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p66Shc in sheep preimplantation embryos: Expression and regulation of oxidative stress through the manganese superoxide dismutase-reactive oxygen species metabolic pathway

  • Tong Zhang;Jiaxin Zhang;Ruilan Li
    • Animal Bioscience
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    • v.36 no.7
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    • pp.1022-1033
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    • 2023
  • Objective: p66Shc, a 66 kDa protein isoform encoded by the proto-oncogene SHC, is an essential intracellular redox homeostasis regulatory enzyme that is involved in the regulation of cellular oxidative stress, apoptosis induction and the occurrence of multiple age-related diseases. This study investigated the expression profile and functional characteristics of p66Shc during preimplantation embryo development in sheep. Methods: The expression pattern of p66Shc during preimplantation embryo development in sheep at the mRNA and protein levels were studied by quantitative real-time polymerase chain reaction (RT-qPCR) and immunofluorescence staining. The effect of p66Shc knockdown on the developmental potential were evaluated by cleavage rate, morula rate and blastocyst rate. The effect of p66Shc deficiency on reactive oxygen species (ROS) production, DNA oxidative damage and the expression of antioxidant enzymes (e.g., catalase and manganese superoxide dismutase [MnSOD]) were also investigated by immunofluorescence staining. Results: Our results showed that p66Shc mRNA and protein were expressed in all stages of sheep early embryos and that p66Shc mRNA was significantly downregulated in the 4-to 8-cell stage (p<0.05) and significantly upregulated in the morula and blastocyst stages after embryonic genome activation (EGA) (p<0.05). Immunofluorescence staining showed that the p66Shc protein was mainly located in the peripheral region of the blastomere cytoplasm at different stages of preimplantation embryonic development. Notably, serine (Ser36)-phosphorylated p66Shc localized only in the cytoplasm during the 2- to 8-cell stage prior to EGA, while phosphorylated (Ser36) p66Shc localized not only in the cytoplasm but also predominantly in the nucleus after EGA. RNAi-mediated silencing of p66Shc via microinjection of p66Shc siRNA into sheep zygotes resulted in significant decreases in p66Shc mRNA and protein levels (p<0.05). Knockdown of p66Shc resulted in significant declines in the levels of intracellular ROS (p<0.05) and the DNA damage marker 8-hydroxy2'-deoxyguanosine (p<0.05), markedly increased MnSOD levels (p<0.05) and resulted in a tendency to develop to the morula stage. Conclusion: These results indicate that p66Shc is involved in the metabolic regulation of ROS production and DNA oxidative damage during sheep early embryonic development.

Chemical profile and antioxidant activity of peel of Yellowball, a novel citrus variety

  • Sun Lee;Seong-Ho Jo;Ji-Hyun An;Seong-man Jeong;Dong-Shin Kim;Sang Suk Kim;Suk Man Park;Su Hyun Yun;Seung-Gab Han;Hyun-Jin Kim
    • Food Science and Preservation
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    • v.30 no.2
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    • pp.235-246
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    • 2023
  • Yellowball (Citrus hybrid cv. Yellowball ) is a new citrus hybrid between Haruka (C. tamurana × natsudaidai ) and Kiyomi (C. unshiu × sinensis) and is known to possess strong antioxidant activity. However, detailed information on the antioxidant components of its peel has not yet been reported. This study evaluated the antioxidant activity of the peel and identified the antioxidant components by fractionating a methanolic extract of Yellowball peels using liquid-liquid extraction with n-hexane, ethyl ether (ether), ethyl acetate (EA), butanol, and water. The phenolic contents and antioxidant activities of the n-hexane, ether, and EA fractions were higher than those of the other fractions, and these fractions were further separated by semi-preparative high-performance liquid chromatography (HPLC). Four antioxidant peaks, EA1, EA2, EA3, and He1, were isolated and analyzed using ultra-performance liquid chromatography-quadrupole-time- of-flight mass spectrometry (UPLC-Q-TOF MS). Sinapoyl glucoside and hesperidin were identified in EA2 and EA3, respectively, and a polymethoxylated flavone (PMF) complex (5-hydroxy-3,6,7,8,3',4'-hexamethoxyflavone, natsudaidain, tetrameth- oxyflavone, and tangeretin) was identified in He1. A compound in EA1 with m/z 223.0246 [M-H] could not be identified and was named unknown2. The antioxidant activity of unknown2 (IC50=69.17 ㎍/mL) was similar to that of Trolox, which was noted as a major antioxidant in Yellowball peel. Further studies on the antioxidant capacity of Yellowball peel are required; however, these results provide a foundation for using Yellowball peel as an antioxidant.

Plasma D-dimer Determination in the Diagnosis of Pulmonary Embolism (폐색전증의 진단에 있어 혈청 D-dimer 측정의 진단적 가치)

  • Lim, Chae-Man;Kim, Hong-Kyu;Choi, Kang-Hyun;Lee, Sand-Do;Koh, Youn-Suck;Kim, Woo-Sung;Jang, Jae-Suck;Kim, Dong-Soon;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.1
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    • pp.69-74
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    • 1996
  • Background: The diagnosis of pulmonary embolism (PE) based on clinical findings is often elusive and therefore requires confirmative diagnostic method. Pulmonary angiography, though the gold standard for the diagnosis of pulmonary embolism, is an invasive method and requires trained personnel and special equipment. Lung V/Q scan, on the other hand, is a noninvasive method but the diagnostic specificity and sensitivity arc not satisfactory in case that the results are either intermediate or low probability scan. Plasma D-dimer is generated when a thrombus is fibrinolysed by plasmin and is known to be increased in various thrombotic disorders. The aim of this study was to investigate the value of the determination of plasma D-dimer level in the diagnosis of pulmonary embolism. Methods: Pulmonary angiography was performed in 17 patients who were clinically suspected to have pulmonary embolism. 9 patients(PE, $56{\pm}13.4$ yrs, M:F=8:1) were diagnosed to have pulmonary embolism by pulmonary angiography. The control group were the 8 patients with negative pulmonary angiography and 13 orthopedic patients with no evidence of pulmonary embolism on scintigraphic and impedance plethysmographic studies(n=21) (non-PE, $54.5{\pm}11.1$ yrs, M:F=11:10). Plasma D-dimer was measured by latex agglutination method in study subjects and the results were analyzed according to the presence or absence of pulmonary embolism. Results: 1) The increased level of plasma D-dimer was more frequently observed in the patients with pulmonary embolism than in the controls(>0.5 mg/L, 8 in PE, 10 in non-PE; <0.5 mg/L, 1 in PE, 11 in non-PE, p=0.049). 2) The diagnostic value of plasma D-dimer level higher than 0.5 mg/L were as follows: sensitivity 88.9%(8/9), specificity 52.4%(11/21), positive predictive value 44.4%(8/18), and negative predictive value 91.7%(11/12). Conclusion: Plasma D-dimer determination showed high sensitivity and negative predictive value in the diagnosis of pulmonary embolism and is therefore thought to be useful in excluding the possibility of pulmonary embolism.

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An Experimental Study on the Required Performances of Roof Concrete Placed in the In-ground LNG Storage Tank (지하식 LNG 저장탱크의 지붕 콘크리트의 요구성능에 관한 실험적 연구)

  • Kwon, Yeong-Ho
    • Journal of the Korea Concrete Institute
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    • v.25 no.3
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    • pp.339-345
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    • 2013
  • This study is to derive from the required performances and the optimum mix proportion of the roof concrete placed in the in-ground LNG storage tank with a capacity of 200000 $m^3$, and propose the actual data for site concrete work. The concrete placing work without sliding and segregation in the fresh concrete condition is very important because the slope of domed roof is varied in the large range by its curvature. Also the control of hydration heat and the strength development at test ages are classified with massive section about 1.4 m thick and considered to the pre-stressing work and removal of air support after concrete placing work. Considering above condition, slump range is selected $100{\pm}25$ mm under the slope $20^{\circ}$ and $150{\pm}25$ mm over the slope $20^{\circ}$ s until 60 minutes of elapsed time. Also, the roof concrete is satisfied with compressive strength range including design strength at 91 days (30 MPa), pre-stressing work at 7 days (10 MPa), air support removal work at 21 days (14 MPa). Replacement ratio of limestone powder is determined by confined water ratio test and main design factors include water-cement ratio (W/C), sand-aggregate ratio and dosage of admixture. As test results, the optimum mix proportion of the roof concrete used low heat cement is as followings. 1) Replacement ratio of limestone powder 25% by confined water ratio test 2) Water-cement ratio 57.8% 3) Sand-aggregate ratio 42.0%. Also, test results for the adiabatic temperature rising test is satisfied with its criteria and shown the lower value compared to preceding storage tank (TK-13, 14). These required performances and the optimum mix proportion is to apply the actual construction work.

Phellinus linteus Extract Regulates Macrophage Polarization in Human THP-1 Cells (상황버섯 추출물의 인간 유래 THP-1 단핵구 세포주의 분극화 조절)

  • Lee, Sang-Yull;Park, Sul-Gi;Yu, Sun-Nyoung;Kim, Ji-Won;Hwang, You-Lim;Kim, Dong-Seob;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.30 no.2
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    • pp.113-121
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    • 2020
  • Macrophages are initiators for regulating a host's defenses to eliminate pathogens and trigger tissue repair. Macrophages are classified into two types: classically (M1) activated macrophages and alternatively (M2) activated macrophages. M1-phenotype macrophages directly or indirectly kill infectious organisms and tumor cells via pro-inflammatory responses, whereas M2-phenotype macrophages remodel wounded tissue through anti-inflammatory responses. In this paper, we investigated how Phellinus linteus hot water extract passed from Diaion HP-20 resin (PLEP) regulates polarization of M1-like or M2-like macrophages in human THP-1 cells. PLEP did not have cytotoxicity at a high concentration of 300 ㎍/ml. We observed morphological alteration of the THP-1 cells, which are stimulated by PLEP, LPS/INF-γ (M1 stimulators) or IL-4/IL13 (M2 stimulators). PLEP exposure induced morphology contiguous with LPS/INF-γ. qPCR was also performed to determine whether PLEP influences M1 or M2 polarization-related genes. M1-phenotype macrophage-specific genes, such as TNF-α, IL-1β, IL-6, IL-8, CXCL10 and CCR7, were enhanced by PLEP in a dose-dependent manner similar to LPS/INF-γ. Conversely, M2-phenotype-specific genes, such as MRC-1, DC-SIGN, CCL17 and CCL22, were suppressed by PLEP. PLEP also significantly up-regulated secretory inflammation cytokines related to M1 polarization of macrophages, including TNFα, IL-1β and IL-6, which was similar to the gene expression. Further, MAPK and NF-κB signaling were increased by treatment with PLEP, resulting in enhancement of cytokine secretion. PLEP might therefore be used as a promising booster of pro-inflammatory responses through M1 polarization of human THP-1 cells.

Report on the External Audits Conducted by Korean Society of Medical Physics (한국의학물리학회 선형가속기 외부 품질관리 실시 현황보고)

  • Huh, Hyun Do;Cho, Kwang Hwan;Cho, Sam Ju;Choi, Sang Hyoun;Kim, Dong Wook;Hwang, Ui-Jung;Kim, Ki Hwan;Min, Chul Kee;Choi, Tae Jin;Oh, Young Kee;Lee, Seoung Jun;Park, Dahl;Park, Sung-Kwang;Ji, Young Hoon
    • Progress in Medical Physics
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    • v.24 no.4
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    • pp.315-322
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    • 2013
  • The aim of this work is to verify the self-quality assurances in medical institutions in Korea through the external audits by the group of experts and have a mutual discussion of the systematic problems. In order to validate the external audits 30 of 80 medical institutions across the nation were picked out considering the regional distribution and the final 25 institutions applied voluntarily to take part in this work. The basic rules were setup that any information of the participants be kept secrete and the measurements be performed with the dosimetry system already verified through intercomparision. The outputs for 2 or more photon beams, the accuracy of gantry rotation and collimator rotation and the poistional accuracy of MLC movement were measured. The findings for the output measurement showed the differences of -0.8%~4.5%, -0.79%~3.01%, and -0.7%~0.07% with respect to that of the verified dosimetry system for the 6MV, 10MV, and 15MV, respectively. For the reference absorbed dose 8 (16%) of 50 photon beams in 25 medical institutions differed 2.0% or greater from the reference value. The coincidences of Field size with x-ray beam and radiation isocenters of Gantry roration and collimator rotation gave the results of within ${\pm}2$ mm for every institute except 2 institutions. The positional accuracy of MLC movement agreed to within ${\pm}1$ mm for every institute. For the beam qualities of 6 MV photon beams kQ values showed the distribution within 0.4% between maximum and minimum. For the protocols 21 institutions (84%) used absorbed dose to water based protocol while 4 insitutions (16%) used air kerma based one. 22 institutions employed the SSD technique while 3 institutions did the SAD one. External audit plays an important role in discovering the systematic problems of self-performing Quality Assurances and having in depth discussion for mutual complementation. Training experts of international level as well as national support system are required so that both the group of experts of medical physicists and government laboratory could perform together periodical and constant external audits.

Metabolites profiling and hypolipidemic/hypocholesterolemic effects of persimmon (Diosyros kaki Thumb.) by different processing procedures: in vitro and in vivo studies (제조방법에 따른 떫은감 (Diosyros kaki Thumb.)의 대사체 프로파일링과 중성지질/콜레스테롤 대사 관련 유전자발현 연구 : in vitro 및 in vivo 연구)

  • Park, Soo-Yeon;Oh, Eun-Kyung;Lim, Yeni;Shin, Ji-Yoon;Jung, Hee-Ah;Park, Song-Yi;Lee, Jin Hee;Choe, Jeong-Sook;Kwon, Oran
    • Journal of Nutrition and Health
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    • v.51 no.4
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    • pp.275-286
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    • 2018
  • Purpose: Our previous study demonstrated that persimmon (Diospyros kaki Thumb.) at different stages of ripening provided different protective effects against high-fat/cholesterol diet (HFD)-induced dyslipidemia in rats. In this study, we compared the metabolites profile and gene expressions related to triglyceride (TG)/cholesterol metabolism in vitro and in vivo after treating with persimmon water extracts (PWE) or tannin-enriched persimmon concentrate (TEP). Methods: Primary and secondary metabolites in test materials were determined by GC-TOF/MS, UHPLC-LTQ-ESI-IT-MS/MS, and UPLC-Q-TOF-MS. The expression of genes related to TG and cholesterol metabolism were determined by RT-PCR both in HepG2 cells stimulated by oleic acid/palmitic acid and in liver tissues obtained from Wistar rats fed with HFD and PWE at 0, 150, 300, and 600 mg/d (experiment I) or TEP at 0, 7, 14, and 28 mg/d (experiment II) by oral gavage for 9 weeks. Results: PLS-DA analysis and heatmap analysis demonstrated significantly differential profiling of metabolites of PWE and TEP according to processing of persimmon powder. In vitro, TEP showed similar hypolipidemic effects as PWE, but significantly enhanced hypocholesterolemic effects compared to PWE in sterol regulatory element-binding protein 2 (SREBP2), HMG-CoA reductase (HMGCR), proprotein convertase subtilisin/kexin type 9 (PCSK9), cholesterol $7{\alpha}-hydroxylase$ (CYP7A1), and low density lipoprotein receptor (LDLR) gene expression. Consistently, TEP and PWE showed similar hypolipidemic capacity in vivo, but significantly enhanced hypocholesterolemic capacity in terms of SREBP2, HMGCR, and bile salt export pump (BSEP) gene expression. Conclusion: These results suggest that column extraction after hot water extraction may be a good strategy to enhance tannins and long-chain fatty acid amides, which might cause stimulation of hypocholesterolemic actions through downregulation of cholesterol biosynthesis gene expression and upregulation of LDL receptor gene expression.

Early Changes after Death of Plaice, Paralichthys olivaceus Muscle -1. Relationship between Early Changes after Death and Temperature Dependency- (넙치(Paralichthys olivaceus)육의 사후 조기 변화 -1. 사후 조기 변화와 온도 의존성의 관계-)

  • KIM Yuck-Yong;CHO Young-Je
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.25 no.3
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    • pp.189-196
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    • 1992
  • To know the extension effect of storage temperature on the pre-rigor period of plaice, Paratichthys olivaceus muscle, the relationship between early changes after death and temperature dependency was studied. Killed plaices instantly with spiking at the brain were stored at $-3^{\circ}C,\;0^{\circ}C,\;5^{\circ}C\;and\;10^{\circ}$ used in studying the changes in rigor Index, ATP and its related compounds, lactate contents and K-value. The most shortest onset time of rigor-mortis and full rigor was revealed in the sample stored at $-3^{\circ}C$ among the all samples, where rigor-mortis began at 4hrs after spiking and maximum tension was attained after 28hrs. However, in case of fresh plaice muscle stored at $10^{\circ}C$, the onset of rigor-mortis and full rigor were retarded to 14hrs and 52hrs after spiking. ATP in sample stored at $5^{\circ}C\;and\;10^{\circ}C$ were decomposed slowly than sample at $0^{\circ}C\;-3^{\circ}C $ and within 35hrs storage. The fastest rate and the maximum content of lactate accumulation were showed in sample stored at $-3^{\circ}$ among the all samples. The correlation coefficient(r) between the rate of rigor mortis and ATP breakdown, rigor mortis and lactate accumulation, and ATP breakdown and lactate accumulation were -0.981946, 0.965044, and -0.964728, respectively. Freshness of $-3^{\circ}C$ stored samples was maintained for the longest time compared with other stored samples. The times reached around $20\%$ of K-value were 240hrs for samples stored at $-3^{\circ}C,\;96hrs\;for\;0^{\circ}C\;samples,\;71hrs\;for\;5^{\circ}C\;samples,\;and\;22hrs\;for\;10^{\circ}C$ samples. Samples stored at $-3^{\circ}C,\;and\;0^{\circ}C$ were showed higher temperature dependency on rate of rigor-mortis, ATP breakdown, and lactate accumulation than $5^{\circ}C\;and\;10^{\circ}C$ stored samples, but those samples have a lower temperature dependency on K-value.

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The Effect of Beef Peptide on Blood Pressure and Serum Lipid Concentration of Spontaneously Hypertensive Rat(SHR) (우육 단백질에서 추출된 펩타이드가 자연발증 고혈압쥐의 혈압과 혈중 지질농도에 미치는 영향)

  • Jang, A.;Cho, Y.J.;Lee, J.I.;Shin, J.H.;Kim, I.S.;Lee, M.
    • Journal of Animal Science and Technology
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    • v.46 no.1
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    • pp.107-114
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    • 2004
  • The purpose of this work was to examine the effect of peptide on blood pressure and serum lipid concentration of spontaneously hypertensive rat(SHR). This peptide was extracted from beef muscle hydrolysates and identified as hexapeptide, V-L-A-Q-Y-K. This peptide showed angiotensin converting enzymc(ACE) inhibition activity in vitro experimentation$(IC_50: I38.34{\mu}\ell$/ml). Diets containing 0.2g. 0.5g. and 1.0g of the peptide per kg body weight were fed to SHR every day for 8 weeks while the control group was ted on a diet and lml of drinking water instead of the peptide. Total cholesterol and LDL-cholesterol concentrations of the treatment groups were lower than those of the control diet feeding group. The significant suppression of systolic blood pressure was shown by increasing the concentration of peptide supplement. especially by 3 weeks of feeding. although it started fluctuating later. These results suggest that the peptide may beneficially affect blood pressure in spontaneously hypertensive rat by the 3-week administration.

Velocity-effective stress response of $CO_2$-saturated sandstones ($CO_2$로 포화된 사암의 속도-유효응력 반응)

  • Siggins, Anthony F.
    • Geophysics and Geophysical Exploration
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    • v.9 no.1
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    • pp.60-66
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    • 2006
  • Three differing sandstones, two synthetic and one field sample, have been tested ultrasonically under a range of confining pressures and pore pressures representative of in-situ reservoir pressures. These sandstones include: a synthetic sandstone with calcite intergranular cement produced using the CSIRO Calcite In-situ Precipitation Process (CIPS); a synthetic sandstone with silica intergranular cement; and a core sample from the Otway Basin Waarre Formation, Boggy Creek 1 well, from the target lithology for a trial $CO_2$ pilot project. Initial testing was carried on the cores at "room-dried" conditions, with confining pressures up to 65 MPa in steps of 5 MPa. All cores were then flooded with $CO_2$, initially in the gas phase at 6 MPa, $22^{\circ}C$, then with liquid-phase $CO_2$ at a temperature of $22^{\circ}C$ and pressures from 7 MPa to 17 MPa in steps of 5 MPa. Confining pressures varied from 10 MPa to 65 MPa. Ultrasonic waveforms for both P- and S-waves were recorded at each effective pressure increment. Velocity versus effective pressure responses were calculated from the experimental data for both P- and S-waves. Attenuations $(1/Q_p)$ were calculated from the waveform data using spectral ratio methods. Theoretical calculations of velocity as a function of effective pressure for each sandstone were made using the $CO_2$ pressure-density and $CO_2$ bulk modulus-pressure phase diagrams and Gassmann effective medium theory. Flooding the cores with gaseous phase $CO_2$ produced negligible change in velocity-effective stress relationships compared to the dry state (air saturated). Flooding with liquid-phase $CO_2$ at various pore pressures lowered velocities by approximately 8% on average compared to the air-saturated state. Attenuations increased with liquid-phase $CO_2$ flooding compared to the air-saturated case. Experimental data agreed with the Gassmann calculations at high effective pressures. The "critical" effective pressure, at which agreement with theory occurred, varied with sandstone type. Discrepancies are thought to be due to differing micro-crack populations in the microstructure of each sandstone type. The agreement with theory at high effective pressures is significant and gives some confidence in predicting seismic behaviour under field conditions when $CO_2$ is injected.