• Title/Summary/Keyword: Embryonic development

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Studies on In Vitro Culture, Freezing and Transfer of Korean Native Cattle Embryos Fertilized In Vitro I. Effect of Co-culture Cells and Growth Factors on In Vitro Development of Korean Native Cattle Embryos Fertilized In Vitro (한우 체외수정란의 체외배양, 동결보존 및 이식에 관한 연구 I. 한우 체외수정란의 체외배양에 대한 공배양세포와 성장인자의 효과)

  • 김일화;손동수;이호준;최선호;양병철;이광원;김경남;장인호
    • Journal of Embryo Transfer
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    • v.11 no.2
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    • pp.111-124
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    • 1996
  • The present study was carried out to investigate the effects of co-culture cells and growth factors on in vitro culture of Korean native cattle(KNC) embryos fertilized in vitro. Two-eight cell embryos were cultured in vitro using 4 types of co-culture cells and 3 growth factors singly or in combination. The results were as follows, In the co-culture of 2~8 cell embryos with bovine oviductal epithelial cell(BOEC), granulosa cell(BGC), uterine epithelial cell(BUEC) and mouse embryonic fibroblast (MEF) monolayers, the developing rate to blastocysts was significantly(P<0.05) higher with BUEC(32.1%) than with MEF(15.3%), BGC(13.2%) and non co-culture control(11.6%). When the morula co-cultured with BOEC for 5 days following in vitro fertilization were co-cultured with BOEC continuously or with BUEC, respectively, the developing rate to blastocysts was higher with BUEC(73.9%) than with BOEC(56.0%). To examine the effects of growth factors on in vitro development of 2~8 cell embryos, epidermal growth factor(EGF), transforming growth factor-$\beta$l(TGF-$\beta$l) and insulin-like growth factor-1(IGF-1) were added singly or in combination to TCM 199 maturation medium with respective concentration. In a addition of each 10, 30 and SOng /rnl EGF, the developing rate to blastocysts was the highest in lOng /ml EGF(25.3%). In addition of each 1, 2 and Sng /mi TGF-$\beta$1, the developing rate to blastocysts was the highest in lng /ml TGF-$\beta$1(28.8%). In addition of each 50, 100ng/ml JGF-l, the developing rate to blastocysts was higher in 100ng/ml IGF-l(16.5%) than in SOng/mi IGF-1(12.9%). When lOng /ml EGF and lng /ml TGF-$\beta$l was added singly or in combination, the developing rate to blastocysts was similar in groups added singly or in combination with EGF and TGF-$\beta$l (23.l~24.6%), although higher than in control(16.7%). In the co-culture of 2~8 cell embryos Wth BOEC + each 10, 30 and 5Ong /rnl EGF, the developing rate to blastocysts was significantly(p<0.05) higher in BOEC + long /ml EGF(32.3%) than in BOEC + 3Ong /ml EGF(18.9%) and BOEC + song /ml EGF(9.7%). In the co-culture of 2~8 cell embryos with BOEC + each 1, 2, Sng /ml TGF-$\beta$l the developing rate to blastocysts was higher in BOEC + Sng/rnl TGF-$\beta$l(28.2%) than in BOEC + lng /ml TGF-$\beta$l(21.7%) and BOEC + 2ng/ml TGF-$\beta$l(21.4%). In summary, higher developing rate to blastocysts were obtained with co-culture of BUEC for co-culture system, with addition of lOng /ml EGF or lng /ml TGF-$\beta$l for growth factor culture system, and with co-culture of BOEC + lOng /ml EGF or BOEC + Sng /ml TGF-$\beta$l for co-culture + growth factor culture system.

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Five Most Common Prognostically Important Fusion Oncogenes are Detected in the Majority of Pakistani Pediatric Acute Lymphoblastic Leukemia Patients and are Strongly Associated with Disease Biology and Treatment Outcome

  • Awan, Tashfeen;Iqbal, Zafar;Aleem, Aamer;Sabir, Noreen;Absar, Muhammad;Rasool, Mahmood;Tahir, Ammara H.;Basit, Sulman;Khalid, Ahmad Mukhtar;Sabar, Muhammad Farooq;Asad, Sultan;Ali, Agha Shabbir;Mahmood, Amer;Akram, Muhammad;Saeed, Tariq;Saleem, Arsalan;Mohsin, Danish;Shah, Ijaz Hussain;Khalid, Muhammad;Asif, Muhammad;Haq, Riazul;Iqbal, Mudassar;Akhtar, Tanveer
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.11
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    • pp.5469-5475
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    • 2012
  • Background and Objectives: Acute lymphoblastic leukemia (ALL) is a complex genetic disease involving many fusion oncogenes (FO) having prognostic significance. The frequency of various FO can vary in different ethnic groups, with important implications for prognosis, drug selection and treatment outcome. Method: We studied fusion oncogenes in 101 pediatric ALL patients using interphase FISH and RT-PCR, and their associations with clinical features and treatment outcome. Results: Five most common fusion genes i.e. BCR-ABL t (22; 9), TCF3-PBX1 (t 1; 19), ETV6-RUNX1 (t 12; 21), MLL-AF4 (t 4; 11) and SIL-TAL1 (del 1p32) were found in 89/101 (88.1%) patients. Frequency of BCR-ABL was 44.5% (45/101). BCR-ABL positive patients had a significantly lower survival ($43.7{\pm}4.24$ weeks) and higher white cell count as compared to others, except patients with MLL-AF4. The highest relapse-free survival was documented with ETV6-RUNX1 (14.2 months) followed closely by those cases in which no gene was detected (13.100). RFS with BCR-ABL, MLL-AF4, TCF3-PBX1 and SIL-TAL1 was less than 10 months (8.0, 3.6, 5.5 and 8.1 months, respectively). Conclusions: This is the first study from Pakistan correlating molecular markers with disease biology and treatment outcome in pediatric ALL. It revealed the highest reported frequency of BCR-ABL FO in pediatric ALL, associated with poor overall survival. Our data indicate an immediate need for incorporation of tyrosine kinase inhibitors in the treatment of BCR-ABL+ pediatric ALL in this population and the development of facilities for stem cell transplantation.

Ecological Risk Assessment of Alachlor using Medaka (Oryzias latipes) and Earthworm (Eisenia fetida) (송사리(Oryzias latipes)와 지렁이(Eisenia fetida)를 이용한 Alachlor의 생태 위해성평가)

  • Lee, Chul-Woo;Kim, Hyun-Mi;Yoon, Jun-Heon;Song, Sang-Hwan;Ryu, Ji-Sung;Kim, Eun-Kyoung;Yang, Chang-Yong;Chung, Young-Hee;Choi, Kyung-Hee;Lee, Moon-Soon
    • Korean Journal of Environmental Biology
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    • v.25 no.1
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    • pp.1-7
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    • 2007
  • Medaka(Oryzias latipes) and earthworm(Eisenia fetida) toxicity tests were carried out and ecological risk assessment in water and soil was performed with national monitoring data. NOEC of alachlor was $100\;{\mu}g\;L^{-1}$ in the medaka early life-stage test. Embryonic development, hatchability and time to hatching of medaka eggs were affected by this chemical. The $LC_{50}$ and NOEC of alachlor were $94.1\;mg\;kg^{-1}\;and\;55.0\;mg\;kg^{-1}$, respectively, in the earthworm acute toxicity test. The environmental monitoring has been carrying out by NIER since 1999. Exposure levels of alachlor in water and soil were $ND{\sim}0.54\;{\mu}g\;L^{-1}\;and\;ND{\sim}0.9\;{\mu}g\;kg^{-1}$, respectively, in national monitoring data which had been performed from 2000 to 2004. The measured water and soil exposure levels were applied to evaluate the environmental risk assessment. The PNEC of alachlor in water and in soil were determined as $1\;{\mu}g\;L^{-1}\;and\;55.0\;{\mu}g\;kg^{-1}$, respectively using the safety factors which were suggested in EU and OECD. The HQs (PEC/PNEC) were determined to be below 1 for both water and soil when the maximum exposure levels ($0.54\;{\mu}g\; L^{-1}$ in water and $0.9\;{\mu}g\;kg^{-1}$ in soil) were applied. Conclusively, our study indicated that there was not significant ecological risk of alachlor in water and soil of our monitoring sites.

Influence of Temperature on the Egg Production and Hatching of Microcotyle sebastis (Monogenea : Microcotylidae), Parasitic on Rockfish, Sebastes schlegeli (수온이 조피볼락에 기생하는 아가미흡충(Microcotyle sebastis)의 산란과 부화에 미치는 영향)

  • Kim, Ki-Hong;Choi, Eun-Seok;Cho, Jae-Bum;Hwang, Yoon-Jung;Park, Soo-Il
    • Journal of fish pathology
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    • v.11 no.2
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    • pp.113-117
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    • 1998
  • The influence of temperature on the rate of egg production and embryonic development of Microcotyle sebastis was investigated to determine the precise time of a second treatment. The survival time of the adults of M. sebastis was inversely proportional to temperature. The number of laid eggs per each replicate during the first 24 h was $39.3{\pm}4.0$ at $10^{\circ}C$, $62.7{\pm}14.2$ at $15^{\circ}C$, $101.0{\pm}5.6$ at $20^{\circ}C$ and $89.0{\pm}11.0$ at $25^{\circ}C$. The time required for egg hatching of M. sebastis was $31.30{\pm}4.88$, $17.52{\pm}3.24$, $11.59{\pm}3.02$ and $10.76{\pm}3.10$ days at 10, 15, 20 and $25^{\circ}C$, respectively. The regression models of the time required for the beginning and 50% point of hatching according to the different temperatures were as follows; Beginning of hatch: D=58.2000-$4.2067{\times}Temp+0.0867{\times}(Temp)^2$ ($P\leq0.01$), 50% of hatch: D=91.3833-$7.5767{\times}Temp+0.1767{\times}(Temp)^2$ ($P\leq0.01$).

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Association with Kinetic Characteristics of sperm in Duroc Boar and the Zygote Arrest 1 gene Polymorphism (g.2540T>C) (Zygote arrest 1 유전자 변이(g.2540T>C)와 두록 정액의 운동학적 특성과의 연관성 분석)

  • Lee, Mi Jin;Ko, Jun Ho;Cho, Kyu Ho;Choi, Tae Jeong;Kim, Yong Min;Kim, Young Sin;Jin, Dong Il;Cho, Eun seok;Kim, Nam Hyung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.9
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    • pp.116-123
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    • 2018
  • The sperm quality is determined by the kinetic characteristics and acrosome integrity of the sperm. In the previous studies, analysis of semen quality had large errors because those experiments by using microscope had been conducted by people. In recent years, the molecular biological methods have been newly developed to complement the previous techniques. The ZAR1 gene is known to be a gene that affects early embryonic development in vertebrates, but there is no study of the association with semen. In this study, we analyzed the association between the kinetic characteristics and ZAR1 single nucleotide polymorphism (SNP) genotype. To detect the SNPs, we performed sequencing using genomic DNA from the whole bloods of Duroc pigs. We identified an SNP in the ZAR1 gene g.2540T>C. ZAR1 SNP genotypeing in 105 pigs revealed that the major and minor alleles were T and C, respectively. After we analyzed the association between the kinetic characteristics of sperm and the ZAR1 SNP genotype, we found a significant association in MOT (p<0.01), VSL (p<0.05) of the kinetic characteristics in the Duroc boars. It was confirmed that the boars with T allele were lower in MOT and VSL than C allele. Therefore, pigs with C allele are judged to be better at the MOT and VSL of semen. Based on these results, ZAR1 SNP genotyping may be a useful molecular biomarker to improve semen quality by applying molecular breeding technology.

Physiological Activity and Nutritional Composition of Pleurotus Species (느타리속 버섯류의 영양성분 및 생리활성)

  • Um, Su-Na;Jin, Gyoung-Ean;Park, Kye-Won;Yu, Young-Bok;Park, Ki-Moon
    • Korean Journal of Food Science and Technology
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    • v.42 no.1
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    • pp.90-96
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    • 2010
  • In this study, the anti-oxidant, anti-tumorigenic, anti-hypertensive, anti-thrombic, anti-diabetic, and anti-inflammatory properties of 18 different species of genus Pleurotus were investigated. In addition, the amino acid, $\beta$-glucan, and polyphenol content were also measured. All species contained more than 20 mg% of polyphenol with the highest contents found in Pleurotus cornucopiae var. citrinopileatus (yellow pleurotus) ($39.13{\pm}0.82\;mg%$). The $\beta$-glucan contents was also the highest in yellow Pleurotus ($37.67{\pm}0.22%$) followed by Won-Hyeong1 (C, $28.75{\pm}0.61%$) and Jang-an PK (A, $27.95{\pm}0.33%$). The yellow Pleurotus exhibited the highest antioxidant activity as assessed by the DPPH scavenging rate with an $IC_{50}$ value of $2.94{\pm}0.44\;mg/mL$. Ethanol extracts from the yellow Pleurotus treated at 1% concentration showed cytotoxic activity up to 36.9% in the human embryonic kidney 293T cell lines. The yellow Pleurotus also showed the highest inhibitory effects on ACE activity ($60.52{\pm}0.2%$). Finally, the yellow Pleurotus exhibited anti-diabetic and anti-inflammatory properties as shown by inhibition of $\alpha$-amyloglucosidase activity ($50.5{\pm}0.8%$) and nitric oxide production ($68.4{\pm}0.3%$). Taken together, our data indicate the yellow pleurotus is a promising functional food ingredients.

Yellowish Friable Embryogenic Callus (YFEC) Production and Plant Regeneration from Immature Embryo Cultures of Domestic Maize Cultivars and Genotypes (Zea may L.) (국내 옥수수 품종 및 계통의 미숙배 배양으로부터 Yellowish Friable Embryogenic 캘러스 (YFEC) 생산과 식물체 재생)

  • Cho Mi-Ae;Park Yun-Ok;Kim Jin-Suck;Park Ki-Jin;Min Hwang-Ki;Liu Jang-Ryol;Choi Pil-Son
    • Journal of Plant Biotechnology
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    • v.32 no.2
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    • pp.117-121
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    • 2005
  • Immature embryos of 3 cultivars (Du Me Chal, Mi Baek Chal, Heug Jeom Chal) and 5 genotypes (HW1, KL103, HW3, HW4, KW7) were cultured on medium containing MS salts, Eriksson's vitamins, 1 mg/L 2,4-dichlorophenoxyacetic acid, 25 mM proline, 100 mg/L casamino acid, 3 mM MES, 1.7 mg/L $AgNO_3$ and 20 g/L sucrose (SIM). Frequency of somatic embryo formation on explant of immature embryos showed in HW1 (45.20%), KL103 (5.75%), HW3 (37.20%), HW4 (30.10%), KW70 (55.20%), Mi Baek Chal (18.74%), Heug Jeom Chal (22.41%), Du Me Chal (36.72%) and Hi II type (<10%), respectively. Yellowish friable embryogenic callus (YFEC) such as type II callus of Hi II genotype only produced from the HW3 and Heug Jeom Chal, whereas other cultivars and genotypes were directly formed somatic embryos with late-embryonic stages or expanded yellowish compact somatic embryo with morphological abnormality. The yellowish friable embryogenic callus (YFEC) could be proliferated on the same medium, which were maintained embryogenic capacity for 6 months over. Upon transfer to first regeneration and second regeneration medium, somatic embryos converted to plantlets at a frequency of approximately 100%. However, the expanded somatic embryos with abnormal morphology were slowly proliferated when subcultured on the same medium, and some of them were degenerated or converted to plantlets at a frequency of approximately 25%. Accordingly, The Heug Jeom Chal and HW3 genotype will be further used for development of high frequency transformation system in domestic maize germplasm.

Reproductive Rate of One-banded Stink Bug, Piezodorus hybneri Linnaeus (Hemiptera: Pentatomidae) in Various Rearing Cages (사육용기의 규격에 따른 가로줄노린재의 증식율)

  • Bae, Soon-Do;Kim, Hyun-Ju;Park, Chung-Gyoo;Lee, Geon-Hwi;Park, Sung-Tae;Song, Yoo-Han
    • Korean journal of applied entomology
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    • v.44 no.4 s.141
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    • pp.293-298
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    • 2005
  • The embryonic and postembryonic developments of Pierodorus hybneri Linnaeus were observed in 5 different rearing cages such as A (Cylindrical, 10 cm in diameter, height of 4 cm), B (Cylindrical, 14.5 cm in diameter, height of 2.8 cm), C (Rectangle, 6.5 by 6.5 cm in $length{\times}breadth$, height of 10 cm), D (Cylindrical, 9 cm in bottom diameter, 11.5 cm in upper diameter, height of 10.8 cm) and E (Cylindrical, 15 cm in diameter, height of 7.5 cm) containing soybean and peanut seeds as feeding food, and sponge-water container under laboratory condition of $24^{\circ}C$ and 16L:8D. Egg duration was 6 days regardless of rearing cages. Hatchability ranged from 63 to 80% with the highest in B (14.5 cm in diameter, 2.8 cm in height) rearing cage. Instar duration was longer from 5 days in 1st instar to about 11 days in 5th instar. Nymphal duration showed 35 to 36 days with'3ut significant difference in rearing cages. Percent emergence was in range from 65 to 82% with the highest in B rearing cage. Adult longevity was 35 to 83 days for male, and 32 to 79 days for female, and was the longest in B rearing cage. Total number of eggs laid by female adult was in range from 38 to 86 with significant difference in rearing cages, and was the most in B rearing cage. Accordingly, the reproductive rate of P. hybneri for 1 generation was within 17 to 56 times, and was the highest in B rearing cage. Therefore, it could be concluded that B cage is most suitable for stable rearing of P. hybneri under laboratory condition.

THE EFFECT OF FIBROBLAST GROWTH FACTOR SIGNALING ON CARTILAGE FORMATION (FGF signaling이 연골 형성에 미치는 영향)

  • Park, Choong-Je;Lee, Sang-Won;Nam, Soon-Hyun;Kim, Young-Jin;Ryoo, Hyhn-Mo;Kim, Hyun-Jung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.30 no.4
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    • pp.643-653
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    • 2003
  • Fibroblast growth factor (FGF) / FGF receptor (FGFR) mediated signaling is required for skeletogenesis in cluding intramembranous and endochondral ossifications Runx2 ($Cbfa1/Pebp2{\alpha}A/AML3$) is an essential transcription factor for osteoblast differentiation and bone formation. Murine calvaria and mandible are concurrently undergoing both intramembranous bone and cartilage formations in the early developmental stage. However the mechanism by which these cartilage formations are regulated remains unclear. To elucidate the effect of FGF signaling on development of cranial sutural cartilage and Meckel's cartilage and to understand the role of Runx2 in these process, we have done both in vivo and in vitro experiments. Alcian blue staining showed that cartilage formation in sagittal suture begins from embryonic stage 16 (E16), Meckel's cartilage formation in mandible from E12. We analyzed by in situ hybridization the characteristics of cartilage cells that type II collagen, not type X collagen, was expressed in sagittal sutural cartilage and Meckel's cartilage. In addition, Runx2 was not expressed in Meckel's cartilage as well as sagittal sutural cartilage, except specific expression pattern only surrounding both cartilages. FGF signaling pathway was further examined in vitro. Beads soaked in FGF2 placed on the sagittal suture and mandible inhibited both sutural and Meckel's cartilage formations. We next examined whether Runx2 gene lies in FGF siganling pathway during regulation of cartilage formation. Beads soaked in FGF2 on sagittal suture induced Runx2 gene expression. These results suggest that FGF signaling inhibits formations of sagittal sutural and Meckel's cartilages, also propose that FGF siganling is involved in the proliferation and differentiation of chondroblasts through regulating the transcription factor Runx2.

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Potential Roles of Hedgehog and Estrogen in Regulating the Progression of Fatty Liver Disease (지방간 진행 조절에 대한 헤지호그와 에스트로겐의 잠재적 역할)

  • Hyun, Jeong-Eun;Jung, Young-Mi
    • Journal of Life Science
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    • v.21 no.12
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    • pp.1795-1803
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    • 2011
  • Non-alcoholic fatty liver disease accompanies the rise in the prevalence of obesity, diabetes and the tendency toward high-fat dietary habits. Specifically, the higher prevalence of non-alcoholic fatty liver disease in men and postmenopausal women seems to be caused by the protective effects of estrogen against liver fibrosis, or lack thereof. There are no effective preventive therapies for liver diseases because the mechanisms underlying the progression of fatty liver diseases to chronic liver diseases and the protective effects of estrogen against fibrogenesis remain unclear. Recently, it has been reported that the hedgehog signaling pathway plays an important role in the progression of chronic liver diseases. Hedgehog, a morphogen regulating embryonic liver development, is expressed in injured livers but not in adult healthy livers. The level of hedgehog expression parallels the stages of liver diseases. Hedgehog induces myofibroblast activation and hepatic progenitor cell proliferation and leads to excessive liver fibrosis, whereas estrogen inhibits the activation of hepatic stellate cells to myofibroblasts and prevents liver fibrosis. Although the mechanism underlying the opposing actions of hedgehog and estrogen on liver fibrosis remain unclear, the suppressive effects of estrogen on the expression of osteopontin, a profibrogenic extracellular matrix protein and cytokine, and the inductive effects of hedgehog on osteopontin transcription suggest that estrogen and hedgehog are associated with liver fibrosis regulation. Therefore, further research on the estrogen-mediated regulatory mechanisms underlying the hedgehog-signaling pathway can identify the mechanism underlying liver fibrogenesis and contribute to developing therapies for preventing the progression of fibrosis to chronic liver diseases.