabolites, butenoic acid (M1) and enanilide (M2) derivatives, act as anti-androgens through actions on the androgen receptor. Although there is growing body of evidence that VCZ's action as an endocrine disrupting chemical (EDC) in male reproductive physiology and pathphysiology, no evidence on the VCZ's EDC action in female is available yet. Previously we found that the prepubertal VCZ exposures could effectively delay the onset of puberty in female rats, suggesting the postponed or weakened activities of hypothalamus-pituitary-ovary (H-P-O) reproductive hormonal axis. The present study was performed to examine whether the VCZ administration affects the transcriptional activities of reproductive hormone-related genes in the same animal model. VCZ (10 mg/kg/day) was administered daily from postnatal day 21 (PND 21) through the day when the first vaginal opening (V.O.) was observed. To determine the transcriptional changes of reproductive hormone-related genes in hypothalamus and pituitary, total RNAs were extracted and applied to the semiquantitative reverse transcription polymerase chain reaction (RT-PCR). As a result, treatment with VCZ significantly lowered the transcriptional activity of nitric oxide synthase-2 (NOS-2) which is known to adjust gonadotropin-releasing hormone (GnRH) secretion in the hypothalamus (p<0.01). Similarly, the mRNA levels of KiSS-1, G protein-coupled receptor 54 (GPR54) and GnRH were significantly decreased in hypothalamus (p<0.01) from VCZ-treated group. As expected, the transcriptional activities of luteinizing hormone-${\beta}$ (LH-${\beta}$) and follicle stimulating hormone-${\beta}$ (FSH-${\beta}$) in the anterior pituitary from VCZ-treated group were also significantly lower than those from the control group. The present study indicates that(i) the inhibitory effect of VCZ exposure on the onset of puberty in immature female rats could be derived from the reduced transcriptional activities of gonadotropin subunits and their upstream modulators such as GnRH and KiSS-1 in hypothalamus-pituitary neuroendocrine axis, and (ii) these inhibitory effects could be mediated by NO signaling pathway.
Objectives: The analysis of heart rate variability (HRV) is a useful non-invasive tool to investigate the autonomic nerve function. Previous studies on the relationship between HRV and depression have been reported controversial results. Similarly, the correlation between the serum lipids and depression is debatable. The purpose of this study is to examine the relationship between heart rate variability, lipid profile and depression. Methods: A total of 42 patients with major depressive disorder (MDD) and 32 age and sex-matched normal subjects who had no previous history of major medical and mental illnesses were recruited for this study. A structured-interview was used to assess the general characteristics and psychiatric illness. HRV measures were assessed by time-domain and frequency-domain analyses. Psychological symptoms were measured using the Hamilton rating scale for anxiety (HAM-A), Hamilton rating scale for depression (HAM-D). In addition, the evaluation for lipid profile was performed by blood test. Results: In serum lipid profile test, MDD group showed higher cholesterol ($197.68{\pm}42.94$ mg/dL vs. $176.85{\pm}34.68$ mg/dL, p=0.044), TG ($139.45{\pm}92.54$ mg/dL vs. $91.4{\pm}65.68$ mg/dL, p=0.018), LDL ($130.03{\pm}33.18$ vs. $106.62{\pm}27.08$, p=0.004) level than normal control group. In HRV time domain analyses, the standard deviation of the NN interval (SDNN) was decreased in MDD group than normal control group, but was not significant ($32.82{\pm}14.33$ ms vs. $40.36{\pm}21.40$ms, p=0.078). ApEn (Approximate Entrophy) was significantly increased in MDD group than normal control group ($1.13{\pm}0.11$ vs. $0.91{\pm}0.18$, p<0.001). ApEn was correlated with LDL level (r=0.277, p=0.028), HAM-D scores (r=0.534, p<0.001) and HAM-A scores (r=0.470, p<0.001). Conclusions: MDD patients showed increased ApEn, one of the HRV measurement. And this ApEn was correlated with LDL, HAM-D and HAM-A scores. In this study, the analysis of ApEn would be a useful test of MDD.
Kim Jeong-Seok;Kim Bum-Soo;Jeun Sin-Soo;Jung So-Lyung;Choe Bo-Young
Investigative Magnetic Resonance Imaging
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v.9
no.1
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pp.43-49
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2005
A few studies have shown that the function of fusiform face area is selectively involved in the perception of faces including a race difference. We investigated the neural substrates of the face-selective region called fusiform face area in the ventral occipital-temporal cortex and same-race memory superiority in the fusiform face area by the event-related fMRI. In our fMRI study, subjects (Oriental-Korean) performed the implicit distinction of the race while they consciously made familiar-judgments, regardless of whether they considered a face as Oriental-Korean or European-American. For race distinction as an implicit task, the fusiform face areas (FFA) and the right parahippocampal gyrus had a greater response to the presentation of Oriental-Korean faces than for the European-American faces, but in the conscious race distinction between Oriental-Korean and European-American faces, there was no significant difference observed in the FFA. These results suggest that different activation in the fusiform regions and right parahippocampal gyrus resulting from superiority of same-race memory could have implicitly taken place by the physiological processes of face recognition.
This study investigated the physiological characteristics of various medicinal plant extracts including Crataegi Fructus (CF), Perilla frutescens Britton var. acuta Kudo (PF), Geranium thunbergii sieb. et Zucc. (GT) and Polygonum multiflorum Thunberg (PM) for use in extending the shelf-life and improving the functional properties of seasoned pork. CF and GT showed a wide spectrum of antimicrobial activities against 5 kinds of Gram (+) bacteria and 5 kinds of Gram (-) bacteria. The antimicrobial activities of the extracts decreased as the heat treatment temperature increased between 60 and $121^{\circ}C$, but their activities remained relatively high following heat treatment. The total phenolic compound contents of the CF, PF, GT, PM extracts were 66.2, 29.0, 96.6 and 13.3 mg/g, respectively. The order of their DPPH radical scavenging activity was GT > CF > PF > PF. The nitrite scavenging activities of the CF, PF and GT extracts at 0.1% concentration were 10.5, 1.6 and 3.8%, respectively. The GT extract (99.7%) showed the highest degree of lipid rancidity inhibition compared to CF (90.3%), PF (88.9%) and PM (41.2%). After heat treatment for 15 min at $121^{\circ}C$, the DPPH radical scavenging activity of CF decreased but it increased in GT and PM. The nitrite scavenging activities of CF and GT decreased after heat treatment for 15 min at $121^{\circ}C$ but remained at relatively high levels. Also, degrees of lipid rancidity inhibition remained relatively high in PF (86.3%) and GT (99.8%) after heat treatment for 15 min at $121^{\circ}C$. These results suggest that CF and GT extracts could be used as ingredients in seasoning sauce to improve the shelf-life and functional properties of seasoned pork.
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