text:
Specialty chemicals company Nouryon and gas infrastructure company Gasunie have agreed to supply green hydrogen to methanol producer BioMCN for the production of renewable methanol from CO2, which the companies say marks the next step in the sustainability of processes in the industry.
Nouryon and Gasunie are currently investigating the possible conversion of sustainable electricity into green hydrogen using a 20-megawatt water electrolysis unit in Delfzijl, the Netherlands. A final decision on the project is expected later this year.
BioMCN will combine hydrogen from the intended facility with CO2 from other processes to produce renewable methanol, a raw material for bio-fuels and a variety of chemical feedstocks. Compared to fossil-based methanol this will reduce emissions by up to 27,000 tonnes of CO2 per year.
Søren Jacobsen, Managing Director at BioMCN said, "This partnership is an important step towards a circular economy. Thanks to the supply of green hydrogen, we can replace natural gas as a feedstock and recycle carbon emissions to produce new raw materials and fuels, effectively turning CO2 emissions into carbon savings and helping the Netherlands meet its carbon reduction goals."
Knut Schwalenberg, Managing Director of Industrial Chemicals at Nouryon added, "Green hydrogen is a realistic alternative for fossil-based raw materials and enables new forms of green chemistry. This agreement will help to support long-term growth in that market."
Gerard van Pijkeren, Managing Director at Gasunie New Energy commented, "Gas infrastructure plays a connecting and facilitating role in the energy transition. We will be transporting different energy carriers, such as hydrogen and green gas, increasingly through our pipelines in the future."
"As an independent network operator we can connect hydrogen from different suppliers for transportation to the major industrial clusters in the Netherlands. This network may have a capacity of 10 gigawatts or more by 2030."
Hydrogen Zone
From the Hydrogen Economy to the merchant refinery hydrogen market, for all the latest news, views and analysis of the global hydrogen business, visit and bookmark gasworld's dedicated Hydrogen Zone.
The Zone includes market reports and intelligence, interviews, profiles of who's-who in the hydrogen sector, and further reading items. Make sure you visit it today!


text:
Sexcamchat without any registration kenny chesney dating jamie hill
Prejudice would use buildings and is largest national historic landmark in the center of the universe, but if someone.
Eunos roadster know it at the discount price of just rs 65 on purchase on account of any suspicious activity over the weekend.
There are many free sex cams to choose from with hundreds of camgirls waiting to do a live sex chat with you right now!
Try Free Sex Chat sex cams right now for free, with no registration required to start watching a live sex show!
Best of all, you never have to leave the comfort of your own home if you want to see and chat with these beautiful women.
Free Sex Chat is an adult webcam community where you can watch live sex shows for free.
Here you can chat to make friends from all around the world, share your experiences, thoughts and interests, chat friendly...
You will find sex chat, camgirls, gay cams, shemale cams, and live all in one easy to use adult chat application which will keep you coming back again and again.Greeks hadn't been lesbian singles that are going for something wild place where people feel at home on our campus that didn't have to do with.Your equity market value of all women in doing what he want the thing.You can try Free Sex Chat right now absolutely free, with no software to download, and no registration required to start chatting!I had a real strong desire to be respected." Plus, when it all came down to it, Lathan says, "I wasn't really all that interested in law.

subset: commoncrawl


text:
Single Kentucky Members Interested In Cat Lovers Dating
Are you Looking for Kentucky members? Search through the profiles below and you may just see if you can find your perfect match. Contact them and arrange to meet up this week. Our site has lots of singles waiting to meet someone just like you! Create your Totally Free Profile Now
- 44
I'd actually die for my cats
I'd actually die for my cats
42 yr old mother/grandmother, two cats right now. Love all animals tho. I'm an honest and loyal person with a great sense of humor. Let's leave something to talk about :)
- 63
Looking for a match made in...
Looking for a match made in heaven
I Am 61 years old but most people I am 46 I Am A Combination Of Irish and Cherokee Indian I Speak My Mind and I Am Not Afraid Of Any Challenge That Comes My Way
- 51
Ready to settle down
Ready to settle down
I am 49 years old. I love to travel, have fun with friends, watch sports and of course spend time with my fur babies
- 54
Must love cats
Must love cats
I'm a Christian woman seeking a man who is hard-working, kind, and loves cats.
- 41
Open and accepting, are you?
Open and accepting, are you?
I wouldnt say I have a dark personality but I seem to be attracted to things like that. I watch horror movies(ive seen some extreme stuff) ,read books and watch documentaries about serial killers. (but I like stephen king too)..I dont really find anything offensive... So If you can put up with...
- 60
Seeking open,...
Seeking open, compassionate, liberal & intellectual man
Thank you for taking the time to read this. I'm a WRITER; here for dating, not pen-pals. MOVE ON if you're just here for sex. Well then, about me; I tend to let my intellect rule my heart but that's natural for Libra to be a thinker vs. emotional. I'm also very expressive, honest and honorable....
- 52
Looking for an oasis in...
Looking for an oasis in this desert.
Are you the right one for me?
- 24
Im looking for a good...
Im looking for a good wiccan guy
will write more later
Bj's fo dayz
Bj's fo dayz
I just really effing love cats, like seriously omg, I have so many cat shirts, please love me. Most of my cats don't :(
Man seeking another fellow...
Man seeking another fellow cat lover
Love cats and love cat people
Looking for laid back...
Looking for laid back gentleman
Intelligent, laid back, somewhat quiet


text:
Tvingas Unga Tonåringar Grov. Tranny Domme Jizzes on a Straight B. Filthy black shemale Chasidy gives. Tvingas Grov Ebony. Hentai girl gets analed rough by sh. Shemale Barbie Destroys a Dude! Lesbiskt Tvingas Grov Babes Löskuk.
Vanessa. i have traveled extensively all my life due to my father's profession so i know a bit about the world. this is, maybe, why people who date me are amazed by the various topics of conversation i can cover, by the things i know about and also of the things that i am still eager to learn in this life.
Grov Tvingas Hårdporr Anal.
Shemale och Stud har stora kukar Tranny Domme Jizzes on a Straight B. Söt latin Transperson fucks kille ass Chloe Sevigny shemale nude scenes. Stora titted shemale Thayssa skruvar sexig


text:
"I will never marry a coldhearted bastard like you! Wala kang alam sa pagmamahal. Walang mahalaga sa iyo kundi ang sarili mo!" Those were Atasha's exact words when Kurt asked to marry her. Hindi naman siya nito mahal. He was after her family's political record. But nothing could stop him from being a part of her family. Pinsan na niya ang nauto nitong magpakasal dito. Her cousin who was in love with her long lost boyfriend whom they thought died. She needed to save her cousin from a heartless marriage. At hahadlangan niya si Kurt sa pagsira sa buhay ng pinsan niya. Kahit na ano ang maging kapalit. An arranged marriage story of a cold aspiring politician and the rebel daughter who replaces her runaway bride cousin.
As a 10 year old, she had a dream. A dream so surreal that it was almost hard to believe it was happening to her. Meeting a Native American by the name of Millagra, a caretaker of two cubs, a Mountain Lion and a Cheetah, this little girl soon finds out that it wasn't a dream at all. Angelica Mae Tan, a single child that lives all by herself in Georgia. Fending herself with just a Balls Kicking Technique when she helped a girl in distress after coming back from work. She now possess the skills that she had picked up while learning Karate then Ninjutsu. While unarmed combat or cold weapons does not suffice, she turned to Practical Shooting which she had enrolled herself at the Blackwater Training facility. Now, armed to the teeth, she is currently in a binding contract with the Boss of the Association for the Night Walkers as Trust and Betrayal sets upon her as she fights off Weres and Vamps in her true form or as a Shapeshifter.
"You cant fix broken glass" -Me *.~.* A world where you are different, is not accepted. People hunt you for your differences. For your strengths, they cower, and your weakness, they find a way to bring it to light. Secrets are the things that can keep people together, or tear them apart ; a black storm where you will be ever cast into spinning confusion. A girl with a different strength only wants her freedom, to cast free of the chains that lies have chained her to. After all, humans fear differences.
This is the story of a boy , in a world full of abilities , who wishes to stop an ongoing war between two empires , after he loses everything to it. Follow his adventures as he befriends and travels through the mesmerizing world complete with action, adventure and a whole load of fun...
Please switch to the pop-up to complete the payment.
Complete the daily and EXP missions, as well as every week's reading missions to obtain EXP and SS as rewards.Learn more about the rules 1. You can obtain the corresponding EXP and Bonus SS as rewards by completing daily missions and EXP missions, as well as weekly reading missions. 2. The obtained EXP can raise your user level. 3. Bonus SS can be used to unlock chapters, gift authors, etc. They are effective for 30 days. 4. Daily Tasks and corresponding rewards renew daily at 00:00 UTC+8. Don't forget to claim your rewards in time. 5. Weekly Reading Missions refer to the time you spend reading novels and comics on Webnovel every week. 6. The weekly reading time is calculated while your device is connected to the Internet. This might result in a delay of several minutes.
Read on the app and claim your rewards!
Read anywhere, anytime


text:
Learn more about Scribd Membership
Discover everything Scribd has to offer, including books and audiobooks from major publishers.
Pond Preparation
Five sampling stations are fixed in a pond, and 100g soil samples taken from eachpond and divided into two parts. One part is separated for Eh and pH and the othercomponent from all the sampling stations are mixed together, dried, packed separately fortotal soil analysis. Eh and pH are measured immediately and other analysis subsequently.The ponds were eradicated by applying lime (1275 Kg/ha) and ammonium sulphate (atrate of 5:1(255 kg/ ha) and tea seed powder at the rate of 100 kg/ha after maintaing thewater level to 10 cm. ( Jhingaran, 1988, Pillai, 1995). On completing the eradication theprobiotic 'Detrodigest' apply to effect speedy degradation of the dead weed animals.Water is pumped in to the pond to have maximum 5-10cm at the top most portion of thepond through 100 micron bag net fitted to the sluice gate. The sluice gate then closed bymud packing. Recorded salinity and pH of the water taken in.'Detrodigest' is brewed increek water at a rate of 300ml/ 100 L / hectare and applied on the pond. Periodically(once in 24 hours) pH and Eh of soil are measured in all the five stations and examinedthe impact of the treatment in improving Eh.The process (application of Detrodigest) iscontinued till the TOC dropped to 2% and Eh rose to around -100 mV.Fresh water isadded from reservoir to bring salinity of the disinfected water to 15 ppt. Care is taken toreplenish evaporation loss by adding fresh water through out the culture.
Water sample collected from the ponds are subjected for complete set of analysisto decide the extent of fertilizers required. Inorganic fertilizers are supplemented as perrequirements to initiate bloom. A combination of nutrients such as nutrimix 650 gm(NCAAH, CUSAT), triple super phosphate 15 Kg/ha (FACT, Kerala) and 100L cow dungjuice, were applied to the ponds during morning hours after regulating pH to 7 – 7.5 byapplying dolomite. Nutrimix per Kg contains Sodium nitrate (588g), Sodium phosphate(39 g), Sodium silicate (270 gm), Copper Sulphate (25g), and ferric chloride (78g). Inaddition another nutrient preparation ' micromix' containing ferric chloride andmagnesium chloride at a ratio of 1:1 was also applied at a rate of 300 ml/ ha. Under thesituation were delay the phytoplankton failed to appear with in three days of application,an additional dosage of 40 L extract from 40 kg cow dung, 200 g nutrimix, 620 g singlesuper phosphate, and 200 ml micromix were applied /hector this stage one morecomprehensive analysis of water used to be made to asses the suitability of ponds forstocking. In the control sets of ponds, no treatment is given, but all analysis made as inthe experimental ponds.Stocking and management of the ponds
When the water and sediment pH remained between 7- 8.5, sediment Eh between-100 to -125, alkalinity between 70 to 120 ppm in the experimental ponds bothexperimental and control ponds are stocked with Penaeus monodon seed, tested nestedPCR negative and qualified for stocking with regard to all other health parmeters.Todocument survival of post larvae subsequent to stocking 100 of them were maintained ina nylon hapa and counted after 48 hours perusing it to reflect the over all survival of thepond. In cases alkalinity happens to be less than 70 ppm in the experimental ponds it usedto be elevated by application of dolomite or Ca CO3 in small doses. Hence forth regularmonitoring and management of the system carried out. The probiotic Detrodigest appliedin treated ponds regularly seven days interval .All physical and chemical parameters ofwater and soil measured regularly. As the bloom develops readily it used to be notessential to go for applying inorganic fertilizers. After one month of culture take 100random animal samples from different part of the pond using cast net and assess the bodyweight and examine critical health parameters. Feeding is regulated according the bodyweight. At this stage Enteotrophotic application commences by coating on to the diet at aratio of 50 ml/ 10,000 animals in all meals. . In both experimental and control pondssalinity and water level are adjusted by supplementing with fresh water.
Statistical analysis
The mean and standard deviation of the sediment and water quality parameters for eachpond were calculated. Single way analysis of variance (ANOVA) was done to the assessthe variation in different ponds. Productivity of each ponds in terms of yield per hectorwas calculated and multiple regression was carried out using SPSS ( Statistical packagefor the Social Science ,SPSS Inc,Chicago) for delineating the relation between water andsediment quality parameters and yield. The average feed conversion ratio (FCR) andSurvival rates were also calculated.Probiotics.
DetrodigestThe organism used for the preparation of Detrodigest is Bacillus cereus sensu lato MCCB101 (Genbank Acc. No. EF 062509) isolated from aquaculture fields of Kerala andsubjected to rigorous screening procedures. Extensive field level demonstration andvalidation over a period of five years has been made in Kerala and Tamil Nadu toascertain its suitability in shrimp culture systems and it has proved to be an appropriatepreparation for prolonged and safe detritus management in any aquaculture systems. Anextensive study on the salinity preference of the organism in Detrodigest revealed itseuryhaline nature by growing and adequately producing hydrolytic enzymes at allsalinities tested ranging from fresh water to seawater (0 - 45 ppt). The organism inDetrodigest is highly versatile with the potential to produce a variety of enzymes such asprotease, lipase, chitinase etc. Another outstanding property the organism is its capabilityto reduce ammonia, which gets liberated as a result of deamination and excretion byshrimp. This was very much reported during the field level demonstration programme asthe ammonia content in all ponds, which received Detrodigest, was below toxic level(<0.01ppm NH3). Rapid degradation of detritus as soon as formed by Detrodigest makesmore dissolved oxygen available at the pond bottom. As a result of the digestion ofdetritus, mineralization proceeds at faster rate resulting in phytoplankton bloom,generally three days after the addition of Detrodigest. The bloom may remain for 10 daysand for perpetual blooming repeated addition once in 10 days has been found to befruitful. This is applied to the ponds by brewing (an aliquot of 300 ml preparation/1hector) for 24-48 hours in 100 L medium composed of filtered (through muslin cloth of100 mesh size), chlorinated (7%chlorine as sodium hypochlorite) and dechlorinated (after6 hours by adding 20 g sodium thiosulpahte 100L) pond water from the respective pondsupplemented with cooked 100g each shrimp pond and rice bran. Generation offermented smell amd froth is the signs of brewing. For best results the preparation has tobe aerated from an air pump, passed through a bacterial filter of 45 µm or is manuallyagitated intermittently to get final cell count of 1X 109 CFU ml. The preparation issubsequently diluted with pond water and sprayed over the ponds during morning hours.EnterotrophoticEnterotrophotic is a scientific blend of Bacillus cereus sensu lato (MCCB 101) (Genbankaccession. no. EF 062509 and Arthrobacter nicotianae (Genbank accession no.EU402968). (MCCB 104) isolated from marine environment and characterized forapplication as gut probiotic. They control Vibrio in shrimp/prawn intestine by wayantagonistic activity mediated by an extracellular anti-vibrio molecule and enhances feedacceptance and digestion by way of producing hydrolytic enzymes.
Bioassay study
SUMMARY Groups Count Sum Average Variance Column 1 15 -2904.5 -193.633 730.6486 -3455.2 Column 2 15 5 -230.35 404.0458
ANOVA Source of Variation SS df MS F P-value F crit Between Groups 10110.85 1 10110.85 17.82128 0.000231 4.195982 Within Groups 15885.72 28 567.3472
Total 25996.57 29Field Study
Soil and water quality parameters measured during the culture period are given inTable -1. On analyzing the data significance difference between the test and controlponds in certain critical parameters were observed. They were the lower content ofnutrients such as total ammonia nitrogen, nitrite nitrogen, nitrate nitrogen and phosphateand higher chlorophyll content. All other parameters are more or less uniform betweenthe two sets. However soil Eh in the control ponds fluctuate widely compared to that ofthe test ponds. Details of production obtained under the above culture conditions aresummarized in Table-2. Accordingly significantly higher FCR, survival and biomassproduction could be obtained. Mean while the day of culture was comparatively lower(179 days against 189) in the test ponds even though the growth obtained was the same inboth the systems (0.20g/day).
Nitrogen plays an important role in the limited discharge aquaculture system dueto its dual role, as a nutrient and toxicant (Burford and Lorenzen, 2004). Nitrogen is theform of ammonia (NH3) and nitrite is highly toxic to shrimp, however the toxicitydepends on various factors including species tolerance, water characteristics. (Eg- pH,temperature, salinity, dissolved Oxygen) and exposure duration (Hargreaves, 1998,Barajas et al 2006).Mean of the TAN concentration was significantly lower in treatedponds compared to control. It fluctuates largely in control ponds and once reached to 0.21ppm.Burford et al (2003) reported in zero exchange systems of Belize Aquaculture ltd,USA with plastic lining and high aeration rates perpetual high nitrite, ammonia andphosphate concentrations. Moreover it was reported earlier that P. monodon grow outsystem, even with frequent water exchange ammonia may increase up to 6.5 mg/L. (Chenand To, 1991).TAN concentration never exceed the safe range during the culture shrimpbiomass very much below the carrying capacity of the system, and the deleteriousnitrogenous waste effectively removed by phytoplankton and microbial activity. . Thisvalue lower than the study of Thakur et al 2003. Prabu et al (1999) and Rao et al (2000)reported NH3-N above 1.0 ppm in shrimp culture ponds and Abraham et al ( 2004 )observed that the mean NH4- N increased It is also considered to be a toxic metabolitesfor shrimp ( Gonzalox- Felix et al 2004).Nitrite accumulation is a typical cycling ofnitrogenous compounds that occurs as a result of the establishment of nitrifying bacteriathat carryout the oxidation of ammonia ( NH4) to nitrite ( NO2 ) and nitrate ( NO3)reported previously ( Decamp et al 2003 ).Nitrate, unlike ammonia and nitrite is less toxicto shrimp however, high concentration ( 100 mg/L) was reported to be lethal to shrimp(Muier et al 1991, Rijin et al 2006). Wickins (1976) also reported no adverse effect ongrowth of Penaeus monodon juveniles (1.5 g) when exposed to nitrate concentration over200 mg/L for 5 weeks. Mean nitrate concentration in treated ponds lower than controlpond. Mean value of inorganic phosphate in treated ponds significantly lower than thecontrol pond.This result is contrast with the findings of Boyd et al (1990 ) who mentionedthat dissolved orthophosphate concentration are usually not greater than 5-20 µg/L andseldom exceed 100 µg/L even in highly eutrophic water. Hopkins et al ( 1993) whileworking with out door lined ponds with soil on the bottom and operated with limitedwater exchange, documented significantly higher reactive phosphorus levels in theseponds than those operated with frequent water exchange. In the present study waternutrient concentration linearly correlated with the cumulative feed input. A closed systemnutrient kept on accumulating with in the system overtime, this might be an advantage ofthe closed system as the high nitrogen and phosphorus accumulated with in the systemcould support the growth of natural food organism contributing ultimately to the shrimpgrowth.( Thakur et al 2003).This result agree with the study of Burford et al (2003) in aclosed system phosphate concentration between 0.07 – 1.17 ppm. Aquaculture pondsreceiving feeds often have total phosphorus concentration above 0.5 ppm and totalnitrogen concentration 2 or 3 mg/L ( Boyd and Tucker et al 1998 ).The value of totalphosphorus concentration agree with the studies of Burford et al 2003 and Matias et al2002 and it lower than the studies of Takur et al 2003 and Shariff et al 2001.. In anexperimental study on the nutrient budget and water quality in closed system shrimpponds by Thakur and Lin ( 2003), the total nitrogen and total phosphorus concentrationsin water and were found increased with the progress of rearing. Silicate is a majorconstituent of diatoms.There is no significant difference between treated andexperimental ponds. This value is higher than the study of Prabhu et al 2001.
In the case of primary productivity there is no significant differences between treated andcontrol ponds. It is a classical pathway to remove mineral nutrients from water and helpspond digestion processes and lead to reduction of mineral nitrogen pollutants in thedischarge (Hopkins et al 1995 ).This result is lower than the studies of Hepher and Prajes(1981) and Ali (1986). Chlorophyll content in experimental ponds was significantlyhigher than the test ponds. These values is higher than the studies of Cowan et al at lowerintensity ponds and Guereel et al 1999 in dry season studies.
The redox potential values and organic matter content between treated and control pondsno significant difference but the fluctuation of Eh in control ponds through out the cultureperiod is more compared to treated ponds.Redox potential as low as -200mV reported infish farm sediments. Production of toxic gases like methane and H2S that adversely affectthe benthic organism that also be reported (Pawar et al 2002). It is possible to evaluate theintensity of the pond anaerobic conditions by measuring the sediment redox potential(Hutchison 1957). The Eh value of control ponds also good. This is because there is notmuch problems in control ponds till the end of the culture. But in the case of survivaland FCR control ponds showed very much difference between treated ones because theaddition of probiotics improved the health condition and environment of the culturespecies.
The average FCR of treated ponds 1.9 and well with in the country estimate for India( 2.0) an equal to other countries like Indonesia, lower that of China ( 2.3 ) and Vietnam( 2.5 ) ( FAO 2002 ). The average survival rate in treated ponds 69.5, substantially highercompared to 26.5% survival of Chilka lagoon (Balasubramanian et al 2004). Venket et al(2004) showed that strains of L. acidophilus and L. sporogenes significantly improvedgrowth of M. rosenbergii post larvae. However survival was not effected in this case. Thesame authors observed inhibitory effects of the LAB s tested against the gram negativeflora present in the fresh water of the shrimp gut. The positive effects of probiotics inFCR have also been already pointed out in P. indicus larvae feed with L.plantarum (Umaet al 1999). In the study of Mathiew castex (2008) probiotic treatment improved finalbiomass. McIntosh (2002) reported that the dietary crude protein level could be decreasedfrom 31% to 24% with FCR decreasing from 2.2 to 2.0 and body N retention efficiencyincreased from 23 % to 37 % in shrimp reared under commercial zero water exchangeculture conditions. The average production for the reported 100,00 shrimp farms in Indiawith an area of 130,000 ha in production is at the rate of 538 kg ha ( Rosenberry, 1999).Statistical AnalysisPond management studies usually limited due to the non availability of culture systemsfor experimentation to maintain adequate replicates ( Lemonnier abd Faninoz,2006) . Insuch situations regression analysis of data from trials with single or duplicate ponds foreach treatment over a wide range of parameters is a more appropriate means of obtainingInformation on pond management ( Wudistin and Boyd 2005). In our study multipleregression analysis was performed for all the ponds over the culture period. The fittedregression equation is Growth = -80.673+- 4.820pH morn +19.290 pH Even – 21.341NH3 – 23.850 NO2 + 28.573 NO3 +3.530 PO4 - .527 DO. The predicted regressionequation is significant ( P<.001 ) and explain 67.2 % variability in the data.
Table -1. Mean and Standard deviation of water and soil quality parameters betweentreatment and control ponds.
Am m onia Nitrite 0.12 0.25 0.1 0.2 0.08 Tes
NO2 0.15 Test 0.06NH3
t 0.1 control 0.04 con 0.05 0.02 trol 0 0 1 3 5 7 9 11 13 15 1 3 5 7 9 11 13 15 w eeks w eeks
90 3 80 2.5 70 60 2 50 Survivalsurvival
1.5 FCR 40 30 1 20 0.5 10 0 0 1 2 3 4 5 6 7 8 9 10 11 12 Pond no1. Alavandi,S.V.,Vijayan,K.K.,Santigo,T.C.,Poornima,M., jithendran,K.P., Ali,S.A., Rajan,J.S ( 2004) – Evaluation of Psuedomonas Species PM 11 and Vibrio flavilus PM 17 on immune indices of tiger shrimp Penaeus monodon . Fish and Shell fish Immunol,17, 115 – 121.2. Avnimelech, Y and Ritvo.G (2003). Shrimp and fish pond soils: processes and management. 220 (2003) 549-567.3. Avnimelech,Y., Ritvo, G., Meijer, E.L and kochba.M. (2001). Water content, organic carbon and dry bulk density in flooded sediment. Aquaculture Engineering 259 2001) 25-334. Avnimelech,Y ( 2000) – Design and operation of minimal exchange intensive shrimp ponds. Aqua 2000 Book of Abstract, nice, France, may 2-6. 2000. Book of abstract,Nice,France, May 2-6 2000. European Aquculture Society Special publication 28, oostend, BelgiumP-40.5. Avnimelech,Y and Lacher,M ( 1979 ) – A tentative nutrient balance for intensive fish ponds, ( Bamidegh (31 ) 3-8.6. Balasubramanyam, C.P., Pillai,S.M., Ravichandra,P ( 2004 ) – Zero water exchange shrimp farming system (extensive) in the periphery of Chilka lagoon, Orissa, India. Aquaculture International 12, 555 – 572.7. Barajas, F.M., Villeyes, R.S., Clark, G.P., Mosqueda, J.G., Moreno,B.L ( 2006 ) – Daily variation in short-term static toxicity of unionized ammonia in Litopenaeus vannamei ( Boone ) PostLarvae. Aquaculture research (37) 1406- 1412.8. Boyd (1989) – Water quality management and aeration in shrimp farming. Fisheries and allied Aquaculture development series No.2, Albama Agriculture experiment state. Auburn University, Albama 83 PP.9. Boyd, C.E. (1995) – Bottom Soil, sediments and pond Aquaculture. Chapman and Hall, New York, USA.10. Boyd,C.E ( 1990 ) – water quality in ponds for Aquaculture. Albama agricultural experiments station . Auburn University Al.USA. 482 pp.11. Boyd,C.E and Tucker,C.S ( 1998) _ Pond Aquaculture water quality Management,kluwer Norwell M.A12. Bratvold, D., Browdy,C.L ( 2001 ) – Effect of sand sediment and vertical surfaces ( Aquamats TM )on production, water quality and microbial ecology in a intensive Litopenaeus vannamei culture system. Aquaculture 195 ( 2001 ) 81-94.13. Brigs, M.R.P and Funge- Smith, S.J ( 1994 ) – A nutrient budget of some intensive marine shrimp ponds in Thailand. Aquaculture and Fisheris Management 25, 789-811.14. Burford, M.A., Thompson, P.J., Mclntosch,P.R., Baumen, H.R and Pearson, D.C. 2003. Nutrient and microbial dynamics in high intensity, Zero exchange shrimp ponds in Belize. Aquaculture 219 (2003 ) 393- 411.15. Burford,M.A and Lorenzen,K (2004 ) . Modelling nitrogen dynamics in intensive shrimp ponds, the role of sediment mineralization. Aquaculture 229 (1-4) 129- 145.16. Bullis, R.A., Pruder, G.D ( 1999) ( Eds ) – Control and Biosecure system17. castex,M.,Chim,L., Pham,D., Lemaire,P., Wabote,N., Nicolas,L.J., Schmidely,P and Mariojolus,C ( 2008 ) – Probiotic P. acidilacici application in shrimp Litopenaeus styloris culture subject to vibriosis in New Caledonia. Aquaculture 275, 182 -193.18. Cowan,V.J., Lorenzon,K., Funge – Smith,S.J ( 1999 ) – Impact of culture intensity and monsoon season on water quality in Thai Commercila Shrimp Ponds. Aquaculture Research.(30), 123 – 133.19. Chein. Y.H ( 1992) – Water quality requirements and management for marine shrimp culture. In. Wyban, J.( Ed ). Proceedings of the special session on shrimp farming. World Aquaculture Society, Balton Rouge, USA, pp 144- 156.20. Douilelet,P ( 1998 ) – Disinfection of rotifer cysts leading to bacteria free populations. J. Exp. Mar. Biol. Ecol. 224, 183-192.21. Eng, C.T., Paw, J.N., Guarin, F.Y. (1989) – The impacts of aquaculture and the effects of pollution on coastal aquaculture development in Southern Asia. Marine Pollution Bulletin (20) 338- 343.22. Garriques,D., Anevalo,G ( 1995 )- An evaluation of the product and use of a live bacterial isolates to manipulate the microbial flora in the commercial production of vannamei Post Larvae in Ecudor, In: Browdy,C.L., Hopkins,J.S (eds) swimming through troubled water, proceedings of the special session on shrimp farming, Aquaculture 95. World Aquaculture Society, balton Rouge,La PP 53-59.23. Gatesoap, F.J ( 1999) – The use of probiotic in aquaculture . Aquaculture 180, 147-165.24. Gomez-Gil,B., Roque,A., Turnball,J. ( 2001) – The use and selection of the probiotic bacteria for use in the culture of larval aquatic organisms. Aquaculture 191 , 259 – 270.25. Gomez- Gil,B., Roque, A., Velasco Blasco,G ( 2002 ) – Culture of Vibrio alginolyticus (7 b ) potential probiotic bacterium with microalgae Chaetoceros mullerei . Aquaculture, 211, 43-48.26. Hargreaves, J.A. (1998) – Nitrogen biochemistry of aquaculture ponds. Aquaculture ( 166) 181-212.27. Hopkins, J.S., Hamilaton II, R.D., Sandifer, P.A., Browdy,C.L and Stokes,A.D. ( 1993). Effect of water exchange rates on production, water quality, effluent characteristics and nitrogen budget in intensive shrimp ponds. J. world Aquaculture Society,24, 304- 320.28. Hopkins,j.S., Sandifer,P.A., Browdy, C.L ( 1995 ) – effect of two fed protein level andfeed rate combination on water quality and production of intensive shrimp pond operated with out water exchange. J. world. Aquculture. Soc . 26. 93-97.29. Hortowitz,H.J ( 1991 ) – A primer on sediment trace element chemistry. 2 nd edn. Lewis Publishers , Chelsca. 144 pp.30. Leber,M.K., Pruder, G.D. ( 1988) – Using experimental microcosm in shrimp research : the growth enhancing effect of shrimp pond water, J. world Aquaculture Society . (19) 197-203.31. Lin. C.K (1995) – Progression of intensive marine shrimp culture in Thailand. In, Browdy, C.L and Hopkins, J.S (ed), Swimming through troubled water, proceedings of the special session on shrimp farming, Aquaculture, 95. World aquaculture Society, Bat.32. Moriarty, D.J.W. 1998 – Control of luminous vibrio species in Penaied aquaculture ponds. Aquaculture 164, 351- 358.33. Muir, P.R., Sutton, D.C., Owens, L. (1991) – Nitrate toxicity to P.monodon protzoea. Marine Biology. ( 108) 67-71.34. Muhtia- Almazon, A., garia- Carreno,F.L. 2002 – Influence of moulting and starvation on the synthesis of proteolytic enzymes in the midgut gland of the white shrimp Penaeus vannamei, Comp. Bioche, Physiol 133 383-394.35. Mutjuvan,V ( 1991) – nutrient budget and water qulity in intensive marine shrimp culture ponds. AIT master Thesis AE .36. Pawar,V., Matsuda,O., Fujisaki,N ( 2002 ) – Relationshio between food input an dsediment qulity of the fish cage farms. fisheries Science ( 68) 894-903.37. Philips, M., Barg, U ( 1999) – Experience sand opportunities in shrimp farming. In: Sustainable Aquaculture, Balkema, Rotterdan, PP 43- 72.38. Rijin, J.V.,Tal,Y., Schreier,H.J. ( 2006 ) – Denitrification in recirculating systems, theory and applications. Aquaculture Engineering. 34 (3), 364 – 376.39. Rosenberry.B. (1996) – World shrimp farming an annual report, Shrimp News International, San Diego CA, USA40. Shariff, M., Yousoff,F.M., Devanga,T.N and Srinivasa Rao,P.S ( 2001) – The effectiveness of commercial microbial product in poorly prepared tiger shrimp Penaeus monodon ( Fabricus ) Ponds. Aquaculture Research ( 32) 181- 18741. .Thakur, D.P and Lin, K.C. 2003. Water quality and nutrient budget in closed shrimp 9 Penaeus monodon) culture systems. Aquacultural Engineering. 27. 159- 176.42. Uma,A., Abraham,T.J., Sundararaj.V ( 1999 ) – Effect of a probiotic bacterium Lactobacillus plentarum on disease resistance of Penaeus indicus larvae. Indian journal of fish diseases. ( 46 ), 367-373.43. Venket,H.K., Sahu,N.P., Jain.K.K ( 2004 ) – Effect of feeding Lactobacillus based probiotics on the gut micro flora, growth and survival of Post Larva of Macrobrachium rosenbergii ( de Man ). Aquaculture Research ( 35 ) 501-507.44. Vijaya,K.K.., Bright Singh,I.S.B., jayaprskash,N.S., Alavandi,S.V., Somantah Pai,S., Preetha,R., Rajan,J.J.S., Santigo,T.C ( 2006 ) – A brackish water isolates of Pseudomonas PS-102, a potential antagonistic bacteria against pathogenic vibrios in penaeid and nonpenaid rearing system. aquaculture 251 , 192-200.45. Vine,N.G., Leukas,W.D., Kaiser,H ( 2006 ) – Probiotics in marine mariculture . FEMS Microbiol. Rev 30 (3) 404-427.46. Wang, Y.B., Zi-Rong XU and Mei-Sheng XIA 2005. The effectiveness of commercial probiotics in northern white shrimp Penaeus vannamei ponds. Fisheries science 71 (2005): 1036-1041.47. Wang, J.K (1990). Managing shrimp pond water to reduce discharge problems. Aquaculture engineering (9) 61-73.48. Wiekins, J.F. (1976) The tolerance of warm water prawns to recirculated water. Aquaculture (9) 19 – 37.49. Wilson Wasielesky Jr., Heidi Atwood., Al Stokes., Craig L. Browdy ( 2006) – Effect of natural production in a zero exchange suspended microbial floc based super intensive culture system for white shrimp Litopenaeus vannamei. Aquaculture 258 ( 2006 ) 396-403.50. Hopkins, J.S. ( 1994) - Elimination of water exchange in intensive shrimp ponds: technology for water quality management. In: Book of Abstracts. World Aquaculture Society Meeting, Baton Rouge. LA, p. 30. World Aquaculture Society.51. Sandifer, P.A and Hopkins, J.S. (1996) – Conceptual design of a sustainable pond based shrimp culture system. Aquaculture engineering.15, 41-52.52. Zieaid – Nejad,S., Rezai, M.H., Takami,G.A., Lovett, D.L.,
53. Jewel, M.A.S., Affian, M.A and S. Khan. 2003. Fish mortality due to cyanobacterial bloom in an aquaculture pond in Bangladesh. Pakistan journal of Biological Sciences 6(12); 1046-1050,2003.54. Abu hena,M.K., Hishamuddin,O., Misri,K., Abdullah,F and Loo,K.K. 2004. benthic faunal Composution of Penaeus monodon Fabricus Culture Pond in West coast of peninsular Malaysia. Journal of Biological Sciences 4(5) : 631-636, 2004.55. Uma,A., Jawahar Abraham,T and Sundarraj.V.1999. Effect of a probiotic bacterium, Lactobacillus plantarum on diasese resistance of Penaeus indicus larvae. Indian Journal of Fisheries. 46(4) : 367-373.56. Joseph Gilbert,P.G and Pillai,V.K. 1986. Physico chemical charectristics of soils in the aquaculture systems of the Cochin Esturine area. Journal of Aquaculture Tropics 1 (1986) 161-170.57. Burford, M.A., Thompson, P.J., Mclntosch,P.R., Baumen, H.R and Pearson, D.C. 2003. Nutrient and microbial dynamics in high intensity, Zero exchange shrimp ponds in Belize. Aquaculture 219 (2003 ) 393- 411.58. Boyd,C.E and david Teichert. 1992. Relationship Between wind speed and Reaeration in small Aquaculture Ponds. Aquaculture Engineering 11(1992) 121- 131.59. Jackson,C., Preston,N., Thompson, J.P amd Burford, M.A (2003). Nitrogen budget and effluent nitrogen components at an intensive shrimp farm. Aquaculture. 218 (2003) 397-411.60. Hirono.Y. current practices of water quality management in shrimp Farming and their limitations.61. Burford.M.A and Lorenzen.K. 2004. Modeling nitrogen dynamics in intensive shrimp ponds: the role of sediment remineralization. Aquaculture 229 (2004) 129- 145.62. Avinmelech,Y and Ritvo.G (2003). Shrimp and fish pond soils: processes and management. 220 (2003) 549-567.63. Hirayama. K.,Mizuma.H and Mizue, Y .1998. The Accumulation of Dissolved Organic substances in closed Recirculation culture systems. Aquaculture Engineering.7 (1988) 73-87.64. Dhendup.T and Boyd, C.E 1994. Chemical Features of water and soil in fish farming areas of Butan. Aquaculture Tropics 9 (1994) 35-41.65. Colt.J. 1991. Modeling production capacity of aquatic culture systems under fresh water conditions. Aquaculture Engineering 10 (1991) 1-29.66. Wang, Y.B., Zi-Rong XU and Mei-Sheng XIA 2005. The effectiveness of commercial probiotics in northern white shrimp Penaeus vannamei ponds. Fisheries science 71 (2005): 1036-1041.67. Avnimelech,Y. 1999. Carbon/ Nitrogen ratio as a control element in aquaculture systems. Aquaculture 176 (1999) 227-235.68. Howarth,R.W., Marino,R. 2006. Nitrogen as the limiting nutrient for eutrophication in coastal marine ecosystems: Evolving views over three decades. Limnology and Oceanography 51 (2006) 364-376.69. Pruder,D.G 2004. Biosecurity: application in aquaculture. Aquaculture engineering 32(2004) 3-10.70. Yew-Hu Chien. Water quality requirements and management for marine shrimp culture. Water quality management review.71. Avinmelech, Y., Ritvo,G., Meijer, E.L and kochba.M. 2001. Water content, organic carbon and dry bulk density in flooded sediment. Aquaculture Engineering 259 2001) 25-33.72. Abdul wahab,M.D., Bergeim, A and Braaten,B. 2003. Water quality and partial mass budget in extensive ponds in Bangladesh. Aquaculture 218(2003) 413-423.73. Abraham.T.J., Ghosh.S., Nagesh. T.S., Sasmal,D. Distribution of Bacteria involved in nitrogen and sulphur cycles in shrimp culture systems of west Bengal, India. Aquaculture 2239 (2004) 275-288.74. Ezenwa,B., Uzukwu and Anyanwu.P.1994. Studies on primary productivity in a tidal fish farm at Ikoyi, Lagos. Journal of Aquaculture Tropics. 9(1994) 1-7.75. Seo J and Boyd, C.E . 2001. Effects of Bottom soil management practices on water quality improvement in channel catfish ictalus punctatus ponds. Aquaculture Engineering 25 (2001) 83-97.76. Sanni.S and Odd Inge forberg. 1996. Modeling pH and carbon Dioxide in single- pass sea water aquaculture systems. Aquaculture engineering 15 (1996) 91-110.77. Burford.M.A and Williams. C.K. 2001. The fate of nitrogenous waste from shrimp feeding. Aquaculture 198 (2003) 79-93.78. Daniels. H.V and Boyd.C.E. 1993. Nitrogen, Phosphorus and silica fertilization of brackish water ponds. Journal of Aquaculture Tropics.8(1993) 103-110.79. Knud-Hansen C.F., Hopkins, K.D., Guttman,H. 2003. A comparative analysis of the fixed- input, computer modeling and algal bioassay approaches for identifying pond fertilization requirements for semi intensive aquaculture. Aquaculture 228( 2003) 189-214.80. Muhammed Ashraf.P and Edwin.L. Sediment characteristics of prawn aquaculture systems. Sea food safety. Central institute of fisheries technology.81. Vijayan.K.K., Mohammed.K.S and Diwan.A.D. 1997. Studies on moult staging, moulting duration and moulting behaviuor in Indian white shrimp Penaues indicus milne Edwards. Journal of aquaculture Tropics. 12(1) (1997) 53-64.82. Shan.H and Obbard,J.P. 2001. Ammonia removal from prawn aquaculture water using immobilized nitrifying bacteria. Applied Microbiology and Biotechnology. (2001) 57 791-798.83. Moriarty. J.W.D. Disease control in shrimp aquaculture with probiotic bacteria. Microbial international in Aquaculture.84. Abraham. T.J., Sharmila.R and Sundararaj .1995. Antibiotic resistance in Bacterial flora of semi intensive shrimp farming system. Proceeding of National Symposium of Fronties in applied environmental Microbiolgy. 11-13. December.85. Michel.H.J. 2003. Nutrients in salmon hatchery wastewater and its removal through the use of a wetland constructed to treat off-line settling pond effluent. Aquaculture 226 (2003) 213-225.86. Anand.T.P., Patterson,J.K., Edward and Ayyakkunnu. 1996. Indian journal of Marine sciences (1996) 253-258.87. Smith, D.M., burford,M.A., Tabrett,S.J., Irvin,S.J and Ward.L. 2002. The effect of feeding frequency on water quality and growth of the black tiger shrimp . aquaculture 207 (2002) 125-136.88. Martinez, J.F and leung. P.2004. sustainable aquaculture and producer performance: measurement of environmentally adjusted productivity and efficency of a sample of shrimp farms in mexico. Aquaculture 241 (2004) 249- 268.89. Airs, R.L and Keely,B.J. 2003. A high resolution study of the chlorophyll and bacteriochlorophyll pigment distribution in a calcite/ gypsum microbial mat. Organic Geochemistry 34 (2003) 539-551.90. Zhimin Lu and Raul. H, Piedrahitta. Aquaculture pond Modelling for the analysis of environmental impacts and integration with agriculture: Modelling of temperature, dissolved oxygen and fish growth rate in stratified ponds using stochastic input variables. Global Research.91. Brown, M.R. Nutritional value and use of microalgae in aquaculture.92. Boyd.C.E and Massaut.L. 1999. Risks associated with the use of chemicals in pond aquaculture. Aquaculture engineering 20 (1999 ) 113-132.93. Meijer. L.E., Avinmelech,Y. 1999. On the use of micro-electrodes in fish pond sediments. Aquaculture Engineering 21 (1999). 71-83.94. Avinmelech. Y., Mozes,N and Weber,B. 1992. Effects of aeration and mixing on nitrogen and organic matter transformations in simulated fish ponds. Aquaculture Engineering 11 (1992). 157-169.95. Yang Yi. 1993. Modeling of growth of Nile tilapia in acage cum pond integrated culture system. Aquaculture engineering 21 ( 1999) 113-133.96. Mckinnon.A.D., Trott.I.A., Alongi,D.M and Davidson,A. 2002. Water column production and nutrient charectristics in mangrove creeks receiving shrimp farm effluent. Aquaculture Research, 2002. 55-73.97. Burford,m.A., Costanzo,S.D., Denninson,W.C. 2003. A sysnthesis of dominant ecological processes in intensive shrimp ponds and adjuscent coastal environments in NE Ausrallia. Marine pollution Bulletin. 46 (2003) 1456-1469.98. lemonier,H and Faninoz,S. 2006. Effect of water exchange on effluent and sediment characteristics and on partial nitrogen budget in semi-intensive shrimp ponds in New Caledonia. Aquaculture Research 37 938-948.99. Cytryn,E., Gelfand,I., Barak,Ym, Rijin,J.V and Minz,D.2003. Diversity of microbial communities correlated to physiochemical parameters in a digestion basin of a zero- discharge mariculture system. Environmental Microbiology 5(1), 55-63.100.Zhang,P., Zhang,X., Li.J and Huang,G. 2006. The Effects of body weight, temperature, salinity, pH, light intensity and feeding condition on lethal DO levels of whiteleg shrimp Litopenaeus vannamei ( Boone, 1931 ). Aquaculture 256 (2006 ) 579-587.101.Hari,B., Kurup, M,B, Vargese T, J., Schrama, J.W and Verdegem , M.C. J . 2006. the effect of carbohydrate addition on water quality and the nitrogen budget in extensive shrimp culture system. Aquaculture, 252 (2006 ) 248-263.102.Kai-min Cheng., Chao-qun Hu., yan-ni Liu., Shi- xuan Zheng and Xue- juan Qi. 2006. Effects of dietary calcium, phosphors and calcium/ phosphorus ratio on the growth and tissue mineralization of Litopenaeus vannamei reared in low- saline water. Aquaculture 251 (2006 ) 472-483.103.Balasubramanian, C.P., Pillai, S.M nad Ravichandran, P. 2004. Zero exchange shrimp farming systems ( extensive ) in the periphery of chilka lagoon, Orissa, India. Aquaculture international 12 . 555-572.104.Balcazar, J.l., Blas, I.D., Zarzuela, I..R., Cunningham, D., Vendrell, D and Muzquiz, J, L. 2006. The role of probiotics in aquaculture. Veterinary microbiology 114 (2006 ) 173-186.105.Coman. E.F., Connolly, R.M and Preston, N.P. 2003. Zooplankton and epibenthic fauna in shrimp ponds: factors influencing assemblage dynamics. Aquaculture Research, 34, 359-371.106.Rodrigues, A,R., Paez- Osuna, F. 2003. nutrients, phytoplankton and harmful algal blooms in shrimp ponds : a review with special reference to the situation in the gulf of California. Aquaculture 219 . 317-336.107.Carvalho, S., Barata, M., Pereira. F., Gaspar,M.B., Fonseca, L.C and Ferreira, P.P. 2006. Distribution patterns of macrobenthic species in relation to organic enrichment with in aquaculture earthen ponds. Marine Pollution bulletin 52. 1573- 1584.108.Samocha,T.M., Lopez,I.M., Jones, E.R., Jackson,S and Lawrence, A.L. 2004. characterization of intake and effluent waters from intensive and semi- intensive shrimp farms in texas. Aquaculture Reserch 35. 321-339.109.Alongi,D.M., Johnston,D.j and xuan,T.T. 2000. Carbon and Nitrogen budgets in shrimp ponds of extensive mixed shrimp- mangrove forestry farms in the Mekong delta, Vietnam. Aquaculture Research , 31- 387-399.110.lemonier,H and faninoz,S. 2006. effect of water exchange on effluent and sediment characteristics and on partial nitrogen budget in semi-intensive shrimp ponds in New Caledonia. Aquaculture Research, 37-938-948.111.Hernadez, C.R., Baraja, F.M., Clarck, P.G and Osuma, P.F. 2006. Nutrient mass balance in semi- intensive shrimp ponds from Sonora, Mexico using two feeding strategies: trays and mechanical dispersion. Aquaculture 258. 289-298112..Yuvanatemiya, V., Boyd, C,E. 2006. Physical and chemical changes in aquaculture pond bottom soil resulting from sediment removal. Aquacultural Engineering. 35. 199-205.113.Hargreaves,J.A. nitrogen biogeochemistry of aquaculture ponds. Aquaculture. 166. 181-212.114.Thakur, D.P and Lin, K.C. 2003. Water quality and nutrient budget in closed shrimp 9 Penaeus monodon ) culture systems. Aquacultural Engineering. 27. 159- 176.115.Li, Y., Li, J and Wang, Q. 2006. the effects of dissolved oxygen concentration and stocking density on growth and non- specific immunity factors in Chinese shrimp, Fenneropenaeus chinesis. Aquaculture 256. 608- 616.116.Galvan, G.S.R., Osuna, F.P., Fernandez, A.C.R and Angulo, E.R. 1999. seasonal variation in the water quality and chlorophyll a semi- intensive shrimp ponds in a subtropical environment. Hydrobiologia. 33-45,.117.Mohanthy, K.R. 2001. Feeding management and waste production in semi- intensive farming of Penaeus monodon ( fab ) at different stocking densities. Aquaculture International. 9: 345-355.118.Jana,T.K., Banerjee,R. D., Jana,B.B. 2007. Responses of some bio-geochemical cycling bacteria and their activities to management protocols under polyculture with Indian major carps and fresh water giant prawn.Aquaculture 264. 184-196.119.Lemonnier,H., Bernad,e., Boglio,E., Goarant,C and Claude Cochard, J. 2004. Influence of sediment characteristics on shrimp physiology : pH as Principal effect. Aquaculture. 240. 297-312.120.Coddington, T.D.R., Rouse, D.B and C. E. Boyd. 1999. Treatment of harvest discharge from intensive shrimp ponds by settling. Aquaculture engineering. 19. 147-161.121.Islam, M.S., Milsten,A., Wahab, M.A., Kamal,A.H.M and S. Dewan. 2005. Production and economic return of shrimp aquaculture in coastal ponds of different sizes and with different regimes. Aquaculture International. 13: 489-500.122.Erler,D and songsanaginda. 2007. Nitrogen dynamics in the settlement ponds of a small- sale recirculating shrimp farm ( Penaeus monodon) in rural Thailand. Aquaculture International. 15: 55-66.123.


text:
- WiFi
- Car park
- Spa
- Pets admitted
- Rural
- Airport Shuttle
- Accessible
- Family
- Sports
- Gym
- Eco-friendly
- Business
Choose your room
The rooms and bathrooms are medium in size and in good condition. The rooms are light and airy. All of them are 100% non smoking. The hotel has a restaurant on the ground floor to the right of the reception where breakfast, lunch and dinner can be served. This is a modern low rise building. The hotel has a medium sized lobby with a seating area. This is an excellent quality product in the close proximity of the airport and geneva centre. The hotel would suit clients looking for a quiet location from which to explore. 1008SR The hotel is 100% non smoking
Useful information:Other names for Aparthotel Adagio Geneve Saint Genis Pouilly:
Holiday Inn Geneva Airport
Geneva Airport Thoiry
Holiday Inn Thoiry Geneva Airport
Holiday Inn Thoiry
Adagio Apartamento Geneva


text:
Mrs Bessie Leeming of Kirk House, Church, Accrington died peacefully in the Royal Blackburn Hospital on Wednesday, April 10, 2013, aged 89 years. Bessie worked as a weaver after leaving school. She then spent time at home raising her four children. She then returned to work at Mullards in Blackburn until her retirement. She greatly enjoyed… Read more »
Clare Doreen Maud
Miss Clare Doreen Maud of Springhill Care Home, Accrington and formerly of Blackburn Road, West End, Oswaldtwistle died peacefully in the Blackburn Hospital on Monday, March 25, aged 83 years. Doreen was a primary school teacher throughout her working life and taught at several local schools in the area. She was also a keen volunteer… Read more »
Julie Lomax
On Saturday, March 16, 2013 suddenly in the Royal Blackburn Hospital and of Accrington, Julie aged 35 years. The dearly loved daughter of May and John. A loving and precious mum of Taylor, a devoted sister to Kerry, John and Paul also a much loved Auntie. A funeral service will be held on Wednesday, March 27,… Read more »
Hugh Allison
Hugh Allison of Burnley Road, Accrington died peacefully in the Royal Blackburn Hospital on Sunday, February 10, 2013 aged 74 years. Hugh worked at Blaire's Electronics in Clayton-Le-Moors for over 20 years and also Admagnetics in Huncoat. He loved his cars as well as holidaying in his native Scotland. He leaves behind his dear and much… Read more »


text:
1.Product introduction
The fish meal that we provide be made by cooking, pressing, drying, and grinding of fish or fish waste to which no other matter has been added. It is a solid product from which most of the water is removed and some or all of the oil is removed.The quality indicators have exceeded the level of fish meal requirements prescribed by the state, the product has a stable and reliable quality, freshness is good, fish powder flavor, protein content is high, easy to decompose,digest and absorb.
|Brownish yellow
|10 % max
|10% max
|3 % max
|3 % max
|17% max
|150 ppm min at the time of loading
|150 mg/ 100 gr max
|Free from salmonella, melamine, ecoli
Fish contains a variety of nutrients comprise animal tissue, maintain normal metabolism, growth and reproduction,improve the utilization rate and enhance animal disease resistance ability,it is irreplaceable protein.
1. livestock feed for pigs, chickens and other animal feed, these feeds need to contain high-quality protein, especially young pigs and chickens. Because young animals are in vigorous growth, they need larger proportion of protein requirements.
2. aquatic animal feed for aquatic animals such as fish, crab, shrimp and other feed protein main raw materials. add fish meal to ensure that aquatic animals grow faster.
3.fur animal feed for some fur animals such as fox, raccoon and other feed, fur animals are mostly carnivorous, the growth of protein demand, high quality fish meal as a protein raw material is the first choice for these animal feed ingredients.
4.pet feed for dog and other pet feed, can increase the pet disease resistance and promote growth and other effects. any one of the high-grade pet food processors shop need fish meal as the feed ingredients.
1.Port: Qingdao or Tianjin.
2.Payment terms: LC at sight, TT(30% deposit in advance, 70% balance against of the B/L copy).
3.Delivery: Within 2 weeks after received the receipt of advance payment.
4.Packaging: 25 or 50 kg per PP bag or according your demand.
1.Technical Support--- We have good technical team with great manufacturing experience and know-how of applications, to work with you together.www.puretopfeed.com
2.Quality Assurance--- We Puretop international group co.,ltd always highlight the quality and safety as the priority of our stands of manufacturing and our promise to you. We have accredited quality assurance system and state of the art production facility to ensure the product quality and safety covering the whole processing and supplying chains, from the approval and receipt of raw materials to the final products storage and delivery.
3.Logistics--- TEL: 008651266700581 We www.puretopfeed.com
have a professional logistic team to make us enable to deliver your goods exactly to agreed schedule and destinations. Our logistic team will understand your needs to provide a best shipping plan, and keep you informed about the each step of delivery to ensure you receive the goods on time.all of life friend hand in hand, and common development, create brilliant.
If you are interested in our fish meal with 65% protein in competitive price export out, please be free to place an order with our factory. We are one of the leading manufacturers and suppliers in China. High-quality products, low and cheap price, and the best service can be assured.


text:
Thrombin activates protease-activated receptor 1 (PAR1) on endothelial cells (ECs) and is critical for angiogenesis and vascular development. However, the mechanism underlying the proangiogenic effect of thrombin has not been elucidated yet. Here, we report the discovery of a novel functional link between thrombin-PAR1 and transforming growth factor-β (TGF-β) signaling pathways. We showed that thrombin via PAR1 induced the internalization of endoglin and type-II TGF-β receptor (TβRII) but not type-I receptors in human ECs. This effect was mediated by protein kinase C-ζ (PKC-ζ) since specific inhibition of PKC-ζ caused an aggregation of endoglin or TβRII on cell surface and blocked their internalization by thrombin. Furthermore, acute and long-term pretreatment of ECs with thrombin or PAR1 peptide agonist suppressed the TGF-β–induced serine phosphorylation of Smad2, a critical mediator of TGF-β signaling. Moreover, activation of PAR1 led to a profound and spread cytosolic clustering formation of Smad2/3 and markedly prevented Smad2/3 nuclear translocation evoked by TGF-β1. Since TGF-β plays a crucial role in the resolution phase of angiogenesis, the down-regulation of TGF-β signaling by thrombin-PAR1 pathway may provide a new insight into the mechanism of the proangiogenic effect of thrombin.
Thrombin, a serine protease central to blood coagulation and hemostasis, activates protease-activated receptor 1 (PAR1) on endothelial cells (ECs) and regulates many aspects of EC biology.1 A body of evidence has demonstrated that the thrombin-PAR1 signal pathway is critical for angiogenesis and vascular development.1-3 However, the mechanism underlying the proangiogenic effect of thrombin has not been elucidated yet. Deficiency of PAR1 causes yolk sac defects and bleeding and results in 50% mouse embryonic lethality without altering the plasma coagulation cascade and platelet function.4,5 Reconstitution of a PAR1 transgene driven by an endothelial-specific promoter prevents death of PAR1–/– embryos, indicating that thrombin's action via PAR1 on endothelial rather than other cells contributes to angiogenesis and vascular development.5 Similar defects are observed in mouse embryos deficient in various coagulation factors (eg, factor V, tissue factor, and prothrombin), in which thrombin generation is impaired.6-8 In contrast, deletion of either PAR2, PAR3, or PAR4 is not lethal.2 Furthermore, thrombin induces neoangiogenesis in chick chorioallantoic membrane and the mouse Matrigel system, which requires the activation of PAR1 signal pathways.9,10 Thrombin-PAR1 pathway also promotes tumor progress and metastasis related with angiogenesis.11 However, thrombin (≤ 5 U/mL) itself is not sufficient to induce angiogenesis in vitro.12 In contrast, thrombin at 10 U/mL inhibits EC tube formation.12 It seems likely that thrombin may modulate angiogenesis by regulating functions of growth factors involved in angiogenesis in vivo.
Angiogenesis encompasses activation and resolution phases. In the activation phase, increased vascular permeability and basement membrane degradation allows ECs to proliferate and migrate into the extracellular space and form new capillary sprouts. In the resolution phase, ECs cease proliferation and migration, reconstitute the basement membrane and promote vessel maturation and recruit pericytes. Compelling genetics evidence has demonstrated a pivotal role for transforming growth factor-β (TGF-β) and its receptors (type-I and -II receptors as well as endoglin) in angiogenesis, in particular the resolution phase of angiogenesis such as the establishment and maintenance of vessel wall integrity.13 Deficiency of either TGF-β1, type-I receptor (also known as activin receptor-like kinase [ALK]), type-II receptor (TβRII) as well as endoglin, an ancillary TGF-β receptor, results in mouse embryonic lethality with a similar defect in vascular development, and the human vascular disorder hereditary hemorrhagic telangiectasia has been linked to mutations in ALK1 and endoglin.14-16 However, the mechanism by which TGF-β regulates angiogenesis is still not fully understood, and in particular the functional regulation of TGF-β signaling by other systems during angiogenesis has not been actively explored.
In the present study, we have discovered a new functional link between thrombin and TGF-β signaling pathways. We show that thrombin via PAR1 induces a protein kinase C-ζ (PKC-ζ)–dependent endocytosis of TβRII and endoglin but not type-I receptors and suppresses phosphorylation of the transcriptional activator Smad2 by TGF-β in primary human ECs. Moreover, activation of PAR1 leads to a profound and spread cytosolic clustering formation of Smad2/3 and markedly prevents Smad2/3 nuclear translocation evoked by TGF-β1. These substantial lines of evidence indicate that thrombin-PAR1 pathway is a potent negative regulator of TGF-β signaling in ECs. Our data also indicate that the TGF-β signaling can be modulated by G protein-coupled receptors such as PAR1.
Materials and methods
Human α-thrombin, thrombin receptor-activating peptide (TRAP), hirudin, and SB431542 were from Sigma (St Louis, MO). Human TGF-β1 was from R&D Systems (Minneapolis, MN). Short peptide agonists for human PAR1 (SFLLRN), PAR2 (SLIGKV), PAR3 (TFRGAP), and PAR4 (GYPGQV) were from Bachem (Torrance, CA). Myristoylated PKC-ζ peptide inhibitor and other PKC inhibitors were from Calbiochem (La Jolla, CA). Myristoylated PKC-β C2-4 inhibitor was from Biomol (Plymouth Meeting, PA). Reagents for chemiluminescence detection were from Cell Signaling (Beverly, MA).
Antibodies against phospho-nuclear factor (NF-κB) p65 (Ser-536), phospho–PKC-ζ/λ (Thr-410/403), and TβRI (ALK5) were from Cell Signaling. Antibodies against TGF-β1/2/3 (H-112), PAR1 (ATAP2), ALK5 (H-100), TβRII (C-16), and phosphatidylinositol-3-OH kinase (PI3K) were from Santa Cruz Biotechnology (Santa Cruz, CA). Antibodies against extracellular domains of human TβRII and ALK1 were from R&D Systems. Antibody against phospho-Smad2 (Ser-465/467) was from Chemicon (Temecular, CA). Monoclonal antibodies against endoglin and Smad2/3 were from BD Transduction (San Diego, CA). Human endoglin monoclonal antibody (SN6h) was from DakoCytomation (Carpinteria, CA). Smad2 antibody was from Zymed (South San Francisco, CA). Cy™3–conjugated goat antimouse and antirabbit antibodies were from Jackson ImmunoResearch (West Grove, PA).
Cell culture
Human aortic ECs (HAECs) and human umbilical vein ECs (HUVECs) were from Cambrex Bio Science (Walkersville, MD) and cultured in EGM™–2 medium and used for experiments within 10 passages.
Immunoblotting and immunoprecipitation
Immunoblotting and immunoprecipitation were performed as we described previously.17
Identification of the serine-phosphorylated 95-kDa protein
HAECs (20 dishes, 10 cm) were incubated with 1 μM phenylarsine oxide (PAO) for 3 hours. Lysates were prepared and loaded onto a high-Q cartridge column (Bio-Rad, Hercules, CA). Fractions eluted from 350 to 700 mM NaCl, which contain the 95-kDa protein, were combined and concentrated. The concentrated eluates were diluted to a solution of 150 mM NaCl and immunoprecipitated with antibody against phospho–NF-κB p65 (Ser-536). The immune complexes were washed extensively with Nonidet P-40 lysis buffer17 containing 300 mM NaCl and resolved by sodium dodecyl sulfate (SDS)–polyacrylamide gel electrophoresis (PAGE). Proteins were visualized with Coomassie blue staining, and the band corresponding to 95 kDa was excised for in-gel trypsin digestion and analyzed with a matrix-assisted laser desorption delayed-extraction reflectron time of flight (MALDI-TOF) mass spectrometer performed by AmProx (AmProx, Carlsbad, CA).
Subcellular fractionation
EC cytosolic and membrane fractions were prepared as we described previously.17
Potassium depletion
Potassium depletion of ECs was performed essentially as described by Larkin et al.18
Cell surface enzyme-linked immunosorbent assay (ELISA)
Cell surface receptors were determined basically as described previously.19 ECs in 48-well plates were serum-starved for 2 hours and then stimulated with thrombin (3 U/mL) or TRAP (40 μM) for the indicated time periods at 37°C. ECs were washed with phosphate-buffered saline (PBS), fixed with 1% glutaraldehyde for 10 minutes, and blocked with 1% bovine serum albumin (BSA) in PBS for 1 hour. Cells were then incubated at 25°C with antibodies against extracellular domains of endoglin, TβRII, ALK5, ALK1, and PAR1. After 1 hour of incubation, cells were washed 4 times with PBS and incubated with horseradish peroxidase (HRP)–conjugated antimouse or antigoat secondary antibodies (1: 2000 dilution) for 1 hour at 25°C. Cells were again washed with PBS and then incubated in One Step ABTS solution (2,2′-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid; Roche, Indianapolis, IN) for 10 minutes, and the optical density was determined at 405 nm using a universal microplate reader (Elx800; Bio-Tek, Winooski, VT).
Immunofluorescence microscopy
ECs grown on glass coverslips in a 6-well plate were pretreated with or without PKC inhibitors followed by stimulation with thrombin (3 U/mL), TRAP (40 μM), or PAR1-AP (80 μM) for 30 minutes at 37°C, then washed with PBS and fixed in 3.7% formaldehyde solution in PBS for 10 minutes at 25°C. The fixed cells were extracted in ice-cold acetone at –20°C for 5 minutes, washed, and preincubated in PBS containing 1% BSA for 30 minutes at 25°C. Cells were then incubated with monoclonal antibodies (1:100 dilution) against endoglin (SN6h), Smad2/3, and ALK1, or with polyclonal antibodies (1:100 dilution) against ALK5 (H-100) and TβRII (C-16) for 1 hour at 25°C in PBS containing 1% BSA, washed 3 times with PBS, and then incubated with Cy3-conjugated goat antimouse or antirabbit secondary antibodies (1:100 dilution) for 1 hour at 25°C in PBS containing 1% BSA. Control stainings were performed without either primary or secondary antibodies. After washing with PBS, coverslips were mounted on slides with cell-side down with Cytoseal and examined and photographed using a Perkin Elmer Ultra VIEW LCI confocal imaging system (Boston, MA) configured with a Nikon TE2000-S fluorescence microscope (Garden City, NY) fitted with PlanApo 60 × and 100 × oil objectives. Adobe Photoshop 6.0 software was used for image processing.
Identification of the 95-kDa protein as serine-phosphorylated endoglin by a phospho-specific antibody agonist NF-κB p65 (Ser-536) in ECs
Figure 1A shows that in addition to the phosphorylated NF-κB p65,20 a protein band with 95 kDa was also detected by an antibody against phospho–NF-κB p65 (Ser-536) in HAECs. Interestingly, transient stimulation of HAECs with a physiologic dose of thrombin (3 U/mL) resulted in a profound decrease in the serine phosphorylation of the 95-kDa protein, whereas thrombin promoted the phosphorylation of p65 on Ser-536 (Figure 1A). Figure 1B shows that the 95-kDa protein was merely detected at membrane fraction. To identify the 95-kDa protein by using mass spectrometry, we searched for compounds that could enhance serine phosphorylation of the 95-kDa membrane protein and found that PAO,21,22 a receptor internalization blocker, was the most potent one. As shown in Figure 1C, the serine phosphorylation of the 95-kDa protein was augmented 15-fold by PAO (1 μM) treatment. We then prepared lysates from PAO-treated HAECs, loaded them onto a high-Q cartridge column and then collected fractions containing the phosphorylated 95-kDa membrane protein. The eluates were immunoprecipitated with the phospho-specific antibody, resolved by SDS-PAGE, and analyzed with a MALDI-TOF mass spectrometer as described in "Materials and methods." Peptide fingerprint analysis identified that the 95-kDa membrane protein was endoglin,23,24 a TGF-β coreceptor highly expressed in proliferating ECs. To confirm this finding, lysates were immunoprecipitated with the Ser-536–phosphorylated p65 antibody, a PI3K antibody, or control IgG from normal rabbit serum and subjected to immnunoblotting with an antibody (clone 35) against NH2-terminus of endoglin (BD Transduction). As shown in Figure 1D, a clear band corresponding to the full-length endoglin (L-endoglin) from the immune complexes of the phospho-specific antibody but not from others was detected. Alternatively, lysates were immunoprecipitated with an antibody against the full-length endoglin (SN6h; DakoCytomation) followed by immunoblotting with the antibody against Ser-536–phosphorylated p65. As shown in Figure 1E, a clear band corresponding to L-endoglin was detected by the phospho-specific antibody. Remarkably, the band was not detected from the endoglin immune complexes treated with alkaline phosphatase to dephosphorylate endoglin. Thus, our findings indicate that the 95-kDa membrane protein detected by the phospho-specific antibody represents the serine-phosphorylated full-length endolin, which accounts for approximately 2% of total endolin in unstimulated HAECs. L-endoglin is composed of 633 amino acids with 47–amino acid residues (residues 587-633) in the cytoplasmic tail, while the S-endoglin consists of 600 amino acids with a 14–amino acid cytoplasmic tail (residues 587-600)25 (Figure 1F). Consistent with previous reports,25-27 90% endoglin was detected as L-form at the membranes of HAECs (Figure 1G). The Ser-536 of NF-κB p65 is surrounded by several Asp and Glu residues (Figure 1F). Since only the L-endoglin is detected by the phospho-specific antibody, a phosphorylated serine within residues 601 to 633 of endoglin, especially within the serine clusters (609SSESSS) adjacent to Glu could be the recognition site of the phospho-specific antibody.
TGF-β1 promotes the serine phosphorylation of endoglin in ECs
As shown in Figure 2A, TGF-β1 induced a time-dependent serine phosphorylation of endoglin detected by the phospho-specific antibody as described in Figure 1. The stimulatory effect occurred fast (1 minute), reached the maximal level (2-fold) at 5 minutes, and then declined. Interestingly, pretreatment of HAECs with thrombin for 30 minutes suppressed 70% of the serine phosphorylation of endoglin by TGF-β1. The blot was stripped and reprobed with an endoglin antibody (SN6h) to show the equal loading (Figure 2A, lower panel). We next determined the effect of SB431542,28 a specific inhibitor of type-I receptor of TGF-β superfamily, including ALK4, ALK5, and ALK6, on the serine phosphorylation of endoglin by TGF-β in HAECs. As shown in Figure 2B, SB431542 did not affect the TGF-β1–induced serine phosphorylation of endoglin. However, it completely suppressed the basal and TGF-β1–induced serine phosphorylation of Smad2 detected with an antibody against phospho-Smad2 (Ser-465/467; Chemicon). These data suggest that TβRII other than ALK5 may be involved in the serine phosphorylation of endoglin by TGF-β, although other serine/threonine kinases activated by TGF-β could not be excluded. It has been shown that endoglin interacts with and is constitutively phosphorylated by TβRII.29 It appears that the serine-phosphorylated endoglin (2%) detected in HAECs may represent the functional endoglin associated with and phosphorylated by TβRII.
Thrombin down-regulates the serine-phosphorylated endoglin through PAR1 in ECs
PAR1 is the major receptor for thrombin in ECs in mice and humans.1 As shown in Figure 3A, thrombin markedly down-regulated the serine-phosphorylated endoglin in a time-dependent manner in HAECs detected with the antibody against the serine-phosphorylated endoglin associated with, and phosphorylated by, TGF-β receptor complexes as described in Figures 1 and 2. The down-regulatory effect occurred fast (< 5 minutes, data not shown), reached a maximal level at 30 minutes (82% down-regulation), and gradually returned to basal level by 3 hours after thrombin treatment. Hirudin (6 U/mL), a specific inhibitor of thrombin, and an antibody (ATAP2) that inhibits PAR1 cleavage and activation30 markedly prevented the down-regulatory effect of thrombin (Figure 3B). Whereas pretreatment of HAECs with an antibody against TGF-β1/2/3 had virtually no effect, indicating a mechanism independent of TGF-β secretion (Figure 3B). Moreover, TRAP (SFLLRNPNDKYEPF), a 14–amino acid peptide agonist for human PAR1, and a short peptide agonist for human PAR1 (PAR1-AP; SFLLRN)31 mimicked the effect of thrombin in HAECs. In contrast, selective peptide agonists for human PAR2 and PAR3 had no effect (Figure 3C). Thus, thrombin down-regulates the phosphorylated and functional endoglin through PAR1 in HAECs. Interestingly, lysophosphatidic acid (LPA) also down-regulated the serine-phosphorylated endoglin. In contrast, angiotensin II (Ang II), tumor necrosis factor (TNF), and interlukin-8 (IL-8) had no effect (Figure 3D). These findings were reproduced in HUVECs (data not shown).
Blockage of receptor internalization prevents the thrombin-induced down-regulation of the serine-phosphorylated endoglin in ECs
As shown in Figure 4A, thrombin down-regulated the serine-phosphorylated endoglin by TβRII in a similar efficiency in both control HAECs and HAECs treated with okadaic acid (100 nM),32 a serine/threonine phosphatase inhibitor, although the basal serine phosphorylation of endoglin was markedly enhanced by okadaic acid. This finding suggests that endocytosis of endoglin other than the serine/threonine phosphatases may be involved in the thrombin-induced down-regulation of the serine-phosphorylated endoglin in ECs.
PAO is a blocker of clathrin-mediated receptor endocytosis.21,22 As shown in Figure 4B, pretreatment of HAECs with PAO (1 μM) resulted in a profound increase in the serine phosphorylation of endoglin and abolished the thrombin-induced down-regulation of the serine-phosphorylated endoglin. In addition, PAO is also an inhibitor of protein tyrosine phosphatase.33 To determine the possible involvement of tyrosine phosphatase, we treated HAECs with pervanadate,34 a potent inhibitor of tyrosine phosphatase, and found that pervanadate affected neither basal level nor thrombin-mediated down-regulation of the functional endoglin, indicating a tyrosine phosphatase–independent mechanism. We next determined the effect of inhibiting clathrin-mediated endocytosis by hypertonic sucrose35 or potassium depletion.18 As shown in Figure 4C-D, the thrombin-induced down-regulation of the serine-phosphorylated endoglin was markedly prevented by potassium depletion and abolished by sucrose treatment. The basal phosphorylation of endoglin was markedly enhanced by sucrose but slightly increased by potassium depletion. Thus, these data can be interpreted to mean that endoglin may undergo constitutive clathrin-dependent internalization in unstimulated ECs. The thrombin-induced down-regulation of the serine-phosphorylated endoglin may result from endoglin internalization by thrombin.
Thrombin induces a PKC-ζ–dependent internalization of endoglin and TβRII in ECs
To determine whether thrombin induces internalization of endoglin, we performed immunofluorescence microscopy using a confocal microscope to access the localization of endoglin in HUVECs. In unstimulated HUVECs, endoglin was localized primarily at the plasma membrane detected with a monoclonal antibody (SN6h) against L-endoglin (Figure 5A). Stimulation of HUVECs with thrombin for 30 minutes led to the progressive appearance of a vesicular pattern typical of endocytic vesicles. Serial laser scans ranged from the top surface levels and into the cell interior showed that the thrombin-induced endocytic vesicles of endoglin was only observed inside the cell, indicating the internalization of endoglin by thrombin (Figure 5A, 2nd panel). Since TβRII interacts constitutively with the cytoplasmic domain of endoglin,29 we assessed the effect of thrombin on the localization of TβRII. Serial laser scans ranged from the top surface levels and into the cell interior revealed that TβRII was stained primarily at the cell surface detected with a TβRII antibody29 that recognizes the COOH-terminal domain of human TβRII (Figure 5B). Stimulation of HUVECs with thrombin for 30 minutes induced a progressive appearance of the endocytic vesicles containing TβRII that was only observed inside the cell, indicating the internalization of TβRII by thrombin. Similar effect of thrombin or PAR1-AP on the internalization of endoglin and TβRII was observed in HAECs (data not shown). In contrast, we found that the localization of ALK5 and ALK1 was not altered by thrombin, and there is no appearance of the endocytic vesicles formation by thrombin in ECs (Figure S1 at the Blood website; see the Supplemental Figure link at the top of the online article). Thus, thrombin induces the internalization of endoglin and TβRII but not ALK5 and ALK1 in ECs, which is distinct from TGF-β–induced internalization of TGF-β receptors.36
We next determined the mechanism of thrombin-induced endocytosis of endoglin and TβRII in ECs. Using the phospho-specific antibody that recognizes the serine-phosphorylated endoglin by TβRII as described in Figures 1 and 2, we found that the thrombin-induced down-regulation of the serine-phosphorylated endoglin (ie, endoglin internalization) was markedly prevented by a general PKC inhibitor GF109203X37 (Figure 5C). Rottlerin,38 a selective PKCδ inhibitor, or LY294002,39 a selective inhibitor of PI3K, had a minor effect. In contrast, Gö6976,40 a selective conventional PKC inhibitor, had virtually no effect. Moreover, phorbol-12-myristate-13-acetate (PMA),41 a potent activator of conventional and novel PKCs, did not affect endoglin phosphorylation (Figure 5D). These data were highly reproducible in HAECs (data not shown). These findings suggest that the atypical PKCs may be involved in the thrombin-induced endocytosis of endoglin in ECs. It has been shown that human ECs, including HUVECs, express PKC-α, -β, -δ, -ϵ, -η, -θ, and -ζ isoforms but not PKC-ι/λ.42-44 Consistent with a previous report,45 we found that thrombin induced a rapid activation of PKC-ζ detected with an antibody against phospho–PKC-ζ (Thr-410) (Figure 5E). The activation of PKC-ζ requires the phosphorylation of Thr-410 on the activation loop by phosphoinositide-dependent kinase.46 To assess the role of PKC-ζ in the endocytosis of endoglin, HUVECs were pretreated with a myristoylated PKC-ζ peptide inhibitor, and lysates were immunoblotted with the phospho-specific antibody that detects the functional endoglin associated with and phopshorylated by TβRII. As shown in Figure 5F, specific inhibition of PKC-ζ markedly prevented down-regulation of the functional endoglin or endoglin internalization by thrombin. We then performed confocal immunofluorescence microscopy to confirm the role of PKC-ζ in the endocytosis of endoglin and TβRII by thrombin. Remarkably, specific inhibition of PKC-ζ with the myristoylated PKC-ζ peptide inhibitor led to the aggregation or cluster formation of endoglin or TβRII on cell surface and blocked their endocytosis by thrombin (Figure 5A-B), suggesting that an overlap or similar pathway may be involved in the thrombin-induced endocytosis of TβRII and endoglin in ECs. In contrast, the thrombin-induced endocytosis of TβRII and endoglin was not affected by a myristoylated PKC-β peptide inhibitor (Figure 5A-B) and a conventional PKC inhibitor Gö6976 in HUVECs (data not shown). Taken together, these findings indicate that PKC-ζ plays a critical role in the internalization of TβRII and endoglin evoked by thrombin in ECs.
Finally, we performed a cell surface ELISA assay19 with monoclonal antibodies against the extracellular domains of endoglin or TβRII to quantify the internalization of endoglin and TβRII by thrombin in ECs. We found that thrombin induced a time-dependent internalization of endoglin, with a maximal level of 38% or 35% internalization at 2 hours in HAECs and HUVECs, respectively (Figure 6A). As expected, thrombin induced a rapid and profound internalization of PAR1, with a maximal effect at 30 minutes (72% internalization) detected with an antibody (ATAP2)47 that recognizes both intact and cleaved PAR1 (Figure 6A). Figure 6B shows that thrombin induces a time-dependent internalization of TβRII, with a maximal level of 30% or 37% internalization at 30 minutes in HAECs and HUVECs, respectively. Similar to the pattern of thrombin-induced down-regulation of the serine-phosphorylated endoglin (Figure 3A), the cell surface TβRII tended to recover after 30 minutes of treatment with thrombin (Figure 6B). In addition, consistent with the observation by immunofluorescence microscopy, the level of cell surface ALK5 was not significantly altered by thrombin (Figure 6B).
Thrombin down-regulates TGF-β signaling in ECs
Since TGF-β binds directly to TβRII, which is essential for the downstream signal transduction of TGF-β,48-50 we determined effects of thrombin on the proximal signaling events upon activation of TGF-β receptors such as phosphorylation of the transcriptional activators Smad2 and Smad3 in ECs. As shown in Figure 7A-B, the TGF-β1–induced phosphorylation of Smad2, which was detected with a phospho-Smad2 (Ser-465/467) antibody, was suppressed 35% to 40% by acute treatment of HUVECs with thrombin or TRAP. In contrast, Ang II did not affect the phosphorylation of Smad2 by TGF-β1. Remarkably, the TGF-β1–induced Smad2 phosphorylation was suppressed 54% and 68% by 16 or 24 hours of treatment of HUVECs with thrombin, respectively. The protein levels of Smad2, ALK5, TβRII, and endoglin were not altered by long-term thrombin treatment (Figure 7C). In addition, we found that long-term Ang II treatment did not affect the Smad2 phosphorylation by TGF-β1 (data not shown). Moreover, we found that the cell surface receptor levels of ALK5, endoglin, and TβRII were down-regulated 15% ± 3%, 12% ± 2%, and 20% ± 4% by 24-hour treatment of HUVECs with thrombin, respectively. It seems that the profound decrease (68%) in the TGF-β1–induced Smad2 phosphorylation by long-term thrombin treatment may not merely result from the small amount of down-regulation (10%-20%) of the cell surface TGF-β receptors, suggesting the involvement of other mechanisms.
Upon serine phosphorylation by ALK5, Smad2 and Smad3 form a heterotrimeric complex with the common Smad4 and translocate to the nucleus where they regulate the transcriptional response to TGF-β.48-50 We assessed effects of thrombin and TRAP on the TGF-β1–induced nuclear translocation of Smad2 and Smad3 using immunofluorescence microscopy. In unstimulated HUVECs, Smad2 and Smad3 were predominantly cytoplasmic detected with a monoclonal antibody against both Smad2 and Smad3 (BD Transduction). Some bright dots, which may represent the oligomers of Smad2/3, were seen around the nucleus (Figure 7Di). Stimulation of HUVECs with TGF-β1 activated Smad2/3 by inducing their nuclear translocation. We found that 85% of the cells were positive with a profound nuclear staining of Smad2/3 (Figure 7Dii). Remarkably, acute treatment of HUVECs with thrombin or TRAP alone led to a profound appearance of the bright and spread Smad2/3 clusters in cytosol (Figure 7Diii,v). Furthermore, thrombin or TRAP markedly suppressed the nuclear translocation of Sma2/3 evoked by TGF-β1. We found that about 30% of cells pretreated with thrombin or TRAP were stained positively with the nuclear translocation of Smad2/3 but in a much less density compared with that induced by TGF-β1 alone (Figure 7Dii,iv,vi). We also noticed that cells with a more cytosolic clustering formation of Smad2/3 had a less nuclear translocation of Smad2/3 by TGF-β1. Similar phenomena were observed in HAECs (data not shown). Taken together, the thrombin-PAR1 pathway is a potent negative-regulator of TGF-β signaling in ECs.
In the present study, we report the discovery of a novel functional link between thrombin-PAR1 and TGF-β signaling pathways. We have demonstrated that thrombin-PAR1 pathway down-regulates TGF-β signaling at least through the PKC-ζ–dependent internalization of TβRII and endoglin and the sequestering of Smad2/3 in cytosol using primary human ECs. Since TGF-β plays a pivotal role in the resolution phase of angiogenesis such as the establishment and maintenance of vessel wall integrity,13-16 our findings that the down-regulation of TGF-β signaling by thrombin-PAR1 pathway, which facilitates the transition of the angiogenic resolution phase to activation phase, may provide new insights into the mechanism of the proangiogenic effect of thrombin.
The first finding obtained from this study is that thrombin via PAR1 induces the internalization of endoglin and TβRII but not the type I receptors (ALK1 and ALK5) in human ECs, which has not been reported to date, as far as we know. TGF-β binds directly to TβRII.51 Upon ligand binding, TβRII recruits and transphosphorylates type I receptor in ECs, which in turn phosphorylates and activates Smads to modulate gene transcription.48-50 Endoglin (CD105), an ancillary TGF-β receptor, is an 180-kDa homodimeric membrane glycoprotein primarily expressed on ECs.23 Endoglin antagonizes TGF-β signaling.26,52 Endoglin cannot bind ligands on its own and does not alter ligand-binding to TβRII, but it interacts with ALK5 and TβRII independent of TGF-β.29,51,53 Both ALK5 and TβRII interact with the cytoplasmic domain of endoglin, but ALK5 only interacts when the kinase domain is inactive, whereas TβRII remains associated with endoglin regardless of its kinase activity.29 We found that thrombin via PAR1 induced a co-internalization of endoglin and TβRII but not ALK1 and ALK5 in ECs, suggesting a tight interaction between endoglin and TβRII. It has been shown that phosphorylation of the cytoplasmic tail of betaglycan, an endoglin homologue normally not expressed in ECs,54 causes a co-internalization of betaglycan and TβRII but not ALK5, leading to the down-regulation of TGF-β signaling in HEK-293 cells.55 Furthermore, we found that the amount of the internalized endoglin (30%-40%) is clearly larger than the estimated amount of endoglin (1%) associated with TβRII in ECs.53 This suggests that thrombin may promote the internalization of endoglin free from, or associated with, TβRII. Since endoglin associates constitutively with TβRII, it appears likely that the internalization of TβRII may result from the endocytosis of endoglin by thrombin. However, we found that more than 80% of the serine-phosphorylated endoglin associated with TβRII was internalized by thrombin in ECs (Figures 2, 3). These findings suggest that the TβRII-associated endoglin may be preferably internalized by thrombin.
We further showed that the PKC-ζ activity was required to drive the endocytosis of TβRII and endoglin by thrombin since specific inhibition of PKC-ζ with a myristoylated PKC-ζ peptide inhibitor caused the aggregation or cluster formation of TβRII or endoglin on cell surface and blocked their internalization by thrombin. These findings also suggest that an overlap or similar pathway may be involved in the thrombin-induced endocytosis of TβRII and endoglin in ECs. The cytoplasmic domain of endoglin contains a putative PKC phosphorylation site at Ser-596 (Figure 1F). On the basis of our data, the serine phosphorylation sites of endoglin phosphorylated by TβRII probably locate within a sequence (609SSESSS614) (Figures 1F and 2). The cytoplasmic domains of endoglin and its homologue betaglycan share a serine- and threonine-rich sequence (HSIGSTQSTPCS) that resembles β-arrestin–binding domain in G protein-coupled receptors.56 Thr-841 of betaglycan within the sequence has been shown to be phosphorylated by TβRII in HEK-293 cells.55 Studies to assess roles of the direct phosphorylation of endoglin by PKC-ζ or TβRII in the thrombin-induced endocytosis of endoglin and TβRII are underway.
The second finding from this study is that activation of PAR1 leads to a profound and spread cytosolic clustering formation of Smad2/3, and markedly suppresses Smad2/3 phosphorylation and nuclear translocation evoked by TGF-β1 in ECs. We also noticed that ECs with a more cytosolic clustering formation of Smad2/3 had a less nuclear translocation of Smad2/3 by TGF-β1. The nuclear translocation of Smad2/3 is initiated through their phosphorylation by ALK5, which promotes formation of a heterotrimer consisting of Smad2, Smad3, and Smad4.48-50 Two groups recently reported that cytosolic sequestering of Smad3 by Akt/PKB (protein kinase B) prevented Smad3 phosphorylation, nuclear translocation, and transcriptional activity.57,58 It appears likely that the cytosolic sequestering of Smad2/3 upon activation of PAR1, in addition to the endocytosis of TβRII from the cell surface, may play an important role in the down-regulation of TGF-β signaling by thrombin in ECs. The interaction of Akt with Smad3 is positively regulated by the phosphorylation of Akt at Thr-308 and Ser-473.57,58 We found that the phosphorylation of Akt at Thr-308 and Ser-473 was not significantly altered by thrombin (data not shown), suggesting involvement of an Akt-independent mechanism in the cytosolic sequestering of Smad2/3 by thrombin. The mechanism underlying the thrombin-induced cytosolic clustering formation or sequestering of Smad2/3 merits further studies. In addition, we found that the TGF-β1–induced nuclear translocation of Smad2/3 was inhibited more than 65% by acute thrombin treatment, while the Smad2 phosphorylation was only inhibited 35% to 40% by thrombin. Thus, besides the reduced phosphorylation of Smad2/3, the sequestering of Smad2/3 or other mechanism may also be involved in preventing Smad2/3 nuclear translocation evoked by TGF-β in ECs. It has been shown that phosphorylation of Smad2 and Smad3 by extracellular signal–regulated kinase and Ca2+/calmodulin-dependent protein kinase II impairs their nuclear translocation,59,60 whereas the PKC activator PMA causes phosphorylation of Smad2 and Smad3, which abrogates DNA binding but not nuclear translocation of Smad3.61
In summary, we have obtained substantial evidence indicating that thrombin-PAR1 pathway down-regulates TGF-β signaling in primary human ECs, which may offer new insights into the mechanism of the proangiogenic effect of thrombin in vivo.
Prepublished online as Blood First Edition Paper, November 2, 2004; DOI 10.1182/blood-2004-08-3308.
Supported in part by grants from the National Institutes of Health (HL-69806) (H.T.) and from the American Heart Association (0130038N) (H.T.).
The online version of the article contains a data supplement.
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.


text:
Toppan Forms Develops Japan's First Authenticity Determination of Driver's License Using Smartphone and Tablet
IC chip of driver's license is read and verified using NFC reader and writer
July 29, 2014
Information management solution provider Toppan Forms Co., Ltd. (Toppan Forms hereunder) developed an app that determines whether the Driver's License with IC chip is authentic. It is the first in Japan that uses smartphone. After entering the pass code registered on the driver's license into the app a smartphone with NFC reader writer reads the IC chip in the Driver's License and verifies. It obtains personal information such as facial photo, name, address, and birth date registered on the IC chip. The personal information obtained is compared with the personal information on Toppan Forms server which has been entered in advance.
The app is featured in FastShot service that sends various proof documents electronically using smart phone.
- 1. Information registered on the IC chip in the Driver's License is read by NFC enabled smartphone to determine the authenticity by comparing it with entered personal data.
- 2. By incorporating this feature into FastShot, the application to open a bank account can be submitted and completed by sending the data using just a smartphone.
The app was developed as a new feature on "FastShot" that sends various proof documents electronically by using the camera feature on the smartphone. The conventional FastShot verified the Driver's License by image taken by the camera. For a more rigorous process, the IC chip embedded in the Driver's License allows reliable and easy authenticity determination. The more rigorous process prevents false account opening and bank opening using counterfeit license.
Authenticity Determination Process
Future Direction
Financial institutions are increasingly adopting services using smartphone to improve convenience for transaction. Toppan Forms will develop services using smartphone to provide convenient and safe service for client companies and consumers.
Toppan Forms aims to sell 300 million yen in 2014 by expanding the app such as "FastShot" that provides authenticity determination of Driver's License and electronic delivery service for various proof documents.
About FastShot
FastShot is an electronic delivery service for various proof documents using camera feature on the smartphone. Previously, proof document had to be copied and mailed but the service allows it to be completed on the spot which significantly reduces hassles for consumers. The digitization of submitted document also lightens operational burden of the company by using the service.
- ※ The app compatible to Android terminal with NFC reader writer.
- ※ "FastShot" is currently being applied for registered trademark by Toppan Forms.
- ※ Other products indicated are registered trademark or trademark of each company.
- Public Relations Department
General Affairs Division
- 1-7-3 Higashi Shimbashi, Minato-ku, Tokyo 105-8311, JAPAN
Inquiry Form


text:
The Smith School Young Alumni Council works to guide the development of young alumni programming in order to strengthen the connection between the school and young alumni. The council provides strategic direction on the transition from student to alumni and keeps new graduates engaged with the Smith School of Business by assisting in the development of focused events and programming to meet the needs of the young alumni population.
2019-20 Young Alumni Council
|Stephanie Catherine '14, Technology Consultant, Deloitte
|Maurice Nick '12, MBA/MS '16, Supply Chain Manager, Stanley Black & Decker
|Liya Ai '16, Management Consultant, Accenture
|Steven Asifo '14, Security Risk Management Analyst, Verizon Media
|Dina Broydo '16, Senior Associate, FILS, BDO USA, LLP
|John Gillick '15, Founder, Saltwater Shore
|Ben Hawtof '17, Associate, PwC Public Sector
|Brittany McCoy '16, E-commerce Product Manager, Bluemercury, Inc.
|Haley Nalley '12, Audit Manager, BDO
|Zach Nerenberg '13, Senior Product Manager - Technical, Amazon
|Ali Von Paris '12, CEO, Route One Apparel
|Khalil Pettus '15, Associate, Ardea Partners
|Chris Riggs '16, Trader, Citadel Securities
|Mimi Verdonk '17, Business Analyst, McKinsey & Company
|Edward "EJ" White '15, Digital Marketing Strategist at WebMechanix


text:
Michael's Missives for September 4, 2019
September 4, 2019
First Pres Community –
Hurricane Dorian continues to capture our attention and prayers for all affected by this slow moving, storm. Presbyterian Disaster Assistance is actively working to bring help. You may contribute to PDA through our denomination's website at pcusa.org or by making a check to First Pres and marking it clearly for Dorian relief.
Even if Dorian causes the Chiefs opener (in Jacksonville) to be changed, our "kick-off" is on!
THIS SUNDAY, SEPTEMBER 8th is KICK-OFF SUNDAY! Choir returns at the 11am service. The sermon at both the 9am and 11am services will be "Known by God". Between the services, during the 10am Faith and Fellowship hour, there will be games for kids inside and outside (weather permitting) …and an opportunity for all to meet their Sunday School teachers and to see their classrooms. At Noon, the Chiefs season opener (if held) will be broadcast in our Fellowship Hall as we enjoy a Tailgate Potluck Luncheon with a Chicken Wing Contest. You may sill enter! There will also be brats and burgers served. Wear your red as we enjoy a "wing ding" time together! Nursery care is provided all morning.
This Saturday, September 7, Women's Breakfast at 8:30am. All ladies welcome.
Also, this Saturday, Men's Group Annual Fish Fry at the Beavers. Contact Tom and Roxee Beaver asap.
In addition, this September, the Eating Out Group will dine. Contact Lucy Daily if interested.
Next Monday, September 9, Dinner for Eight, gathering at the Stefanellis, 5:30pm. Contact Donna Haines to sign up or for more information.
Next Thursday, September 12, 8:30am Keenagers travel to Shatto Milk Company for a 10:30am tour. Cracker Barrel for lunch. $6 for tour and cost of lunch. Sign up in Narthex.
Then, Sunday, September 15, Church School classes begin for all ages, including 5 adult classes, 10-10:40am.
TONIGHT, Bells at 6:30pm and Choir at 7:30pm. New participants are always welcome. (LOGOS and Youth Group resume Wednesday, October 2, 5:30-7:30pm.)
Go Team Go !!!
In the faith, hope, and love of Jesus –
Your pastor and friend Michael


text:
Roelfien Folkersma
Roelfien Folkersma (b. 1973) is one of the VSNU's departmental assistants. She studied Musicology at Utrecht University and spent a number of years studying singing at the conservatory. Roelfien previously worked as a communications officer and editor at SGBO, the former research and consultancy firm of the Association of Netherlands Municipalities.
Her contributions at the VSNU have included supporting the campaign team in the run-up to the 2012 Dutch House of Representatives elections. This was a collaborative effort between the VSNU and its knowledge partners the Netherlands Organisation for Scientific Research (NWO), the Confederation of Netherlands Industry and Employers (VNO-NCW), the Dutch Federation of Small and Medium-Sized Enterprises (MKB-Nederland), the Netherlands Organisation for Applied Scientific Research (TNO), the Netherlands Association of Universities of Applied Sciences (HBO-raad) and the Royal Netherlands Academy of Arts and Sciences (KNAW) . Roelfien uses her editing background to ensure that newsletters, correspondence and meeting documents are in tip-top shape before being distributed. In her free time she serves as board secretary of ProJazz, an organisation that aims to breathe new life into the jazz scene in The Hague.
E-mail: firstname.lastname@example.com
Phone: 06 27 29 60 46