Induction of podoplanin by transforming growth factor-β in human fibrosarcoma

AbstractPodoplanin/aggrus is increased in tumors and its expression was associated with tumor malignancy. Podoplanin on cancer cells serves as a platelet-aggregating factor, which is associated with the metastatic potential. However, regulators of podoplanin remain to be determined. Transforming growth factor-β (TGF-β) regulates many physiological events, including tumorigenesis. Here, we found that TGF-β induced podoplanin in human fibrosarcoma HT1080 cells and enhanced the platelet-aggregating-ability of HT1080. TGF-β type I receptor inhibitor (SB431542) and short hairpin RNAs for Smad4 inhibited the podoplanin induction by TGF-β. These results suggest that TGF-β is a physiological regulator of podoplanin in tumor cells

Platelets Strongly Induce Hepatocyte Proliferation with IGF-1 and HGF In Vitro

Background. It is well known that platelets have athrombotic effect. However, platelets play an importantrole not only in hemostasis but also in woundhealing and tissue regeneration. Platelets have beenreported to accumulate in the liver and promote liverregeneration after an extended hepatectomy, but themechanism is unclear. The present study was designedto clarify the mechanism by which plateletshave a direct proliferative effect on hepatocytes invitro.Materials and methods. Hepatocytes obtained frommale BALB/c mice by collagenase digestion and immortalizedhepatocytes (TLR2) were used. To elucidatethe mechanism of the proliferative effect of platelets,DNA synthesis of hepatocytes was measuredunder various conditions and the related cellular signalswere analyzed. Chromatographic analysis wasalso performed to clarify which elements of plateletshave mitogenic activity.Results. DNA synthesis significantly increased in thehepatocytes cultured with platelets (P < 0.001). However,when the platelets and hepatocytes were separated,the platelets did not have a proliferative effect.Whole disrupted platelets, the supernatant fraction,and fresh isolated platelets had a similar proliferativeeffect, while the membrane fraction did not. After theaddition of platelets, both Akt and extracellularsignal-regulated kinases ERK1/2 were activated, butextracellular signal-regulated kinase STAT3 was not activated. Some mitogenic fractions were obtainedfrom the platelet extracts by gel exclusion chromatography;the fractions were rich in hepatocyte growthfactor and IGF-1.Conclusions. Direct contact between platelets andhepatocytes was necessary for the proliferative effect.The direct contact initiated signal transduction involvedin growth factor activation. Hepatocyte growthfactor, vascular endothelial growth factor, and insulin-like growth factor-1, rather than platelet-derivedgrowth factor, mainly contributed to hepatocyteproliferation

Postnatal lethality and chondrodysplasia in mice lacking both chondroitin sulfate N-acetylgalactosaminyltransferase-1 and -2

Chondroitin sulfate (CS) is a sulfated glycosaminoglycan (GAG) chain. In cartilage, CS plays important roles as the main component of the extracellular matrix (ECM), existing as side chains of the major cartilage proteoglycan, aggrecan. Six glycosyltransferases are known to coordinately synthesize the backbone structure of CS; however, their in vivo synthetic mechanism remains unknown. Previous studies have suggested that two glycosyltransferases, Csgalnact1 (t1) and Csgalnact2 (t2), are critical for initiation of CS synthesis in vitro. Indeed, t1 single knockout mice (t1 KO) exhibit slight dwarfism and a reduction in CS content in cartilage compared with wild-type (WT) mice. To reveal the synergetic roles of t1 and t2 in CS synthesis in vivo, we generated systemic single and double knockout (DKO) mice and cartilage-specific t1 and t2 double knockout (Col2-DKO) mice. DKO mice exhibited postnatal lethality, whereas t2 KO mice showed normal size and skeletal development. Col2-DKO mice survived to adulthood and showed severe dwarfism compared with t1 KO mice. Histological analysis of epiphyseal cartilage from Col2-DKO mice revealed disrupted endochondral ossification, characterized by drastic GAG reduction in the ECM. Moreover, DKO cartilage had reduced chondrocyte proliferation and an increased number of apoptotic chondrocytes compared with WT cartilage. Conversely, primary chondrocyte cultures from Col2-DKO knee cartilage had the same proliferation rate as WT chondrocytes and low GAG expression levels, indicating that the chondrocytes themselves had an intact proliferative ability. Quantitative RT-PCR analysis of E18.5 cartilage showed that the expression levels of Col2a1 and Ptch1 transcripts tended to decrease in DKO compared with those in WT mice. The CS content in DKO cartilage was decreased compared with that in t1 KO cartilage but was not completely absent. These results suggest that aberrant ECM caused by CS reduction disrupted endochondral ossification. Overall, we propose that both t1 and t2 are necessary for CS synthesis and normal chondrocyte differentiation but are not sufficient for all CS synthesis in cartilage

Symbol Nomenclature for Graphical Representations of Glycans

ISSN:0959-665

Enhancement of metastatic ability by ectopic expression of ST6GalNAcI on a gastric cancer cell line in a mouse model

ST6GalNAcI is a sialyltransferase responsible for the synthesis of sialyl Tn (sTn) antigen which is expressed in a variety of adenocarcinomas including gastric cancer especially in advanced cases, but the roles of ST6GalNAcI and sTn in cancer progression are largely unknown. We generated sTn-expressing human gastric cancer cells by ectopic expression of ST6GalNAcI to evaluate metastatic ability of these cells and prognostic effect of ST6GalNAcI and sTn in a mouse model, and identified sTn carrier proteins to gain insight into the function of ST6GalNAcI and sTn in gastric cancer progression. A green fluorescent protein-tagged human gastric cancer cell line was transfected with ST6GalNAcI to produce sTn-expressing cells, which were transplanted into nude mice. STn-positive gastric cancer cells showed higher intraperitoneal metastatic ability in comparison with sTn-negative control, resulting in shortened survival time of the mice, which was mitigated by anti-sTn antibody administration. Then, sTn-carrying proteins were immunoprecipitated from culture supernatants and lysates of these cells, and identified MUC1 and CD44 as major sTn carriers. It was confirmed that MUC1 carries sTn also in human advanced gastric cancer tissues. Identification of sTn carrier proteins will help understand mechanisms of metastatic phenotype acquisition of gastric cancer cells by ST6GalNAcI and sTn

BioHackathon series in 2011 and 2012: penetration of ontology and linked data in life science domains

The application of semantic technologies to the integration of biological data and the interoperability of bioinformatics analysis and visualization tools has been the common theme of a series of annual BioHackathons hosted in Japan for the past five years. Here we provide a review of the activities and outcomes from the BioHackathons held in 2011 in Kyoto and 2012 in Toyama. In order to efficiently implement semantic technologies in the life sciences, participants formed various sub-groups and worked on the following topics: Resource Description Framework (RDF) models for specific domains, text mining of the literature, ontology development, essential metadata for biological databases, platforms to enable efficient Semantic Web technology development and interoperability, and the development of applications for Semantic Web data. In this review, we briefly introduce the themes covered by these sub-groups. The observations made, conclusions drawn, and software development projects that emerged from these activities are discussed

Serum Wisteria Floribunda Agglutinin-Positive Mac-2 Binding Protein Values Predict the Development of Hepatocellular Carcinoma among Patients with Chronic Hepatitis C after Sustained Virological Response

Measurement of Wisteria floribundaagglutinin-positive human Mac-2 binding protein (WFA+-M2BP) in serum was recently shown to be a noninvasive method to assess liver fibrosis. The aim of this study was to evaluate the utility of serum WFA+-M2BP values to predict the development of hepatocellular carcinoma (HCC) in patients who achieved a sustained virological response (SVR) by interferon treatment. For this purpose, we retrospectively analyzed 238 patients with SVR who were treated with interferon in our department. Serum WFA+-M2BP values were measured at pre-treatment (pre-Tx), post-treatment (24 weeks after completion of interferon; post-Tx), the time of HCC diagnosis, and the last clinical visit. Of 238 patients with SVR, HCC developed in 16 (6.8%) patients. The average follow-up period was 9.1 years. The cumulative incidence of HCC was 3.4% at 5 years and 7.5% at 10 years. The median pre-Tx and post-Tx WFA+-M2BP values were 1.69 (range: 0.28 to 12.04 cutoff index (COI)) and 0.80 (range: 0.17 to 5.29 COI), respectively. The WFA+-M2BP values decreased significantly after SVR (P 60 years), sex (male), pre-Tx platelet count ( 2.0 COI) were associated with the development of HCC after SVR. Conclusion: Post-Tx WFA+-M2BP (> 2.0 COI) is associated with the risk for development of HCC among patients with SVR. The WFA+-M2BP values could be a new predictor for HCC after SVR

Lycenchelys melanostomias Toyoshima 1983

Lycenchelys melanostomias Toyoshima, 1983 (Japanese name: Ohotsuku-hebigenge) (Figs. 21–25; Table 6) Lycenchelys sp.: Shiogaki, 1982: 23 (species list). Lycenchelys melanostomias Toyoshima, 1983: 271, 333, figs. 25–27, pl. 157 (original description, type locality: southern Okhotsk Sea, Hokkaido Island, Japan); Toyoshima, 1984: 293, pl. 274-A (brief description); Toyoshima, 1985: 170, figs. 6–7, 26–27, 31, tables 1, 4 (description); Hatooka, 1993: 902, unnumbered fig. (key to species); Anderson, 1994: 113, 117 (species list); Amaoka et al., 1995: 240, pl. 402 (brief description); Koyanagi, 1997: 538, fig. 3 (brief description); Hatooka, 2000: 1032, unnumbered fig. (key to species); Hatooka, 2002: 1032, unnumbered fig. (key to species); Anderson & Fedorov, 2004: 18 (species list); Imamura et al., 2004: 84, figs. 1–3, table 1 (synonymy of Lycenchelys melanostomias and Lycenchelys brevimaxillaris); Shiogaki et al., 2004: 71 (species list); Imamura et al., 2005: 1, figs. 1–3, table 1 (synonymy of Lycenchelys melanostomias and Lycenchelys brevimaxillaris); Shinohara & Anderson, 2007: 64 (key to species); Kitagawa et al., 2008: 95, unnumbered fig. (brief description); Shinohara et al., 2009: 724 (species list); Amaoka et al., 2011: 315, unnumbered fig. (brief description); Balushkin et al., 2011: 1026 (species list); Hatooka, 2013: 1226, 2078, unnumbered fig. (key to species); Nakabo & Hirashima, 2015: 217 (species list and etymology of scientific name). Lycenchelys brevimaxillaris: Toyoshima, 1985: 174, figs. 6–7, 29–30, 31, table 1 (original description, type locality: off Aomori Prefecture, Pacific coast of Honshu, Japan); Hatooka, 1993: 902, unnumbered fig. (key to species); Anderson, 1994: 117 (species list); Imamura, 1998: 31, fig. 9 (brief description); Hatooka, 2000: 1033, unnumbered fig. (key to species); Hatooka, 2002: 1033, unnumbered fig. (key to species); Anderson & Fedorov, 2004: 16 (species list); Imamura et al., 2004: 84, figs. 1–3, table 1 (synonymy of Lycenchelys melanostomias and Lycenchelys brevimaxillaris); Shiogaki et al., 2004: 71 (species list); Imamura et al., 2005: 1, figs. 1–3, table 1 (synonymy of Lycenchelys melanostomias and Lycenchelys brevimaxillaris). Materials examined Holotype: HUMZ 77572, male, 182.3 mm SL, Kitami-Yamato Bank, Okhotsk Sea (44°19.5’N, 145°01’E), 915–925 m depth, 11 Oct. 1978; Other specimens (41 specimens, 119.3–227.6 mm SL): HUMZ 189817 (holotype of Lycenchelys brevimaxillaris), female, 185.8 mm SL, off Aomori Prefecture, Tohoku District, northwestern Pacific (41°13’N, 141°44’E), 690–750 m depth, 18 Jan. 1982; HUMZ 152380, 152384, 152404–05, 152411, 157545, 177188, 178572–75, 182565, 192724, 192804, 4 males and 10 females, 124.0– 195.1 mm SL, Tohoku District, northwestern Pacific; HUMZ 177263, 192453, 205157, 228057, 228067–68, 228070, 228073, 228089, 5 males and 4 females, 119.3–193.2 mm SL, eastern Hokkaido Island, northwestern Pacific; HUMZ 120346, 121158, 121161, 121461, 124054–55, 124116, 126101, 126215–16, 126219–22, 126230, 126359, 126361, 10 males and 7 females, 120.6–227.6 mm SL, northeastern Hokkaido Island, Okhotsk Sea. Diagnosis. Vertebrae 22–25 + 93–102 = 117–124; head length 11.6–15.0% SL; interorbital pore 1; occipital pores usually 2; postorbital pores usually 4; suborbital pores 6–7 + 2–3; preoperculomandibular pores usually 9; vomerine teeth 3–10; palatine teeth 2–11, usually arranged in single row (sometimes 1–2 rows); opercular flap well developed; pelvic-fin base positioned posterior to lower edge of gill opening; lateral line complete and positioned ventrally; scales absent on pectoral fin and its base; body uniformly grayish-brown when fresh. Description. Counts and proportional measurements in Table 6. ...Continued next page Body very elongate, cross section oval anteriorly, compressed laterally near tail; its width at anal-fin origin 3.8–7.0 (5.0)% SL. Head short, ovoid; dorsal profile of head sloping extremely gently from posterior edge of eye to above opercular flap. Head of males slightly longer than of females in adults. Snout short, 74.4–131.0% of eye diameter (eye damaged in holotype). Eye ovoid, moderately large. Interorbital space narrow, width 12.7–33.8% of eye diameter (eye damaged in holotype). Nostril tube short, not reaching upper lip when depressed. Mouth subterminal. Posterior edge of upper jaw reaching vertical through posterior part of eye in adult males, reaching vertical through anterior margin of pupil in females and juveniles. Labial lobe of lower jaw tending to be more developed in large males than in females and juveniles (labial lobe of lower jaw damaged in holotype). Teeth on jaws sharp; upper jaw with 2–3 rows anteriorly, 1 or 1–2 rows posteriorly (1); anteriormost teeth larger than other teeth; lower jaw with 2–5 irregular rows anteriorly, 1 or 1–2 rows posteriorly (1); vomerine and palatine teeth small and conical; vomerine teeth irregularly arranged; palatine teeth usually in single row, but sometimes 1–2 rows (no data for holotype). Lower edge of gill opening slightly above lower end of pectoral-fin base. Opercular flap well developed. Gill rakers short and triangular (Fig. 23). Pseudobranch filaments short. Lateral line deciduous, complete and positioned ventrally; originating posterior to last postorbital pore and terminating on tail. Scales small and cycloid, present on body and tail, except for area around pelvic fin. Scales covering basal portions of dorsal and anal fins anteriorly; extent of scaled areas gradually increasing posteriorly, except at margins. Head, nape, pectoral axilla, pectoral fin, pectoral-fin base and area around pelvic fin without scales. Dorsal-fin origin posterior to vertical through pectoral-fin base; 1st dorsal-fin pterygiophore between neural spines of 2nd to 5th (between 2nd and 3rd) vertebrae. Anal-fin origin below 17th to 20th (18th) dorsal-fin ray; 1st anal-fin pterygiophore posterior to parapophysis of ultimate or penultimate (penultimate) abdominal vertebra. Last dorsal-fin pterygiophore between neural spines of 2nd to 5th (between 2nd and 3rd) preural vertebrae. Last analfin pterygiophore between hemal spines of 3rd to 5th (between 3rd and 4th) preural vertebrae. Caudal fin with 1–3 (2) epural, 3–4 (4) upper hypural and 3–4 (3) lower hypural rays. Pectoral fin moderately short, not quite reaching middle of abdomen; its posterior margin rounded dorsally and having notches ventrally. Upper end of pectoral-fin base about on lateral midline of body. Pelvic fin short; its base posterior to lower edge of gill opening; its posterior margin reaching about vertical through pectoral-fin base. Head pores well developed and distinct. Nasal pores 2; anterior pore in front of nostril tube, posterior pore above line of 1st suborbital pore (Fig. 22A, B). Postorbital pores usually 4, rarely 5 (5); when 4, distance between 1st and 2nd pores longest of those between adjacent pores; when 5, 1 additional pore present between 1st and 2nd pores (Fig. 22A, B). Suborbital pores 8–10 (unknown for holotype), 6 or 7 pores located below eye and remaining 2 or 3 pores on ascending part of suborbital canal behind eye; 5th pore below posterior margin of pupil; last pore of those below eye located posterior to vertical through posterior margin of eye (Fig. 22A). Preoperculomandibular pores usually 9 (9), rarely 8; 4 on lower jaw, 2 at junction of lower jaw and preopercle, and 3 on preopercle; 2 pores at junction of lower jaw and preopercle united into 1 pore in some specimens and preoperculomandibular pore series counted as 8; 2nd and 3rd pores united into 1 pore on right side of HUMZ 126361 and series counted as 8; last preoperculomandibular pore located posterior to lower part of eye (Fig. 22A, C). One interorbital pore located on dorsal midline anterior to middle of eyes (Fig. 22B). Occipital pores usually 2 (2), usually positioned on either side of dorsal midline; only 1 pore on right side in HUMZ 182565; occipital pore(s) located anterior to 3rd postorbital pore (Fig. 22B). Two additional unnamed pores present in interorbital space on either side in line with postorbital pores, behind posterior to posterior margin of pupil only in HUMZ 178574 (Fig. 22D). Color in alcohol. Color of holotype (Fig. 24) unknown owing to head, body, and fins lacking skin. Long preserved holotype of L. brevimaxillaris with brownish head, pectoral fin and vertical fins, uniformly grayish brown body and gray abdomen. Other recently preserved specimens with dark brown head, pectoral fin and vertical fins, uniformly pale brown body and blackish abdomen. Color when fresh (based on color photograph of HUMZ 124054; Fig. 21). Head and pectoral fin black; body and vertical fins uniformly grayish brown; margin of vertical fins slightly darker and abdomen pale purplish. Distribution. Southern Okhotsk Sea, off northwestern Pacific coast of eastern Hokkaido Island and in the northwestern Pacific from Aomori to Ibaraki prefectures, at depths of 425–1440 m (Shiogaki, 1982; Toyoshima, 1983, 1984, 1985; Hatooka, 1993, 2000, 2002, 2013; Anderson, 1994; Amaoka et al., 1995, 2011; Koyanagi, 1997; Imamura, 1998; Anderson & Fedorov, 2004; Imamura et al., 2004, 2005; Shiogaki et al., 2004; Shinohara & Anderson, 2007; Kitagawa et al., 2008; Shinohara et al., 2009; this study). Size. The largest specimen examined during this study measured 227.6 mm SL (230.8 mm TL), about equal to the previously recorded maximum length of 23 cm TL (Hatooka, 2013). Remarks. Lycenchelys melanostomias is similar to L. hippopotamus, L. makushok and L. rassi in having more than 100 total vertebrae, 1 interorbital pore, 1–2 occipital pores, 4–5 postorbital pores, a single lateral line positioned ventrally and no distinct spots or blotches on the body (vs. lacking this combination of characters in other species of Lycenchelys) (e.g., Toyoshima, 1983, 1985; Fedorov & Andriashev, 1993; Anderson, 1995; Imamura et al., 2004; Shinohara & Anderson, 2007; this study). See Remarks under accounts of L. hippopotamus, L. makushok and L. rassi for detailed comparisons of L. melanostomias with each. Two additional pores behind the posterior margin of the pupil were found in 1 specimen of L. melanostomias (Fig. 22D). These pores have not been previously described for L. melanostomias, although similar pores are known in Lycenchelys parini Fedorov, 1995 (Fig. 25). Lycenchelys melanostomias also resembles L. parini in having similar counts, proportional measurements and arrangements of head pores (e.g., dorsal-fin rays 111–120 vs. 118, and anal-fin rays 98–107 vs. 105, head length 11.6–15.0 vs. 12.2% SL, postorbital pores usually 4 vs. 4 and suborbital pores 8–10 vs. 9 in L. melanostomias respectively; Table 6) (Fedorov, 1995a; Imamura et al., 2004, 2005; this study). However, L. melanostomias is easily separable from L. parini in having the lateral line positioned ventrally rather than midlaterally on the side (Fedorov, 1995a).Published as part of Kawarada, Shumpei, Imamura, Hisashi, Narimatsu, Yoji & Shinohara, Gento, 2020, Taxonomic revision of the genus Lycenchelys (Osteichthyes: Zoarcidae) in Japanese waters, pp. 1-66 in Zootaxa 4762 (1) on pages 27-32, DOI: 10.11646/zootaxa.4762.1.1, http://zenodo.org/record/374369