Organic components from scallop shell increase expression of keratinocyte growth factor in human skin fibroblast

We observed previously that scallop shells contain organic components that have various useful applications for skin. The organic components from scallop shell (scallop shell extract) enhance the turnover rate of the rat skin epidermal layer and increase the rate of recovery of UVB-induced injury to rat dorsal skin. We investigated the effect of scallop shell extract on the expression of keratinocyte growth factor (KGF) from human skin fibroblasts because it has been reported that KGF promotes wound healing and reduces the generation of UVB-induced free radicals in skin. Scallop shell extract increased the mRNA expression level of KGF in skin fibroblasts and the secretion of KGF from the fibroblasts at the protein level. These results suggest that the scallop shell extract may promote recovery from UVB-induced injury in skin through an increase in the secretion of KGF from the fibroblast cell.     

Scallop shell extract promotes recovery from UV-B-induced damage in rat skin epidermal layer

ABSTRACT:   It has been previously reported that scallop shell extract can protect keratinocyte cells against UV-B-induced damage in vitro by acting as an antioxidant and a growth promoter. To establish the growth promoting activity of scallop shell extract, the effect was investigated in vivo. Exposure of rat dorsal skin to UV-B caused the formation of erythema and eschar. Either the scallop shell extract or vehicle (water) was applied once a day for 5 days to the injured dorsal area and the area of the erythema and eschar was estimated using National Institute of Health (NIH) image software. Wound healing of the erythema and eschar was clearly promoted after application of scallop shell extract when compared to application of the vehicle control. Histological studies indicated that scallop shell extract promoted the recovery of the epidermal layer. These results suggest that scallop shell extract activates rat keratinocyte cells and promotes the turnover of skin stratum corneum. This conclusion was also supported by measurement of the turnover rate of the stratum corneum. Therefore, scallop shell extract may be effective in protecting skin against UV-B.     

Photoprotective activity of scallop shell water-extract in keratinocyte cells

ABSTRACT:   The photoprotective activity of the scallop shell water-extract was investigated using cultured rat skin keratinocyte cells. It was found that the scallop shell water-extract protected keratinocyte cells against ultraviolet (UV)-B-induced damage. The scallop shell water-extract had protective effects against hydrogen peroxide (H₂O₂) in skin keratinocyte cells and also showed 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity and lipid peroxidation inhibitory activity. In addition, the scallop shell water-extract promoted proliferation of keratinocyte cells. The scallop shell water-extract may protect skin keratinocyte cells against UV-B-induced damage through two factors; antioxidant activity and growth-promoting activity in skin keratinocyte cells. These results suggest utilization of the scallop shell water-extract as materials for protecting skin.     

Glycoproteins isolated from scallop shells inhibit differentiation of 3T3-L1 preadipocyte cells

We previously demonstrated that the organic components isolated from scallop shells (scallop shell extract) inhibit the differentiation of 3T3-L1 preadipocyte cells. In this study, we show that a pronase-treated scallop shell extract inhibited differentiation, but degradation of sugar chains in the scallop shell extract with trifluoromethane sulfonic acid resulted in the loss of the differentiation-inhibiting activity, suggesting that the sugar chain modifications were responsible for the inhibitory activity. To identify the bioactive substance in the scallop shell extract, we isolated glycoproteins from the scallop shell extract via affinity chromatography using concanavalin A (Con A), wheat germ agglutinin, lens culinaris agglutinin (LCA), and ricinus communis agglutinin. LCA and Con A binding fractions inhibited the differentiation of 3T3-L1 preadipocyte cells. In addition, a glycoprotein with a molecular weight of 16 kDa that was purified from the LCA binding fraction reduced lipid accumulation in 3T3-L1 cells during differentiation. The glycoprotein inhibited differentiation-associated mitotic clonal expansion and suppressed the expression of an adipocyte-specific protein, fatty acid binding protein. These results suggest that the sugar chains of glycoproteins in the scallop shell extract inhibit the differentiation of 3T3-L1 preadipocyte cells.     

Identification of a resistant protein from scallop shell extract and its bile acid-binding activity

In this study, we showed that feeding rats the organic extract of scallop shells (scallop shell extract) caused a decrease in the weights of white adipose tissues in rats fed a high-fat diet. In addition, the cholesterol concentration in the serum of rats that received a diet containing scallop shell extract was significantly lower than that in the serum of rats on the control diet. Feeding this scallop shell extract to rats increased the fecal weight as well as the fecal excretion of bile acids. The amino acid composition of the feces from rats fed the scallop shell extract was different from that of feces from rats fed the control diet, and treatment of the extract with pepsin and pancreatin identified a protein with a molecular weight of 90 kDa (90-kDa protein) as one of the indigestible proteins. Interestingly the 90-kDa protein was found to be identical to a free radical-scavenging protein we previously identified and showed the ability to bind bile acids. These results suggest that indigestible proteins (resistant proteins) in the scallop shell extract, including the 90-kDa protein, inhibit the absorption of bile acid by binding to it and cause increased excretion of fecal bile acid, which subsequently may decrease the serum cholesterol level.     

Culture of mantle epithelial cells expressing shell matrix proteins from scallop Patinopecten yessoensis

ABSTRACT:   In molluscs, mantle epithelial cells secrete organic matrix proteins to form shells. In this study, we established a culture of mantle epithelial cells by using the mantle pallial layer of scallops. We aimed to identify the mantle epithelial cells expressing scallop shell matrix proteins and establish a culture system of epithelial cells. After the mantle pallial layer was carefully isolated from the mantle tissue, explant culture was performed at 4°C. Most cells that migrated from the explant tissue were round cells. Most of the adhered cells retained round morphology, while some of the cells adhered to the dish and showed morphology similar to that of epithelial-like and fibroblast-like cells. When the cultured cells were immunostained with a polyclonal antibody against the shell matrix protein, the antibody recognized many of the adhered cells. An estimation of the number of epithelial cells revealed that approximately 70% of the adhered cells were epithelial cells. This is the first report to describe epithelial cells in cultured mantle cells, which express shell matrix proteins. This culture system may be a useful method for characterization of the mantle epithelial cells.     

cDNA clonings of shell matrix proteins from scallop shell

ABSTRACT:   A cDNA encoding the matrix shell protein from scallop (MSP-2) has been cloned. Comparison of the sequence of MSP-2 with the GenBank database revealed the sequence displays a high degree of homology with MSP-1, which was recently identified in the shell of the scallop Patinopecten yessoensis . Matrix shell protein-2 contains only 323 amino acids, while MSP-1 comprises 824 amino acids. The structural difference between MSP-1 and MSP-2 is mainly the presence or absence of the tandem repeat arrangement. In addition, multiple isoforms of MSP-2 were found. These isoforms share a high degree of sequence similarity to each other, suggesting that the MSP-2, MSP-2 isoforms and MSP-1 may constitute the same family. The MSP-2 isoforms in scallop shell may be responsible for the formation of molluscan shells.     

Purification and characterization of a matrix shell protein from the shell of scallop <Emphasis Type="Italic">Patinopecten yessoensis</Emphasis>

ABSTRACT:   A new protein named MSP-SC (matrix shell protein from scallop) with a molecular weight of 14 kDa was isolated from the shell of a scallop, Patinopecten yessoensis , using gel filtration column chromatography, ion exchange column chromatography, and reverse phase C4 column chromatography. A comparison of the known protein sequences with the N-terminal sequence of MSP-SC showed that the protein sequence of MSP-SC was novel. Immunohistochemistry using a polyclonal antibody against MSP-SC showed that MSP-SC is expressed in the mantle pallial cell layer but not in the muscle tissue, and showed a punctate distribution along the horizontal calcified layer in the shell. The isolated MSP-SC inhibited the formation of calcium carbonate (CaCO₃) crystals in a dose-dependent manner. The CaCO₃ crystals grown in the presence of a lower concentration of MSP-SC were much larger and aggregated when compared with those formed in the absence of MSP-SC. In addition, the crystal had a radial and not cubical morphology. These results suggest that MSP-SC regulates the formation and the morphology of CaCO₃ crystals in the shell. Moreover, its ability to aggregate CaCO₃ crystals and its localization along the horizontal calcified layer in the shell suggest that MSP-SC may serve to connect the CaCO₃ layers in the scallop shell.     

Association of zinc with connective tissue in the digestive tract of common carp

ABSTRACT:   Zinc (Zn) concentration in the digestive tract of common carp is always >10 times higher than most animal tissues. In a previous paper, it was reported that this high Zn came from a 43 kDa Zn-binding membrane protein. In this present study it was further found that in the digestive tract of common carp, Zn content was closely associated with the amounts of extracellular macromolecules. The higher the Zn content, the more there is of collagen and glycosaminoglycans. An indirect immunoperoxidase staining method using antibody against the 43 kDa Zn-binding protein, or the fibroblast marker (Thy 1.1 protein) was applied to the sections of digestive tract of fish. It was found that the Zn-binding protein in the digestive tract of common carp is mainly located in the connective tissue of its lamina propria and submucosal layer. Connective tissue cells, probably fibroblasts, hold the 43 kDa Zn-binding protein. In the common carp Zn might be bound to the external side of the fibroblast. The present finding may have a significant meaning on extending the studies of Zn in biology to the field of Zn with extracellular matrix.     

Properties of scallop mantle collagen: its content,tissue distribution and thermal behavior

ABSTRACT:   To obtain fundamental information for the effective use of scallop Patinopecten yessoensis mantle, which is one of the underutilized marine resources. Some properties of collagen contained in the mantle were examined by chemical and histochemical techniques. Collagen content in the mantle varied annually, ranging from 0.98 to 1.72% of wet tissue, 7.7 to 12.6% of dry tissue and 13.5 to 26.5% of total protein, being relatively in high level of collagen content of invertebrate muscles. Collagen fiber was densely distributed in the inner connective tissue matrix of the mantle pallial, in contrast to the inner fold part which was rich in muscle fibers. The collagen contained in the crude collagen fraction (residue after alkali extraction), prepared from the mantle, was revealed to have considerably low solubility on hot-water extraction, constantly less than 20% of the total collagen at the temperatures in the range of 20–90°C.     

海湾扇贝外套膜创伤修复过程的组织学和组织化学观察

Ｍａｌｌｏｒｙ‘ｓ三色染色显示海湾扇贝外套膜创伤的组织学修复过程,外套切割３小时、６小时、伤口处涌出血淋巴细胞,有非颗粒变形细胞与颗粒变形细胞两种构成。切割２４小时伤口内出现大量的成纤维细胞,至切割后８天,伤愈组织内出现少量成肌细胞,１６天,愈合物内肌纤维增多,并可见到血窦形成,切割３２天,肌纤维含量丰富,但这些肌纤维并未形成明显的肌肉束。组织化学观察表明,高碘酸Ｓｃｈｉｆｆ反应发现血淋巴细胞质内     

Gelation of low salt soluble proteins from scallop adductor muscle in relation to its freshness

ABSTRACT: The low salt extract from scallop striated adductor muscle contained extra proteins in addition to sarcoplasmic proteins and was turned into a gel upon standing or immediately by the addition of Ca²⁺. The amount of extra protein, which was composed of a large amount of actin and a small amount of myosin, decreased with a decrease in adenosine triphosphate (ATP) content in the muscle during storage at 5°C. Actin was easily extracted from scallop myofibrils in low salt solution with ATP at 1 mM or above. Furthermore, ATP was required to induce the gelation of the low salt extract. A visual observation of the gelation of low salt extract is therefore a simple and easy method to inspect prerigor state of scallop adductor muscle.     

Possible degradation of type I collagen in relation to yellowtail muscle softening during chilled storage

ABSTRACT:   In this paper, the detection of type I collagen degradation during the softening phenomenon of yellowtail muscle, was examined. Acid soluble collagen was isolated from dorsal ordinary muscle at death and after 24-h chilled storage. In the abundant ratio of subunit components, an increase in β12 chain (5.4 points) and a decrease in components with molecular weights larger than γ chain (7.0 points) after 24-h chilled storage, was found. Type I collagen was detected in the alkali-soluble fraction by SDS-PAGE. Its amount calculated from hydroxyproline contents in alkali-soluble fraction was increased from 0.097 mg/g muscle to 0.155 mg/g muscle during 24-h storage. The increased alkali-soluble collagen (0.058 mg/g muscle) was about 1.4% of whole collagen. These results suggest that a slight decomposition of type I collagen of yellowtail muscle may occur and subsequently becomes alkali-soluble corresponding to postmortem softening.     

Solubilization of type I and V collagens in Japanese flounder muscle during chilled storage

ABSTRACT:   The post-mortem changes of type I and V collagens in Japanese flounder muscle during chilled storage were examined. The muscle softened significantly within 12 h under chilled storage. Both type I and V collagens isolated by ion-exchange chromatographies showed no remarkable changes in band patterns even after 24 h of storage, suggesting that degraded collagen molecules may be removed from the collagen fraction by a conventional preparation method. In contrast, type I and V collagen molecules were detected by Western blotting with each specific antibody in a saline extract and gradually increased during chilled storage. These results suggest that both type I and V collagens may participate in the post-mortem softening of fish muscle.     

桑沟湾不同区域养殖栉孔扇贝的固碳速率

测定桑沟湾深水区、浅水区栉孔扇贝固碳量,并进行固碳速率的标准化处理,增加了与陆地生态系统固碳率的可比性,分析了栉孔扇贝在不同养殖区的固碳速率及其主要控制因素。研究显示,对于同一养殖种类,深水区生物固碳的速率比浅水区高两倍。不同区域,贝类壳碳及软体部中碳的含量没有显著性差异,导致区域性差异的主要原因是由于生长速度、养殖密度及存活率的不同而导致单位面积的产量存在差异。养殖栉孔扇贝的固碳速率可与森林相媲美。另外,贝类的养殖活动与浅海生态系统的碳循环之间关系复杂,需要加强贝类的摄食、呼吸、生物沉积、钙化等整个生理生态学过程研究。     

Effect of increased water recirculation rate on algal supply and post-larval performance of scallop (Pecten maximus) reared in a partial open and continuous feeding system

In a commercial scallop hatchery spat production depends on a culture system which ensures high survival and good growth. Reuse of water with algae may increase the food exploitation and hence reduce the costs. Post-larvae of great scallop (Pecten maximus) were studied in a commercial hatchery using a partial open and continuous feeding tank system. Three different water recirculation rates (67, 83 and 92%) were tried out in two experiments with post-larvae originating from three spawning groups of ages between 43 and 57 days post-spawn, 316–886 μm shell-height and 1.1–9.6 μg ash-free dry weight. The post-larvae were held in sieves in tanks of 2500 l where a downwelling flow was maintained by airlifts. New water with a mix of monocultured algae was continuously added to the tanks at algal concentrations of 10 and 15 cells μl⁻¹ in experiment 1 (groups 1 and 2) and 2 (group 3), respectively. The algal supply to each sieve was reduced along with increased recirculation rate, but was kept between 6 and 13 cells μl⁻¹. Generally no significant differences in survival, growth or chemical content were found between the three recirculation rates, while few differences were found between and within groups. Large variation in survival was found between and within groups (1–81%). Highest survival was found in experiment 1, and where post-larvae from two settlements were used, the first settlement survived better than the second. The daily growth ranged from 15 to 62 μm shell-height and from 0.3 to 2.6 μg ash-free dry weight. The scallop post-larvae could well be reared at all three recirculation rates studied as an increase from 67 to 92% did not seem to affect the post-larval performance seriously. The algal supply, however, had to be compensated by an increasing number of cells (>10 cells μl⁻¹) when increasing the recirculation rate.     

不同规格虾夷扇贝捕后耐干露特性比较

为探究商品虾夷扇贝规格与其活品耐藏特性之间的关联,对春季上市虾夷扇贝的规格及质量进行分组,再将各组置于4 ℃下湿布覆盖进行3 d冷却干露贮藏,分别于0、1、2 d和3 d采样进行生化代谢指标分析。试验结果显示,市售春季虾夷扇贝依据壳长大致可分成(105.8±1.5) mm、(98.4± 2.3 ) mm及(90.5±1.7) mm 3个规格组,各规格单贝质量相差20~30 g;(98.4±2.3) mm组扇贝的肥满度及闭壳肌出成率较其他两组高,(90.5±1.7) mm组三磷酸腺苷初始含量最高,表明其捕后胁迫耐力好于较大规格扇贝,(90.5±1.7) mm组扇贝同样表现出更好的耐干露特性,干露3 d 三磷酸腺苷含量和核苷酸能荷均明显优于其他两组;糖原含量各组无明显差异。此外,各组扇贝冷却干露2 d后存活率均为100%,干露至第3 d各组开始出现死亡,其中(90.5±1.7) mm组扇贝耐干露特性最好;各组闭壳肌pH与体腔液pH差异不显著;氨基酸分析结果发现,甘氨酸和精氨酸是扇贝闭壳肌游离氨基酸中含量最丰富的氨基酸,其余游离氨基酸变化趋势不明显;体腔液含氮物中,氨含量较高,其余氨基酸含量均较低并在干露期间无明显变化趋势。     

Effect of ethanolic extract of Zingiber officinale on growth performance and mucosal immune responses in rainbow trout (Oncorhynchus mykiss)

This study was conducted to examine the effect of ethanolic ginger (Zingiber officinale) extract on growth performance and some skin mucus immune parameters in rainbow trout (Oncorhynchus mykiss). A total of 600 rainbow trout fingerling (10.07 ± 0.21 g) were randomly assigned to four groups in triplicate and fed for 45 days with 1, 2.5 and 5 g kg^?1 of ginger extract and with unsupplemented basal diet as the control. The total protein content and lysozyme, alkaline phosphatase and protease activities of skin mucus were investigated following 15, 30 and 45 days of feeding. Results showed that administration of 2.5 and 5 g kg^?1 of ethanolic ginger extract for 45 days improved weight gain, feed utilization efficiency and feed conversion ratio of O. mykiss (P < 0.05). Significantly higher (P < 0.05) skin mucus lysozyme, alkaline phosphatase and protease activity and protein level were observed in the 2.5 and 5 g kg^?1 ginger extract groups on the 30th and 45th day compared to the controls. The findings suggest that ethanolic extract of ginger enhances growth performance and skin mucus immune parameters of rainbow trout.     

Evaluation of antioxidant properties and biofunctions of polar,nonpolar, and water-soluble fractions extracted from gonad and body wall of the sea urchin Tripneustes gratilla

This study was performed to compare the various bioactivities of water-soluble, polar, and nonpolar extracts from the edible part (gonad) and waste (body wall) of the sea urchin Tripneustes gratilla. During antioxidant activity screening, these three extracts from both gonad and body wall showed various antioxidant activities in different tests. During human skin fibroblast (CCD966SK) viability and collagen screening, only water-soluble and polar extracts from gonad and body wall were observed to promote cell viability, while the water-soluble gonad extract promoted the collagen-generating activity of CCD966SK cells. The water-soluble and polar extracts from gonad and body wall showed proliferative activity towards different cell types. In contrast, the nonpolar extracts of gonad and body wall exerted antiproliferative effects on most tumor cells. These results indicate that the bioactivities of sea urchin extracts depend on the part of the urchin that the extract was taken from as well as the extraction method used.