Protective Effect of Topically Applied Polypeptide from Chlamys farreri Against Ultraviolet Radiation-Induced Chronic Skin Damage in Guinea Pig

INTRODUCTIONSolarultraviolet(UV)radiationisknowntobeamajoretiologicalriskofphotoaging.AsUVC(2 0 0nm -2 90nm)isabsorbedentirelybythestratosphericozonelayer,sunlightontheearthsur faceiscomposedofUVA (3 2 0nm -4 0 0nm)andUVB (2 90nm -3 2 0nm)only.UVAradiationenergyaccountsformorethan 90 %ofsolarUV ;andpenetratesthebasallayerofepidermismoreefficientlythanUVB (Brulsetal.,1984) .However,atsimilardosages,UVBismuchmoremuta genicthanUVA (Drobetskyetal.,1995) .Furthermore ,humanskinisexp…     

Effects of polypeptide from<Emphasis Type="Italic">Chlamys farreri</Emphasis> on amino acid content in guinea pig skin irradiated by chronic ultraviolet A and B

We examined the effects of polypeptide fromChlamys farreri (PCF) on the amount of hydroxyproline in guinea pig skin irradiated by chronic ultraviolet A (UVA) and ultraviolet B (UVB) radiation. PCF was applied locally before repeated exposure of guinea pig to UVA and UVB. The contents of hydroxyproline and other amino acids in guinea pig skin were determined by automatic amino acid analyzer. Our results showed that: (1) long-time UVA and UVB radiation can reduce dramatically the amounts of hydroxyproline, aspartic acid, threonine, glycine, phenylalanine and lysine in guinea pig skin in comparison with the control group (P<0.05); (2) Compared with model group, pre-treatment with 5% and 20% PCF prior to UVA and UVB radiation can inhibit the decline of amino acids content in guinea pig skin in a dose-dependent manner (P<0.05). As the decrease of hydroxyproline, glycine and lysine contents in the skin directly reflexes type I collagen degeneration, our results indicated that the chronic application of PCF can protect skin type I collagen against UV radiation, and thus protect skin from photoaging. Project No. 39970638 supported by the NSFC, and also supported by the Science and Technology Bureau of Qingdao (No: 2001-28-50).     

Effects of polypeptide from Chlamys farreri on amino acid con-tent in guinea pig skin irradiated by chronic ultraviolet A and B

1 INTRODUCTION Long-term exposure of the skin to ultraviolet (UV) radiation results in degenerative processes involved in photoaging and photocarcinogenesis (Differy, 1987). As UVC is absorbed entirely by the stratospheric ozone layer and does not reach the earth抯 surface, sunlight is composed of UVA (320 nm 400 nm) and UVB (290 nm 320 nm). Human skin is exposed constantly to both UVA and UVB radiation, it is, therefore, important to protect the skin against UVA and UVB-induce…     

The polypeptide in Chlamys farreri can protect human dermal fibroblasts from ultraviolet B damage

To investigate the effect of polypeptide fromChlamys farreri (PCF) on NHDFin vitro, we modeled oxidative damage on normal human dermal fibroblasts (NHDF) exposed to ultraviolet B (UVB). In this study, 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) and lactate dehydrogenase (LDH) were tested to measure cell viability. Enzymes including superoxide dismutase (SOD), glutathione peroxidase (GSH-PX), catalase (CAT) and xanthine oxidase (XOD) were determined biochemically. Total antioxidative capacity (T-AOC) and anti-superoxide anion capacity (A-SAC) were also determined. Ultrastructure of fibroblasts was observed under transmission electron microscope. The results showed that: UVB (1.176×10⁻⁴ J/cm²) suppressed the growth of fibroblasts and the introduction of PCF (0.25%–1%) before UVB reduced the suppression in a concentration-dependent manner. PCF could enhance the activities of SOD, GSH-PX and T-AOC as well as A-SAC. Also PCF could inhibit XOD activity, while it did not affect CAT activity. Ultrastructure of fibroblasts were damaged after UVB irradiation, concentration-dependent PCF reduced the destructive effect of UVB on cells. These results indicated that PCF can protect human dermal fibroblasts from being harmed by UVB irradiation via its antioxidant proerty. This work was supported by the National Natural Science Foundation of China (No. 39970638) and the Science and Technology Bureau of Qingdao (No. 2001-28-50).     

Free radical scavenging abilities of polypeptide from Chlamys farreri

1 INTRODUCTION Active oxygen free radicals can damage cell structure, even kill the cells, causing aging and cer- tain serious diseases (Bors et al., 1989). Usually, the production and scavenging of active oxygen free radicals are in balance in healthy hu…     

Effects of benzo(a)pyrene exposure on the antioxidant enzyme activity of scallop <Emphasis Type="Italic">Chlamys farreri</Emphasis>

Scallop Chlamys farreri was exposed to different concentrations of benzo(a)pyrene (BaP) (0.5 μg/L, 1.0 μg/L, 10.0 μg/L and 50.0 μg/L) for 30 days in seawater. The 7-ethoxyresorufin O-deethylase (EROD) activity was significantly induced, and increased with the increasing BaP concentration. The glutathione-S-transferase (GST), superoxide dismutase (SOD), catalase (CAT), Glutathione peroxidase (GPx) activities increased in short time at low concentration of BaP, and was significantly depressed at high concentrations. Scallop gill was more sensitive to BaP than the digestive gland, and the digestive gland was the main tissue to deal with oxyradicals. The contents of malondialdehyde (MDA) increased with the exposure time and there was a positive correlation (concentration-effect) between the MDA content and the concentration of BaP. The biomarkers determined in this experiment had important roles in detoxification, and showed great potential as biomarkers for oxidative stress. Controlled laboratory experiments designed to simulate field exposure scenarios are particularly useful in ascertaining biomarkers suitable for use with complex contaminant mixtures in the marine environment. Supported by the Technology Development Program of Shandong (No. 2008GG1005010) and the Program of Introducing Talents of Discipline of Universities (111 Project, No. B08049)     

Polypeptide from Chlamys farreri inhibits UVB-induced apoptosis of HaCaT cells via iNOS/NO and HSP90

Polypeptide from Chlamys farreri(PCF) is a novel marine bioactive product that was isolated from the gonochoric Chinese scallop Chlamys farreri,and was found to be an effective antioxidant in our recent studies.In this study,we investigated the effect of PCF on ultraviolet B(UVB)-induced apoptosis of HaCaT cells and the intracellular signaling pathways involved.Pretreatment with the inducible nitric oxide synthase(iNOS) inhibitor S-methylisothiourea sulfate inhibited UVB-induced apoptosis,indicating that iN...     

CYTOLOGICAL COMPARISON OF GAMETOGENESIS OF SCALLOPS, ARGOPECTEN IRRADIANS AND CHLAMYS FARRERI

Histological characteristics of gametogenesis of two kinds of scallops, gonochoric Chinese scallop, Chlamys farreri and hermaphroditic bay scallop, Argopecten irradians were investigated in this study. Spermatogenesis in C. farreri has different developmental stages: spermatogonia, primary spermatocyte, second spermatocyte, spermatid and spermatozoon. A large number of same developmental stage spermatic cells converge at a definite area of the testis. Premeiotic, previtellogenic and vitellogenic oocytes can be found during oogenesis in C. farreri, where oocyte distribution is obviously irregular. The A. irradians gonad consists of two different parts in one individual: one part functions as testis, the other as ovary. Between these two parts is a special appearance area, where a large number of spermatic cells are bound with two layers of acellular substance with many oocytes in it.     

Studies on the antitumor activity and isolation of the glycoprotein from the gonad ofChlamys (Azumapecten) farreri

After being degreased with acetone, the female gonad ofChlamys (Azumapecten) farreri was extracted with water, and then subjected to DEAE—cellulose column chromatography and purified by Sephadex G-100 chromatography to obtain a pure female gonad glycoprotein (FGGP). The IR spectra, electrophoresis and chemical analysis of FGGP indicated that it was a kind of acid glycoprotein and that the content of total protein and sugar were 54.8% and 40%, respectively. Pharmacological tests showed that FGGP could inhibit (inhibition rate of 40.87%) the growth of Sarcoma 180 of mice.     

A polypeptide from Chlamys farreri inhibits UVB-induced HaCaT cells apoptosis via the Apaf-1/caspase-9 and Smac/XIAP signaling pathway

A novel marine active polypeptide(PCF),isolated from the gonochoric Chinese scallop,Chlamys farreri,has potential antioxidant and anti-apoptotic activity against ultraviolet irradiation.We investigated whether UVB-induced HaCaT cell apoptosis occurs via the mitochondrial pathways Apaf-1/caspase-9 and Smac/XIAP/caspase-3.We then investigated the molecular mechanisms controlling the anti-apoptotic effect of PCF.Pre-treatment with PCF and caspase-9 inhibitor significantly inhibited UVB-induced apoptosis in HaC...     

IMPACT OF STARVATION ON SURVIVAL，MEAT CONDITION AND METABOLISM OF CHLAMYS FARRERI

The effects of 60-day starvation on survival rate. condition index (CI) , changes ofnutrient composition of different tissues, respiration and excretion of scallop ChlartnCs farreri were studiedin laboratory from Oct. 17 to Dec. 15 ,1997. Two groups (control and starvation with 200 individualseach) were cultured in two 2 m3 tanks, with 31 to 32 salinity water at 1 7 “C . Starvation effects were mea-sured alter 1 0, 20, 40 and 60 days. There was no mass mortality of scallops cf the two tanks and survivalrates of the control and starvation groups were 93 .5 % and 92.0 %, respectively. Starvation had strongeffect on the meat condition of the scallops, especially after 10 days; when relative lipid percentagedropped sharply while relative protein percentage increased. The impact of starvation on the oxygen consumption mtc (OCR) and the ammonia-N cxcretion mtc (AER) was obvious.     

Polyculture of scallopChlamys farreri and kelpLaminaria japonica in Sungo Bay

Several polyculture models of scallopChlamys farreri and kelpLaminaria japonica currently employed in Sungo Bay and other parts of northern China are described in this paper. Economic benefits of different polyculture models are analysed based on the growth rate, culture density and market price. In addition, site selection, critical environmental conditions and polyculture problems are discussed in detail. This project was funded by the International Development and Research Centre (IDRC), Canada, and the National Committee of Science and Technology of China.     

Variations in retention efficiency of bivalves to different concentrations and organic content of suspended particles

Retention efficiencies (RE) of scallop (Chlamys farreri), oyster (Crassostrea gigas) and mussel (Mytilus edulis) in a flow-through system were measured to understand the short-term response to various particle and organic matter concentrations. By comparing the RE of C. farreri with that of C. gigas and M. edulis, we gained further knowledge on the feeding physiological characteristics of C. farreri and ascertained the possible cause of high summer mortalities of this species. The experimental feeding conditions included natural differences in the abundance and composition of suspended seston, as well as conditions in which seston abundance and composition were manipulated by adding natural silt or cultured microalgae. The results show that in natural sea water, the minimum particle size for maximal retention in M. edulis, C. gigas and C. farreri was approximately 4, 6, and 8 μm, respectively; the RE of 2-μm (equivalent spherical diameter) particles was 17%, 19%, and 8%, respectively; and the relative standardized RE was 58%, 49%, and 18%, respectively. In C. gigas and M. edulis, the minimal particle size for maximal retention did not change with food quality (organic content). C. farreri was more sensitive to fluctuations in particle concentration and organic content. With particle concentration increase, the minimal particle size for maximal retention in scallop shifted to large particles and the RE for 2-μm particle decreased from 8% in natural seawater to 1.6%–6% in silt-enriched groups. With organic content increase, the minimal particle size for maximal retention shifted from 8 to 5 μm in natural seawater.Variation in RE of C. farreri with food conditions and the relationship between lower RE and smaller particle size may hamper C. farreri from food taking due to the decrease in the size distribution of phytoplankton in Sungo Bay.     

Isozyme expression of Chinese and Japanese populations of Chlamys farreri and their reciprocal hybrids

Chinese and Japanese population ofChlamys farreri and their reciprocal hybrids were surveyed in isozyme variability at 13 loci by polyacrytamide gel electrophoresis (PAGE). Isozyme banding patterns indicated these hybrids were diploid. Loci that were observed as being monomorphic in inbred populations ofC. farreri were also found to be monomorphic in filial progeny; loci that observed to be polymorphic in parental type generations were also polymorphic in hybrid generations. Differences existed among allelic frequency of the four types of cross. Within the reciprocal hybrids the expression of malic enzyme (ME) isozyme was sufficient to distinguishing individual hybrids because of the band, Rf=0.38. However, there were no noticeable variations among all the samples to differentiate one from another. Inbreeding was likely to be the main problem in aquaculture. The introduction of new broodstock can improve the genetic diversity. Hybrid vigor has manifested to a certain extent in the present study. Supported by the National “973” Program (G1999012008) and “863” Program (2002AA626020).     

Survival, growth and immune activity of scallop Chlamys farreri cultured at different depths in Haizhou Bay (Yellow Sea, China) during hot season

Survival, growth and immune response of the scallop, Chlamys farreri, cultured in lantern nets at five different depths (2, 5, 10, 15, and 20 m below the sea surface) were studied in Haizhou Bay during the hot season (summer and autumn) of 2007. Survival and growth rates were quantified bimonthly. Immune activities in hemolymph (superoxide dismutase (SOD) and acid phosphatase (ACP)) were measured to evaluate the health of scallops at the end of the study. Environmental parameters at the five depths were also monitored during the experiment. Mortalities mainly occurred during summer. Survival of scallops suspended at 15 m (78.0%) and 20 m (86.7%) was significantly higher than at 2 m (62.9%), 5 m (60.8%) or 10 m (66.8%) at the end of the study. Mean shell height grew significantly faster at 10 m (205.0 μm/d) and 20 m (236.9 μm/d) than at 2, 5 or 15 m in summer (July 9 to September 1); however, shell growth rate at 20 m was significantly lower than at the other four depths in autumn (September 2 to November 6). In contrast to summer, scallops at 5 m grew faster (262.9 μm/d) during autumn. The growth of soft tissue at different depths showed a similar trend to the shell. Growth rates of shell height and soft tissue were faster in autumn than in summer, with the exception of shell height at 20 m. SOD activity of scallops increased with depth, and ACP activity was significantly higher at 15 and 20 m than at other depths, which suggests that scallops were healthier near the bottom. Factors explaining the depth-related mortality and growth of scallops are also discussed. We conclude that the mass mortality of scallop, C. farreri, during summer can be prevented by moving the culture area to deeper water and yield can be maximized by suspending the scallops in deep water during summer and then transferring them to shallow water in autumn.     

Tetraploid induction by inhibiting mitosis I in scallopChlamys farreri

Tetraploid induction was carried out by inhibiting mitosis I in fertilized eggs ofChlamys farreri. Mitosis I was blocked with cold shock (5–7°C), Cytochalasin B (0.75 mg/L) and 6-dimethylaminopurine (6-DMAP) (60–75 mg/L) when 60% fertilized eggs released polar body II at 20°C. At 4-cells embryo stage, the ploidy was determined by counting chromosome number. In control groups, most embryos were diploids (72.22%) and aneuploids (24.78%). In Cytochalasin B, cold shock and 6-DMAP treated groups, tetraploids were respectively 10.51%, 4.08%, and 13.34%; aneuploids were 43.10%, 35.93% and 29.16%, and triploids were 7.84%, 8.52% and 18.33%. At D-larva stage, ploidy was determined by flow cytometry (FCM). The ploidy analysis of day 2 larvae showed diploids in control group and also in three treated groups. Juvenile scallops (0.2–0.3cm) which were harvested in two control groups and two CB treated groups were all diploids through checking ploidy individually by FCM. Contribution No. 3648 from the Institute of Oceanology, Chinese Academy of Sciences. Contribution No. 238 from the Experimental Marine Biology Laboratory, Chinese Academy of Sciences. Project funded by grant (No. 819-01-07) from the Chinese Science and Technology Ministry and also by “Hundred Talents Plan” grant from the Chinese Academy of Sciences.     

Effects of different combinations of cryoprotectants on spermatozoon vitality of Chinese scallop,Chlamys farreri,during low temperature equilibrium

The effects of membrane penetrating anti—freezing agents (MPAAs), DMSO (dimethyl sulfoxide), glycerol, EG (ethylene glycol) and methanol in combination with different cryoprotective additives such as carbohydrates, macromoleules and inorganic compounds on the spermatozoon vitality of Chinese scallop,Chlamys farreri, during 1 h 0 °C equilibrium were investigated. When only MPAAs existed, the detrimental effects of different MPAAs ranked in the following order: DMSO<methanol<EG<glycerol. When carbohydrates were added into MPAAs solution, 5% glucose caused larger decrease of spermatozoon vitality than 2.4% lactose. 5% glucose or 2.4% lactose in 7.5% glycerol caused complete damage. 10% yolk was best in maintaining the spermatozoon vitality except when used in combination with 10% methanol. 10% milk significantly decreased spermatozoon vitality in EG and methanol and enhanced its vitality in glycerol, but did not significantly influence it in DMSO. Glycine MPAAs ranked in the following order. 10% yolk <10% yolk +2.4% lactose <2.4% lactose <2.0% citrate <10% milk <10% milk +2.0% citrate <5% glucose <80mmol/L glycine +55 mmol/L NaHCO₃. Contribution No. 2434 from the Institute of Oceanology, Chinese Academy of Sciences.     

Effects of reduced salinity on oxygen consumption and ammonia-N excretion ofChlamys farreri

Effects of reduced salinity on the oxygen consumption rate (OCR) and the ammonia-N excretion rate (AER) of scallopChlamys farreri (3.2–5.9 cm in shell height, 0.147–1.635 g in soft tissue dry weight) were studied in laboratory from March 21, 1997 to April 16, 1997. Under the controlled conditions of reduced salinity from 31.5 to 15.0 and ambient temperature 17°C and 23°C, the concentrations of dissolved oxygen and ammonia-N were determined by the Winkle method and the hypobromite method, respectively. Results showed that with controlled reduced salinity, the mean values of the OCR were 2.17 mg/(g.h) at 17°C, and 2.86 mg/(g.h) at 23°C and that the mean values of the AER were 178.0 μg/(g.h) at 17°C and 147.0 μg/(g.h) at 23°C. The OCR and the AER decreased with reducing salinity from 31.5 to 15.0 both at 17°C and 23°C. The effects of reduced salinity on the OCR and the AER of scallopC. farreri could be represented by the allometric equation and the exponential equation, respectively. Contribution No. 3295 from the Institute of Oceanology, Chinese Academy of Sciences. Project supported by the National Ministry of Science and Technology of China and by the Chinese Academy of Sciences, Grant No. 96-922-02-04 and KZ951-A1-102-02.     

Development of an in situ loop-mediated isothermal amplification technique for chromosomal localization of DNA sequences

In situ loop-mediated isothermal amplification (in situ LAMP) combines in situ hybridization and loop-mediated isothermal amplification (LAMP) techniques for chromosomal localization of DNA sequences. In situ LAMP is a method that is generally more specific and sensitive than conventional techniques such as fluorescence in situ hybridization (FISH), primed in situ labeling (PRINS), and cycling primed in situ labeling (C-PRINS). Here, we describe the development and application of in situ LAMP to identify the chromosomal localization of DNA sequences. To benchmark this technique, we successfully applied this technique to localize the major ribosomal RNA gene on the chromosomes of the Zhikong scallop (Chlamys farreri).     

Chitosan oligosaccharide-Ca complex accelerates the depuration of cadmium from Chlamys ferrari

This study investigated the effect of a chitosan oligosaccharide-Ca complex (COS-Ca) on the depuration of cadmium (Cd) from Chlamys ferrari. After exposure to 0.5 mg L-1 CdCl2 for 3 or 7 d, the scallops were treated by COS-Ca prior to determina-tion of Cd, calcium (Ca) and zinc (Zn) contents, Cd distribution in organs, malondialdehyde (MDA) content and antioxidant variables. Results showed that COS-Ca reduced Cd content in the viscera of the scallops, with highest Cd depuration rate (47%) observed on day 3. The COS-Ca concentration substantially affected Cd depuration, and the exposure to 8.75 mg L-1 COS-Ca led to significantly higher Cd depuration rate compared with those of lower COS-Ca concentrations (1.75, 3.5, 5.25, and 7.00 mg L-1). Distribution analysis of Cd in scallop organs indicated that COS-Ca significantly reduced Cd content in the kidney throughout the 5-d experiment, as well as in the gill during the early stage of Cd depuration. In addition, COS-Ca treatment decreased glutathione peroxidase (GSH-Px) activity and MDA content while increasing superoxide dismutase (SOD) and catalase (CAT) activities on different days. Our work suggested COS-Ca complex treatment as an effective method for acceleration of Cd depuration from Cd-contaminated bivalves.