Sample records for pacific cod trypsin
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76 FR 2084 - North Pacific Fishery Management Council; Public Meetings
Federal Register 2010, 2011, 2012, 2013, 2014
2011-01-12
... Amendment(T). 6. Aleutian Island (AI) Pacific Cod Issues: Discussion paper on BS&AI Pacific cod Split take action as necessary; Initial review AI Pacific cod processing Sideboards; (postponed). 7. BSAI Crab.... AI Pacific Cod. 4. BSAI Crab Management. 5. Miscellaneous Groundfish. 6. Economic SAFE review. 7...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-04-15
... is opening directed fishing for Pacific cod by catcher vessels less than 60 feet (18.3 meters) length... meters) length overall using hook-and-line or pot gear in the BSAI under Sec. 679.20(d)(1)(iii) on March... 400 metric tons of Pacific cod remain in the 2010 Pacific cod apportionment for catcher vessels less...
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Phenotypic flexibility of digestive system in Atlantic cod (Gadus morhua).
Blier, P U; Dutil, J-D; Lemieux, H; Bélanger, F; Bitetera, L
2007-02-01
This study examined the restoration of the digestive capacity of Atlantic cod (Gadus morhua Linnaeus) following a long period of food deprivation. Fifty cod (48 cm, 1 kg) were food-deprived for 68 days and then fed in excess with capelin (Mallotus villosus Müller) on alternate days. Ten fish were sampled after 0, 2, 6, 14 and 28 days and the mass of the pyloric caeca, intestine and carcass determined. Two metabolic enzymes (cytochrome c oxidase and citrate synthase) were assayed in white muscle, pyloric caeca and intestine, and trypsin activity was measured in the pyloric caeca. A delay of 14 days was required before body mass started to increase markedly, whereas most of the increase in mass of both the pyloric caeca and intestine relative to fish length occurred earlier in the experiment. By day 14, the activities of trypsin and citrate synthase in the pyloric caeca as well as citrate synthase in the intestine had reached maxima. The growth of the digestive tissues and restoration of their metabolic capacities thus occur early upon refeeding and are likely required for recovery growth to take place. The phenotypic flexibility of the cod digestive system is therefore remarkable: increases in trypsin activity and size of pyloric caeca resulted in a combined 29-fold increase in digestive capacity of the fish during the refeeding period. Our study suggests that Atlantic cod are able to cope with marked fluctuations in food availability in their environment by making a rapid adjustment of their digestive capacity as soon as food availability increases.
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Wu, Wenfei; Li, Bafang; Hou, Hu; Zhang, Hongwei; Zhao, Xue
2017-12-13
A calcium-chelating peptide is considered to have the ability to improve calcium absorption. In this study, Pacific cod skin gelatin hydrolysates treated with trypsin for 120 min exhibited higher calcium-chelating activity. Sequential chromatography, involving hydroxyapatite affinity chromatography and reversed phase high performance liquid chromatography, was used for the purification of calcium-chelating peptides. Two novel peptides with the typical characteristics of collagen were sequenced as GDKGESGEAGER and GEKGEGGHR based on LC-HRMS/MS, which showed a high affinity to calcium. Calcium-peptide complexation was further characterized by ESI-MS (MS and MS/MS) and FTIR spectroscopy. The results showed that the complexation of the two peptides with calcium was conducted mainly at the ratio of 1 : 1. The amino terminal group and the peptide bond of the peptide backbone as well as the amino group of the lysine side chain and the carboxylate of the glutamate side chain were the possible calcium binding sites for the two peptides. Meanwhile, several amino acid side chain groups, including the hydroxyl (Ser) and carboxylate (Asp) of GDKGESGEAGER and the imine (His) of GEKGEGGHR, were crucial in the complexation. The arginine residue in GEKGEGGHR also participated in the calcium coordination. Additionally, several active fragments with calcium-chelating activity were obtained using MS/MS spectra, including GDKGESGEAGE, GEAGER, GEK, EKG and KGE. This study suggests that gelatin-derived peptides have the potential to be used as a calcium-chelating ingredient to combat calcium deficiency.
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-06-28
...-AY53 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod Allocations in the Gulf of Alaska... the uncertainty regarding the distribution of Pacific cod catch, enhance stability among the sectors... available for public review and comment. The groundfish fisheries in the exclusive economic zone of the Gulf...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-11-15
.... 0910131363-0087-02] RIN 0648-XA038 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod in the Bering Sea and Aleutian Islands Management Area AGENCY: National Marine Fisheries Service (NMFS...: NMFS is prohibiting retention of Pacific cod in the Bering Sea and Aleutian Islands Management Area...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-03-09
... Using Jig Gear in the Central Regulatory Area of the Gulf of Alaska AGENCY: National Marine Fisheries...; closure. SUMMARY: NMFS is prohibiting directed fishing for Pacific cod by vessels using jig gear in the... season allowance of the 2012 Pacific cod total allowable catch apportioned to vessels using jig gear in...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-02-09
... Using Pot Gear in the Western Regulatory Area of the Gulf of Alaska AGENCY: National Marine Fisheries...; closure. SUMMARY: NMFS is prohibiting directed fishing for Pacific cod by vessels using pot gear in the... season allowance of the 2012 Pacific cod total allowable catch apportioned to vessels using pot gear in...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-02-14
... Using Pot Gear in the Central Regulatory Area of the Gulf of Alaska AGENCY: National Marine Fisheries...; closure. SUMMARY: NMFS is prohibiting directed fishing for Pacific cod by vessels using pot gear in the... season allowance of the 2012 Pacific cod total allowable catch apportioned to vessels using pot gear in...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-01-25
.... 0810141351-9087-02] RIN 0648-XT95 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod by Vessels Participating in the Amendment 80 Limited Access Fishery in Bering Sea and Aleutian Islands Management Area... management area (BSAI). This action is necessary to prevent exceeding the 2010 Pacific cod total allowable...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-09-06
.... 111207737-2141-02] RIN 0648-XC205 Fisheries of the Exclusive Economic Zone Off Alaska; Reallocation of Pacific Cod in the Western Regulatory Area of the Gulf of Alaska Management Area AGENCY: National Marine... Alaska management area. This action is necessary to allow the 2012 total allowable catch of Pacific cod...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-03-25
... Catcher Vessels Greater Than or Equal to 50 Feet (15.2 Meters) Length Overall Using Hook-and-Line Gear in...: NMFS is prohibiting directed fishing for Pacific cod by catcher vessels (CVs) greater than or equal to... of the 2013 Pacific cod total allowable catch apportioned to CVs greater than or equal to 50 feet (15...
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Kortner, Trond M; Overrein, Ingrid; Oie, Gunvor; Kjørsvik, Elin; Bardal, Tora; Wold, Per-Arvid; Arukwe, Augustine
2011-10-01
We have profiled the expression of twelve genes, in order to provide an overview on the molecular ontogeny of digestive capability with the associated endocrine control during Atlantic cod (Gadus morhua) larval development. Enzyme activity levels for the key digestive enzyme, trypsin, was also measured. Specifically, transcripts for trypsin, amylase, lipolytic enzymes: bile salt activated lipase (BAL), phospholipase A2 (PLA2) and Acyl CoA dehydrogenase (ACADM), regulatory peptides: neuropeptide Y (NPY), orexin (OX) cholecystokinin (CCK) and cocaine and amphetamine-related transcript (CART), the somatotropic factors: growth hormone (GH), preprosomatostatin 1 (PPSS1) and thyroid hormone receptors (TRα and TRβ) were analyzed using quatitative (real-time) polymerase chain reaction (qPCR). Trypsin and BAL mRNA levels peaked at approximately day 17 and 25 post-hatch, respectively, and thereafter displayed a decreasing pattern until metamorphosis. GH mRNA levels decreased moderately from 3 to 33dph, and thereafter, an increase was observed until 46dph. TRα mRNA levels showed a fluctuating pattern peaking at day 39 post-hatch. TRβ mRNA levels were too low to obtain quantitative measurements. Amylase mRNA slightly increased from day 3 to 17 post-hatch, and thereafter showed a steady decrease until day 60. Interestingly, PLA2 mRNA expression showed a consistent increase throughout the study period, indicating an increasingly important role during larval development. Overall, data from this study indicate that cod larvae show differential developmental mode of expression patterns for key genes and endocrine factors that regulate digestive capability, growth and development. These data are discussed in relation to larval trypsin enzyme activity and previous reports for other teleost species. Copyright © 2011 Elsevier Inc. All rights reserved.
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-04-15
... gear to catcher vessels less than 60 feet (18.3 meters) length overall (LOA) using hook-and-line or pot... vessels will not be able to harvest 400 mt of the B season apportionment of the 2010 Pacific cod TAC...)(iii)(A), NMFS apportions 400 mt of Pacific cod from the B season jig gear apportionment to catcher...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-08-26
... is opening directed fishing for Pacific cod by catcher vessels less than 60 feet (18.3 meters) length... catcher vessels less than 60 feet (18.3 meters) LOA using hook-and-line or pot gear specified for the BSAI... 679. NMFS closed directed fishing for Pacific cod by catcher vessels less than 60 feet (18.3 meters...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-03-07
.... 101126522-0640-2] RIN 0648-XB062 Pacific Cod by Catcher Vessels Less Than 50 Feet (15.2 Meters) Length... (CVs) less than 50 feet (15.2 meters (m)) in length overall (LOA) using hook-and-line gear in the... season allowance of the 2012 Pacific cod total allowable catch apportioned to CVs less than 50 feet (15.2...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-09-11
... closure. SUMMARY: NMFS is opening directed fishing for Pacific cod by catcher vessels less than 60 feet... Pacific cod allocated to catcher vessels less than 60 feet LOA using hook-and-line or pot gear in the BSAI... vessels less than 60 feet LOA using hook-and-line or pot gear in the BSAI under Sec. 679.20(d)(1)(iii) on...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-05-02
... closure. SUMMARY: NMFS is opening directed fishing for Pacific cod by catcher vessels less than 60 feet... Pacific cod allocated to catcher vessels less than 60 feet LOA using hook-and-line or pot gear in the BSAI... less than 60 feet LOA using hook-and-line or pot gear in the BSAI under Sec. 679.20(d)(1)(iii) on March...
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NASA Astrophysics Data System (ADS)
Copeman, L.; Laurel, B.; Spencer, M. L.; Iseri, P.; Sremba, A. L.
2016-02-01
Climate change impacts on Arctic ecosystems will largely be determined by temperature-dependent bioenergetics of resident and invading forage fish species. In this study, we experimentally measured total lipids and lipid class storage in the liver and muscle of juvenile Arctic gadids (Arctic cod, Boreogadus saida and saffron cod, Eleginus gracilis) and two North Pacific gadids (walleye pollock, Gadus chalcogrammus and Pacific cod, Gadus macrocephalus). Experiments were conducted over a 6-wk period across five temperatures (0, 5, 9, 16 and 20 °C) at the Hatfield Marine Science Center in Newport, OR, USA. Results indicated clear physiological differences among species in terms of temperature-dependent growth and lipid storage. Arctic cod exhibited highest growth and lipid storage (27 mg/g WW) at the coldest temperature (0 °C) compared to the other gadids, with near maximum growth at 5 °C and onset of mortality above 9 °C. In contrast, saffron cod growth rates steadily increased at temperatures beyond 16 °C, but lipid storage was low overall with only slightly higher lipid storage at warm temperatures (10 to 17 mg/g WW). Both walleye pollock and Pacific cod showed a domed response with increased lipid storage and growth at intermediate temperatures (9 - 12°C) and reduced growth and lipid storage at cold and warm maxima. We did not observe a trade-off between growth rate and lipid accumulation in any species. These results suggest that saffron cod can thrive in a warming Arctic but will be energetically inferior as a prey item to the more temperature-sensitive Arctic cod. Alternatively, North Pacific gadids can energetically resemble Arctic cod at warmer temperatures and could theoretically be an important prey item if their range extends northward with continued climate change.
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-11-29
... miles of shore adjacent to the Bering Sea and Aleutian Islands management area (BSAI). The affected... Cod Fishing in the Parallel Fishery in the Bering Sea and Aleutian Islands Management Area AGENCY... limits access by Federally permitted vessels to the parallel fishery for Pacific cod in three ways. First...
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Code of Federal Regulations, 2014 CFR
2014-10-01
...% of the TAC. Pacific ocean perch 96.1% of the TAC. Pelagic shelf rockfish 89.6% of the TAC. Central GOA Pacific cod 4.4% of the TAC. Pacific ocean perch Subject to regulations in subpart G to this part... regulations in subpart G to this part. Western GOA Pacific cod 2.0% of the TAC. Pacific ocean perch 99.4% of...
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Code of Federal Regulations, 2011 CFR
2011-10-01
...% of the TAC. Pacific ocean perch 96.1% of the TAC. Pelagic shelf rockfish 89.6% of the TAC. Central GOA Pacific cod 4.4% of the TAC. Pacific ocean perch Subject to regulations in subpart G to this part... regulations in subpart G to this part. Western GOA Pacific cod 2.0% of the TAC. Pacific ocean perch 99.4% of...
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Code of Federal Regulations, 2010 CFR
2010-10-01
...% of the TAC. Pacific ocean perch 96.1% of the TAC. Pelagic shelf rockfish 89.6% of the TAC. Central GOA Pacific cod 4.4% of the TAC. Pacific ocean perch Subject to regulations in subpart G to this part... regulations in subpart G to this part. Western GOA Pacific cod 2.0% of the TAC. Pacific ocean perch 99.4% of...
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Code of Federal Regulations, 2012 CFR
2012-10-01
...% of the TAC. Pacific ocean perch 96.1% of the TAC. Pelagic shelf rockfish 89.6% of the TAC. Central GOA Pacific cod 4.4% of the TAC. Pacific ocean perch Subject to regulations in subpart G to this part... regulations in subpart G to this part. Western GOA Pacific cod 2.0% of the TAC. Pacific ocean perch 99.4% of...
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Code of Federal Regulations, 2013 CFR
2013-10-01
...% of the TAC. Pacific ocean perch 96.1% of the TAC. Pelagic shelf rockfish 89.6% of the TAC. Central GOA Pacific cod 4.4% of the TAC. Pacific ocean perch Subject to regulations in subpart G to this part... regulations in subpart G to this part. Western GOA Pacific cod 2.0% of the TAC. Pacific ocean perch 99.4% of...
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75 FR 20344 - North Pacific Fishery Management Council; Public Meeting
Federal Register 2010, 2011, 2012, 2013, 2014
2010-04-19
...) groundfish plan teams will meet via teleconference May 6, 2010, 12:30 p.m. Alaska Standard Time (AST) to review proposals for models to be considered for inclusion in the GOA and BSAI Pacific cod assessments... INFORMATION: Agenda: Review proposals for models Pacific cod stock assessments. The agenda is posted on the...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-12-01
... GOA Pacific cod TACs among gear and operation type, based primarily on historical dependency and catch... 83 sector allocations are based on historical dependency, each sector's retained catch history of the...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-09-30
... authorizing a designated vessel to catch and process Pacific cod in the BSAI hook-and-line fisheries to use... (MLOA) to 220 feet (67 m) on LLP licenses authorizing vessels to catch and process Pacific cod with hook... cod with hook-and-line and pot gear in the BSAI to increase the MLOA on the LLP license to 220 feet...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-03-11
... Department of Fish and Game (ADF&G) through emergency order under the authority of State regulations at 5 AAC... Pacific cod among ten fishery sectors created in Amendment 67, to better reflect the historical dependency...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-05-02
... catch of Pacific cod to be harvested. DATES: Effective April 30, 2011, through 2400 hrs, Alaska local... gear is 207 mt for the period 2400 hrs, A.l.t., January 1, 2011, through 1200 hrs, A.l.t., June 10...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-03-10
... Catcher Vessels Less Than 60 Feet (18.3 m) Length Overall Using Hook-and-Line or Pot Gear in the Bering... prohibiting directed fishing for Pacific cod by catcher vessels less than 60 feet (18.3 m) length overall (LOA...
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NASA Astrophysics Data System (ADS)
Adlerstein, Sara A.; Trumble, Robert J.
1998-03-01
Mortality of discarded Pacific halibut bycatch from Pacific cod fisheries in the Bering Sea leads to significant losses in the halibut setline and in the Pacific cod fisheries. The commercial halibut fishery loses yield because of catch limit reductions to compensate the resource for lost spawning potential and because halibut killed as bycatch will not be available for subsequent harvest, and the cod fisheries may lose harvest if they reach a bycatch mortality limit before reaching allowed catch. In this study, significant differences in Pacific halibut bycatch rates and associated yield losses were found among months and areas of the Bering Sea in the longline and trawl fisheries for Pacific cod in 1990-1992. Bycatch rates were usually highest in late spring and early summer and in areas close to the Unimak Pass. With the exception of 1992, yield loss in the longline fishery was around 1 kg per kg of bycatch mortality, irrespective of where or when bycatch occurred. In the trawl fishery, loss of halibut yield varied from 1 to 4 kg per kg of bycatch mortality. Highest halibut net yield losses per tonne of groundfish harvest usually coincided with highest bycatch rates. When both fisheries operated in one area, trawl bycatch often imposed higher yield losses than longline bycatch, despite lower bycatch rates. Bycatch was affected by the strong 1987 halibut year class. Highest bycatch and yield loss rates occurred in the trawl fishery in 1990 and 1991 when the population was dominated by halibut age-3 and -4, and in the longline fishery in 1992 as fish reached age-5.
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-06-12
... in the BSAI exclusive economic zone according to the Fishery Management Plan for Groundfish of the.... 111213751-2102-02] RIN 0648-XC064 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod for... Management Area AGENCY: National Marine Fisheries Service (NMFS), National Oceanic and Atmospheric...
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Tillner, Robert; Rønnestad, Ivar; Harboe, Torstein; Ueberschär, Bernd
2013-11-01
In order to maximize protein digestion, the release of enzymes into the gut lumen is closely controlled by a regulatory loop. Cholecystokinin (CCK) is among the enteric hormones that play a key role in the control of digestive enzyme secretion, but its role in first-feeding larvae is still unclear and may differ between species. However, in all marine fish larvae that have not developed a stomach by first-feeding, trypsin is the most important proteolytic enzyme. In order to examine the regulation and feedback mechanisms in the gut of larval cod, we therefore studied the interactions between cholecystokinin and tryptic enzyme activity following the administration of solutions containing test substances directly into the gut. We tube-fed a single dose of physiological saline solution containing either CCK, CCK antagonist, trypsin inhibitor, phytohemagglutinin (PHA; a possible trigger for the digestive response) or physiological saline alone, while a further control group was left untreated. We then followed the response in CCK and tryptic enzyme activity for 0.5-8h after the administration. We performed the experiment on larvae at 26day post first-feeding, which is before the stomach has evolved and the size of the larvae allows easier handling. Individual larvae were analyzed for CCK and tryptic enzyme activity using radioimmunoassay and fluorimetric techniques respectively. Both factors varied over time in the untreated control group, possibly due to an endogenous daily rhythm. The higher CCK levels at 4h and 8h in the saline-injected group may be caused by reflexes initiated by distension of the gut. An increase in tryptic enzyme activity after injection of CCK supports the hypothesis that this hormone plays a part in the release of pancreatic enzymes in larval cod at this developmental stage. However, administration of a CCK antagonist and a trypsin inhibitor did not reveal conclusive results, probably due to the relatively low concentrations used. The response in tryptic activity in the PHA group was similar to the administration of CCK, pointing towards a stimulatory effect of PHA on the proteolytic enzyme capacity of cod larvae. © 2013.
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-03-14
..., sablefish, deep-water flatfish, rex sole, Pacific ocean perch, northern rockfish, shortraker rockfish... cod, shallow-water flatfish, arrowtooth flounder, flathead sole, ``other rockfish'' in the Eastern GOA...) guideline harvest levels (GHLs) for Pacific cod so that the ABC is not exceeded. The shallow-water flatfish...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-06-04
... DEPARTMENT OF COMMERCE National Oceanic and Atmospheric Administration 50 CFR Part 679 [Docket No. 0910131363-0087-02] RIN 0648-XW75 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod for American Fisheries Act Catcher Processors Using Trawl Gear in the Bering Sea and Aleutian Islands...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-10-26
... DEPARTMENT OF COMMERCE National Oceanic and Atmospheric Administration 50 CFR Part 679 [Docket No. 101126521-0640-02] RIN 0648-XA791 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod by Catcher/Processors Using Pot Gear in the Bering Sea and Aleutian Islands Management Area AGENCY: National...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-02-23
... DEPARTMENT OF COMMERCE National Oceanic and Atmospheric Administration 50 CFR Part 679 [Docket No. 0810141351-9087-02] RIN 0648-XU52 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod for American Fisheries Act Catcher-Processors Using Trawl Gear in the Bering Sea and Aleutian Islands...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-08-04
... DEPARTMENT OF COMMERCE National Oceanic and Atmospheric Administration 50 CFR Part 679 [Docket No. 101126521-0640-2] RIN 0648-XA616 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod for American Fisheries Act Catcher/Processors Using Trawl Gear in the Bering Sea and Aleutian Islands...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-01-26
... DEPARTMENT OF COMMERCE National Oceanic and Atmospheric Administration 50 CFR Part 679 [Docket No. 0910131363-0087-02] RIN 0648-XA176 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod by Catcher/Processors Using Pot Gear in the Bering Sea and Aleutian Islands Management Area AGENCY: National...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-04-20
... Catcher Vessels Using Trawl Gear in the Bering Sea and Aleutian Islands Management Area AGENCY: National... catcher vessels using trawl gear in the Bering Sea and Aleutian Islands Management Area (BSAI). This... Pacific cod allocated to catcher vessels using trawl gear in the BSAI. DATES: Effective 1200 hrs, Alaska...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-02-26
... DEPARTMENT OF COMMERCE National Oceanic and Atmospheric Administration 50 CFR Part 679 [Docket No. 0810141351-9087-02] RIN 0648-XU65 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod by Catcher/Processors Using Pot Gear in the Bering Sea and Aleutian Islands Management Area AGENCY: National Marine...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-01-25
... DEPARTMENT OF COMMERCE National Oceanic and Atmospheric Administration 50 CFR Part 679 [Docket No. 101126521-0640-02] RIN 0648-XA955 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod by Catcher/Processors Using Pot Gear in the Bering Sea and Aleutian Islands Management Area AGENCY: National...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-02-23
... DEPARTMENT OF COMMERCE National Oceanic and Atmospheric Administration 50 CFR Part 679 [Docket No. 101126521-0640-02] RIN 0648-XB028 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod for American Fisheries Act Catcher/Processors Using Trawl Gear in the Bering Sea and Aleutian Islands...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-04-06
... DEPARTMENT OF COMMERCE National Oceanic and Atmospheric Administration 50 CFR Part 679 [Docket No. 0910131363-0087-02] RIN 0648-XV66 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod for American Fisheries Act Catcher Processors Using Trawl Gear in the Bering Sea and Aleutian Islands...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-03-30
... DEPARTMENT OF COMMERCE National Oceanic and Atmospheric Administration 50 CFR Part 679 [Docket No. 111213751-2102-02] RIN 0648-XB138 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod for American Fisheries Act Catcher/Processors Using Trawl Gear in the Bering Sea and Aleutian Islands...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-09-27
... DEPARTMENT OF COMMERCE National Oceanic and Atmospheric Administration 50 CFR Part 679 [Docket No. 0910131363-0087-02] RIN 0648-XZ27 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod by Catcher/Processors Using Pot Gear in the Bering Sea and Aleutian Islands Management Area AGENCY: National Marine...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-02-01
... DEPARTMENT OF COMMERCE National Oceanic and Atmospheric Administration 50 CFR Part 679 [Docket No. 111213751-2102-02] RIN 0648-XC465 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod by Catcher/Processors Using Pot Gear in the Bering Sea and Aleutian Islands Management Area AGENCY: National...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-04-14
... Catcher Vessels Using Trawl Gear in the Bering Sea and Aleutian Islands Management Area AGENCY: National... catcher vessels using trawl gear in the Bering Sea and Aleutian Islands Management Area (BSAI). This... Pacific cod allocated to catcher vessels using trawl gear in the BSAI. DATES: Effective 1200 hrs, Alaska...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-03-14
... who also had significant participation in, or dependence on, the GOA Pacific cod fishery to maintain... vessels that demonstrated minimal participation in, or dependence on, the Bering Sea snow crab fishery and sufficient participation in, or dependence on, the GOA Pacific cod fishery from 1996 through 2000. Non-AFA...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-09-14
.... 0910131363-0087-02] RIN 0648-XZ01 Fisheries of the Exclusive Economic Zone Off Alaska; Reallocation of Pacific cod in the Bering Sea and Aleutian Islands Management Area AGENCY: National Marine Fisheries... Sea and Aleutian Islands management area. These actions are necessary to allow the 2010 total...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-02-08
.... 111213751-2102-02] RIN 0648-XC481 Fisheries of the Exclusive Economic Zone Off Alaska; Reallocation of Pacific Cod in the Bering Sea and Aleutian Islands Management Area AGENCY: National Marine Fisheries... in the Bering Sea and Aleutian Islands management area. This action is necessary to allow the A...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-03-07
.... 101126521-0640-02] RIN 0648-XA260 Fisheries of the Exclusive Economic Zone Off Alaska; Reallocation of Pacific Cod in the Bering Sea and Aleutian Islands Management Area AGENCY: National Marine Fisheries... in the Bering Sea and Aleutian Islands management area. This action is necessary to allow the A...
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Code of Federal Regulations, 2012 CFR
2012-10-01
...) Bogoslof Pacific cod exemption area. (1) All catcher vessels less than 60 ft (18.3 m) LOA using jig or hook... Pacific cod have been caught by catcher vessels less than 60 ft (18.3 m) LOA using jig or hook-and-line... using jig or hook-and-line gear in the exemption area by notification published in the Federal Register...
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Code of Federal Regulations, 2014 CFR
2014-10-01
...) Bogoslof Pacific cod exemption area. (1) All catcher vessels less than 60 ft (18.3 m) LOA using jig or hook... Pacific cod have been caught by catcher vessels less than 60 ft (18.3 m) LOA using jig or hook-and-line... using jig or hook-and-line gear in the exemption area by notification published in the Federal Register...
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Code of Federal Regulations, 2013 CFR
2013-10-01
...) Bogoslof Pacific cod exemption area. (1) All catcher vessels less than 60 ft (18.3 m) LOA using jig or hook... Pacific cod have been caught by catcher vessels less than 60 ft (18.3 m) LOA using jig or hook-and-line... using jig or hook-and-line gear in the exemption area by notification published in the Federal Register...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-10-27
... Octopus in the Bering Sea and Aleutian Islands Management Area AGENCY: National Marine Fisheries Service... catch of octopus by vessels using pot gear to fish for Pacific cod the BSAI. DATES: Effective 1200 hrs... and management measures prevent overfishing. The 2011 octopus overfishing level in the BSAI is 528...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-04-18
... Catcher Vessels Using Trawl Gear in the Bering Sea and Aleutian Islands Management Area AGENCY: National... using trawl gear in the Bering Sea and Aleutian Islands management area (BSAI). This action is necessary... fishing for Pacific cod by catcher vessels using trawl gear in the BSAI. After the effective date of this...
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50 CFR Table 5 to Part 679 - Steller Sea Lion Protection Areas Pacific Cod Fisheries Restrictions
Code of Federal Regulations, 2010 CFR
2010-10-01
... 50 Wildlife and Fisheries 9 2010-10-01 2010-10-01 false Steller Sea Lion Protection Areas Pacific Cod Fisheries Restrictions 5 Table 5 to Part 679 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE (CONTINUED) FISHERIES OF THE EXCLUSIVE ECONOMIC ZONE OFF ALASKA Pt....
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50 CFR Table 5 to Part 679 - Steller Sea Lion Protection Areas Pacific Cod Fisheries Restrictions
Code of Federal Regulations, 2011 CFR
2011-10-01
... 50 Wildlife and Fisheries 11 2011-10-01 2011-10-01 false Steller Sea Lion Protection Areas Pacific Cod Fisheries Restrictions 5 Table 5 to Part 679 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE (CONTINUED) FISHERIES OF THE EXCLUSIVE ECONOMIC ZONE OFF ALASKA Pt....
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-12-06
... in the Gulf of Alaska exclusive economic zone according to the Fishery Management Plan for Groundfish.... 120918468-3111-02] RIN 0648-XC976 Fisheries of the Exclusive Economic Zone Off Alaska; Reallocation of Pacific Cod in the Central Regulatory Area of the Gulf of Alaska Management Area AGENCY: National Marine...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-12-06
.... 120918468-3111-02] RIN 0648-XC975 Fisheries of the Exclusive Economic Zone Off Alaska; Reallocation of Pacific Cod in the Western Regulatory Area of the Gulf of Alaska Management Area AGENCY: National Marine... the Western Regulatory Area of the Gulf of Alaska management area (GOA). This action is necessary to...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-09-10
... in the Gulf of Alaska exclusive economic zone according to the Fishery Management Plan for Groundfish.... 120918468-3111-02] RIN 0648-XC856 Fisheries of the Exclusive Economic Zone Off Alaska; Reallocation of Pacific Cod in the Western Regulatory Area of the Gulf of Alaska Management Area AGENCY: National Marine...
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50 CFR Table 5 to Part 679 - Steller Sea Lion Protection Areas Pacific Cod Fisheries Restrictions
Code of Federal Regulations, 2014 CFR
2014-10-01
... 50 Wildlife and Fisheries 13 2014-10-01 2014-10-01 false Steller Sea Lion Protection Areas Pacific Cod Fisheries Restrictions 5 Table 5 to Part 679 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE (CONTINUED) FISHERIES OF THE EXCLUSIVE ECONOMIC ZONE OFF ALASKA Pt....
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50 CFR Table 5 to Part 679 - Steller Sea Lion Protection Areas Pacific Cod Fisheries Restrictions
Code of Federal Regulations, 2012 CFR
2012-10-01
... 50 Wildlife and Fisheries 13 2012-10-01 2012-10-01 false Steller Sea Lion Protection Areas Pacific Cod Fisheries Restrictions 5 Table 5 to Part 679 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE (CONTINUED) FISHERIES OF THE EXCLUSIVE ECONOMIC ZONE OFF ALASKA Pt....
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50 CFR Table 5 to Part 679 - Steller Sea Lion Protection Areas Pacific Cod Fisheries Restrictions
Code of Federal Regulations, 2013 CFR
2013-10-01
... 50 Wildlife and Fisheries 13 2013-10-01 2013-10-01 false Steller Sea Lion Protection Areas Pacific Cod Fisheries Restrictions 5 Table 5 to Part 679 Wildlife and Fisheries FISHERY CONSERVATION AND MANAGEMENT, NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION, DEPARTMENT OF COMMERCE (CONTINUED) FISHERIES OF THE EXCLUSIVE ECONOMIC ZONE OFF ALASKA Pt....
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Meyers, T.R.; Sullivan, J.; Emmenegger, E.; Follett, J.; Short, S.; Batts, W.N.; Winton, J.R.
1992-01-01
Ulcerative slun tissues from 2 Pacific cod Gadus rnacrocephalus caught in Prince William Sound, Alaska, USA, were examined for virus by Fish Pathology staff within the F.R.E.D. Division of the Alaska Department of Fish and Game. Six days after inoculation of Epitheliorna papulosum cyprini (EPC) cells at 14"C, diffuse rounding and lifting of cells from the monolayers suggestive of cytopathlc effect became visible in the lower sample dilutions. Ultrastructural examinations of affected EPC cells showed rhabdovirus particles within cytoplasmic vacuoles and on the cell surface membranes. Virus isolates from both cod were subsequently confirmed as viral hemorrhagic septicemia virus (VHSV) by serum neutralizabon and immunoblot assay. This is the first VHSV isolated from Pacific cod, which represents a new host species for the virus. Histologically, cod skin ulcers appeared to be caused by a foreign-body-type inflammatory response to foci of protozoa resembling X cells that also had plasmodial stages. Whether the rhabdovirus was incidental to the slun lesion or played a role in its etiology remains to be determined. The possible relationship between thls virus and the recent occurrences of VHSV in anadromous salmoruds from Washington State, USA, is discussed.
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A colostrum trypsin inhibitor gene expressed in the Cape fur seal mammary gland during lactation.
Pharo, Elizabeth A; Cane, Kylie N; McCoey, Julia; Buckle, Ashley M; Oosthuizen, W H; Guinet, Christophe; Arnould, John P Y
2016-03-01
The colostrum trypsin inhibitor (CTI) gene and transcript were cloned from the Cape fur seal mammary gland and CTI identified by in silico analysis of the Pacific walrus and polar bear genomes (Order Carnivora), and in marine and terrestrial mammals of the Orders Cetartiodactyla (yak, whales, camel) and Perissodactyla (white rhinoceros). Unexpectedly, Weddell seal CTI was predicted to be a pseudogene. Cape fur seal CTI was expressed in the mammary gland of a pregnant multiparous seal, but not in a seal in its first pregnancy. While bovine CTI is expressed for 24-48 h postpartum (pp) and secreted in colostrum only, Cape fur seal CTI was detected for at least 2-3 months pp while the mother was suckling its young on-shore. Furthermore, CTI was expressed in the mammary gland of only one of the lactating seals that was foraging at-sea. The expression of β-casein (CSN2) and β-lactoglobulin II (LGB2), but not CTI in the second lactating seal foraging at-sea suggested that CTI may be intermittently expressed during lactation. Cape fur seal and walrus CTI encode putative small, secreted, N-glycosylated proteins with a single Kunitz/bovine pancreatic trypsin inhibitor (BPTI) domain indicative of serine protease inhibition. Mature Cape fur seal CTI shares 92% sequence identity with Pacific walrus CTI, but only 35% identity with BPTI. Structural homology modelling of Cape fur seal CTI and Pacific walrus trypsin based on the model of the second Kunitz domain of human tissue factor pathway inhibitor (TFPI) and porcine trypsin (Protein Data Bank: 1TFX) confirmed that CTI inhibits trypsin in a canonical fashion. Therefore, pinniped CTI may be critical for preventing the proteolytic degradation of immunoglobulins that are passively transferred from mother to young via colostrum and milk. Copyright © 2015 Elsevier B.V. All rights reserved.
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-03-12
... TACs for 2010 and 2011 that are equal to ABCs for pollock, deep-water flatfish, rex sole, sablefish... recommended TACs for 2010 and 2011 that are less than the ABCs for Pacific cod, flathead sole, shallow-water... that the sum of all State and Federal water Pacific cod removals from the GOA not exceed ABC...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-10-15
... Vessels Using Pot Gear in the Central Regulatory Area of the Gulf of Alaska AGENCY: National Marine... gear in the Central Regulatory Area of the Gulf of Alaska (GOA). This action is necessary to prevent exceeding the 2012 Pacific cod total allowable catch apportioned to vessels using pot gear in the Central...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-02-19
.... 0810141351-9087-02] RIN 0648-XU36 Fisheries of the Exclusive Economic Zone Off Alaska; Pacific Cod by Catcher/Processors Using Hook-and-Line Gear in the Bering Sea and Aleutian Islands Management Area AGENCY: National... using hook-and-line gear in the Bering Sea and Aleutian Islands management area (BSAI). This action is...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-03-01
.... 1801 et seq. Dated: February 26, 2013. Kara Meckley, Acting Deputy Director, Office of Sustainable... hours. This action is necessary to fully use the A season allowance of the 2013 Pacific cod total...: Effective 1200 hours, Alaska local time (A.l.t.), March 1, 2013, through 1200 hours, A.l.t., March 3, 2013...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-01-24
... Catcher Vessels Greater Than or Equal To 60 Feet (18.3 Meters) Length Overall Using Pot Gear in the Bering... prohibiting directed fishing for Pacific cod by pot catcher vessels greater than or equal to 60 feet (18.3... allowable catch (TAC) specified for pot catcher vessels greater than or equal to 60 feet (18.3 m) LOA in the...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-08-28
... gear and catcher vessels greater than 60 feet (18.3 meters) length overall (LOA) using hook-and-line gear to catcher vessels less than 60 feet (18.3 meters) LOA using hook-and-line or pot gear in the...)(iii)(A), NMFS apportions 2,500 mt of Pacific cod to catcher vessels less than 60 feet (18.3 meters(m...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-03-25
... Catcher Vessels Less Than 50 Feet (15.2 Meters) Length Overall Using Hook-and-Line Gear in the Central... directed fishing for Pacific cod by catcher vessels (CVs) less than 50 feet (15.2 meters (m)) in length... catch apportioned to CVs less than 50 feet (15.2 m) LOA using hook-and-line gear in the Central...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-11-26
... Catcher Vessels Less Than 60 Feet (18.3 m) Length Overall Using Hook-and-Line or Pot Gear in the Bering.... SUMMARY: NMFS is prohibiting directed fishing for Pacific cod by catcher vessels less than 60 feet (18.3 m... allocated to catcher vessels less than 60 feet LOA using hook-and-line or pot gear in the BSAI. DATES...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-03-28
... Catcher Vessels Less Than 60 feet (18.3 meters) Length Overall Using Jig or Hook-and-Line Gear in the... less than 60 feet (18.3 meters (m)) length overall (LOA) using jig or hook-and-line gear in the... action is necessary to prevent exceeding the limit of Pacific cod for catcher vessels less than 60 feet...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-08-31
... gear, catcher vessels greater than 60 feet (18.3 meters) length overall (LOA) using hook-and-line gear, and catcher vessels using trawl gear to catcher vessels less than 60 feet (18.3 meters) LOA using hook... than 60 feet (18.3 meters(m)) LOA using hook-and- line or pot gear. The 2012 Pacific cod TAC specified...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-02-22
... Catcher Vessels Less Than 60 Feet (18.3 Meters) Length Overall Using Hook-and-Line or Pot Gear in the...: NMFS is prohibiting directed fishing for Pacific cod by catcher vessels less than 60 feet (18.3 meters... catch (TAC) specified for catcher vessels less than 60 feet (18.3 m) LOA using hook-and-line or pot gear...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-02-08
... Catcher Vessels Less Than 60 Feet (18.3 Meters) Length Overall Using Hook-and-Line or Pot Gear in the...: NMFS is prohibiting directed fishing for Pacific cod by catcher vessels less than 60 feet (18.3 meters... catch (TAC) specified for catcher vessels less than 60 feet (18.3 m) LOA using hook-and-line or pot gear...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-01-23
... Catcher Vessels Greater Than or Equal To 60 Feet (18.3 Meters) Length Overall Using Pot Gear in the Bering... prohibiting directed fishing for Pacific cod by pot catcher vessels greater than or equal to 60 feet (18.3... allowable catch (TAC) specified for pot catcher vessels greater than or equal to 60 feet (18.3 m) LOA in the...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-02-02
... Vessels Greater Than or Equal to 60 Feet (18.3 Meters) Length Overall Using Pot Gear in the Bering Sea and... directed fishing for Pacific cod by pot catcher vessels greater than or equal to 60 feet (18.3 meters (m...) specified for pot catcher vessels greater than or equal to 60 feet (18.3 m) LOA in the BSAI. DATES...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-05-21
... Vessels Less Than 60 feet (18.3 m) Length Overall Using Hook-and-Line or Pot Gear in the Bering Sea and... directed fishing for Pacific cod by catcher vessels less than 60 feet (18.3 m) length overall (LOA) using... exempt from review under Executive Order 12866. Authority: 16 U.S.C. 1801 et seq. Dated: May 18, 2010...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-01-24
... Catcher Vessels Greater Than or Equal to 60 Feet (18.3 Meters) Length Overall Using Pot Gear in the Bering... prohibiting directed fishing for Pacific cod by pot catcher vessels greater than or equal to 60 feet (18.3... catch (TAC) specified for pot catcher vessels greater than or equal to 60 feet (18.3 m) LOA in the BSAI...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-03-23
.... 111207737-2141-02] RIN 0648-XB112 Pacific Cod by Catcher Vessels Greater Than or Equal to 50 Feet (15.2... catcher vessels (CVs) greater than or equal to 50 feet (15.2 meters (m)) in length overall (LOA) using... to CVs greater than or equal to 50 feet (15.2 m) LOA using hook-and-line gear in the Central...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-03-30
... Vessels Less Than 60 feet (18.3 m) Length Overall Using Hook-and-Line or Pot Gear in the Bering Sea and... directed fishing for Pacific cod by catcher vessels less than 60 feet (18.3 m) length overall (LOA) using... than 60 feet LOA using hook-and-line or pot gear in the BSAI. DATES: Effective 1200 hrs, Alaska local...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-08-31
... using trawl gear and catcher vessels greater than or equal to 60 feet (18.3 meters (m)) length overall (LOA) using hook-and-line gear to catcher vessels less than 60 feet (18.3 m) LOA using hook-and-line or... catcher vessels less than 60 feet (18.3 m) LOA using hook-and-line or pot gear. The 2011 Pacific cod total...
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50 CFR 660.60 - Specifications and management measures.
Code of Federal Regulations, 2011 CFR
2011-10-01
... land more fish than allowed under the new trip limit. This means that, unless otherwise announced in...; Pacific whiting; lingcod; Pacific cod; spiny dogfish; cabezon in Oregon and California and “other fish” as..., California scorpionfish, leopard shark, soupfin shark, finescale codling, Pacific rattail (grenadier...
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75 FR 20566 - North Pacific Fishery Management Council; Public Meeting
Federal Register 2010, 2011, 2012, 2013, 2014
2010-04-20
... review proposals for models to be considered for inclusion in the GOA and BSAI Pacific cod assessments... physically accessible to people with disabilities. Requests for sign language interpretation or other...
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Shibata, Mami; Mekuchi, Miyuki; Mori, Kazuki; Muta, Shigeru; Chowdhury, Vishwajit Sur; Nakamura, Yoji; Ojima, Nobuhiko; Saitoh, Kenji; Kobayashi, Takanori; Wada, Tokio; Inouye, Kiyoshi; Kuhara, Satoru; Tashiro, Kosuke
2016-06-01
Bluefin tuna are high-performance swimmers and top predators in the open ocean. Their swimming is grounded by unique features including an exceptional glycolytic potential in white muscle, which is supported by high enzymatic activities. Here we performed high-throughput RNA sequencing (RNA-Seq) in muscles of the Pacific bluefin tuna (Thunnus orientalis) and Pacific cod (Gadus macrocephalus) and conducted a comparative transcriptomic analysis of genes related to energy production. We found that the total expression of glycolytic genes was much higher in the white muscle of tuna than in the other muscles, and that the expression of only six genes for glycolytic enzymes accounted for 83.4% of the total. These expression patterns were in good agreement with the patterns of enzyme activity previously reported. The findings suggest that the mRNA expression of glycolytic genes may contribute directly to the enzymatic activities in the muscles of tuna.
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-02-26
... Vessels Less Than 60 feet (18.3 m) Length Overall Using Jig or Hook-and-Line Gear in the Bogoslof Pacific... less than 60 feet (18.3 m) length overall (LOA) using jig or hook-and-line gear in the Bogoslof Pacific...
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50 CFR 679.64 - Harvesting sideboard limits in other fisheries.
Code of Federal Regulations, 2010 CFR
2010-10-01
.../processors be calculated? Except for Aleutian Islands pollock and BSAI Pacific cod, the Regional... group in which a TAC is specified for an area or subarea of the BSAI as follows: (i) Aleutian Islands Pacific ocean perch. (A) The Aleutian Islands Pacific ocean perch harvest limit will be equal to the 1996...
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NASA Astrophysics Data System (ADS)
Farley, Edward V.; Heintz, Ron A.; Andrews, Alex G.; Hurst, Thomas P.
2016-12-01
The revised Oscillating Control Hypothesis for the Bering Sea suggests that recruitment of groundfish is linked to climatic processes affecting seasonal sea ice that, in turn, drives the quality and quantity of prey available to young fish for growth and energy storage during their critical life history stages. We test this notion for age-0 (juvenile) Pacific cod (Gadus macrocephalus) by examining the variability in size, diet, and energetic condition during warm (2003-2005), average (2006), and cool (2007-2011) climate states in the eastern Bering Sea. Juvenile cod stomachs contained high proportions of age-0 walleye pollock (by wet weight) during years with warm sea temperatures with a shift to euphausiids and large copepods during years with cool sea temperatures. Juvenile cod were largest during years with warm sea temperatures and smallest during years with cool sea temperatures. However, energetic status (condition) of juvenile cod was highest during years with cool sea temperatures. This result is likely linked to the shift to high quality, lipid-rich prey found in greater abundance on the shelf and in the stomach contents of juvenile cod during cool years. Our examination of juvenile cod size, diet, and energetic status provided results that are similar to those from studies on juvenile pollock, suggesting that the common mechanisms regulating gadid recruitment on the eastern Bering Sea shelf are climate state, prey quality and quantity, and caloric density of gadids prior to winter.
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50 CFR Table 1d to Part 660... - At-Sea Whiting Fishery Annual Set-Asides, 2011
Code of Federal Regulations, 2012 CFR
2012-10-01
... Set-Asides, 2011 Species of species complex Set-aside (mt) Lingcod 6 Pacific Cod 5 Pacific Whiting Allocation a Sablefish N. of 36° 50 Sablefish S. of 36° NA PACIFIC OCEAN PERCH Allocation a WIDOW ROCKFISH... California scorpionfish NA Cabezon (off CA only) NA Other Fish 520 Longnose Skate 5 Pacific Halibut 10 b a...
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50 CFR Table 2d to Part 660... - At-Sea Whiting Fishery Annual Set-Asides, 2012 and beyond
Code of Federal Regulations, 2012 CFR
2012-10-01
... Annual Set-Asides, 2012 and beyond Species or species complex Set-aside (mt) Lingcod 6 Pacific Cod 5 Pacific Whiting Allocation a Sablefish N. of 36° 50 Sablefish S. of 36° NA PACIFIC OCEAN PERCH Allocation... Pacific Halibut 10 b a/ See Table 2.b., to Subpart C, for the at-sea whiting allocations for these species...
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NASA Astrophysics Data System (ADS)
Wang, Xueying; Shi, Xuehui; Liu, Yifan; Yu, Daode; Guan, Shuguang; Liu, Qinghua; Li, Jun
2016-07-01
The present study evaluated the effects of chilled storage and cryopreservation on sperm motion characteristics, antioxidant enzyme activities, and lipid peroxidation in the Pacific cod Gadus macrocephalus. Sperm motility and the activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase (Gr), and lipid peroxidation (measured via malondialdehyde (MDA) content) were determined after the milt was stored at 4°C for 12 h, cryopreserved without cryoprotectant in 12% propylene glycol (PG), cryopreserved in 12% PG+0.1 mol/L trehalose, or cryopreserved in 12% PG spermatozoa but centrifuged to decant the supernatant prior to cryopreservation (only sperm cells were cryopreserved). After chilled storage or cryopreservation, the SOD, CAT and GPx activities were reduced in sperm cells and increased in seminal plasma in almost all treatments; sperm motility parameters were also decreased. However, the addition of trehalose into the cryoprotectant could significantly improve the postthaw sperm quality as revealed by the sperm average path velocity. This improvement might be attributed to the function of trehalose in scavenging reactive oxygen species. Chilled storage and cryopreservation had significant effects on sperm motion characteristics, antioxidant enzyme activities, and lipid peroxidation in the Pacific cod.
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50 CFR Table 11 to Part 679 - BSAI Retainable Percentages
Code of Federal Regulations, 2014 CFR
2014-10-01
... 15 20 2 20 141 Pacific Ocean perch 20 20 20 20 35 35 20 20 20 20 35 15 7 15 20 2 20 152/151... at § 679.2, except for Pacific ocean perch; and northern, shortraker, and rougheye rockfish. 4 The... Squid Aggregatedforage fish 7 Otherspecies 4 110 Pacific cod 20 na 5 20 20 35 35 20 20 20 20 1 1 2 5 20...
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50 CFR Table 11 to Part 679 - BSAI Retainable Percentages
Code of Federal Regulations, 2013 CFR
2013-10-01
... 15 20 2 20 141 Pacific Ocean perch 20 20 20 20 35 35 20 20 20 20 35 15 7 15 20 2 20 152/151... at § 679.2, except for Pacific ocean perch; and northern, shortraker, and rougheye rockfish. 4 The... Squid Aggregatedforage fish 7 Otherspecies 4 110 Pacific cod 20 na 5 20 20 35 35 20 20 20 20 1 1 2 5 20...
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Genetic comparison of viral hemorrhagic septicemia virus isolates from North America and Europe
Oshima, K.H.; Higman, K.H.; Arakawa, C.K.; de Kinkelin, P.; Jorgensen, P.E.V.; Meyers, T.R.; Winton, J.R.
1993-01-01
Viral hemorrhagic septicemia virus (VHSV) is the cdusative agent of a serious rhabdoviral d~sease of rainbow trout Oncorhynchus myklss in Europe The first isolation of the vlrus in North Amenca occurred In the fall of 1988 when it was recovered from adult chinook 0 tshawytscha and coho 0 klsutch salmon returning to 2 hatcher~es in the state of Washington, USA The following year, VHSV was isolated from adult coho salmon at 2 other hatcher~es in northwestern Washington In 1990 and 1991, VHSV was recovered from Pacific cod Gadus macrocephalus caught in Pnnce Willlam Sound, Alaska Genetic vanation among the 4 isolates from salmon and the 1990 ~solate from Pacific cod was determ~ned uslng T1 nbonuclease finqerprlnt~ng In addition, 4 d~verse isolates from Europe were lncluded for companson The North Amencan isolates of VHSV formed a slngle fingerprint group In which the 4 isolates from salmonids were h~ghly similar to each other and the isolate from Pacific cod was related but less s~milar The 4 European ~solates which included an isolate from Atlantic cod G morhua, formed a second fingerpnnt group The genetic d~vers~ty among the isolates within each fingerpnnt group was estimated to be less than 5 % whlle the North Amencan and European strains of the virus were judged to differ by more than 5% The results indicate that the North Amerlcan isolates of VHSV are not of European ongln and that the virus may be enzootic wlthin the manne environment.
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-08-26
... trawl catcher vessels to catcher vessels less than 60 feet (18.3 meters) length overall (LOA) using hook... cod from the C season trawl catcher vessel apportionment to catcher vessels less than 60 feet (18.3... vessels less than 60 feet (18.3 m) LOA using hook-and-line or pot gear. Classification This action...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-03-08
... Catcher Vessels Less Than 60 Feet (18.3 m) Length Overall Using Jig or Hook-and-Line Gear in the Bogoslof... less than 60 feet (18.3 m) length overall (LOA) using jig or hook-and-line gear in the Bogoslof Pacific...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-12-10
... trend for pollock, Pacific cod, deep-water flatfish, Pacific ocean perch, northern rockfish, shortraker..., shallow-water flatfish, rex sole, arrowtooth flounder, flathead sole, rougheye rockfish, demersal shelf... include the TACs for shallow-water flatfish in the West Yakutat and Southeast Outside Districts of the GOA...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-04-18
... DEPARTMENT OF COMMERCE National Oceanic and Atmospheric Administration RIN 0648-XA361 Fisheries of the Exclusive Economic Zone Off Alaska; Catch Accounting in the Longline Catcher/Processor Pacific Cod... longliners in the Bering Sea and Aleutian Islands to support different catch accounting methods for Pacific...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-02-26
... halibut PSC trawl limits between the trawl gear deep-water and the shallow-water species fishery... for pollock, sablefish, deep-water flatfish, rex sole, Pacific ocean perch, northern rockfish... less than the ABCs for Pacific cod, shallow-water flatfish, arrowtooth flounder, flathead sole, ``other...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-07-13
... Fisheries Act (AFA) trawl catcher/processor sector (otherwise known as the Amendment 80 sector... catcher/processors. Hook-and-line catcher/processors are allocated 48.7 percent of the annual BSAI Pacific... harvest of Pacific cod by hook-and-line catcher/processors, although this is one of the major groundfish...
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50 CFR 660.140 - Shorebased IFQ Program.
Code of Federal Regulations, 2010 CFR
2010-10-01
... Groundfish Species 2.7 Lingcod—coastwide 2.5 Pacific Cod 12.0 Pacific whiting (shoreside) 10.0 Sablefish: N....1 CANARY ROCKFISH 4.4 Chilipepper Rockfish 10.0 BOCACCIO 13.2 Splitnose Rockfish 10.0 Yellowtail....0 Minor Rockfish South: Shelf Species 9.0 Slope Species 6.0 Dover sole 2.6 English Sole 5.0 Petrale...
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USDA-ARS?s Scientific Manuscript database
The objectives of this study were to investigate the nutritional value of skate meal (SM) and black cod viscera meal (BCVM) from Alaska and to ascertain their suitability as replacements for commercial pollock fishmeal in diets for Pacific threadfin (Polydactylus sexfilis). Test diets were made by r...
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Code of Federal Regulations, 2011 CFR
2011-10-01
... 90 10 Aleutian Islands Pacific ocean perch 543 All years 98 2 542 2008 95 5 2009 and all future years 90 10 541 2008 95 5 2009 and all future years 90 10 Pacific cod BSAI All years 13.4 N/A Rock sole...
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Code of Federal Regulations, 2013 CFR
2013-10-01
... 90 10 Aleutian Islands Pacific ocean perch 543 All years 98 2 542 2008 95 5 2009 and all future years 90 10 541 2008 95 5 2009 and all future years 90 10 Pacific cod BSAI All years 13.4 N/A Rock sole...
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Code of Federal Regulations, 2012 CFR
2012-10-01
... 90 10 Aleutian Islands Pacific ocean perch 543 All years 98 2 542 2008 95 5 2009 and all future years 90 10 541 2008 95 5 2009 and all future years 90 10 Pacific cod BSAI All years 13.4 N/A Rock sole...
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Code of Federal Regulations, 2014 CFR
2014-10-01
... 90 10 Aleutian Islands Pacific ocean perch 543 All years 98 2 542 2008 95 5 2009 and all future years 90 10 541 2008 95 5 2009 and all future years 90 10 Pacific cod BSAI All years 13.4 N/A Rock sole...
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Viral hemorrhagic septicemia virus in North America
Meyers, Theodore R.; Winton, James R.
1995-01-01
The first detections of viral hemorrhagic septicemia virus (VHSV) in North America were in Washington State from adult coho (Oncorhynchus kisutch) and chinook (O. tshawytscha) salmon in 1988. Subsequently, VHSV was isolated from adult coho salmon returning to hatcheries in the Pacific Northwest in 1989, 1991 and 1994. These isolates represented a strain of VHSV that was genetically different from European VHSV as determined by DNA sequence analysis and T1 ribonuclease fingerprinting. The North American strain of VHSV was also isolated from skin lesions of Pacific cod (Gadus macrocephalus) taken from Prince William Sound (PWS), Alaska in 1990, 1991 and 1993. In 1993 and 1994, the virus was isolated from Pacific herring (Clupea harengus pallasi) in Alaskan waters of PWS, Kodiak Island, Auke Bay and Port Frederick. During 1993 and 1994 the herring fishery in PWS failed from a probable complex of environmental stressors but VHSV isolates were associated with hemorrhages of the skin and fins in fish that returned to spawn. Also in 1993 and 1994, VHSV was isolated from apparently healthy stocks of herring in British Columbia, Canada and Puget Sound, Washington. Thus, the North American strain of VHSV is enzootic in the Northeastern Pacific Ocean among Pacific herring stocks with Pacific cod serving as a secondary reservoir. Although the North American strain of the virus appears to be moderately pathogenic for herring, causing occasional self-limiting epizootics, it was shown to be relatively avirulent for several species of salmonids. Pacific herring are common prey for cod and salmon and were most probably the source of the VHSV isolates from the adult salmon returning to spawn in rivers or at hatcheries in Washington State. Compelling circumstances involving the European isolates of VHSV suggest that this strain of the virus also is enzootic among marine fish in the Atlantic Oean. The highly pathogenic nature of the European strain of VHSV for salmonid fish may be the result of the exposure of rainbow trout (O. mykiss), an introduced species, in a stressful environment of intensive culture and the high rate of mutation inherent in all rhabdoviruses. Consequently, we recommend that efforts be made to eradicate the North American strain of VHSV when detected in live salmonids to reduce the possibility of its evolution into a more virulent salmonid virus.
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Summer distributions of forage fish in the eastern Bering Sea
NASA Astrophysics Data System (ADS)
Parker-Stetter, Sandra L.; Horne, John K.; Farley, Edward V.; Barbee, David H.; Andrews, Alexander G.; Eisner, Lisa B.; Nomura, Jennifer M.
2013-10-01
Juvenile and small adult fish, typically called forage fish, are an important but poorly studied part of the eastern Bering Sea (EBS) ecosystem. Acoustic and trawl data from a non-target survey were used to evaluate distributions of capelin (Mallotus villosus), age-0 Pacific cod (Gadus macrocephalus), and age-0 walleye pollock (Theragra chalcogramma, herein pollock) in 2006-2010. Within the surface zone (15 m to ~35 m, evaluated 2006-2010), capelin occurred throughout the EBS but primarily in the middle shelf. Capelin were also present in the midwater zone (~35 m to 0.5 m off-bottom) in both 2009 and 2010 (evaluated 2009-2010), and resulted in water column zone (15 m to 0.5 m off-bottom) distributions that differed from surface zone characterizations. Age-0 Pacific cod occurred primarily in the surface zone of the middle and outer shelf regions in all years. As midwater and surface zone age-0 Pacific cod were often coincident, water column zone distributions in 2009-2010 were similar to surface zone distributions. Age-0 pollock were found in the EBS surface zone in all years, primarily in the middle and outer shelf regions. High densities of age-0 pollock occurred in the midwater zone in 2006-2007 and 2009-2010. Water column zone distributions of age-0 pollock were similar to surface zone distributions in 2006-2007, but differed in 2009-2010 due to low numbers of age-0 pollock in the surface zone and presence of high densities in the midwater zone of the outer shelf region. While general patterns in capelin distribution in the surface zone were similar between the present and the previous studies, the acoustic-trawl characterization suggested that capelin densities were high in the middle shelf region. As expected, surface zone distributions of age-0 Pacific cod and age-0 pollock were similar to previous characterizations. Observed high densities of midwater age-0 pollock have not been described by previous studies. Annual abundance indices based on bottom or surface trawl data alone will not be sufficient in all years. Data and conclusions from non-target surveys may be constrained compared to dedicated survey efforts, but can provide baseline distributions, potential abundance indices, and insight for planning future research.
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-10-31
... Administrator has also determined that this unharvested amount is unlikely to be harvested through the hierarchy.... Immediate notification is necessary to allow for the orderly conduct and efficient operation of this fishery...
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NASA Astrophysics Data System (ADS)
Miller, Jessica A.; DiMaria, Ruth A.; Hurst, Thomas P.
2016-12-01
Effective and sustainable management depends on knowledge of spawning locations and their relative contributions to marine fish populations. Pacific cod (Gadus macrocephalus) in the southeastern Bering Sea aggregate at discrete spawning locations but there is little information on patterns of larval dispersal and the relative contribution of specific spawning areas to nursery habitats. Age-0 Pacific cod from two cohorts (2006 and 2008) were examined to address the following questions: (1) does size, age, and otolith chemistry vary among known capture locations; (2) can variation in elemental composition of the otolith cores (early larval signatures) be used to infer the number of chemically distinct sources contributing to juvenile recruits in the Bering Sea; and (3) to what extent are juvenile collection locations represented by groups of fish with similar chemical histories throughout their early life history? Hierarchical cluster (HCA) and discriminant function analyses (DFA) were used to examine variation in otolith chemistry at discrete periods throughout the early life history. HCA identified five chemically distinct groups of larvae in the 2006 cohort and three groups in 2008; however, three sources accounted for 80-100% of the juveniles in each year. DFA of early larval signatures indicated that there were non-random spatial distributions of early larvae in both years, which may reflect interannual variation in regional oceanography. There was also a detectable and substantial level of coherence in chemical signatures within groups of fish throughout the early life history. The variation in elemental signatures throughout the early life history (hatch to capture) indicates that otolith chemical analysis could be an effective tool to further clarify larval sources and dispersal, identify juvenile nursery habitats, and estimate the contributions of juvenile nursery habitats to the adult population within the southeastern Bering Sea.
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Yakubu, Abdul-Aziz; Li, Nianpeng; Conrad, Jon M; Zeeman, Mary-Lou
2011-07-01
Overfishing, pollution and other environmental factors have greatly reduced commercially valuable stocks of fish. In a 2006 Science article, a group of ecologists and economists warned that the world may run out of seafood from natural stocks if overfishing continues at current rates. In this paper, we explore the interaction between a constant proportion harvest policy and recruitment dynamics. We examine the discrete-time constant proportion harvest policy discussed in Ang et al. (2009) and then expand the framework to include stock-recruitment functions that are compensatory and overcompensatory, both with and without the Allee effect. We focus on constant proportion policies (CPPs). CPPs have the potential to stabilize complex overcompensatory stock dynamics, with or without the Allee effect, provided the rates of harvest stay below a threshold. If that threshold is exceeded, CPPs are known to result in the sudden collapse of a fish stock when stock recruitment exhibits the Allee effect. In case studies, we analyze CPPs as they might be applied to Gulf of Alaska Pacific halibut fishery and the Georges Bank Atlantic cod fishery based on harvest rates from 1975 to 2007. The best fit models suggest that, under high fishing mortalities, the halibut fishery is vulnerable to sudden population collapse while the cod fishery is vulnerable to steady decline to zero. The models also suggest that CPP with mean harvesting levels from the last 30 years can be effective at preventing collapse in the halibut fishery, but these same policies would lead to steady decline to zero in the Atlantic cod fishery. We observe that the likelihood of collapse in both fisheries increases with increased stochasticity (for example, weather variability) as predicted by models of global climate change. Copyright © 2011 Elsevier Inc. All rights reserved.
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-08-27
... this unharvested amount is unlikely to be harvested through the hierarchy set forth in Sec. 679.20(a)(7... orderly conduct and efficient operation of this fishery, to allow the industry to plan for the fishing...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-09-25
... harvested through the hierarchy set forth in Sec. 679.20(a)(7)(iii)(A). Therefore, in accordance with Sec... operation of this fishery, to allow the industry to plan for the fishing season, and to avoid potential...
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50 CFR Table 28c to Part 679 - Allocation of Rockfish Secondary Species
Code of Federal Regulations, 2013 CFR
2013-10-01
... ZONE OFF ALASKA Pt. 679, Table 28c Table 28c to Part 679—Allocation of Rockfish Secondary Species For... sector . . . Pacific cod 3.81% N/A Sablefish 6.78% 3.51% Rougheye rockfish N/A 58.87% Shortraker rockfish...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-06-15
... Fisheries Service (NMFS), National Oceanic and Atmospheric Administration (NOAA), Commerce. ACTION: Proposed... carry one observer and use a motion-compensated scale to weigh Pacific cod before it is processed. The... and catch reporting requirements that apply to catcher/processors not subject to this proposed action...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-10-17
... published on September 26, 2012, modifying equipment and operational requirements for freezer longliners... Pacific cod at sea with hook-and-line gear in the Bering Sea and Aleutian Islands Management Area (BSAI... modifying equipment and operational requirements for freezer longliners published September 26, 2012, NMFS...
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Katsuki, Kota; Seto, Koji; Noguchi, Takuro; Sonoda, Takeshi; Kim, JuYong
2012-10-01
Diatom fossils from core sediments and living diatoms from water samples of Notoro Lagoon in northern Japan were examined to evaluate natural climate effects on lagoon environmental changes. In 1974, the artificial inlet was excavated. Immediately after, the anoxic bottom water in Notoro Lagoon began to disappear due to an increasing water exchange rate. However, chemical oxygen demand (COD) in the bottom water of Notoro Lagoon gradually increased, with fluctuations, during the last 30 years. In addition, the dominant diatom assemblages in Notoro Lagoon shifted to ice-related and spring bloom taxa after the excavation. The dominant taxa of each year in the sediment core were also strongly related to the timing of lagoon ice melting. This is because the COD in Notoro Lagoon was affected by the deposited volume of blooming diatoms, which was controlled by the duration of ice cover and the timing of ice discharge to the Okhotsk Sea likely due to an air pressure pattern change over the northern North Pacific like the Pacific Decadal Oscillation (PDO). Copyright © 2012 Elsevier Ltd. All rights reserved.
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Energy Content of Arctic Forage Fish
NASA Astrophysics Data System (ADS)
Vollenweider, J.; Heintz, R.; Callahan, M.; Robertson, A.; Barton, M. B.; Sousa, L.; Mueter, F. J.; Mosher, S.; Moran, J.; Logerwell, E.; Boswell, K. M.; Sformo, T.; Farley, E.
2016-02-01
Global changes in climate conditions are most extreme in high latitudes and have the potential to restructure Arctic marine fish assemblages. Of concern is the ability for fish to adapt to changing habitats, the potential for range expansion from lower latitudes, and resultant introduction of competitors and/or predators. Bioenergetic models are useful tools to understand potential cascading trophic effects, and fish energy density is a key parameter in these models. We present energy density (kJ/g dry mass) values for 28 Arctic fish species collected in the Chukchi and Beaufort Seas between 2005 and 2014. Pacific herring (Clupea pallasii) was the most energy-rich species (25.0 kJ/g), followed by two species of primary importance to subsistence people, Arctic cisco (Coregonus autumnalis) and Bering cisco (Coregonus laurettae) (24.9 kJ/g and 23.5 kJ/g, respectively). The abundant and ecologically important Arctic cod (Boreogadus saida) was the most energy-rich gadid, averaging 22.7 kJ/g and having 6% more energy than walleye pollock (Theragra chalcogramma) (21.5 kJ/g) and 11% more energy than saffron cod (Eleginus gracilis) (20.5 kJ/g). In general, pelagic species such as juvenile Pacific salmon (Oncorhynchus sp) had the highest energy content, whereas benthic species such as sculpin (Cottidae), flatfish (Pleuronectidae), eelblennies (Stichaeidae) and eelpouts (Zoarcidae) were amongst the lowest. The high energy content of Arctic cod and their ubiquitous distribution in the water column makes them valuable prey for both demersal and pelagic predators.
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Proximate composition and energy density of some North Pacific forage fishes
van Pelt, Thomas I.; Piatt, John F.; Lance, Brian K.; Roby, Daniel D.
1997-01-01
Mature pelagic forage fish species (capelin, sand lance, squid) had greater lipid concentrations than juvenile age-classes of large demersal and pelagic fish species (walleye pollock, Pacific cod, Atka mackerel, greenling, prowfish, rockfish, sablefish). Myctophids preyed on by puffins have at least twice as much lipid per gram compared to mature capelin, sand lance and squid, and an order of magnitude greater lipid concentrations than juvenile forage fish. Energy density of forage fishes was positively correlated with lipid content, and negatively correlated with water, ash-free lean dry mass (mostly protein), and ash contents.
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-03-28
... mt (1,102,311 lbs) of GOA Pacific cod from 1996 through 2000, and the vessel's catch history... submit confidential business information or otherwise sensitive or protected information. NMFS will... allocation of crab QS was based on a qualifying harvest history in a CR Program fishery. Each QS allocation...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-12-08
... for 2011 and 2012 that are equal to proposed ABCs for pollock, deep-water flatfish, rex sole... certain species: Pacific cod, flathead sole, shallow-water flatfish, arrowtooth flounder, and other... that the ABC is not exceeded. The flathead sole, shallow-water flatfish, and arrowtooth flounder TACs...
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Code of Federal Regulations, 2014 CFR
2014-10-01
... of the ex-vessel production value of all post-reduction Longline Subsector landings, the fee will be... percent of the ex-vessel Pacific cod revenues, a standardized additional fee will be assessed. The... revenue records and NMFS conversion factors for pollock, arrowtooth flounder, Greenland turbot, skate...
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Code of Federal Regulations, 2012 CFR
2012-10-01
... of the ex-vessel production value of all post-reduction Longline Subsector landings, the fee will be... percent of the ex-vessel Pacific cod revenues, a standardized additional fee will be assessed. The... revenue records and NMFS conversion factors for pollock, arrowtooth flounder, Greenland turbot, skate...
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Code of Federal Regulations, 2013 CFR
2013-10-01
... of the ex-vessel production value of all post-reduction Longline Subsector landings, the fee will be... percent of the ex-vessel Pacific cod revenues, a standardized additional fee will be assessed. The... revenue records and NMFS conversion factors for pollock, arrowtooth flounder, Greenland turbot, skate...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-01-03
... to the 2013 Bering Sea and Aleutian Islands Pollock, Atka Mackerel, and Pacific Cod Total Allowable... is adjusting the 2013 total allowable catch (TAC) amounts for the Bering Sea and Aleutian Islands....), December 28, 2012, until the effective date of the final 2013 and 2014 harvest specifications for BSAI...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-01-03
... to the 2013 Gulf of Alaska Pollock and Pacific Cod Total Allowable Catch Amounts AGENCY: National...: Temporary rule; inseason adjustment; request for comments. SUMMARY: NMFS is adjusting the 2013 total... (A.l.t.), December 28, 2012, until the effective date of the final 2013 and 2014 harvest...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-07-26
... resources within State waters, and the Commissioner of the Alaska Department of Fish and Game (ADF&G) opens... and Game (ADF&G) toward a GHL, which limits catch in the fishery in a manner similar to management of... and operation types, based primarily on historical dependency and use by each sector, while also...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-03-01
.... The Council recommended TACs for 2011 and 2012 that are equal to ABCs for pollock, deep-water flatfish... less than the ABCs for Pacific cod, flathead sole, shallow-water flatfish, arrowtooth flounder, other...-water flatfish, and arrowtooth flounder TACs are set to conserve the halibut PSC limit for use in other...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-04-02
... Catcher Vessels Using Trawl Gear in the Central Regulatory Area of the Gulf of Alaska AGENCY: National... (CVs) using trawl gear in the Central Regulatory Area of the Gulf of Alaska (GOA). This action is... apportioned to CVs using trawl gear in the Central Regulatory Area of the GOA. DATES: Effective 1200 hrs...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-03-30
... Vessels Using Trawl Gear in the Bering Sea and Aleutian Islands Management Area AGENCY: National Marine... using trawl gear in the Bering Sea and Aleutian Islands management area (BSAI). This action is necessary... for catcher vessels using trawl gear in the BSAI. DATES: Effective 1200 hrs, Alaska local time (A.l.t...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-02-20
... Catcher Vessels Using Trawl Gear in the Western Regulatory Area of the Gulf of Alaska AGENCY: National... (CVs) using trawl gear in the Western Regulatory Area of the Gulf of Alaska (GOA). This action is... apportioned to CVs using trawl gear in the Western Regulatory Area of the GOA. DATES: Effective 1200 hours...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-04-01
... Vessels Using Trawl Gear in the Bering Sea and Aleutian Islands Management Area AGENCY: National Marine... using trawl gear in the Bering Sea and Aleutian Islands management area (BSAI). This action is necessary... for catcher vessels using trawl gear in the BSAI. DATES: Effective 1200 hrs, Alaska local time (A.l.t...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-03-05
... Catcher Vessels Using Trawl Gear in the Bering Sea and Aleutian Islands Management Area AGENCY: National... using trawl gear in the Bering Sea and Aleutian Islands management area (BSAI). This action is necessary... vessels using trawl gear in the BSAI. After the effective date of this closure the maximum retainable...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-02-27
... Vessels Using Trawl Gear in the Western Regulatory Area of the Gulf of Alaska AGENCY: National Marine... (CVs) using trawl gear in the Western Regulatory Area of the Gulf of Alaska (GOA). This action is... apportioned to CVs using trawl gear in the Western Regulatory Area of the GOA. DATES: Effective 1200 hrs...
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-04-05
... Catcher Vessels Using Trawl Gear in the Bering Sea and Aleutian Islands Management Area AGENCY: National... using trawl gear in the Bering Sea and Aleutian Islands management area (BSAI). This action is necessary... for catcher vessels using trawl gear in the BSAI. DATES: Effective 1200 hrs, Alaska local time (A.l.t...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-03-27
... Catcher Vessels Using Trawl Gear in the Central Regulatory Area of the Gulf of Alaska AGENCY: National... (CVs) using trawl gear in the Central Regulatory Area of the Gulf of Alaska (GOA). This action is... apportioned to CVs using trawl gear in the Central Regulatory Area of the GOA. DATES: Effective 1200 hrs...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-03-16
... Vessels Using Trawl Gear in the Bering Sea and Aleutian Islands Management Area AGENCY: National Marine... using trawl gear in the Bering Sea and Aleutian Islands management area (BSAI). This action is necessary... for catcher vessels using trawl gear in the BSAI. DATES: Effective 1200 hrs, Alaska local time (A.l.t...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-03-12
... Catcher Vessels Using Trawl Gear in the Bering Sea and Aleutian Islands Management Area AGENCY: National... using trawl gear in the Bering Sea and Aleutian Islands management area (BSAI). This action is necessary... catcher vessels using trawl gear in the BSAI. After the effective date of this closure the maximum...
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NASA Astrophysics Data System (ADS)
Holsman, Kirstin K.; Ianelli, James; Aydin, Kerim; Punt, André E.; Moffitt, Elizabeth A.
2016-12-01
Multi-species statistical catch at age models (MSCAA) can quantify interacting effects of climate and fisheries harvest on species populations, and evaluate management trade-offs for fisheries that target several species in a food web. We modified an existing MSCAA model to include temperature-specific growth and predation rates and applied the modified model to three fish species, walleye pollock (Gadus chalcogrammus), Pacific cod (Gadus macrocephalus) and arrowtooth flounder (Atheresthes stomias), from the eastern Bering Sea (USA). We fit the model to data from 1979 through 2012, with and without trophic interactions and temperature effects, and use projections to derive single- and multi-species biological reference points (BRP and MBRP, respectively) for fisheries management. The multi-species model achieved a higher over-all goodness of fit to the data (i.e. lower negative log-likelihood) for pollock and Pacific cod. Variability from water temperature typically resulted in 5-15% changes in spawning, survey, and total biomasses, but did not strongly impact recruitment estimates or mortality. Despite this, inclusion of temperature in projections did have a strong effect on BRPs, including recommended yield, which were higher in single-species models for Pacific cod and arrowtooth flounder that included temperature compared to the same models without temperature effects. While the temperature-driven multi-species model resulted in higher yield MBPRs for arrowtooth flounder than the same model without temperature, we did not observe the same patterns in multi-species models for pollock and Pacific cod, where variability between harvest scenarios and predation greatly exceeded temperature-driven variability in yield MBRPs. Annual predation on juvenile pollock (primarily cannibalism) in the multi-species model was 2-5 times the annual harvest of adult fish in the system, thus predation represents a strong control on population dynamics that exceeds temperature-driven changes to growth and is attenuated through harvest-driven reductions in predator populations. Additionally, although we observed differences in spawning biomasses at the accepted biological catch (ABC) proxy between harvest scenarios and single- and multi-species models, discrepancies in spawning stock biomass estimates did not translate to large differences in yield. We found that multi-species models produced higher estimates of combined yield for aggregate maximum sustainable yield (MSY) targets than single species models, but were more conservative than single-species models when individual MSY targets were used, with the exception of scenarios where minimum biomass thresholds were imposed. Collectively our results suggest that climate and trophic drivers can interact to affect MBRPs, but for prey species with high predation rates, trophic- and management-driven changes may exceed direct effects of temperature on growth and predation. Additionally, MBRPs are not inherently more conservative than single-species BRPs. This framework provides a basis for the application of MSCAA models for tactical ecosystem-based fisheries management decisions under changing climate conditions.
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Abookire, Alisa A.; Piatt, John F.
2005-01-01
Forage fishes were sampled with a mid-water trawl in lower Cook Inlet, Alaska, USA, from late July to early August 1996 to 1999. We sampled 3 oceanographically distinct areas of lower Cook Inlet: waters adjacent to Chisik Island, in Kachemak Bay, and near the Barren Islands. In 163 tows using a mid-water trawl, 229437 fishes with fork length <200 mm were captured. More than 39 species were captured in lower Cook Inlet, but Pacific sand lance Ammodytes hexapterus, juvenile Pacific herring Clupea pallasi, and juvenile walleye pollock Theragra chalcogramma comprised 97.5% of the total individuals. Both species richness and species diversity were highest in warm, low-salinity, weakly stratified waters near Chisik Island. Kachemak Bay, which had thermohaline values between those found near Chisik Island and the Barren Islands, had an intermediate value of species richness. Species richness was lowest at the Barren Islands, an exposed region that regularly receives oceanic, upwelled water from the Gulf of Alaska. Non-metric multidimensional scaling (NMDS) was used to compute axes of species composition based on an ordination of pairwise site dissimilarities. Each axis was strongly rank-correlated with unique groups of species and examined separately as a function of environmental parameters (temperature, salinity, depth), area, and year. Oceanographic parameters accounted for 41 and 12% of the variability among forage fishes indicated by Axis 1 and Axis 2, respectively. Axis 1 also captured the spatial variability in the upwelled area of lower Cook Inlet and essentially contrasted the distribution of species among shallow, nearshore (sand lance, herring) and deep, offshore (walleye pollock) habitats. Axis 2 captured the spatial variability in forage fish communities from the north (Chisik Island) to the south (Barren Islands) of lower Cook Inlet and essentially contrasted a highly diverse community dominated by salmonids and osmerids (warmer, less saline) with a fish community dominated by Pacific sand lance (colder, more saline). Axis 3 reflected the negative spatial association of capelin Mallotus villosus and Pacific cod Gadus macrocephalus. Correlations of year with Axes 1 and 3 indicate that from 1996 to 1999 the forage fish community significantly decreased in lipid-poor gadids (walleye pollock and Pacific cod), and significantly increased in lipid-rich species such as Pacific sand lance, Pacific herring, and capelin.
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Abookire, Alisa A.; Piatt, John F.
2005-01-01
Forage fishes were sampled with a mid-water trawl in lower Cook Inlet, Alaska, USA, from late July to early August 1996 to 1999. We sampled 3 oceanographically distinct areas of lower Cook Inlet: waters adjacent to Chisik Island, in Kachemak Bay, and near the Barren Islands. In 163 tows using a mid-water trawl, 229 437 fishes with fork length < 200 mm were captured. More than 39 species were captured in lower Cook Inlet, but Pacific sand lance Ammodytes hexapterus, juvenile Pacific herring Clupea pallasi, and juvenile walleye pollock Theragra chalcogramma comprised 97.5% of the total individuals. Both species richness and species diversity were highest in warm, low-salinity, weakly stratified waters near Chisik Island. Kachemak Bay, which had thermohaline values between those found near Chisik Island and the Barren Islands, had an intermediate value of species richness. Species richness was lowest at the Barren Islands, an exposed region that regularly receives oceanic, upwelled water from the Gulf of Alaska. Non-metric multidimensional scaling (NMDS) was used to compute axes of species composition based on an ordination of pairwise site dissimilarities. Each axis was strongly rank-correlated with unique groups of species and examined separately as a function of environmental parameters (temperature, salinity, depth), area, and year. Oce??anographie parameters accounted for 41 and 12% of the variability among forage fishes indicated by Axis 1 and Axis 2, respectively. Axis 1 also captured the spatial variability in the upwelled area of lower Cook Inlet and essentially contrasted the distribution of species among shallow, nearshore (sand lance, herring) and deep, offshore (walleye pollock) habitats. Axis 2 captured the spatial variability in forage fish communities from the north (Chisik Island) to the south (Barren Islands) of lower Cook Inlet and essentially contrasted a highly diverse community dominated by salmonids and osmerids (warmer, less saline) with a fish community dominated by Pacific sand lance (colder, more saline). Axis 3 reflected the negative spatial association of capelin Mallotus villosus and Pacific cod Gadus macrocephalus. Correlations of year with Axes 1 and 3 indicate that from 1996 to 1999 the forage fish community significantly decreased in lipid-poor gadids (walleye pollock and Pacific cod), and significantly increased in lipid-rich species such as Pacific sand lance, Pacific herring, and capelin. ?? Inter-Research 2005.
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Federal Register 2010, 2011, 2012, 2013, 2014
2011-05-23
... gear to catcher vessels less than 60 feet (18.3 meters) length overall using hook-and-line or pot gear... catcher vessel apportionment to catcher vessels less than 60 feet length overall (LOA) using hook-and-line... feet LOA using hook-and-line or pot gear. Classification This action responds to the best available...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-12-05
... greater than or equal to 60 feet length overall (LOA) using pot gear to hook-and-line catcher/processors, pot catcher/processors, and catcher vessels less than 60 feet LOA using hook-and-line or pot gear in... total allowable catch (TAC) specified for catcher vessels greater than or equal to 60 feet length...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-11-13
... Vessels Using Jig Gear in the Central Regulatory Area of the Gulf of Alaska AGENCY: National Marine... vessels using jig gear in the Central Regulatory Area of the Gulf of Alaska (GOA). This action is... gear in the Central Regulatory Area of the GOA. DATES: Effective 1200 hrs, Alaska local time (A.l.t...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-01-22
.../Processors Using Trawl Gear in the Western Regulatory Area of the Gulf of Alaska AGENCY: National Marine... (C/Ps) using trawl gear in the Western Regulatory Area of the Gulf of Alaska (GOA). This action is... apportioned to C/Ps using trawl gear in the Western Regulatory Area of the GOA. DATES: Effective 1200 hours...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-02-13
... Vessels Using Pot Gear in the Central Regulatory Area of the Gulf of Alaska AGENCY: National Marine... gear in the Central Regulatory Area of the Gulf of Alaska (GOA). This action is necessary to prevent... pot gear in the Central Regulatory Area of the GOA. DATES: Effective 1200 hours, Alaska local time (A...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-02-01
... Vessels Using Pot Gear in the Western Regulatory Area of the Gulf of Alaska AGENCY: National Marine... using pot gear in the Western Regulatory Area of the Gulf of Alaska (GOA). This action is necessary to... vessels using pot gear in the Western Regulatory Area of the GOA. DATES: Effective 1200 hrs, Alaska local...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-04-23
... Catcher/Processors Using Hook-and-line Gear in the Western Regulatory Area of the Gulf of Alaska AGENCY.../processors (C/Ps) using hook-and-line gear in the Western Regulatory Area of the Gulf of Alaska (GOA). This... catch apportioned to C/Ps using hook-and-line gear in the Western Regulatory Area of the GOA. DATES...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-04-22
... Catcher/Processors Using Trawl Gear in the Central Regulatory Area of the Gulf of Alaska AGENCY: National... (C/Ps) using trawl gear in the Central Regulatory Area of the Gulf of Alaska (GOA). This action is... apportioned to C/Ps using trawl gear in the Central Regulatory Area of the GOA. DATES: Effective 1200 hours...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-02-17
.../Processors Using Trawl Gear in the Western Regulatory Area of the Gulf of Alaska AGENCY: National Marine... (C/Ps) using trawl gear in the Western Regulatory Area of the Gulf of Alaska (GOA). This action is... apportioned to C/Ps using trawl gear in the Western Regulatory Area of the GOA. DATES: Effective 1200 hrs...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-02-28
.../Processors Using Hook-and-Line Gear in the Central Regulatory Area of the Gulf of Alaska AGENCY: National... (C/Ps) using hook-and-line gear in the Central Regulatory Area of the Gulf of Alaska (GOA). This... catch apportioned to C/Ps using hook-and-line gear in the Central Regulatory Area of the GOA. DATES...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-10-30
... Vessels Using Pot Gear in the Central Regulatory Area of the Gulf of Alaska AGENCY: National Marine... vessels using pot gear in the Central Regulatory Area of the Gulf of Alaska (GOA). This action is... gear in the Central Regulatory Area of the GOA. DATES: Effective 1200 hrs, Alaska local time (A.l.t...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-09-05
... Catcher/Processors Using Trawl Gear in the Central Regulatory Area of the Gulf of Alaska AGENCY: National... (C/Ps) using trawl gear in the Central Regulatory Area of the Gulf of Alaska (GOA). This action is... gear in the Central Regulatory Area of the GOA. DATES: Effective 1200 hours, Alaska local time (A.l.t...
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-09-14
... 709 West 9th Street, Room 420A, Juneau, AK. Comments must be submitted by one of the above methods to... the GOA. In 1997, NMFS increased the MRAs for pollock and Pacific cod from zero to 5 percent when... arrowtooth flounder fishery from zero to 20 percent for deep-water flatfish, rex sole, flathead sole, shallow...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-08-29
....S.C. 1801 et seq. Dated: August 26, 2013. Kelly Denit, Acting Deputy Director, Office of Sustainable... Islands Management Area (BSAI). This action is necessary to fully use the 2013 total allowable catch of.... DATES: Effective 1200 hrs, Alaska local time (A.l.t.), August 26, 2013, through 2400 hrs, A.l.t...
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NASA Astrophysics Data System (ADS)
Sreenivasan, Ashwin; Heintz, Ron
2016-10-01
Pacific cod (Gadus macrocephalus) are generalist predators in the Gulf of Alaska (GOA), and are an important predator on other commercially important species. Efficient management of this species can benefit by knowing how these fish adapt to changing environmental conditions, with a focus on how growth and condition are affected by changes in temperature and diet. We conducted a feeding study to understand the relationship between growth, ration, and temperature, and how these factors interact to affect energy allocation strategies. Since growth and condition of juveniles can determine recruitment into the population, this study focused on growth and consumption of age 1+Pacific cod held over 4 temperature treatments (4 °C, 8 °C, 12 °C, and 16 °C) and 3 ration levels (unlimited ration, medium ration, and low ration). We also compared cellular nucleic acid (RNA/DNA) ratios, an instantaneous growth index, total-body lipid, and proximate composition between fish. At 4 °C, 8 °C, and 12 °C, fish at medium and low rations had higher growth rates relative to fish at high rations. Higher food consumption appears to negatively affect digestive ability, assimilation efficiency, and nutrient utilization. RNA/DNA was clearly correlated with growth rates at 4 °C and 8 °C, but this relationship did not hold at higher temperatures. A secondary growth study was conducted to test the reliability of the growth/consumption models derived from the main growth study. Temperature influenced energy reserves (lipid) while tissue growth (protein) was influenced by ration level. Average lipid values were higher at 4 °C than at 8 °C or 12 °C, suggesting a predisposition to heightened lipid synthesis at colder temperatures. Longer durations of warmer water temperature in the GOA could consequently affect energy allocation strategies, with dietary changes in the field potentially amplifying this effect in cold and warm years. This energy allocation strategy could be detrimental with warmer temperatures predicted in the GOA.
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-03-30
... under Executive Order 12866. Authority: 16 U.S.C. 1801 et seq. Dated: March 26, 2012. Emily H. Menashes... the keyword search. Locate the document you wish to comment on from the resulting list and click on... governing fishing by U.S. vessels in accordance with the FMP appear at subpart H of 50 CFR part 600 and 50...
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-11-18
... Catcher Vessels Greater Than or Equal to 60 Feet (18.3 Meters) Length Overall Using Pot Gear in the Bering... to 60 feet (18.3 meters (m)) length overall (LOA) in the Bering Sea and Aleutian Islands management... (TAC) specified for pot catcher vessels greater than or equal to 60 feet (18.3 m) LOA in the BSAI...
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Meyers, T.R.; Short, S.; Lipson, K.; Batts, W.N.; Winton, J.R.; Wilcock, J.; Brown, E.
1994-01-01
Only one-third of the Pacific hernng Clupea harengus pallasi expected to spawn in Prince William Sound (PWS), Alaska, USA, in sprlng 1993 were observed. Of these herring, 15 to 43 '% had external ulcers or subdermal hemorrhages of the skln and fins. A rhabdovirus identified as the North American strain of viral hemorrhagic septicemia virus (VHSV) was isolated from affected herring and 1 Pacific cod Gadus macrocephalus with skin lesions from PWS, and from herring with similar lesions collected near Kodiak Island. No other pathogens were detected in the herring examined. Although VHSV may have been responsible for the skin lesions, there was no confirmed mass herring mortality observed in PWS; hence the actual cause of the reduced herring numbers is still unknown. The same strain of VHSV was subsequently isolated from captive juvenile herring collected from Auke Bay, Alaska, near Juneau, from herring in British Columbia, Canada, and from Puget Sound, Washington, USA. These findings suggest the virus is an opportunistic pathogen that is widely indigenous to Pacific herring populations in the Pacific Northwest and that herring are a significant marine reservoir for North American VHSV.
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-02-14
... gear to catcher vessels less than 60 feet (18.3 meters) length overall using hook-and-line or pot gear... apportionment to catcher vessels less than 60 feet (18.3 meters(m)) length overall (LOA) using hook-and-line or... and 6,445 mt to catcher vessels less than 60 feet (18.3 m) LOA using hook-and-line or pot gear...
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Federal Register 2010, 2011, 2012, 2013, 2014
2013-11-20
...) LOA using hook- and-line or pot gear, American Fisheries Act (AFA) catcher/processors (C/P), Amendment 80 (A80) C/Ps, C/P vessels using pot gear, and C/P vessels using hook-and-line gear in the Bering Sea... meters) LOA using hook-and-line or pot gear, AFA C/P vessels, A80 C/P vessels, C/P vessels using pot gear...
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NASA Astrophysics Data System (ADS)
Pershing, A. J.; Alexander, M. A.; Hernandez, C.; Kerr, L. A.; Le Bris, A.; Mills, K.; Nye, J. A.; Record, N.; Scannell, H. A.; Scott, J. D.; Sherwood, G. D.; Thomas, A. C.
2016-02-01
Climate change is altering conditions in all marine ecosystems, but the pace of change is not uniform. Rapid changes in environmental conditions pose a challenge for resource management, especially when available tools or policies assume the environment is stationary. Between 2004 and 2013, the Gulf of Maine and northwest Atlantic Shelf warmed at a rate that few large marine ecosystems have ever experienced. This warming was associated with a northward shift in the Gulf Stream and with Atlantic Multidecadal Oscillation and Pacific Decadal Oscillation. The unprecedented warming led to reduced recruitment and enhanced mortality of Atlantic cod. Fisheries management has built-in feedbacks designed to reduce quotas as populations decline, but the management process could not keep pace with the rapid temperature-related changes in the Gulf of Maine cod stock. Future recovery of this fishery now depends on both sound management and favorable temperatures. The experience in the Gulf of Maine highlights the need to incorporate environmental factors into resource management and to build resiliency in coupled social-ecological systems. It also highlights a need for scientific and policy guidance for managing species threatened by future warming.
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Wilson, Robert E.; Sage, George K.; Sonsthagen, Sarah A.; Gravley, Megan C.; Menning, Damian; Talbot, Sandra L.
2017-01-01
The Arctic cod (Boreogadus saida) is an abundant marine fish that plays a vital role in the marine food web. To better understand the population genetic structure and the role of natural selection acting on the maternally-inherited mitochondrial genome (mitogenome), a molecule often associated with adaptations to temperature, we analyzed genetic data collected from 11 biparentally-inherited nuclear microsatellite DNA loci and nucleotide sequence data from from the mitochondrial DNA (mtDNA) cytochrome b (cytb) gene and, for a subset of individuals, the entire mitogenome. In addition, due to potential of species misidentification with morphologically similar Polar cod (Arctogadus glacialis), we used ddRAD-Seq data to determine the level of divergence between species and identify species-specific markers. Based on the findings presented here, Arctic cod across the Pacific Arctic (Bering, Chukchi, and Beaufort Seas) comprise a single panmictic population with high genetic diversity compared to other gadids. High genetic diversity was indicated across all 13 protein-coding genes in the mitogenome. In addition, we found moderate levels of genetic diversity in the nuclear microsatellite loci, with highest diversity found in the Chukchi Sea. Our analyses of markers from both marker classes (nuclear microsatellite fragment data and mtDNA cytb sequence data) failed to uncover a signal of microgeographic genetic structure within Arctic cod across the three regions, within the Alaskan Beaufort Sea, or between near-shore or offshore habitats. Further, data from a subset of mitogenomes revealed no genetic differentiation between Bering, Chukchi, and Beaufort seas populations for Arctic cod, Saffron cod (Eleginus gracilis), or Walleye pollock (Gadus chalcogrammus). However, we uncovered significant differences in the distribution of microsatellite alleles between the southern Chukchi and central and eastern Beaufort Sea samples of Arctic cod. Finally, using ddRAD-Seq data, we identified species-specific markers and in conjunction with mitogenome data, identified an Arctic cod x Polar cod hybrid in western Canadian Beaufort Sea. Overall, the lack of genetic structure among Arctic cod within the Bering, Chukchi and Beaufort seas of Alaska is concordant with the absence of geographic barriers to dispersal and typical among marine fishes. Arctic cod may exhibit a genetic pattern of isolation-by-distance, whereby populations in closer geographic proximity are more genetically similar than more distant populations. As this signal is only found between our two fartherest localities, data from populations elsewhere in the species’ global range are needed to determine if this is a general characteristic. Further, tests for selection suggested a limited role for natural selection acting on the mitochondrial genome of Arctic cod, but do not exclude the possibility of selection on genes involved in nuclear-mitogenome interactions. Unlike previous genetic assessment of Arctic cod sampled from the Chukchi Sea, the high levels of genetic diversity found in Arctic cod assayed in this study, across regions, suggests that the species in the Beaufort and Chukchi seas does not suffer from low levels of genetic variation, at least at neutral genetic markers. The large census size of Arctic cod may allow this species to retain high levels of genetic diversity. In addition, we discovered the presence of hybridization between Arctic and Polar cod (although low in frequency). Hybridization is expected to occur when environmental changes modify species distributions that result in contact between species that were previously separated. In such cases, hybridization may be an evolutionary mechanism that promotes an increase in genetic diversity that may provide species occupying changing environments with locally-adapted genotypes and, therefore, phenotypes. Natural selection can only act on the standing genetic variation present within a population. Therefore, given its higher levels of genetic diversity in combination with a large population size, Arctic cod may be resilient to current and future environmental change, as high genetic diversity is expected to increase opportunities for positive selection to act on genetic variants beneficial in different environments, regardless of the source of that genetic variation.
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Aggregation of trypsin and trypsin inhibitor by Al cation.
Chanphai, P; Kreplak, L; Tajmir-Riahi, H A
2017-04-01
Al cation may trigger protein structural changes such as aggregation and fibrillation, causing neurodegenerative diseases. We report the effect of Al cation on the solution structures of trypsin (try) and trypsin inhibitor (tryi), using thermodynamic analysis, UV-Visible, Fourier transform infrared (FTIR) spectroscopic methods and atomic force microscopy (AFM). Thermodynamic parameters showed Al-protein bindings occur via H-bonding and van der Waals contacts for trypsin and trypsin inhibitor. AFM showed that Al cations are able to force trypsin into larger or more robust aggregates than trypsin inhibitor, with trypsin 5±1 SE (n=52) proteins per aggregate and for trypsin inhibitor 8.3±0.7 SE (n=118). Thioflavin T test showed no major protein fibrillation in the presence of Al cation. Al complexation induced more alterations of trypsin inhibitor conformation than trypsin. Copyright © 2017 Elsevier B.V. All rights reserved.
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Federal Register 2010, 2011, 2012, 2013, 2014
2010-03-12
... near future. For 2011, the model produces an ABC 15,000 mt higher than the proposed ABC. The SSC...,110,000 815,000 295,000 AI 19,000 19,000 0 19,000 19,000 0 Bogoslof 50 10 40 50 10 40 Pacific cod BSAI 168,780 193,030 -24,250 207,580 193,030 14,550 Sablefish BS 2,790 2,520 270 2,500 2,520 -20 AI 2,070 2...
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Marcogliese, David J; Jacobson, Kym C
2015-01-01
Parasites have been considered as natural biological tags of marine fish populations in North America for almost 75 years. In the Northwest Atlantic, the most studied species include Atlantic cod (Gadus morhua), Atlantic herring (Clupea harengus) and the redfishes (Sebastes spp.). In the North Pacific, research has centred primarily on salmonids (Oncorhynchus spp.). However, parasites have been applied as tags for numerous other pelagic and demersal species on both the Atlantic and Pacific coasts. Relatively few studies have been undertaken in the Arctic, and these were designed to discriminate anadromous and resident salmonids (Salvelinus spp.). Although rarely applied in fresh waters, parasites have been used to delineate certain fish stocks within the Great Lakes-St Lawrence River basin. Anisakid nematodes and the copepod Sphyrion lumpi frequently prove useful indicators in the Northwest Atlantic, while myxozoan parasites prove very effective on the coast and open seas of the Pacific Ocean. Relative differences in the ability of parasites to discriminate between fish stocks on the Pacific and Atlantic coasts may be due to oceanographic and bathymetric differences between regions. Molecular techniques used to differentiate populations and species of parasites show promise in future applications in the field.
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Johnston, I A; Altringham, J D
1985-09-01
Single fast fibres were isolated from the myotomal muscles of icefish (Chaenocephalus aceratus Lönnberg, Antarctica), North Sea Cod (Gadus morhua L.) and Pacific Blue Marlin (Makaira nigricans Wakiya, Hawaii). Fibres were chemically skinned with the non-ionic detergent Brij-58. Maximum tensions (Po, kN m-2) developed at the characteristic body temperature of each species are 231 for icefish (-1 degree C), 187 for cod (8 degrees C) and 156 for marlin (20 degrees C). At 0 degree C Po is 7 times higher for fibres from the icefish than from the marlin. Fibres from icefish and cod failed to relax completely following activations at temperatures above approximately 12 degrees C. The resultant post-contraction force is associated with a proportional increase in stiffness, suggesting the formation of a population of Ca-insensitive cross bridges. At 10 degrees C there is little interspecific variation in unloaded contraction velocity (Vmax) among the three species. Vmax (muscle lengths s-1) at normal body temperatures are 0.9 for icefish (-1 degree C), 1.0 for cod (8 degrees C) and 3.4 for marlin (20 degrees C). The force-velocity (P-V) relationship becomes progressively more curved with increasing temperature for all three species. Maximum power output for the fast muscle fibres from the Antarctic species at -1 degree C is around 60% of that of the tropical fish at 20 degrees C. Evolutionary temperature compensation of muscle power output appears largely to involve differences in the ability of cross bridges to generate force.
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Machado, Suzy Wider; de Oliveira, Caio Fernando Ramalho; Zério, Neide Graciano; Parra, José Roberto Postali; Macedo, Maria Lígia Rodrigues
2017-08-01
Plant protease inhibitors (PIs) are elements of a common plant defense mechanism induced in response to herbivores. The fall armyworm, Spodoptera frugiperda, a highly polyphagous lepidopteran pest, responds to various PIs in its diet by expressing genes encoding trypsins. This raises the question of whether the PI-induced trypsins are also inhibited by other PIs, which we posed as the hypothesis that Inga laurina trypsin inhibitor (ILTI) inhibits PI-induced trypsins in S. frugiperda. In the process of testing our hypothesis, we compared its properties with those of selected PIs, soybean Kunitz trypsin inhibitor (SKTI), Inga vera trypsin inhibitor (IVTI), Adenanthera pavonina trypsin inhibitor (ApTI), and Entada acaciifolia trypsin inhibitor (EATI). We report that ILTI is more effective in inhibiting the induced S. frugiperda trypsins than SKTI and the other PIs, which supports our hypothesis. ILTI may be more appropriate than SKTI for studies regarding adaptive mechanisms to dietary PIs. © 2017 Wiley Periodicals, Inc.
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Proteolytic enzymes in seawater: contribution of prokaryotes and protists
NASA Astrophysics Data System (ADS)
Obayashi, Y.; Suzuki, S.
2016-02-01
Proteolytic enzyme is one of the major catalysts of microbial processing of organic matter in biogeochemical cycle. Here we summarize some of our studies about proteases in seawater, including 1) distribution of protease activities in coastal and oceanic seawater, 2) responses of microbial community and protease activities in seawater to organic matter amending, and 3) possible contribution of heterotrophic protists besides prokaryotes to proteases in seawater, to clarify cleared facts and remaining questions. Activities of aminopeptidases, trypsin-type and chymotrypsin-type proteases were detected from both coastal and oceanic seawater by using MCA-substrate assay. Significant activities were detected from not only particulate (cell-associated) fraction but also dissolved fraction of seawater, especially for trypsin-type and chymotrypsin-type proteases. Hydrolytic enzymes in seawater have been commonly thought to be mainly derived from heterotrophic prokaryotes; however, it was difficult to determine actual source organisms of dissolved enzymes in natural seawater. Our experiment with addition of dissolved protein to subtropical oligotrophic Pacific water showed drastically enhancement of the protease activities especially aminopeptidases in seawater, and the prokaryotic community structure simultaneously changed to be dominant of Bacteroidetes, indicating that heterotrophic bacteria were actually one of the sources of proteases in seawater. Another microcosm experiment with free-living marine heterotrophic ciliate Paranophrys marina together with an associated bacterium showed that extracellular trypsin-type activity was mainly attributed to the ciliate. The protist seemed to work in organic matter digestion in addition to be a grazer. From the results, we propose a system of organic matter digestion by prokaryotes and protists in aquatic environments, although their actual contribution in natural environments should be estimated in future studies.
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Martínez-Lagos, R; Tovar-Ramírez, D; Gracia-López, V; Lazo, J P
2014-06-01
The leopard grouper is an endemic species of the Mexican Pacific with an important commercial fishery and good aquaculture potential. In order to assess the digestive capacity of this species during the larval period and aid in the formulation of adequate weaning diets, this study aimed to characterize the ontogeny of digestive enzymes during development of the digestive system. Digestive enzymes trypsin, chymotrypsin, acid protease, leucine-alanine peptidase, alkaline phosphatase, aminopeptidase N, lipase, amylase and maltase were quantified in larvae fed live prey and weaned onto a formulated microdiet at 31 days after hatching (DAH) and compared with fasting larvae. Enzyme activity for trypsin, lipase and amylase were detected before the opening of the mouth and the onset of exogenous feeding, indicating a precocious development of the digestive system that has been described in many fish species. The intracellular enzyme activity of leucine-alanine peptidase was high during the first days of development, with a tendency to decrease as larvae developed, reaching undetectable levels at the end of the experimental period. In contrast, activities of enzymes located in the intestinal brush border (i.e., aminopeptidase and alkaline phosphatase) were low at the start of exogenous feeding but progressively increased with larval development, indicating the gradual maturation of the digestive system. Based on our results, we conclude that leopard grouper larvae possess a functional digestive system at hatching and before the onset of exogenous feeding. The significant increase in the activity of trypsin, lipase, amylase and acid protease between 30 and 40 DAH suggests that larvae of this species can be successfully weaned onto microdiets during this period.
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Fernandes, Telmo J R; Costa, Joana; Oliveira, M Beatriz P P; Mafra, Isabel
2017-09-01
This work aimed to exploit the use of DNA mini-barcodes combined with high resolution melting (HRM) for the authentication of gadoid species: Atlantic cod (Gadus morhua), Pacific cod (Gadus macrocephalus), Alaska pollock (Theragra chalcogramma) and saithe (Pollachius virens). Two DNA barcode regions, namely cytochrome c oxidase subunit I (COI) and cytochrome b (cytb), were analysed in silico to identify genetic variability among the four species and used, subsequently, to develop a real-time PCR method coupled with HRM analysis. The cytb mini-barcode enabled best discrimination of the target species with a high level of confidence (99.3%). The approach was applied successfully to identify gadoid species in 30 fish-containing foods, 30% of which were not as declared on the label. Herein, a novel approach for rapid, simple and cost-effective discrimination/clustering, as a tool to authenticate Gadidae fish species, according to their genetic relationship, is proposed. Copyright © 2017 Elsevier Ltd. All rights reserved.
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Larsen, Anett K; Kristiansen, Kurt; Sylte, Ingebrigt; Seternes, Ole-Morten; Bang, Berit E
2013-07-20
Salmon trypsin is shown to increase secretion of the pro-inflammatory cytokine interleukin (IL)-8 from human airway epithelial cells through activation of PAR-2. Secretion of IL-8 induced by king crab trypsin is observed in a different concentration range compared to salmon trypsin, and seems to be only partially related to PAR-2 activation. This report aim to identify differences in the molecular structure of king crab trypsin (Paralithodes camtschaticus) compared to salmon (Salmo salar) and bovine trypsin (Bos taurus) that might influence the ability to activate protease-activated receptor-2 (PAR-2). During purification king crab trypsin displayed stronger binding capacity to the anionic column used in fast protein liquid chromatography compared to fish trypsins, and was identified as a slightly bigger molecule. Measurements of enzymatic activity yielded no obvious differences between the trypsins tested. Molecular modelling showed that king crab trypsin has a large area with strong negative electrostatic potential compared to the smaller negative areas in bovine and salmon trypsins. Bovine and salmon trypsins also displayed areas with strong positive electrostatic potential, a feature lacking in the king crab trypsin. Furthermore we have identified 3 divergent positions (Asp196, Arg244, and Tyr247) located near the substrate binding pocket of king crab trypsin that might affect the binding and cleavage of PAR-2. These preliminary results indicate that electrostatic interactions could be of importance in binding, cleavage and subsequent activation of PAR-2.
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Wang, Yaping; Zhang, Guowen; Wang, Langhong
2015-01-14
Dimethyl phthalate (DMP) is widely used as a plasticizer in industrial processes and has been reported to possess potential toxicity to the human body. In this study, the interaction between DMP and trypsin in vitro was investigated. The results of fluorescence, UV–vis, circular dichroism, and Fourier transform infrared spectra along with cyclic voltammetric measurements indicated that the remarkable fluorescence quenching and conformational changes of trypsin resulted from the formation of a DMP–trypsin complex, which was driven mainly by hydrophobic interactions. The molecular docking and trypsin activity assay showed that DMP primarily interacted with the catalytic triad of trypsin and led to the inhibition of trypsin activity. The dimensions of the individual trypsin molecules were found to become larger after binding with DMP by atomic force microscopy imaging. This study offers a comprehensive picture of DMP–trypsin interaction, which is expected to provide insights into the toxicological effect of DMP.
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Kaur, Gurvir; Tripathi, S K
2015-01-05
The paper presents the interactions between trypsin and water soluble cadmium selenide (CdSe) quantum dots investigated by spectrophotometric methods. CdSe quantum dots have strong ability to quench the intrinsic fluorescence of trypsin by a static quenching mechanism. The quenching has been studied at three different temperatures where the results revealed that electrostatic interactions exist between CdSe quantum dots and trypsin and are responsible to stabilize the complex. The Scatchard plot from quenching revealed 1 binding site for quantum dots by trypsin, the same has been confirmed by making isothermal titrations of quantum dots against trypsin. The distance between donor and acceptor for trypsin-CdSe quantum dot complexes is calculated to be 2.8 nm by energy transfer mechanisms. The intrinsic fluorescence of CdSe quantum dots has also been enhanced by the trypsin, and is linear for concentration of trypsin ranging 1-80 μl. All the observations evidence the formation of trypsin-CdSe quantum dot conjugates, where trypsin retains the enzymatic activity which in turn is temperature and pH dependent. Copyright © 2014 Elsevier B.V. All rights reserved.
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Proteinases during Early Development of the Pacific Whiteleg Shrimp Penaeus vannamei.
Hernandez-Cortes, Patricia; Rivera-Pérez, Crisalejandra; García-Carreño, Fernando; Martínez-Alarcón, Diana
2017-02-01
During shrimp larval development, changes occur in molecular components. Enzyme activity and mRNA expression of proteinases were assayed in Penaeus vannamei during larval development, which consists of 5 nauplius stages, 3 protozoeal stages, 3 mysis stages, and 12 postlarval stages. Trypsin activity reached a maximum at the beginning of postlarval stages 1 and 2, and significantly decreased in subsequent postlarval stages. Chymotrypsin activity increased at the third protozoeal stage, then significantly decreased in subsequent stages. Identification of proteinase by mass spectrometry and inhibitors allowed us to track their appearance in zymograms and to distinguish between isoenzymes. Chymotrypsin BI and BII had a distinguishing pattern of appearance during larval development, which could compensate for the reduction in trypsin activity. The mRNA content of isotrypsin 21, chymotrypsin 1, and zinc proteinase was differentially expressed in larvae. Zinc proteinase and chymotrypsin 1 mRNA were expressed at a basal content at the beginning of the protozoeal stages, increased by the end of the mysis stages and onward, while isotrypsin 21 mRNA had a peak at mysis stage 3. Transcript changes reflect transcriptional regulation of the proteinases tested. Proteinase mRNA in tissues, other than the digestive gland, suggests potentially different roles besides digestion during ontogeny.
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Stimulatory effects of bombesin on plasma trypsin release and exocrine pancreatic secretion in dogs.
Kiriyama, S; Hayakawa, T; Kondo, T; Shibata, T; Kitagawa, M; Sakai, Y; Sobajima, H; Ikei, N; Kodaira, T; Hamaoka, T
1990-01-01
We examined the effect of bombesin on plasma trypsin release and exocrine pancreatic secretion in dogs. Bombesin significantly increased plasma immunoreactive trypsin (IRT). Atropine significantly inhibited the response of plasma IRT to bombesin. Pancreatic trypsin secretion was also increased by bombesin, as well as bicarbonate and protein outputs. Atropine failed to inhibit pancreatic trypsin secretion. In conclusion, bombesin has a stimulatory effect on plasma trypsin release mediated by a cholinergic mechanism and different from pancreatic secretion.
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Fish trypsins: potential applications in biomedicine and prospects for production.
Jesús-de la Cruz, Kristal; Álvarez-González, Carlos Alfonso; Peña, Emyr; Morales-Contreras, José Antonio; Ávila-Fernández, Ángela
2018-04-01
In fishes, trypsins are adapted to different environmental conditions, and the biochemical and kinetic properties of a broad variety of native isoforms have been studied. Proteolytic enzymes remain in high demand in the detergent, food, and feed industries; however, our analysis of the literature showed that, in the last decade, some fish trypsins have been studied for the synthesis of industrial peptides and for specific biomedical uses as antipathogenic agents against viruses and bacteria, which have been recently patented. In addition, innovative strategies of trypsin administration have been studied to ensure that trypsins retain their properties until they exert their action. Biomedical uses require the production of high-quality enzymes. In this context, the production of recombinant trypsins is an alternative. For this purpose, E. coli -based systems have been tested for the production of fish trypsins; however, P. pastoris -based systems also seem to show great potential in the production of fish trypsins with higher production quality. On the other hand, there is a lack of information regarding the specific structures, biochemical and kinetic properties, and characteristics of trypsins produced using heterologous systems. This review describes the potential uses of fish trypsins in biomedicine and the enzymatic and structural properties of native and recombinant fish trypsins obtained to date, outlining some prospects for their study.
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NASA Astrophysics Data System (ADS)
Parker-Stetter, Sandra; Urmy, Samuel; Horne, John; Eisner, Lisa; Farley, Edward
2016-12-01
Hypotheses on the factors affecting forage fish species distributions are often proposed but rarely evaluated using a comprehensive suite of indices. Using 24 predictor indices, we compared competing hypotheses and calculated average models for the distributions of capelin, age-0 Pacific cod, and age-0 pollock in the eastern Bering Sea from 2006 to 2010. Distribution was described using a two stage modeling approach: probability of occurrence ("presence") and density when fish were present. Both local (varying by location and year) and annual (uniform in space but varying by year) indices were evaluated, the latter accounting for the possibility that distributions were random but that overall presence or densities changed with annual conditions. One regional index, distance to the location of preflexion larvae earlier in the year, was evaluated for age-0 pollock. Capelin distributions were best predicted by local indices such as bottom depth, temperature, and salinity. Annual climate (May sea surface temperature (SST), sea ice extent anomaly) and wind (June wind speed cubed) indices were often important for age-0 Pacific cod in addition to local indices (temperature and depth). Surface, midwater, and water column age-0 pollock distributions were best described by a combination of local (depth, temperature, salinity, zooplankton) and annual (May SST, sea ice anomaly, June wind speed cubed) indices. Our results corroborated some of those in previous distribution studies, but suggested that presence and density may also be influenced by other factors. Even though there were common environmental factors that influenced all species' distributions, it is not possible to generalize conditions for forage fish as a group.
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Cheng, Mengxia; Chen, Zilin
2017-08-01
A trypsin immobilized enzyme microreactor was successfully prepared in capillary for studying enzyme kinetics of trypsin and online screening of trypsin inhibitors from traditional Chinese medicine through capillary electrophoresis. Trypsin was immobilized on the inner wall at the inlet of the capillary treated with polydopamine. The rest of the capillary was used as a separation channel. The parameters including the separation efficiency and the activity of immobilized trypsin were comprehensively evaluated. Under the optimal conditions, online screening of trypsin inhibitors each time can be carried out within 6 min. The Michaelis-Menten constant of immobilized trypsin was calculated to be 0.50 mM, which indicated high affinity of the immobilized trypsin for the substrate. The half-maximal inhibitory concentration of known inhibitor of benzamidine hydrochloride hydrate as a model inhibitor was 13.32 mM. The proposed method was successfully applied to screen trypsin inhibitors from 15 compounds of traditional Chinese medicine. It has been found that baicalin showed inhibitory potency. Molecular docking study well supported the experimental result by exhibiting molecular interaction between enzyme and inhibitors. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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Kumari, Rakhi; Gupta, Subodh; Singh, Arvind R; Ferosekhan, S; Kothari, Dushyant C; Pal, Asim Kumar; Jadhao, Sanjay Balkrishna
2013-01-01
Exogenous proteolytic enzyme supplementation is required in certain disease conditions in humans and animals and due to compelling reasons on use of more plant protein ingredients and profitability in animal feed industry. However, limitations on their utility in diet are imposed by their pH specificity, thermolabile nature, inhibition due to a variety of factors and the possibility of intestinal damage. For enhancing the efficacy and safety of exogenous trypsin, an efficient chitosan (0.04%) nanoencapsulation-based controlled delivery system was developed. An experiment was conducted for 45 days to evaluate nanoencapsulated trypsin (0.01% and 0.02%) along with 0.02% bare trypsin and 0.4% chitosan nanoparticles against a control diet on productive efficiency (growth rate, feed conversion and protein efficiency ratio), organo-somatic indices, nutrient digestibility, tissue enzyme activities, hematic parameters and intestinal histology of the fish Labeo rohita. All the synthesized nanoparticles were of desired characteristics. Enhanced fish productive efficiency using nanoencapsulated trypsin over its bare form was noticed, which corresponded with enhanced (P<0.01) nutrient digestibility, activity of intestinal protease, liver and muscle tissue transaminases (alanine and aspartate) and dehydrogenases (lactate and malate), serum blood urea nitrogen and serum protein profile. Intestinal tissues of fish fed with 0.02% bare trypsin showed broadened, marked foamy cells with lipid vacuoles. However, villi were healthier in appearance with improved morphological features in fish fed with nanoencapsulated trypsin than with bare trypsin, and the villi were longer in fish fed with 0.01% nanoencapsulated trypsin than with 0.02% nanoencapsulated trypsin. The result of this premier experiment shows that nanoencapsulated trypsin mimics zymogen-like proteolytic activity via controlled release, and hence the use of 0.01% nanoencapsulated trypsin (in chitosan nanoparticles) over bare trypsin can be favored as a dietary supplement in animals and humans.
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Otsuka, Makoto; Fukui, Yuya; Ozaki, Yukihiro
2009-03-01
The purpose of this study was to evaluate the enzymatic stability of colloidal trypsin powder during heating in a solid-state by using Fourier transform infrared (FT-IR) spectra with chemoinformatics and generalized two-dimensional (2D) correlation spectroscopy. Colloidal crystalline trypsin powders were heated using differential scanning calorimetry. The enzymatic activity of trypsin was assayed by the kinetic degradation method. Spectra of 10 calibration sample sets were recorded three times with a FT-IR spectrometer. The maximum intensity at 1634cm(-1) of FT-IR spectra and enzymatic activity of trypsin decreased as the temperature increased. The FT-IR spectra of trypsin samples were analyzed by a principal component regression analysis (PCR). A plot of the calibration data obtained was made between the actual and predicted trypsin activity based on a two-component model with gamma(2)=0.962. On the other hand, a 2D method was applied to FT-IR spectra of heat-treated trypsin. The result was consistent with that of the chemoinformetrical method. The results for deactivation of colloidal trypsin powder by heat-treatment indicated that nano-structure of crystalline trypsin changed by heating reflecting that the beta-sheet was mainly transformed, since the peak at 1634cm(-1) decreased with dehydration. The FT-IR chemoinformetrical method allows for a solid-state quantitative analysis of the bioactivity of the bulk powder of trypsin during drying.
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Trypsin from the pyloric caeca of bluefish (Pomatomus saltatrix).
Klomklao, Sappasith; Benjakul, Soottawat; Visessanguan, Wonnop; Kishimura, Hideki; Simpson, Benjamin K
2007-12-01
Trypsin was purified from the pyloric caeca of bluefish (Pomatomus saltatrix) by ammonium sulfate precipitation, acetone precipitation and soybean trypsin inhibitor-Sepharose 4B affinity chromatography. Bluefish trypsin migrated as a single band using both sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and native-PAGE and had a molecular mass of 28 kDa. The optima pH and temperature for the hydrolysis of benzoyl-dl-arginine-p-nitroanilide (BAPNA) were 9.5 and 55 degrees C, respectively. The enzyme was stable over a broad pH range (7 to 12), but was unstable at acidic pH, and at temperatures greater than 40 degrees C. The enzyme was inhibited by specific trypsin inhibitors: soybean trypsin inhibitor (SBTI), N-p-tosyl-l-lysine chloromethyl ketone (TLCK) and the serine protease inhibitor phenylmethyl sulfonylfluoride (PMSF). CaCl2 partially protected trypsin against activity loss at 40 degrees C, but NaCl (0 to 30%) decreased the activity in a concentration dependent manner. The N-terminal amino acid sequence of trypsin was determined as IVGGYECKPKSAPVQVSLNL and was highly homologous to other known vertebrate trypsins.
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do Nascimento Campos Ribeiro, Joycellane Alline; Serquiz, Alexandre Coellho; dos Santos Silva, Priscila Fabíola; Barbosa, Patrícia Batista Barra Medeiros; Sampaio, Tarcísio Bruno Montenegro; de Araújo, Raimundo Fernandes; de Oliveira, Adeliana Silva; Machado, Richele Janaina Araújo; Maciel, Bruna Leal Lima; Uchôa, Adriana Ferreira; dos Santos, Elizeu Antunes; de Araújo Morais, Ana Heloneida
2015-01-01
OBJECTIVES: Seeds are excellent sources of proteinase inhibitors, some of which may have satietogenic and slimming actions. We evaluated the effect of a trypsin inhibitor from Tamarindus indica L. seeds on weight gain, food consumption and cholecystokinin levels in Wistar rats. METHODS: A trypsin inhibitor from Tamarindus was isolated using ammonium sulfate (30–60%) following precipitation with acetone and was further isolated with Trypsin-Sepharose affinity chromatography. Analyses were conducted to assess the in vivo digestibility, food intake, body weight evolution and cholecystokinin levels in Wistar rats. Histological analyses of organs and biochemical analyses of sera were performed. RESULTS: The trypsin inhibitor from Tamarindus reduced food consumption, thereby reducing weight gain. The in vivo true digestibility was not significantly different between the control and Tamarindus trypsin inhibitor-treated groups. The trypsin inhibitor from Tamarindus did not cause alterations in biochemical parameters or liver, stomach, intestine or pancreas histology. Rats treated with the trypsin inhibitor showed significantly elevated cholecystokinin levels compared with animals receiving casein or water. CONCLUSION: The results indicate that the isolated trypsin inhibitor from Tamarindus reduces weight gain by reducing food consumption, an effect that may be mediated by increased cholecystokinin. Thus, the potential use of this trypsin inhibitor in obesity prevention and/or treatment should be evaluated. PMID:25789523
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Ribeiro, Joycellane Alline do Nascimento Campos; Serquiz, Alexandre Coellho; Silva, Priscila Fabíola dos Santos; Barbosa, Patrícia Batista Barra Medeiros; Sampaio, Tarcísio Bruno Montenegro; Araújo Junior, Raimundo Fernandes de; Oliveira, Adeliana Silva de; Machado, Richele Janaina Araújo; Maciel, Bruna Leal Lima; Uchôa, Adriana Ferreira; Santos, Elizeu Antunes dos; Morais, Ana Heloneida de Araújo
2015-02-01
Seeds are excellent sources of proteinase inhibitors, some of which may have satietogenic and slimming actions. We evaluated the effect of a trypsin inhibitor from Tamarindus indica L. seeds on weight gain, food consumption and cholecystokinin levels in Wistar rats. A trypsin inhibitor from Tamarindus was isolated using ammonium sulfate (30-60%) following precipitation with acetone and was further isolated with Trypsin-Sepharose affinity chromatography. Analyses were conducted to assess the in vivo digestibility, food intake, body weight evolution and cholecystokinin levels in Wistar rats. Histological analyses of organs and biochemical analyses of sera were performed. The trypsin inhibitor from Tamarindus reduced food consumption, thereby reducing weight gain. The in vivo true digestibility was not significantly different between the control and Tamarindus trypsin inhibitor-treated groups. The trypsin inhibitor from Tamarindus did not cause alterations in biochemical parameters or liver, stomach, intestine or pancreas histology. Rats treated with the trypsin inhibitor showed significantly elevated cholecystokinin levels compared with animals receiving casein or water. The results indicate that the isolated trypsin inhibitor from Tamarindus reduces weight gain by reducing food consumption, an effect that may be mediated by increased cholecystokinin. Thus, the potential use of this trypsin inhibitor in obesity prevention and/or treatment should be evaluated.
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Effects of bisphenol S on the structures and activities of trypsin and pepsin.
Wang, Yan-Qing; Zhang, Hong-Mei
2014-11-19
The effects of bisphenol S on the structures and activities of trypsin and pepsin were investigated by various methods like UV-visible absorbance, fluorescence, circular dichroism, and molecular docking. The secondary and tertiary structures of trypsin and pepsin were altered by bisphenol S binding, which resulted in the loosening of the skeletons of trypsin and pepsin. In addition, bisphenol S induced microenvironmental changes around tyrosine and tryptophan residues of trypsin and pepsin. The activity experimental results showed that the activity of pepsin decreases obviously with the increasing concentration of BPS, while the activity of trypsin does not change remarkably. The binding and thermodynamic parameters obtained by molecular docking and fluorescence spectroscopy showed that the bindings of bisphenol S to trypsin and pepsin were spontaneous processes and hydrogen bonding and hydrophobic interactions played a vital role in stabilizing the bisphenol S-trypsin and bisphenol S-pepsin complexes. The binding constants (K(A)) of bisphenol S with trypsin were 7.42 × 10(4) (298 K) and 5.91 × 10(4) L/mol (310 K), and those of pepsin were 5.78 × 10(4) (298 K) and 4.44 × 10(4) L/mol (310 K). Moreover, there was one main kind of binding site for bisphenol S on trypsin or pepsin.
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Singh, Arvind R.; Ferosekhan, S.; Kothari, Dushyant C.; Pal, Asim Kumar; Jadhao, Sanjay Balkrishna
2013-01-01
Exogenous proteolytic enzyme supplementation is required in certain disease conditions in humans and animals and due to compelling reasons on use of more plant protein ingredients and profitability in animal feed industry. However, limitations on their utility in diet are imposed by their pH specificity, thermolabile nature, inhibition due to a variety of factors and the possibility of intestinal damage. For enhancing the efficacy and safety of exogenous trypsin, an efficient chitosan (0.04%) nanoencapsulation-based controlled delivery system was developed. An experiment was conducted for 45 days to evaluate nanoencapsulated trypsin (0.01% and 0.02%) along with 0.02% bare trypsin and 0.4% chitosan nanoparticles against a control diet on productive efficiency (growth rate, feed conversion and protein efficiency ratio), organo-somatic indices, nutrient digestibility, tissue enzyme activities, hematic parameters and intestinal histology of the fish Labeo rohita. All the synthesized nanoparticles were of desired characteristics. Enhanced fish productive efficiency using nanoencapsulated trypsin over its bare form was noticed, which corresponded with enhanced (P<0.01) nutrient digestibility, activity of intestinal protease, liver and muscle tissue transaminases (alanine and aspartate) and dehydrogenases (lactate and malate), serum blood urea nitrogen and serum protein profile. Intestinal tissues of fish fed with 0.02% bare trypsin showed broadened, marked foamy cells with lipid vacuoles. However, villi were healthier in appearance with improved morphological features in fish fed with nanoencapsulated trypsin than with bare trypsin, and the villi were longer in fish fed with 0.01% nanoencapsulated trypsin than with 0.02% nanoencapsulated trypsin. The result of this premier experiment shows that nanoencapsulated trypsin mimics zymogen-like proteolytic activity via controlled release, and hence the use of 0.01% nanoencapsulated trypsin (in chitosan nanoparticles) over bare trypsin can be favored as a dietary supplement in animals and humans. PMID:24040333
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Interaction of methotrexate with trypsin analyzed by spectroscopic and molecular modeling methods
NASA Astrophysics Data System (ADS)
Wang, Yanqing; Zhang, Hongmei; Cao, Jian; Zhou, Qiuhua
2013-11-01
Trypsin is one of important digestive enzymes that have intimate correlation with human health and illness. In this work, the interaction of trypsin with methotrexate was investigated by spectroscopic and molecular modeling methods. The results revealed that methotrexate could interact with trypsin with about one binding site. Methotrexate molecule could enter into the primary substrate-binding pocket, resulting in inhibition of trypsin activity. Furthermore, the thermodynamic analysis implied that electrostatic force, hydrogen bonding, van der Waals and hydrophobic interactions were the main interactions for stabilizing the trypsin-methotrexate system, which agreed well with the results from the molecular modeling study.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Jun, Seung-Hyun; Chang, Mun Seock; Kim, Byoung Chan
2010-09-15
The construction of a trypsin reactor in a chromatography column for rapid and efficient protein digestion in proteomics is described. Electrospun and alcohol-dispersed polymer nanofibers were used for the fabrication of highly stable trypsin coating, which was prepared by a two-step process of covalent attachment and enzyme crosslinking. In a comparative study with the trypsin coatings on asspun and non-dispersed nanofibers, it has been observed that a simple step of alcohol dispersion improved not only the enzyme loading but also the performance of protein digestion. In-column digestion of enolase was successfully performed in less than twenty minutes. By applying themore » alcohol dispersion of polymer nanofibers, the bypass of samples was reduced by filling up the column with well-dispersed nanofibers, and subsequently, interactions between the protein and the enzymes were improved yielding more complete and reproducible digestions. Regardless of alcohol-dispersion or not, trypsin coating showed better digestion performance and improved performance stability under recycled uses than covalently-attached trypsin. The combination of highly stable trypsin coating and alcoholdispersion of polymer nanofibers has opened up a new potential to develop a trypsin column for on-line and automated protein digestion.« less
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Soares, Tatiane S; Watanabe, Renata M O; Lemos, Francisco J A; Tanaka, Aparecida S
2011-12-10
Trypsin-like enzymes play an important role in the Aedes aegypti digestive process. The trypsin-like enzymes present in adults were characterized previously, but little is known about trypsins in larvae. In the present work, we identified one of the trypsin enzymes from Ae. aegypti larval midgut using a library of trypsin gene fragments, which was the sequence known as AAEL005607 from the Ae. aegypti genome. Quantitative PCR analysis showed that AAEL005607 was transcribed in all larval instars, but it was not present in adult midgut. In order to confirm transcription data, the trypsin-like enzymes from 4th instar larvae of Ae. aegypti midgut were purified and sequenced. Purified trypsin showed identity with the amino-terminal sequence of AAEL005607, AAEL005609 and AAEL005614. These three trypsins have high amino acids identity, and could all be used as a template for the design of inhibitors. In conclusion, for the first time, digestive enzymes of 4th larval instar of Ae. aegypti were purified and characterized. The knowledge of digestive enzymes present in Ae. aegypti larvae may be helpful in the development of a larvicide. Copyright © 2011 Elsevier B.V. All rights reserved.
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Ní Ógáin, Orla; Tajber, Lidia; Corrigan, Owen I; Healy, Anne Marie
2012-09-01
The aim of this study was to determine if spray-drying could successfully produce microparticles containing the model protein trypsin in a form suitable for inhalation. Trypsin was spray-dried with raffinose from a methanol : n-butyl acetate solvent system (MeOH : BA). The solvent system was then adjusted to include water, and trypsin was co-spray-dried with raffinose, trehalose or hydroxpropyl-β-cyclodextrin. The spray-dried products were characterised by SEM, XRD, DSC, TGA and FTIR. Protein biological activity and in-vitro deposition of trypsin : excipient nanoporous/nanoparticulate microparticles (NPMPs) was also assessed. The inclusion of water in a MeOH : BA solvent system allowed for the successful production of NPMPs of trypsin : excipient by spray-drying. Trypsin formulated as trypsin : excipient NPMPs retained biological activity on processing and showed no deterioration in activity or morphological characteristics when stored with desiccant at either 4 or 25°C. Hydroxpropyl-β-cyclodextrin showed advantages over the sugars in terms of producing powders with appropriate density and with greater physical stability under high-humidity conditions. Fine particle fractions of between 41 and 45% were determined for trypsin : excipient NPMPs. NPMPs of trypsin : excipient systems can be produced by spray-drying by adjustment of the solvent system to allow for adequate solubility of trypsin. © 2012 The Authors. JPP © 2012 Royal Pharmaceutical Society.
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A thermostable trypsin inhibitor with antiproliferative activity from small pinto beans.
Chan, Yau Sang; Zhang, Yanbo; Sze, Stephen Cho Wing; Ng, Tzi Bun
2014-08-01
Small pinto bean is a cultivar of Phaseolus vulgaris. It produces a 16-kDa trypsin inhibitor that could be purified using anion exchange and size chromatography. Q-Sepharose, Mono Q and Superdex 75 columns were employed for the isolation process. Small pinto bean trypsin inhibitor demonstrated moderate pH stability (pH 2-10) and marked heat stability, with its trypsin inhibitory activity largely retained after exposure to 100 °C for half an hour. The activity was abolished in the presence of dithiothreitol, in a dose-dependent manner, implying that disulfide bonds in small pinto bean trypsin inhibitor are crucial for the activity. The trypsin inhibitor showed a blocked N-terminus. The trypsin inhibitor only slightly inhibited the viability of breast cancer MCF7 and hepatoma HepG2 cells at 125 μM.
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Bovine Pancreatic Trypsin Inhibitor-Trypsin Complex as a Detection System for Recombinant Proteins
NASA Astrophysics Data System (ADS)
Borjigin, Jimo; Nathans, Jeremy
1993-01-01
Bovine pancreatic trypsin inhibitor (BPTI) binds to trypsin and anhydrotrypsin (an enzymatically inactive derivative of trypsin) with affinities of 6 x 10-14 and 1.1 x 10-13 M, respectively. We have taken advantage of the high affinity and specificity of this binding reaction to develop a protein tagging system in which biotinylated trypsin or biotinylated anhydrotrypsin is used as the reagent to detect recombinant fusion proteins into which BPTI has been inserted. Two proteins, opsin and growth hormone, were used as targets for insertional mutagenesis with BPTI. In each case, both domains of the fusion protein appear to be correctly folded. The fusion proteins can be specifically and efficiently detected by biotinylated trypsin or biotinylated anhydrotrypsin, as demonstrated by staining of transfected cells, protein blotting, affinity purification, and a mobility shift assay in SDS/polyacrylamide gels.
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Johnson, S C; Ewart, K V; Osborne, J A; Delage, D; Ross, N W; Murray, H M
2002-09-01
The salmon louse, Lepeophtheirus salmonis, is a marine ectoparasitic copepod that infects salmonid fishes. We are studying the interactions between this parasite and its salmonid hosts, as it is a common cause of disease in both wild and farmed stocks of salmon. In this paper, we report on the cloning and sequencing of seven trypsin-like enzymes from a cDNA library prepared from whole body preadult female and male L. salmonis. The predicted trypsin activation peptides are 23 or 24 residues in length, considerably longer than previously reported activation peptides of other animals. Differences in the putative signal and activation peptide sequences of the trypsin isoforms suggest that these forms differ in their regulation and function. The calculated molecular weights of the trypsins range from 23.6 to 23.7 kDa. There are eight cysteine residues, which suggest the presence of four disulfide bridges. These trypsins are very similar (>or=46% aa identity) to other crustacean trypsins and insect hypodermins. Using in situ hybridization techniques trypsinogen expression could be identified in all three cell types of the midgut.
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Plasma trypsin in chronic pancreatitis and pancreatic adenocarcinoma.
Adrian, T E; Besterman, H S; Mallinson, C N; Pera, A; Redshaw, M R; Wood, T P; Bloom, S R
1979-10-01
We have used a simple and precise radioimmunoassay to measure trypsin in human plasma. Fasting plasma trypsin concentrations were extremely low in patients with chronic pancreatitis with steatorrhoea (5 +/- 2 ng/ml) when compared to healthy controls (86 +/- 7 ng/ml, p less than 0.001). In patients with chronic pancreatitis but no steatorrhoea basal plasma trypsin levels were similar to those of the normal controls (99 +/- 25 ng/ml). A small but significant postprandial rise in plasma trypsin concentrations was observed in normal subjects (mean increment 15 +/- 4%, p less than 0.005, paired t test) but was absent in patients with chronic pancreatitis with steatorrhoea. In contrast to exocrine deficient chronic pancreatitis, other malabsorptive conditions associated with steatorrhoea (active coeliac disease and acute tropical sprue) demonstrated mean fasting trypsin concentrations similar to controls. Patients with adenocarcinoma of the pancreas had basal trypsin concentrations similar to healthy subjects as did patients with adenocarcinoma of the stomach, colon, rectum, brochus, and breast. In some cases measurement of plasma trypsin may be of help in the differential diagnosis of steatorrhoea.
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Salar, Safoura; Mehrnejad, Faramarz; Sajedi, Reza H; Arough, Javad Mohammadnejad
2017-10-01
Herein, we investigated the effect of the chitosan nanoparticles (CsNP) on the structure, dynamics, and activity of trypsin. The enzyme activity in complex with the nanoparticles slightly increased, which represents the interactions between the nanoparticles and the enzyme. The kinetic parameters of the enzyme, K m and k cat , increased after adding the nanoparticles, resulting in a slight increase in the catalytic efficiency (k cat /K m ). However, the effect of the nanoparticles on the kinetic stability of trypsin has not exhibited significant variations. Fluorescence spectroscopy did not show remarkable changes in the trypsin conformation in the presence of the nanoparticles. The circular dichroism (CD) spectroscopy results also revealed the secondary structure of trypsin attached to the nanoparticles slightly changed. Furthermore, we used molecular dynamics (MD) simulation to find more information about the interaction mechanisms between the nanoparticles and trypsin. The root mean square deviation (RMSD) of Cα atoms results have shown that in the presence of the nanoparticles, trypsin was stable. The simulation and the calculation of the binding free energy demonstrate that the nonpolar interactions are the most important forces for the formation of stable nanoparticle-trypsin complex. This study has explicitly elucidated that the nanoparticles have not considerable effect on the trypsin. Copyright © 2017. Published by Elsevier B.V.
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Manira, Maarof; Khairul Anuar, Khairoji; Seet, Wan Tai; Ahmad Irfan, Abd Wahab; Ng, Min Hwei; Chua, Kien Hui; Mohd Heikal, Mohd Yunus; Aminuddin, Bin Saim; Ruszymah, Bt Hj Idrus
2014-03-01
Animal-derivative free reagents are preferred in skin cell culture for clinical applications. The aim of this study was to compare the performance and effects between animal-derived trypsin and recombinant trypsin for skin cells culture and expansion. Full thickness human skin was digested in 0.6 % collagenase for 6 h to liberate the fibroblasts, followed by treatment with either animal-derived trypsin; Trypsin EDTA (TE) or recombinant trypsin; TrypLE Select (TS) to liberate the keratinocytes. Both keratinocytes and fibroblasts were then culture-expanded until passage 2. Trypsinization for both cell types during culture-expansion was performed using either TE or TS. Total cells yield was determined using a haemocytometer. Expression of collagen type I, collagen type III (Col-III), cytokeratin 10, and cytokeratin 14 genes were quantified via RT-PCR and further confirmed with immunocytochemical staining. The results of our study showed that the total cell yield for both keratinocytes and fibroblasts treated with TE or TS were comparable. RT-PCR showed that expression of skin-specific genes except Col-III was higher in the TS treated group compared to that in the TE group. Expression of proteins specific to the two cell types were confirmed by immunocytochemical staining in both TE and TS groups. In conclusion, the performance of the recombinant trypsin is comparable with the well-established animal-derived trypsin for human skin cell culture expansion in terms of cell yield and expression of specific cellular markers.
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Zamani, Abbas; Benjakul, Soottawat
2016-02-01
Fish proteases, especially trypsin, could be used to prepare fish protein hydrolysates with antioxidative activities. In this study, trypsin from the pyloric caeca of unicorn leatherjacket was purified by ammonium sulfate precipitation and soybean trypsin inhibitor (SBTI)-Sepharose 4B affinity chromatography. Hydrolysate from Indian mackerel protein isolate with different degrees of hydrolysis (20, 30 and 40% DH) was prepared using the purified trypsin, and antioxidative activities (1,1-diphenyl-2-picrylhydrazyl and 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) radical-scavenging activities, ferric-reducing antioxidant power and ferrous-chelating activity) of the hydrolysate were determined. Trypsin was purified 26.43-fold with a yield of 13.43%. The purified trypsin had a molecular weight (MW) of 23.5 kDa and optimal activity at pH 8.0 and 55 °C. It displayed high stability in the pH range of 6.0-11.0 and was thermally stable up to 50 °C. Both SBTI (0.05 mmol L(-1)) and N-p-tosyl-L-lysine-chloromethylketone (5 mmol L(-1)) completely inhibited trypsin activity. Antioxidative activities of the hydrolysate from Indian mackerel protein isolate increased with increasing DH up to 40% (P < 0.05). Based on sodium dodecyl sulfate polyacrylamide gel electrophoresis, the hydrolysate with 40% DH had a MW lower than 6.5 kDa. The purified protease from unicorn leatherjacket pyloric caeca was identified as trypsin based on its ability to hydrolyze a specific synthetic substrate and the response to specific trypsin inhibitors. The purified trypsin could hydrolyze Indian mackerel protein isolate, and the resulting hydrolysate exhibited antioxidative activity depending on its DH. © 2015 Society of Chemical Industry.
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ENSO events in the northern Gulf of Alaska, and effects on selected marine fisheries
Bailey, K.M.; Macklin, S.A.; Reed, R.K.; Brodeur, R.D.; Ingraham, W.J.; Piatt, John F.; Shima, M.; Francis, R.C.; Anderson, P.J.; Royer, T.C.; Hollowed, A.; Somerton, D.A.; Wooster, W.S.
1995-01-01
The 1991-93 El Nino-Southern Oscillation (ENSO) event first appeared in the northern Gulf of Alaska in autumn 1991 with warm sea-surface temperatures. In winter 1992, there were pulses of increased sea level and anomalous circulation. El Nino conditions persisted at least through summer 1993. The effects of this ENSO event on major groundfish species and Pacific herring in the northern Gulf of Alaska were examined and compared with the effects of previous ENSO events. There is little evidence that the 1991-93 or 1982-83 ENSO events affected landings of walleye pollock, Pacific cod, Pacific halibut, or arrowtooth flounder. Some changes in distribution of groundfish species were observed in 1993, but the effect was similar to changes observed in non-ENSO warm years. In general, warm ocean conditions have a positive effect on recruitment of northern stocks, but ENSO events appear to have an inconsistent effect on year-class strength within species and among different species. For example, strong year classes of halibut and arrowtooth flounder sometimes, but not always, coincide with ENSO events; ENSO events are associated with moderate to weak year classes of cod and pollock. However, post-ENSO warm years often are associated with strong recruitment of many groundfish species. Major changes have occurred in the Gulf of Alaska ecosystem since 1977. The influence of the 1976 ENSO event in precipitating these changes and the role of the frequency or strength of subsequent El Nino events is presently unknown. Herring and other stocks of small pelagic fishes may be more affected by ENSO events. In particular, decreased catches, recruitment, and weight-at-age of herring are sometimes associated with ENSO events. Furthermore, a variety of seabirds which feed mostly on pelagic forage fishes or the pelagic juvenile stages of groundfish suffered widespread mortalities and breeding failures in the Gulf of Alaska during the ENSO years of 1983 and 1993. These effects on seabirds were also observed over a wider geographic range, from California to the western Bering Sea.
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Miles, A. Keith; Ricca, Mark A.; Anthony, Robert G.; Estes, James A.
2009-01-01
Organochlorines were examined in liver and stable isotopes in muscle of fishes from the western Aleutian Islands, Alaska, in relation to islands or locations affected by military occupation. Pacific cod (Gadus macrocephalus), Pacific halibut (Hippoglossus stenolepis), and rock greenling (Hexagrammos lagocephalus) were collected from nearshore waters at contemporary (decommissioned) and historical (World War II) military locations, as well as at reference locations. Total (Σ) polychlorinated biphenyls (PCBs) dominated the suite of organochlorine groups (ΣDDTs, Σchlordane cyclodienes, Σother cyclodienes, and Σchlorinated benzenes and cyclohexanes) detected in fishes at all locations, followed by ΣDDTs and Σchlordanes; dichlorodiphenyldi-chloroethylene (p,p′DDE) composed 52 to 66% of ΣDDTs by species. Organochlorine concentrations were higher or similar in cod compared to halibut and lowest in greenling; they were among the highest for fishes in Arctic or near Arctic waters. Organochlorine group concentrations varied among species and locations, but ΣPCB concentrations in all species were consistently higher at military locations than at reference locations. Moreover, all organochlorine group concentrations were higher in halibut from military locations than those from reference locations. A wide range of molecular weight organochlorines was detected at all locations, which implied regional or long-range transport and deposition, as well as local point-source contamination. Furthermore, a preponderance of higher-chlorinated PCB congeners in fishes from contemporary military islands implied recent exposure. Concentrations in all organochlorine groups increased with δ15N enrichment in fishes, and analyses of residual variation provided further evidence of different sources of ΣPCBs and p,p′DDE among species and locations.
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Using Trypsin & Soybean Trypsin Inhibitor to Teach Principles of Enzyme Kinetics
ERIC Educational Resources Information Center
Howard, David R.; Herr, Julie; Hollister, Rhiannon
2006-01-01
Trypsin and soybean trypsin inhibitor (Kunitz inhibitor) can be used in a relatively simple and inexpensive student exercise to demonstrate the usefulness of enzyme kinetics. The study of enzyme kinetics is essential to biology because enzymes play such a crucial role in the biochemical pathways of all living organisms. The data from enzyme…
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In vitro assessment of phthalate acid esters-trypsin complex formation.
Chi, Zhenxing; Zhao, Jing; Li, Weiguo; Araghi, Arash; Tan, Songwen
2017-10-01
In this work, interactions of three phthalate acid esters (PAEs), including dimethyl phthalate (DMP), diethyl phthalate (DEP) and dibutyl phthalate (DBP), with trypsin have been studied in vitro, under simulated physiological conditions using multi-spectroscopic techniques and molecular modeling. The results show that these PAEs can bind to the trypsin, forming trypsin-PAEs complexes, mainly via hydrophobic interactions, with the affinity order of DMP > DEP > DBP. Binding to the PAEs is found to result in molecular deformation of trypsin. The modeling results suggest that only DBP can bind with the amino acid residues of the catalytic triad and S1 binding pocket of trypsin, leading to potential competitive enzyme inhibition. Copyright © 2017 Elsevier Ltd. All rights reserved.
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Melrose, J; Taylor, T K; Ghosh, P
1997-06-01
Trypsin inhibitory proteins of low buoyant density (p < or = 1.35 g/mL) fractions were prepared by CsCl density gradient ultracentrifugation of 4 M guanidinium hydrochloride extracts of lumbar beagle and greyhound annulus fibrosus and nucleus pulposus from animals aged 1 to 6 years. Affinity blotting with biotinylated trypsin was used to identify active trypsin inhibitory protein species; these species were also identified immunologically by Western blotting using antibodies against bovine pancreatic trypsin inhibitor (BPTI), and human inter-alpha-trypsin inhibitor (ITI). None of the trypsin inhibitory species evident in Western blots were reactive with anti-human alpha1-proteinase inhibitor (alpha-1-PI), alpha2-macroglobulin or secretory leucocyte proteinase inhibitor. The greyhound intervertebral disc samples generally had higher levels of active trypsin inhibitor species per unit weight of tissue extracted, and a more extensive range of inhibitor species. Inhibitor species of 30, 32, 34 kDa were identified in both beagle and greyhound intervertebral disc samples; these species were generally most prominent in the annulus fibrosus samples. In contrast, the nucleus pulposus samples contained relatively large trypsin inhibitor species; the anti-BPTI detected an inhibitor species of approximately 85-90 kDa; anti-ITI detected species of 120-250 kDa; biotinylated trypsin detected species of 60-110 kDa. A small molecular mass trypsin inhibitor species of 6 kDa, which was of similar mobility to BPTI, was also detected in annulus fibrosus samples; however, this species did not react with anti-BPTI.
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Macedo, Maria Lígia Rodrigues; Freire, Maria das Graças Machado; Franco, Octávio Luiz; Migliolo, Ludovico; de Oliveira, Caio Fernando Ramalho
2011-02-01
Digestive endoprotease activities of the coconut palm weevil, Homalinotus coriaceus (Coleoptera: Curculionidae), were characterized based on the ability of gut extracts to hydrolyze specific synthetic substrates, optimal pH, and hydrolysis sensitivity to protease inhibitors. Trypsin-like proteinases were major enzymes for H. coriaceus, with minor activity by chymotrypsin proteinases. More importantly, gut proteinases of H. coriaceus were inhibited by trypsin inhibitor from Inga laurina seeds. In addition, a serine proteinase inhibitor from I. laurina seeds demonstrated significant reduction of growth of H. coriaceus larvae after feeding on inhibitor incorporated artificial diets. Dietary utilization experiments show that 0.05% I. laurina trypsin inhibitor, incorporated into an artificial diet, decreases the consumption rate and fecal production of H. coriaceus larvae. Dietary utilization experiments show that 0.05% I. laurina trypsin inhibitor, incorporated into an artificial diet, decreases the consumption rate and fecal production of H. coriaceus larvae. We have constructed a three-dimensional model of the trypsin inhibitor complexed with trypsin. The model was built based on its comparative homology with soybean trypsin inhibitor. Trypsin inhibitor of I. laurina shows structural features characteristic of the Kunitz type trypsin inhibitor. In summary, these findings contribute to the development of biotechnological tools such as transgenic plants with enhanced resistance to insect pests. Copyright © 2010 Elsevier Inc. All rights reserved.
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Wu, Nan; Wang, Siming; Yang, Ye; Song, Jiayi; Su, Ping; Yang, Yi
2018-07-01
A novel type of trypsin capillary microreactor was developed based on a DNA-directed immobilization (DDI) technique applied to a fused-silica capillary modified with polyamidoamine (PAMAM) dendrimers. Trypsin binding to the inner wall of the capillary was confirmed by confocal laser scanning microscopy. The properties of the trypsin-DNA conjugated, PAMAM-modified capillary microreactor were investigated by monitoring hydrolysis of Nα-benzoyl- L -arginine ethyl ester. Through the hybridization and dehybridization of the DNA, the inner wall of the capillary functionalized with trypsin can be regenerated, thus indicating the renewability of this enzyme microreactor. In addition, these results demonstrated that introduction of PAMAM enabled higher amounts of trypsin to be immobilized, markedly improving the enzymolysis efficiency, compared with traditional modified capillaries. The digestion performance of the trypsin capillary microreactor was further evaluated by digesting cytochrome C, and a peptide numbers of 8, and a sequence coverage of 59% were obtained. This renewable and efficient immobilized trypsin capillary microreactor combines advantages of both DDI technology and PAMAM, and is potentially adaptable to high-throughput enzyme assays in biochemical and clinical research. Copyright © 2018. Published by Elsevier B.V.
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Zhao, Mingzhi; Wu, Feilin; Xu, Ping
2015-12-01
Trypsin is one of the most important enzymatic tools in proteomics and biopharmaceutical studies. Here, we describe the complete recombinant expression and purification from a trypsinogen expression vector construct. The Sus scrofa cationic trypsin gene with a propeptide sequence was optimized according to Escherichia coli codon-usage bias and chemically synthesized. The gene was inserted into pET-11c plasmid to yield an expression vector. Using high-density E. coli fed-batch fermentation, trypsinogen was expressed in inclusion bodies at 1.47 g/L. The inclusion body was refolded with a high yield of 36%. The purified trypsinogen was then activated to produce trypsin. To address stability problems, the trypsin thus produced was acetylated. The final product was generated upon gel filtration. The final yield of acetylated trypsin was 182 mg/L from a 5-L fermenter. Our acetylated trypsin product demonstrated higher BAEE activity (30,100 BAEE unit/mg) than a commercial product (9500 BAEE unit/mg, Promega). It also demonstrated resistance to autolysis. This is the first report of production of acetylated recombinant trypsin that is stable and suitable for scale-up. Copyright © 2015 Elsevier Inc. All rights reserved.
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Macedo, Maria Lígia R; Diz Filho, Eduardo B S; Freire, Mariadas Graças M; Oliva, Maria Luiza V; Sumikawa, Joana T; Toyama, Marcos H; Marangoni, Sérgio
2011-01-01
The present paper describes the purification, characterization and determination of the partial primary structure of the first trypsin inhibitor isolated from the family Sapindaceae. A highly stable, potent trypsin inhibitor (SSTI) was purified to homogeneity. SDS-PAGE analysis revealed that the protein consists of a two-polypeptide chain with molecular masses of approximately 15 and 3 kDa. The purified inhibitor inhibited bovine trypsin at a 1:1 M ratio. Kinetic analysis revealed that the protein is a competitive inhibitor with an equilibrium dissociation constant of 10⁻⁹ M for trypsin. The partial NH₂- terminal sequence of 36 amino acids in SSTI indicates homology with other members of the trypsin-inhibitor family from different sources. This inhibitor is highly stable in the presence of denaturing agents. SSTI showed significant inhibitory activity against trypsin-like proteases present in the larval midgut on Anagasta kuehniella, Corcyra cephalonica, Diatreae saccharalis and Anticarsia gemmatalis.
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The sequence and X-ray structure of the trypsin from Fusarium oxysporum.
Rypniewski, W R; Hastrup, S; Betzel, C; Dauter, M; Dauter, Z; Papendorf, G; Branner, S; Wilson, K S
1993-06-01
The trypsin from Fusarium oxysporum is equally homologous to trypsins from Streptomyces griseus, Streptomyces erythraeus and to bovine trypsin. A DFP (diisopropylfluorophosphate) inhibited form of the enzyme has been crystallized from 1.4 M Na2SO4, buffered with citrate at pH 5.0-5.5. The crystals belong to space group P2(1) with cell parameters a = 33.43 A, b = 67.65 A, c = 39.85 A and beta = 107.6 degrees. There is one protein molecule in the asymmetric unit. X-ray diffraction data to a resolution of 1.8 A were collected on film using synchrotron radiation. The structure was solved by molecular replacement using models of bovine and S. griseus trypsins and refined to an R-factor of 0.141. The overall fold is similar to other trypsins, with some insertions and deletions. There is no evidence of the divalent cation binding sites seen in other trypsins. The covalently bound inhibitor molecule is clearly visible.
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Mechanism of cinnamic acid-induced trypsin inhibition: A multi-technique approach
NASA Astrophysics Data System (ADS)
Zhang, Hongmei; Zhou, Qiuhua; Cao, Jian; Wang, Yanqing
2013-12-01
In order to investigate the association of the protease trypsin with cinnamic acid, the interaction was characterized by using fluorescence, UV-vis absorption spectroscopy, molecular modeling and an enzymatic inhibition assay. The binding process may be outlined as follows: cinnamic acid can interact with trypsin with one binding site to form cinnamic acid-trypsin complex, resulting in inhibition of trypsin activity; the spectroscopic data show that the interaction is a spontaneous process with the estimated enthalpy and entropy changes being -8.95 kJ mol-1 and 50.70 J mol-1 K-1, respectively. Noncovalent interactions make the main contribution to stabilize the trypsin-cinnamic acid complex; cinnamic acid can enter into the primary substrate-binding pocket and alter the environment around Trp and Tyr residues.
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Lyu, Junchen; Liu, Yuan; An, Tianchen; Liu, Yujun; Wang, Manchuriga; Song, Yanting; Zheng, Feifei; Wu, Dan; Zhang, Yingxia; Deng, Shiming
2015-05-01
A proteinaceous inhibitor against trypsin was isolated from the seeds of Artocarpus heterophyllus Lam. by successive ammonium sulfate precipitation, ion-exchange, and gel-filtration chromatography. The trypsin inhibitor, named as AHLTI (A. heterophyllus Lam. trypsin inhibitor), consisted of a single polypeptide chain with a molecular weight of 28.5 kDa, which was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel-filtration chromatography. The N-terminal sequence of AHLTI was DEPPSELDAS, which showed no similarity to other known trypsin inhibitor sequence. AHLTI completely inhibited bovine trypsin at a molar ratio of 1:2 (AHLTI:trypsin) analyzed by native polyacrylamide gel electrophoresis, inhibition activity assay, and gel-filtration chromatography. Moreover, kinetic enzymatic studies were carried out to understand the inhibition mechanism of AHLTI against trypsin. Results showed that AHLTI was a competitive inhibitor with an equilibrium dissociation constant (Ki) of 3.7 × 10(-8) M. However, AHLTI showed weak inhibitory activity toward chymotrypsin and elastase. AHLTI was stable over a broad range of pH 4-8 and temperature 20-80°C. The reduction agent, dithiothreitol, had no obvious effect on AHLTI. The trypsin inhibition assays of AHLTI toward digestive enzymes from insect pest guts in vitro demonstrated that AHLTI was effective against enzymes from Locusta migratoria manilensis (Meyen). These results suggested that AHLTI might be a novel trypsin inhibitor from A. heterophyllus Lam. belonging to Kunitz family, and play an important role in protecting from insect pest. © The Author 2015. Published by ABBS Editorial Office in association with Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences.
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Mechanism of cinnamic acid-induced trypsin inhibition: a multi-technique approach.
Zhang, Hongmei; Zhou, Qiuhua; Cao, Jian; Wang, Yanqing
2013-12-01
In order to investigate the association of the protease trypsin with cinnamic acid, the interaction was characterized by using fluorescence, UV-vis absorption spectroscopy, molecular modeling and an enzymatic inhibition assay. The binding process may be outlined as follows: cinnamic acid can interact with trypsin with one binding site to form cinnamic acid-trypsin complex, resulting in inhibition of trypsin activity; the spectroscopic data show that the interaction is a spontaneous process with the estimated enthalpy and entropy changes being -8.95 kJ mol(-1) and 50.70 J mol(-1) K(-1), respectively. Noncovalent interactions make the main contribution to stabilize the trypsin-cinnamic acid complex; cinnamic acid can enter into the primary substrate-binding pocket and alter the environment around Trp and Tyr residues. Copyright © 2013 Elsevier B.V. All rights reserved.
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Modifications in trypsin digestion protocol for increasing the efficiency and coverage.
Syal, Kirtimaan; Tadala, Raghu
2015-01-01
Standard trypsin digestion protocol of proteins followed by MALDI-MS analysis has been realized as an important tool for the identification and characterization of proteins. In this article, we proposed the elimination of the step of 'staining/de-staining of gel pieces' in in-gel digestion protocol in order to improve the efficiency of trypsin digestion. Coomassie dye is known to interfere with digestion of proteins by trypsin and the procedure of staining-de-staining could result in loss of photoaffinity probe, post translational modifications and catalytic activities of enzymes. Further, we studied parameters like hydrophobicity and isoelectric point, and attempted to quantitatively relate it to the efficiency of trypsin digestion. We suggest that properties of proteins should be considered and trypsin digestion protocol should be appropriately modified as per sequence and other information.
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de Jong, A J; Klamer, M; Lamers, C B
1987-01-01
This study was undertaken to determine the effect of atropine and somatostatin, two inhibitors of intraduodenal pancreatic enzyme secretion, on bombesin-stimulated release of plasma immunoreactive trypsin in 6 healthy volunteers. Infusion of 5 ng/kg.min bombesin during 30 min induced significant increases in plasma trypsin from 206 +/- 20 to 334 +/- 44 ng/ml (p less than 0.01). Atropine (15 ng/kg as i.v. bolus followed by 5 ng/kg.h) had no influence on the bombesin-stimulated increase in plasma immunoreactive trypsin (207 +/- 20 to 326 +/- 54 ng/ml). Somatostatin (125 micrograms as i.v. bolus followed by 125 micrograms/h) also failed to inhibit the plasma trypsin response to bombesin (207 +/- 18 to 663 +/- 166 ng/ml). These results point to major differences in the regulation of plasma and intraduodenal trypsin secretion.
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Pontual, Emmanuel Viana; Napoleão, Thiago Henrique; Dias de Assis, Caio Rodrigo; de Souza Bezerra, Ranilson; Xavier, Haroudo Satiro; Navarro, Daniela Maria do Amaral Ferraz; Coelho, Luana Cassandra Breitenbach Barroso; Paiva, Patrícia Maria Guedes
2012-03-01
Aedes aegypti control is crucial to reducing dengue fever. Aedes aegypti larvae have developed resistance to organophosporous insecticides and the use of natural larvicides may help manage larval resistance by increasing elements in insecticide rotation programs. Here, we report on larvicidal activity of Moringa oleifera flower extract against A. aegypti L(1), L(2), L(3), and L(4) as well as the effect of flower extract on gut trypsin and whole-larval acetylcholinesterase from L(4.) In addition, the heated flower extract was investigated for larvicidal activity against L(4) and effect on larval gut trypsin. Moringa oleifera flower extract contains a proteinaceous trypsin inhibitor (M. oleifera flower trypsin inhibitor, MoFTI), triterpene (β-amyrin), sterol (β-sitosterol) as well as flavonoids (kaempferol and quercetin). Larvicidal activity was detected against L(2), L(3), and L(4) (LC(50) of 1.72%, 1.67%, and 0.92%, respectively). Flower extract inhibited L(4) gut trypsin (MoFTI K(i) = 0.6 nM) and did not affect acetylcholinesterase activity. In vivo assay showed that gut trypsin activity from L(4) treated with M. oleifera flower extract decreased over time (0-1,440 min) and was strongly inhibited (98.6%) after 310 min incubation; acetylcholinesterase activity was not affected. Thermal treatment resulted in a loss of trypsin inhibitor and larvicidal activities, supporting the hypothesis that flower extract contains a proteinaceous trypsin inhibitor that may be responsible for the deleterious effects on larval mortality. © 2012 Wiley Periodicals, Inc.
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NASA Astrophysics Data System (ADS)
Song, Wei; Yu, Zehua; Hu, Xinxin; Liu, Rutao
2015-02-01
Studies on the effects of environmental pollutants to protein in vitro has become a global attention. Hydrogen peroxide (H2O2) is used as an effective food preservative and bleacher in industrial production. The toxicity of H2O2 to trypsin was investigated by multiple spectroscopic techniques and the molecular docking method at the molecular level. The intrinsic fluorescence of trypsin was proved to be quenched in a static process based on the results of fluorescence lifetime experiment. Hydrogen bonds interaction and van der Waals forces were the main force to generate the trypsin-H2O2 complex on account of the negative ΔH0 and ΔS0. The binding of H2O2 changed the conformational structures and internal microenvironment of trypsin illustrated by UV-vis absorption, fluorescence, synchronous fluorescence, three-dimensional (3D) fluorescence and circular dichroism (CD) results. However, the binding site was far away from the active site of trypsin and the trypsin activity was only slightly affected by H2O2, which was further explained by molecular docking investigations.
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Investigation on potential enzyme toxicity of clenbuterol to trypsin
NASA Astrophysics Data System (ADS)
Chai, Jun; Xu, Qifei; Dai, Jinping; Liu, Rutao
2013-03-01
Clenbuterol (CLB) is a kind of β2-adrenergic agonists which was illegally used as feed additives nowadays. The toxic interaction of CLB with trypsin, an important digestive enzyme, was studied in vitro using multi-spectroscopic methods and molecular modeling methods. CLB was proved to bind with trypsin in S1 pocket, forming a complex driven by the dominant force of H-bond. The binding constant was calculated to be 1.79887 × 105 L mol-1 at 289 K and 0.32584 × 105 L mol-1 at 310 K, respectively. The skeleton of trypsin became loosened and unfolded with the amino residues microenvironment changed. The secondary and tertiary structure of trypsin also varied. Molecular modeling studies illustrated specific display of the binding information and explained most of the experiment phenomena. The binding site of CLB induced the fluorescence quenching as well as inhibition of enzyme activity of trypsin. The study confirmed that CLB had potential toxicity on both the structure and function of trypsin and the effects enhanced with the increasing concentration of CLB.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Page, Michael J.; Carrell, Christopher J.; Di Cera, Enrico
2008-05-28
Some trypsin-like proteases are endowed with Na{sup +}-dependent allosteric enhancement of catalytic activity, but this important mechanism has been difficult to engineer in other members of the family. Replacement of 19 amino acids in Streptomyces griseus trypsin targeting the active site and the Na{sup +}-binding site were found necessary to generate efficient Na{sup +} activation. Remarkably, this property was linked to the acquisition of a new substrate selectivity profile similar to that of factor Xa, a Na{sup -} activated protease involved in blood coagulation. The X-ray crystal structure of the mutant trypsin solved to 1.05 {angstrom} resolution defines the engineeredmore » Na{sup +} site and active site loops in unprecedented detail. The results demonstrate that trypsin can be engineered into an efficient allosteric protease, and that Na+ activation is interwoven with substrate selectivity in the trypsin scaffold.« less
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Xu, Xiao-yan; Nie, Xiao-cui; Ma, Hai-ying; Song, Guo-qing; Zhang, Xiao-tong; Jin, Yu-nan; Yu, Yan-qiu
2015-04-01
Flow cytometry method (FCM) is a generally accepted tool to analyze apoptosis. Although apoptosis assay kit was applied by many companies, the manufacturers were not consistent with whether using Trypsin with EDTA to collect the adherent cells. In another words, the influence of EDTA on apoptotic ratio is not clear. In this work, we compared the proportion of apoptotic cells with EDTA or EDTA-free Trypsin treatment by FCM. We concluded that Trypsin with or without EDTA has little influence on the proportion of apoptotic cells. In addition, we found that the ratio of necrosis and apoptosis was different in cells collected by scraping. WAVE2 protein was analyzed as a typical example for movement related protein. WAVE2 expression is elevated in the EDTA Trypsin treated group, compared with EDTA-free Trypsin treatment and scrapping group. © The Author(s) 2014.
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Mercury in tunas and blue marlin in the North Pacific Ocean.
Drevnick, Paul E; Brooks, Barbara A
2017-05-01
Models and data from the North Pacific Ocean indicate that mercury concentrations in water and biota are increasing in response to (global or hemispheric) anthropogenic mercury releases. In the present study, we provide an updated record of mercury in yellowfin tuna (Thunnus albacares) caught near Hawaii that confirms an earlier conclusion that mercury concentrations in these fish are increasing at a rate similar to that observed in waters shallower than 1000 m. We also compiled and reanalyzed data from bigeye tuna (Thunnus obesus) and blue marlin (Makaira nigricans) caught near Hawaii in the 1970s and 2000s. Increases in mercury concentrations in bigeye tuna are consistent with the trend found in yellowfin tuna, in both timing and magnitude. The data available for blue marlin do not allow for a fair comparison among years, because mercury concentrations differ between sexes for this species, and sex was identified (or reported) in only 3 of 7 studies. Also, mercury concentrations in blue marlin may be insensitive to modest changes in mercury exposure, because this species appears to have the ability to detoxify mercury. The North Pacific Ocean is a region of both relatively high rates of atmospheric mercury deposition and capture fisheries production. Other data sets that allow temporal comparisons in mercury concentrations, such as pacific cod (Gadus macrocephalus) in Alaskan waters and albacore tuna (Thunnus alalunga) off the US Pacific coast, should be explored further, to aid in understanding human health and ecological risks and to develop additional baseline knowledge for assessing changes in a region expected to respond strongly to reductions in anthropogenic mercury emissions. Environ Toxicol Chem 2017;36:1365-1374. © 2017 SETAC. © 2017 SETAC.
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Bassan, Juliana Cristina; de Souza Bezerra, Thaís Milena; Peixoto, Guilherme; da Cruz, Clariana Zanutto Paulino; Galán, Julián Paul Martínez; Vaz, Aline Buda dos Santos; Garrido, Saulo Santesso; Filice, Marco; Monti, Rubens
2016-01-01
In this study, trypsin (Enzyme Comission 3.4.21.4) was immobilized in a low cost, lignocellulosic support (corn cob powder—CCP) with the goal of obtaining peptides with bioactive potential from cheese whey. The pretreated support was activated with glyoxyl groups, glutaraldehyde and IDA-glyoxyl. The immobilization yields of the derivatives were higher than 83%, and the retention of catalytic activity was higher than 74%. The trypsin-glyoxyl-CCP derivative was thermally stable at 65 °C, a value that was 1090-fold higher than that obtained with the free enzyme. The trypsin-IDA-glyoxyl-CCP and trypsin-glutaraldehyde-CCP derivatives had thermal stabilities that were 883- and five-fold higher, respectively, then those obtained with the free enzyme. In the batch experiments, trypsin-IDA-glyoxyl-CCP retained 91% of its activity and had a degree of hydrolysis of 12.49%, while the values for trypsin-glyoxyl-CCP were 87% and 15.46%, respectively. The stabilized derivative trypsin-glyoxyl-CCP was also tested in an upflow packed-bed reactor. The hydrodynamic characterization of this reactor was a plug flow pattern, and the kinetics of this system provided a relative activity of 3.04 ± 0.01 U·g−1 and an average degree of hydrolysis of 23%, which were suitable for the production of potentially bioactive peptides. PMID:28773482
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A low-cost method for visible fluorescence imaging.
Tarver, Crissy L; Pusey, Marc
2017-12-01
A wide variety of crystallization solutions are screened to establish conditions that promote the growth of a diffraction-quality crystal. Screening these conditions requires the assessment of many crystallization plates for the presence of crystals. Automated systems for screening and imaging are very expensive. A simple approach to imaging trace fluorescently labeled protein crystals in crystallization plates has been devised, and can be implemented at a cost as low as $50. The proteins β-lactoglobulin B, trypsin and purified concanavalin A (ConA) were trace fluorescently labeled using three different fluorescent probes: Cascade Yellow (CY), Carboxyrhodamine 6G (CR) and Pacific Blue (PB). A crystallization screening plate was set up using β-lactoglobulin B labeled with CR, trypsin labeled with CY, ConA labeled with each probe, and a mixture consisting of 50% PB-labeled ConA and 50% CR-labeled ConA. The wells of these plates were imaged using a commercially available macro-imaging lens attachment for smart devices that have a camera. Several types of macro lens attachments were tested with smartphones and tablets. Images with the highest quality were obtained with an iPhone 6S and an AUKEY Ora 10× macro lens. Depending upon the fluorescent probe employed and its Stokes shift, a light-emitting diode or a laser diode was used for excitation. An emission filter was used for the imaging of protein crystals labeled with CR and crystals with two-color fluorescence. This approach can also be used with microscopy systems commonly used to observe crystallization plates.
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França, Renata Cristina da Penha; Assis, Caio Rodrigo Dias; Santos, Juliana Ferreira; Torquato, Ricardo José Soares; Tanaka, Aparecida Sadae; Hirata, Izaura Yoshico; Assis, Diego Magno; Juliano, Maria Aparecida; Cavalli, Ronaldo Olivera; Carvalho, Luiz Bezerra de; Bezerra, Ranilson Souza
2016-10-15
A thermostable alkaline peptidase was purified from the processing waste of cobia (Rachycentron canadum) using bovine pancreatic trypsin inhibitor (BPTI) immobilized onto Sepharose. The purified enzyme had an apparent molecular mass of 24kDa by both sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrometry. Its optimal temperature and pH were 50°C and 8.5, respectively. The enzyme was thermostable until 55°C and its activity was strongly inhibited by the classic trypsin inhibitors N-ρ-tosyl-l-lysine chloromethyl ketone (TLCK) and benzamidine. BPTI column allowed at least 15 assays without loss of efficacy. The purified enzyme was identified as a trypsin and the N-terminal amino acid sequence of this trypsin was IVGGYECTPHSQAHQVSLNSGYHFC, which was highly homologous to trypsin from cold water fish species. Using Nα-benzoyl-dl-arginine ρ-nitroanilide hydrochloride (BApNA) as substrate, the apparent km value of the purified trypsin was 0.38mM, kcat value was 3.14s(-1), and kcat/km was 8.26s(-1)mM(-1). The catalytic proficiency of the purified enzyme was 2.75×10(12)M(-1) showing higher affinity for the substrate at the transition state than other fish trypsin. The activation energy (AE) of the BApNA hydrolysis catalyzed by this enzyme was estimated to be 11.93kcalmol(-1) while the resulting rate enhancement of this reaction was found to be approximately in a range from 10(9) to 10(10)-fold evidencing its efficiency in comparison to other trypsin. This new purification strategy showed to be appropriate to obtain an alkaline peptidase from cobia processing waste with high purification degree. According with N-terminal homology and kinetic parameters, R. canadum trypsin may gathers desirable properties of psychrophilic and thermostable enzymes. Copyright © 2016 Elsevier B.V. All rights reserved.
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COTTRELL, GRAEME S.; AMADESI, SILVIA; PIKIOS, STELLA; CAMERER, ERIC; WILLARDSEN, J. ADAM; MURPHY, BRETT R.; CAUGHEY, GEORGE H.; WOLTERS, PAUL J.; COUGHLIN, SHAUN R.; PETERSON, ANDERS; KNECHT, WOLFGANG; POTHOULAKIS, CHARALABOS; BUNNETT, NIGEL W.; GRADY, EILEEN F.
2008-01-01
Background & Aims We studied the role of protease-activated receptor 2 (PAR2) and its activating enzymes, trypsins and tryptase, in Clostridium difficile toxin A (TxA)-induced enteritis. Methods We injected TxA into ileal loops in PAR2 or dipeptidyl peptidase I (DPPI) knockout mice or in wild-type mice pretreated with tryptase inhibitors (FUT-175 or MPI-0442352) or soybean trypsin inhibitor. We examined the effect of TxA on expression and activity of PAR2 and trypsin IV messenger RNA in the ileum and cultured colonocytes. We injected activating peptide (AP), trypsins, tryptase, and p23 in wild-type mice, some pretreated with the neurokinin 1 receptor antagonist SR140333. Results TxA increased fluid secretion, myeloperoxidase activity in fluid and tissue, and histologic damage. PAR2 deletion decreased TxA-induced ileitis, reduced luminal fluid secretion by 20%, decreased tissue and fluid myeloperoxidase by 50%, and diminished epithelial damage, edema, and neutrophil infiltration. DPPI deletion reduced secretion by 20% and fluid myeloperoxidase by 55%. In wild-type mice, FUT-175 or MPI-0442352 inhibited secretion by 24%−28% and tissue and fluid myeloperoxidase by 31%−71%. Soybean trypsin inhibitor reduced secretion to background levels and tissue myeloperoxidase by up to 50%. TxA increased expression of PAR2 and trypsin IV in enterocytes and colonocytes and caused a 2-fold increase in Ca2+ responses to PAR2 AP. AP, tryptase, and trypsin isozymes (trypsin I/II, trypsin IV, p23) caused ileitis. SR140333 prevented AP-induced ileitis. Conclusions PAR2 and its activators are proinflammatory in TxA-induced enteritis. TxA stimulates existing PAR2 and up-regulates PAR2 and activating proteases, and PAR2 causes inflammation by neurogenic mechanisms. PMID:17570216
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Flow Cytometry of Human Primary Epidermal and Follicular Keratinocytes
Gragnani, Alfredo; Ipolito, Michelle Zampieri; Sobral, Christiane S; Brunialti, Milena Karina Coló; Salomão, Reinaldo; Ferreira, Lydia Masako
2008-01-01
Objective: The aim of this study was to characterize using flow cytometry cultured human primary keratinocytes isolated from the epidermis and hair follicles by different methods. Methods: Human keratinocytes derived from discarded fragments of total skin and scalp hair follicles from patients who underwent plastic surgery in the Plastic Surgery Division at UNIFESP were used. The epidermal keratinocytes were isolated by using 3 different methods: the standard method, upon exposure to trypsin for 30 minutes; the second, by treatment with dispase for 18 hours and with trypsin for 10 minutes; and the third, by treatment with dispase for 18 hours and with trypsin for 30 minutes. Follicular keratinocytes were isolated using the standard method. Results: On comparing the group treated with dispase for 18 hours and with trypsin for 10 minutes with the group treated with dispase for 18 hours and with trypsin for 30 minutes, it was observed that the first group presented the largest number of viable cells, the smallest number of cells in late apoptosis and necrosis with statistical significance, and no difference in apoptosis. When we compared the group treated with dispase for 18 hours and with trypsin for 10 minutes with the group treated with trypsin, the first group presented the largest number of viable cells, the smallest number of cells in apoptosis with statistical significance, and no difference in late apoptosis and necrosis. When we compared the results of the group treated with dispase for 18 hours and with trypsin for 10 minutes with the results for follical isolation, there was a statistical difference in apoptosis and viable cells. Conclusion: The isolation method of treatment with dispase for 18 hours and with trypsin for 10 minutes produced the largest number of viable cells and the smallest number of cells in apoptosis/necrosis. PMID:18350110
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Flow cytometry of human primary epidermal and follicular keratinocytes.
Gragnani, Alfredo; Ipolito, Michelle Zampieri; Sobral, Christiane S; Brunialti, Milena Karina Coló; Salomão, Reinaldo; Ferreira, Lydia Masako
2008-02-19
The aim of this study was to characterize using flow cytometry cultured human primary keratinocytes isolated from the epidermis and hair follicles by different methods. Human keratinocytes derived from discarded fragments of total skin and scalp hair follicles from patients who underwent plastic surgery in the Plastic Surgery Division at UNIFESP were used. The epidermal keratinocytes were isolated by using 3 different methods: the standard method, upon exposure to trypsin for 30 minutes; the second, by treatment with dispase for 18 hours and with trypsin for 10 minutes; and the third, by treatment with dispase for 18 hours and with trypsin for 30 minutes. Follicular keratinocytes were isolated using the standard method. On comparing the group treated with dispase for 18 hours and with trypsin for 10 minutes with the group treated with dispase for 18 hours and with trypsin for 30 minutes, it was observed that the first group presented the largest number of viable cells, the smallest number of cells in late apoptosis and necrosis with statistical significance, and no difference in apoptosis. When we compared the group treated with dispase for 18 hours and with trypsin for 10 minutes with the group treated with trypsin, the first group presented the largest number of viable cells, the smallest number of cells in apoptosis with statistical significance, and no difference in late apoptosis and necrosis. When we compared the results of the group treated with dispase for 18 hours and with trypsin for 10 minutes with the results for follical isolation, there was a statistical difference in apoptosis and viable cells. The isolation method of treatment with dispase for 18 hours and with trypsin for 10 minutes produced the largest number of viable cells and the smallest number of cells in apoptosis/necrosis.
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Braia, Mauricio Javier; Loureiro, Dana Belén; Tubio, Gisela; Romanini, Diana
2015-12-01
Protein-polyelectrolyte complexes are very interesting systems since they can be applied in many long-established and emerging areas of biotechnology. From nanotechnology to industrial processing, these complexes are used for many purposes: to build multilayer particles for biosensors; to entrap and deliver proteins for pharmaceutical applications; to isolate and immobilize proteins. The enteric copolymer poly(methacrylic acid-co-methyl methacrylate) 1:2 (MMA) has been designed for drug delivery although its chemical properties allow to use it for other applications. Understanding the interaction between trypsin and this polymer is very important in order to optimize the mechanism of formation of this complex for different biotechnological applications.The formation of the trypsin-MMA complex was studied by spectroscopy and isothermal titration calorimetry. Structural analysis of trypsin was carried out by catalytic activity assays, circular dichroism and differential scanning calorimetry. Isothermal titration calorimetry experiments showed that the insoluble complex contains 12 trypsin molecules per MMA molecule at pH 5 and they interact with high affinity to form insoluble complexes. Both electrostatic and hydrophobic forces are involved in the formation of the complex. The structure of trypsin is not affected by the presence of MMA, although it interacts with some domains of trypsin affecting its thermal denaturation as seen in the differential scanning calorimetry experiments. Its catalytic activity is not altered. Dynamic light scattering demonstrated the presence of a soluble trypsin-copolymer complex at pH 5 and 8. Turbidimetric assays show that the insoluble complex can be dissolved by low ionic strength and/or pH in order to obtain free native trypsin. Copyright © 2015 Elsevier B.V. All rights reserved.
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Interaction of sodium benzoate with trypsin by spectroscopic techniques
NASA Astrophysics Data System (ADS)
Mu, Yue; Lin, Jing; Liu, Rutao
2011-12-01
The toxicity of sodium benzoate to trypsin was investigated by fluorescence spectroscopy, synchronous fluorescence spectroscopy, UV-visible absorption spectroscopy and circular dichroism (CD) spectroscopy under mimic physiological conditions. Sodium benzoate could unfold trypsin by decreasing the β-sheet structure, which leads to more exposure of internal amino acid groups and the obvious intrinsic fluorescence quenching with the rising concentration of sodium benzoate. The results of spectroscopic measurements indicated that sodium benzoate changed the internal microenvironment of trypsin and induced the alteration of the whole molecule, which were performed toxic effects on the organism. Trypsin and sodium benzoate interacted with each other to produce a substance by van der Waals forces and hydrogen bond, the model of which was shown by AutoDock software.
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Trypsin-like Proteins of the Fungi as Possible Markers of Phytopathogenicity
USDA-ARS?s Scientific Manuscript database
Sequences of peptidases with conserved motifs around the active site residues that are characteristic of trypsins (similar to trypsin peptidases, STP) were obtained from publicly available fungal genomes and related databases. Among the 74 fungal genomes, 30 species of parasitic Ascomycota contained...
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21 CFR 866.5890 - Inter-alpha trypsin inhibitor immunological test system.
Code of Federal Regulations, 2013 CFR
2013-04-01
... HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunological Test... measure by immunochemical techniques the inter-alpha trypsin inhibitor (a protein) in serum and other body fluids. Measurement of inter-alpha trypsin inhibitor may aid in the diagnosis of acute bacterial...
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21 CFR 866.5890 - Inter-alpha trypsin inhibitor immunological test system.
Code of Federal Regulations, 2011 CFR
2011-04-01
... HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunological Test... measure by immunochemical techniques the inter-alpha trypsin inhibitor (a protein) in serum and other body fluids. Measurement of inter-alpha trypsin inhibitor may aid in the diagnosis of acute bacterial...
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21 CFR 866.5890 - Inter-alpha trypsin inhibitor immunological test system.
Code of Federal Regulations, 2012 CFR
2012-04-01
... HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunological Test... measure by immunochemical techniques the inter-alpha trypsin inhibitor (a protein) in serum and other body fluids. Measurement of inter-alpha trypsin inhibitor may aid in the diagnosis of acute bacterial...
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21 CFR 866.5890 - Inter-alpha trypsin inhibitor immunological test system.
Code of Federal Regulations, 2014 CFR
2014-04-01
... HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunological Test... measure by immunochemical techniques the inter-alpha trypsin inhibitor (a protein) in serum and other body fluids. Measurement of inter-alpha trypsin inhibitor may aid in the diagnosis of acute bacterial...
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Structural Insights into the Role of the Cyclic Backbone in a Squash Trypsin Inhibitor*
Daly, Norelle L.; Thorstholm, Louise; Greenwood, Kathryn P.; King, Gordon J.; Rosengren, K. Johan; Heras, Begoña; Martin, Jennifer L.; Craik, David J.
2013-01-01
MCoTI-II is a head-to-tail cyclic peptide with potent trypsin inhibitory activity and, on the basis of its exceptional proteolytic stability, is a valuable template for the design of novel drug leads. Insights into inhibitor dynamics and interactions with biological targets are critical for drug design studies, particularly for protease targets. Here, we show that the cyclization and active site loops of MCoTI-II are flexible in solution, but when bound to trypsin, the active site loop converges to a single well defined conformation. This finding of reduced flexibility on binding is in contrast to a recent study on the homologous peptide MCoTI-I, which suggested that regions of the peptide are more flexible upon binding to trypsin. We provide a possible explanation for this discrepancy based on degradation of the complex over time. Our study also unexpectedly shows that the cyclization loop, not present in acyclic homologues, facilitates potent trypsin inhibitory activity by engaging in direct binding interactions with trypsin. PMID:24169696
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Fang, Evandro Fei; Ng, Tzi Bun
2015-04-01
Nephelium lappaceum L., commonly known as "rambutan," is a typical tropical tree and is well known for its juicy and sweet fruit which has an exotic flavor. Chemical studies on rambutan have led to the identification of various components such as monoterpene lactones and volatile compounds. Here, a 22.5-kDa trypsin inhibitor (N . lappaceum trypsin inhibitor (NLTI)) was isolated from fresh rambutan seeds using liquid chromatographical techniques. NLTI reduced the proteolytic activities of both trypsin and α-chymotrypsin. Dithiothreitol reduced the trypsin inhibitory activity of NLTI at a concentration of 1 mM, indicating that an intact disulfide bond is essential to the activity. NLTI inhibited HIV-1 reverse transcriptase with an IC50 of 0.73 μM. In addition, NLTI manifested a time- and dose-dependent inhibitory effect on growth in many tumor cells. NLTI is one of the few trypsin inhibitors with nitric oxide-inducing activity and may find application in tumor therapy.
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Action of trypsin on structural changes of collagen fibres from sea cucumber (Stichopus japonicus).
Liu, Zi-Qiang; Tuo, Feng-Yan; Song, Liang; Liu, Yu-Xin; Dong, Xiu-Ping; Li, Dong-Mei; Zhou, Da-Yong; Shahidi, Fereidoon
2018-08-01
Trypsin, a representative serine proteinase, was used to hydrolyse the collagen fibres from sea cucumber (Stichopus japonicus) to highlight the role of serine proteinase in the autolysis of sea cucumber. Partial disaggregation of collagen fibres into collagen fibrils upon trypsin treatment occurred. The trypsin treatment also caused a time-dependent release of water-soluble glycosaminoglycans and proteins. Therefore, the degradation of the proteoglycan bridges between collagen fibrils might account for the disaggregation of collagen fibrils. For trypsin-treated collagen fibres (72 h), the collagen fibrils still kept their structural integrity and showed characteristic D-banding pattern, and the dissolution rate of hydroxyproline was just 0.21%. Meanwhile, Fourier transform infrared analysis showed the collagen within trypsin-treated collagen fibres (72 h) still retaining their triple-helical conformation. These results suggested that serine proteinase participated in the autolysis of S. japonicus body wall by damaging the proteoglycan bridges between collagen fibrils and disintegrating the latter. Copyright © 2018 Elsevier Ltd. All rights reserved.
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Salmon trypsin stimulates the expression of interleukin-8 via protease-activated receptor-2
DOE Office of Scientific and Technical Information (OSTI.GOV)
Larsen, Anett K.; Department of Pharmacology, Institute of Medical Biology, University of Tromso, Tromso; Seternes, Ole-Morten
2008-08-01
In this study, we focus on salmon trypsin as an activator of inflammatory responses in airway cells in vitro. The rationale behind the investigation is that salmon industry workers are exposed to aerosols containing enzymes, which are generated during industrial processing of the fish. Knowing that serine proteases such as trypsin are highly active mediators with diverse biological activities, the stimulation of nuclear factor-kappa B (NF-{kappa}B) and interleukin (IL)-8 and the role of protease-activated receptors (PAR) in inflammatory signal mediation were investigated. Protease-activated receptors are considered important under pathological situations in the human airways, and a thorough understanding of PAR-inducedmore » cellular events and their consequences in airway inflammation is necessary. Human airway epithelial cells (A549) were exposed to trypsin isolated from fish (Salmo salar), and we observed that purified salmon trypsin could generate secretion of IL-8 in a concentration-dependent manner. Furthermore, we demonstrate that PAR-2 activation by salmon trypsin is coupled to an induction of NF-{kappa}B-mediated transcription using a PAR-2 transfected HeLa cell model. Finally, we show that the release of IL-8 from A549 following stimulation with purified salmon trypsin is mediated through activation of PAR-2 using specific small interfering RNAs (siRNAs). The results presented suggest that salmon trypsin, via activation of PAR-2, might influence inflammation processes in the airways if inhaled in sufficient amounts.« less
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Iskandar, Viska I; Sasaki, Yutaka; Yoshino, Naoto; Abubakar, Raden Z R; Sato, Shigehiro; Muraki, Yasushi
2018-02-01
A cell-based vaccine production method for influenza virus may be an effective and more rapid alternative to egg-based systems. For high-yield virus production, the effect of bovine, porcine, fungal, and recombinant trypsins on influenza A/H1N1 virus replication in MDCK SI-6 cells (SI-6 cells), a novel MDCK cell line developed by our research group, was examined. SI-6 cells infected with influenza A/H1N1 virus were incubated in the presence of four trypsin types at various concentrations, and virus yields in the culture medium were evaluated by a hemagglutination (HA) assay. Virus growth was most efficient in the presence of bovine and porcine trypsins. An analysis of the optimized concentration and definitive HA titer of each trypsin by Gaussian distribution revealed that comparable high virus yields (166.1 and 164.2 HAU/50μl) were obtained at the optimized concentrations of bovine (0.4μg/ml) and porcine (2.1μg/ml) trypsins, respectively, the yields of which were significantly higher than that of fungal and recombinant trypsins. We conclude that bovine and porcine trypsins are suitable for influenza A/H1N1 virus replication in SI-6 cells. This result complements our previous study and suggests the possible application of SI-6 cells to the development of cell-based influenza vaccines. Copyright © 2017 Elsevier B.V. All rights reserved.
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Antonini, E; Ascenzi, P; Bolognesi, M; Menegatti, E; Guarneri, M
1983-04-25
The formation of the bovine beta-trypsin-bovine basic pancreatic trypsin inhibitor (Kunitz) (BPTI) complex was monitored, making use of three different signals: proflavine displacement, optical density changes in the ultraviolet region, and the loss of the catalytic activity. The rates of the reactions indicated by the three different signals were similar at neutral pH, but diverged at low pH. At pH 3.50, proflavine displacement precedes the optical density changes in the ultraviolet and the loss of enzyme activity by several orders of magnitude in time (Antonini, E., Ascenzi, P., Menegatti, E., and Guarneri, M. (1983) Biopolymers 22, 363-375). These data indicated that the bovine beta-trypsin-BPTI complex formation is a multistage process and led to the prediction that, at pH 3.50, BPTI addition to the bovine beta-trypsin-proflavine complex would remove proflavine inhibition and the enzyme would recover transiently its catalytic activity before being irreversibly inhibited by completion of BPTI binding. The kinetic evidences, by completion of BPTI binding. The kinetic evidences, here shown, verified this prediction, indicating that during the bovine beta-trypsin-BPTI complex formation one transient intermediate occurs, which is not able to bind proflavine but may bind and hydrolyze the substrate. Thus, the observed peculiar catalytic behavior is in line with the proposed reaction mechanism for the bovine beta-trypsin-BPTI complex formation, which postulates a sequence of distinct polar and apolar interactions at the contact area.
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Infectious rotavirus enters cells by direct cell membrane penetration, not by endocytosis
DOE Office of Scientific and Technical Information (OSTI.GOV)
Kaljot, K.T.; Shaw, R.D.; Greenberg, H.B.
1988-04-01
Rotaviruses are icosahedral viruses with a segmented, double-stranded RNA genome. They are the major cause of severe infantile infectious diarrhea. Rotavirus growth in tissue culture is markedly enhanced by pretreatment of virus with trypsin. Trypsin activation is associated with cleavage of the viral hemagglutinin (viral protein 3 (VP3); 88 kilodaltons) into two fragments (60 and 28 kilodaltons). The mechanism by which proteolytic cleavage leads to enhanced growth is unknown. To determine whether trypsin treatment affected rotavirus internalization, the authors studied the kinetics of entry of infectious rhesus rotavirus (RRV) into MA104 cells. Trypsin-activated RRV was internalized with a half-time ofmore » 3 to 5 min, while nonactivated virus disappeared from the cell surface with a half-time of 30 to 50 min. In contrast to trypsin-activated RRV, loss of nonactivated RRV from the cell surface did not result in the appearance of infection, as measured by plaque formation. Purified trypsin-activated RRV added to cell monolayers at pH 7.4 mediated {sup 51}Cr, ({sup 14}C)choline, and ({sup 3}H)inositol released from prelabeled MA104 cells. This release could be specifically blocked by neutralizing antibodies to VP3. These results suggest that MA104 cell infection follows the rapid entry of trypsin-activated RRV by direct cell membrane penetration. Cell membrane penetration of infectious RRV is initiated by trypsin cleavage of VP3. Neutralizing antibodies can inhibit this direct membrane penetration.« less
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Chi, Liang; Liu, Qinghua; Xu, Shihong; Xiao, Zhizhong; Ma, Daoyuan; Li, Jun
2015-10-01
Trypsin is an important serine protease that is considered to be involved in digestion of protein in teleost fish. Nevertheless, studies on trypsin/trypsinogen in fish embryos are very limited. In this study, the trypsinogen of turbot (Scophthalmus maximus) (tTG) was identified and the expression patterns and activity of trypsinogen/trypsin were investigated. The results showed that the tTG mRNA was evenly distributed in the oocytes and was also expressed along the yolk periphery in early embryos. At later embryo stages and 1 days after hatching (dph), the tTG mRNA concentrated at the alimentary tract and head. Quantitative expression analysis showed that the tTG transcripts decreased after fertilization until the gastrula stage, then increased with the embryo and larvae development. This result was also confirmed by the specific activity analysis of trypsin and in-situ-hybridization (ISH). All of the results indicated that tTG in early embryo stages was maternally derived and expressed by itself after gastrula stages. Additionally, location of tTG mRNA in embryos and larvae was investigated; we considered that trypsin may have multiple functions during the embryo development process. Based on our results regarding trypsinogen in embryos and early development, we concluded that the trypsin/trypsinogen in turbot embryos was inherited from a maternal source and we suggested that trypsin in early development has multiple functions in the process of development. Copyright © 2015 Elsevier Inc. All rights reserved.
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21 CFR 866.5890 - Inter-alpha trypsin inhibitor immunological test system.
Code of Federal Regulations, 2010 CFR
2010-04-01
... HUMAN SERVICES (CONTINUED) MEDICAL DEVICES IMMUNOLOGY AND MICROBIOLOGY DEVICES Immunological Test Systems § 866.5890 Inter-alpha trypsin inhibitor immunological test system. (a) Identification. An inter... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Inter-alpha trypsin inhibitor immunological test...
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Walsh, J.J.
1976-01-01
Temporal changes of the Plymouth herring, Atlanto-Scandian herring, Norweigian cod, New York menhaden, Maine lobster, California sardine, anchovy, and red crab, and Japanese herring and sardine are considered in relation to oscillations of Peruvian anchovy and guano bird populations in response to variations of wind strength, of atmospheric and sea surface temperature anomalies, and of current speed for the Eastern Tropical Pacific. It is suggested that marine communities, either off Peru or throughout the ocean, respond in a similar manner to global oscillations at the climatic and El Nino scales by geographical relocation of their centers of abundance. It ismore » further suggested that these two longer scales of variability are minor perturbations of marine ecosystems in comparison with an interaction of overfishing and natural oscillations at the event scale of variability, i.e., that the failure of most of the world's clupeid fisheries may be linked to imposition of this additional stress and the local perturbations of the larval drift of an organism on a time scale of days to weeks.« less
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THE INACTIVATION OF DILUTE SOLUTIONS OF CRYSTALLINE TRYPSIN BY X-RADIATION
McDonald, Margaret R.
1954-01-01
The activity of dilute solutions of crystalline trypsin is destroyed by x-rays. The inactivation is an exponential function of the radiation dose. The reaction yield of inactivation is independent of the intensity at which the radiation is delivered or the quality of the x-rays. The reaction yield increases with increasing concentration of trypsin, varying from 0.06 to 0.7 micromoles per liter per 1000 r for trypsin solutions ranging from 1 x 10–7 to 2 x 10–4 M. PMID:13192318
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Interaction of sodium benzoate with trypsin by spectroscopic techniques.
Mu, Yue; Lin, Jing; Liu, Rutao
2011-12-01
The toxicity of sodium benzoate to trypsin was investigated by fluorescence spectroscopy, synchronous fluorescence spectroscopy, UV-visible absorption spectroscopy and circular dichroism (CD) spectroscopy under mimic physiological conditions. Sodium benzoate could unfold trypsin by decreasing the β-sheet structure, which leads to more exposure of internal amino acid groups and the obvious intrinsic fluorescence quenching with the rising concentration of sodium benzoate. The results of spectroscopic measurements indicated that sodium benzoate changed the internal microenvironment of trypsin and induced the alteration of the whole molecule, which were performed toxic effects on the organism. Trypsin and sodium benzoate interacted with each other to produce a substance by van der Waals forces and hydrogen bond, the model of which was shown by AutoDock software. Copyright © 2011 Elsevier B.V. All rights reserved.
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Lin, Y J; Harada, S; Loten, E G; Smith, R M; Jarett, L
1992-01-01
H35 hepatoma cells were treated with trypsin to abolish insulin binding and insulin-stimulated receptor kinase activity. Insulin was, however, internalized by fluid-phase endocytosis in trypsin-treated cells. Furthermore, nuclear accumulation of insulin was similar in control and trypsin-treated hepatoma cells. Northern blot analysis revealed insulin increased g33 and c-fos mRNA concentrations identically in control and trypsin-treated cells but had no effect on beta 2-microglobulin mRNA. Actinomycin D treatment prior to or after insulin addition demonstrated that insulin increased gene transcription and had no effect on mRNA degradation. These studies suggest that the accumulation of intact insulin in cell nuclei may be directly involved in the increased transcription of immediate-early genes. Images PMID:1409684
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Chen, Yingzhuang; Wu, Minghuo; Wang, Keyi; Chen, Bo; Yao, Shouzhuo; Zou, Hanfa; Nie, Lihua
2011-11-04
A novel thiol-ene "click" strategy for the preparation of monolithic trypsin microreactor was proposed. The hybrid organic-inorganic monolithic capillary column with ene-functionality was fabricated by sol-gel process using tetramethoxysilane (TMOS) and γ-methacryloxypropyltrimethoxysilane (γ-MAPS) as precursors. The disulfide bonds of trypsin were reduced to form free thiol groups. Then the trypsin containing free thiol groups was attached on the γ-MAPS hybrid monolithic column with ene-functionality via thiol-ene click chemistry to form a trypsin microreactor. The activity of the trypsin microreactor was characterized by detecting the substrate (Nα-p-tosyl-L-arginine methyl ester hydrochloride, TAME) and the product (Nα-p-tosyl-L-arginine, TA) with on-line capillary zone electrophoresis. After investigating various synthesizing conditions, it was found that the microreactor with poly(N,N'-methylenebisacrylamide) as spacer can deliver the highest activity, yielding a rapid reaction rate. After repeatedly sampling and analyzing for 100 times, the monolithic trypsin microreactor still remained 87.5% of its initial activity. It was demonstrated that thiol-ene "click" strategy for the construction of enzyme microreactor is a promising method for the highly selective immobilization of proteins under mild conditions, especially enzymes with free thiol radicals. Copyright © 2011 Elsevier B.V. All rights reserved.
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Souza, Joyce; Garcia, Juberlan; Neves, Renata H; Machado-Silva, José Roberto; Maldonado, Arnaldo
2013-12-01
Trypsin and bile salts have been identified as important triggers for excystation of Echinostoma metacercariae. Although excystation in trematodes is a well-known phenomenon, some morphological developmental changes remain to be elucidated. In order to gain further insight into the in vitro development of metacercariae, we assayed different cultivating conditions: 0.5% trypsin and 0.5% bile salts; 1% trypsin and 1% bile salts; 1% trypsin and 0.5% bile salts; 0.5% bile salts; or 0.5% trypsin. By means of light microscopy and confocal microscopy, we characterized each encysted, activated, breached and excysted stage based on the morphological features. However, breached and excysted stages were not revealed in both bile salts and trypsin-free medium. Excretory concretions (25 ± 3.9) were visualized within excretory tubules, close to the ventral sucker and genital anlage. The oral sucker armed with spines and digestive system was similar to those of adult worms. The reproductive system is composed of a genital anlage and the cirrus sac primordium. In short, trypsin and bile salts associated were fundamental for the in vitro metacercariae excystation of Echinostoma paraensei. This article presents the first detailed information of all stages of metacercariae excystation obtained through light and confocal microscopy. Copyright © 2013. Published by Elsevier Inc.
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Mogaki, Rina
2015-01-01
Water-soluble bioadhesive polymers bearing multiple guanidinium ion (Gu+) pendants at their side-chain termini (Gluen–BA, n = 10 and 29) that were conjugated with benzamidine (BA) as a trypsin inhibitor were developed. The Gluen–BA molecules are supposed to adhere to oxyanionic regions of the trypsin surface, even in buffer, via a multivalent Gu+/oxyanion salt-bridge interaction, such that their BA group properly blocks the substrate-binding site. In fact, Glue10–BA and Glue29–BA exhibited 35- and 200-fold higher affinities for trypsin, respectively, than a BA derivative without the glue moiety (TEG–BA). Most importantly, Glue10–BA inhibited the protease activity of trypsin 13-fold more than TEG–BA. In sharp contrast, mGlue27–BA, which bears 27 Gu+ units along the main chain and has a 5-fold higher affinity than TEG–BA for trypsin, was inferior even to TEG–BA for trypsin inhibition. PMID:28706668
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Niu, Canfang; Yang, Peilong; Luo, Huiying; Huang, Huoqing; Wang, Yaru; Yao, Bin
2017-08-30
Susceptibility to proteases usually limits the application of phytase. We sought to improve the pepsin and trypsin resistance of YeAPPA from Yersinia enterocolitica and YkAPPA from Y. kristensenii by optimizing amino acid polarity and charge. The predicted pepsin/trypsin cleavage sites F89/K226 in pepsin/trypsin-sensitive YeAPPA and the corresponding sites (F89/E226) in pepsin-sensitive but trypsin-resistant YkAPPA were substituted with S and H, respectively. Six variants were produced in Pichia pastoris for catalytic and biochemical characterization. F89S, E226H, and F89S/E226H elevated pepsin resistance and thermostability and K226H and F89S/K226H improved pepsin and trypsin resistance and stability at 60 °C and low pH. All the variants increased the ability of the proteins to hydrolyze phytate in corn meal by 2.6-14.9-fold in the presence of pepsin at 37 °C and low pH. This study developed a genetic manipulation strategy specific for pepsin/trypsin-sensitive phytases that can improve enzyme tolerance against proteases and heat and benefit the food and feed industry in a cost-effective way.
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Oterino, J; Sánchez Toranzo, G; Zelarayán, L; Ajmat, M T; Bonilla, F; Bühler, M I
2006-05-01
During activation of amphibian eggs, cortical granule exocytosis causes elaborate ultrastructural changes in the vitelline envelope. These changes involve modifications in the structure of the vitelline envelope and formation of a fertilization envelope (FE) that can no longer be penetrated by sperm. In Bufo arenarum, as the egg traverses the oviduct, the vitelline envelope is altered by a trypsin-like protease secreted by the oviduct, which induces an increased susceptibility of the vitelline envelope to sperm lysins. Full-grown oocytes of B. arenarum, matured in vitro by progesterone, are polyspermic, although cortical granule exocytosis seems to occur within a normal chronological sequence. These oocytes can be fertilized with or without trypsin treatment, suggesting that the vitelline envelope is totally sperm-permeable. Vitelline envelopes without trypsin treatment cannot retain either gp90 or gp96. This suggests that these glycoproteins are involved in the block to polyspermy and that trypsin treatment of matured in vitro oocytes before insemination is necessary to enable vitelline envelopes to block polyspermy. The loss of the binding capacity in vitelline envelopes isolated from B. arenarum oocytes matured in vitro with trypsin treatment and activated by electric shock suggests that previous trypsin treatment is a necessary step for sperm block to occur. When in vitro matured oocytes were incubated with the product of cortical granules obtained from in vitro matured oocytes (vCGP), vitelline envelopes with trypsin treatment were able to block sperm entry. These oocytes exhibited the characteristic signs of activation. These results support the idea that B. arenarum oocytes can be activated by external stimuli and suggest the presence of unknown oocyte surface receptors linked to the activation machinery in response to fertilization. Electrophoretic profiles obtained by SDS-PAGE of solubilized vitelline envelopes from oocytes matured in vitro revealed the conversion of gp40 (in vitro matured oocytes, without trypsin treatment) to gp38 (ascribable to trypsin activity or cortical granule product activity, CGP) and the conversion of gp70 to gp68 (ascribable to trypsin activity plus CGP activity). Taking into account that only the vitelline envelopes of in vitro matured oocytes with trypsin treatment and activated can block sperm entry, we may suggest that the conversion of gp70 to gp68 is related to the changes associated with sperm binding.
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Trophodynamics of some PFCs and BFRs in a western Canadian Arctic marine food web.
Tomy, Gregg T; Pleskach, Kerri; Ferguson, Steve H; Hare, Jonathon; Stern, Gary; Macinnis, Gordia; Marvin, Chris H; Loseto, Lisa
2009-06-01
The trophodynamics of per- and polyfluorinated compounds and bromine-based flame retardants were examined in components of a marine food web from the western Canadian Arctic. The animals studied and their relative trophic status in the food web, established using stable isotopes of nitrogen (delta15N), were beluga (Delphinapterus leucas) > ringed seal (Phoca hispida) > Arctic cod (Boreogadus saida) > Pacific herring (Clupea pallasi) approximately equal to Arctic cisco (Coregonus autumnalis) > pelagic amphipod (Themisto libellula) > Arctic copepod (Calanus hyperboreus). For the brominated diphenyl ethers, the lipid adjusted concentrations of the seven congeners analyzed (Sigma7BDEs: -47, -85, -99, -100, -153, -154, and -209) ranged from 205.4 +/- 52.7 ng/g in Arctic cod to 2.6 +/- 0.4 ng/g in ringed seals. Mean Sigma7BDEs concentrations in Arctic copepods, 16.4 ng/g (n = 2, composite sample), were greater than those in the top trophic level (TL) marine mammals and suggests that (i) Arctic copepods are an important dietary component that delivers BDEs to the food web and (ii) because these compounds are bioaccumulative, metabolism and depletion of BDE congeners in top TL mammals is an important biological process. There were differences in the concentration profiles of the isomers of hexabromocyclododecane (HBCD) in the food web. The most notable difference was observed for beluga, where the alpha-isomer was enriched (accounting for approximately 90% of the SigmaHBCD body burden), relative to its primary prey species, Arctic cod, where the alpha-isomer accounted for only 20% of the SigmaHBCD body burden (beta: 4% and gamma: 78%). For the C8-C11 perfluorinated carboxylic acids, the trophic magnification factors (TMFs) were all greater than unity and increased with increasing carbon chain length. PFOS and its neutral precursor, PFOSA, also had TMF values greater than one. There were also pronounced differences in the PFOSA to PFOS ratio in ringed seal (0.04) and in beluga (1.4) and suggests that, in part, there are differences in the efficacy of biotransforming PFOSA by whale and seal top predators that both preferentially feed on Arctic cod.
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Complete amino acid sequence of the myoglobin from the Pacific sei whale, Balaenoptera borealis.
Jones, B N; Rothgeb, T M; England, R D; Gurd, F R
1979-04-25
The complete amino acid sequence of the major component myoglobin from Pacific sei whale, Balaenoptera borealis, was determined by specific cleavage of the protein to obtain large peptides which are readily degraded by the automatic sequencer. The acetimidated apomyoglobin was selectively cleaved at its two methionyl residues with cyanogen bromide and at its three arginyl residues by trypsin. From the sequence analysis of four of these peptides and the apomyoglobin, over 75% of the covalent structure of the protein was obtained. The remainder of the primary structure was determined by the sequence analysis of peptides that resulted from further digestion of the amino-terminal and central cyanogen bromide fragments. The amino-terminal fragment was specifically cleaved at its two tryptophanyl residues with N-chlorosuccinimide and the central cyanogen bromide fragment was cleaved at its glutamyl residues with staphylococcal protease and at its single tyrosyl residue with N-bromosuccinimide. The primary structure of this myoglobin proved identical with that from the gray whale but differs from that of the finback whale at four positions, from that of the minke whale at three positions and from the myoglobin of the humpback whale at one position. The above sequence identities and differences reflect the close taxonomic relationship of these five species of Cetacea.
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Brzezinski, P; Andréasson, L E
1995-06-06
Reaction centers from Rhodobacter sphaeroides R-26 were treated with trypsin in the dark and during illumination (in the charge-separated state). Trypsination resulted in a time-dependent modification of the reaction centers, reflected in changes in the charge recombination rate, in the inhibition of QA- to QB electron transfer, and eventually to inhibition of charge separation. Comparisons of centers with ubiquinone or anthraquinone in the QA site, in which the charge recombination pathways are different, indicate that trypsination affects charges close to the QA(-)-binding site. Studies of light-induced voltage changes from moving charges in reaction centers incorporated in lipid layers on a Teflon film, a technique which allows the discrimination of effects on donor and acceptor sides, indicate that the acceptor side is preferentially degraded by trypsin in the dark. Tryptic digestion during illumination generally resulted in a marked strengthening and acceleration of the effects seen already during dark treatment, but new effects were also detected in gel electrophoretic peptide patterns, in optical spectra, and in the kinetic measurements. Optical kinetic measurements revealed that the donor side of the reaction centers became susceptible to modification by trypsin during illumination as seen in the value of the binding constant for soluble cytochrome c2 which increased by a factor of 2, whereas it was much less affected after trypsination of reaction centers in the dark. The influence of illumination on the rate and mode by which trypsin acts on reaction centers indicates that changes in the protein conformation follow charge separation.(ABSTRACT TRUNCATED AT 250 WORDS)
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Hu, Zhengyan; Zhao, Liang; Zhang, Hongyan; Zhang, Yi; Wu, Ren'an; Zou, Hanfa
2014-03-21
Proteins interacting with nanoparticles would form the protein coronas on the surface of nanoparticles in biological systems, which would critically impact the biological identities of nanoparticles and/or result in the physiological and pathological consequences. The enzymatic digestion of protein corona was the primary step to achieve the identification of protein components of the protein corona for the bottom-up proteomic approaches. In this study, the investigation on the tryptic digestion of protein corona by the immobilized trypsin on a magnetic nanoparticle was carried out for the first time. As a comparison with the usual overnight long-time digestion and the severe self-digestion of free trypsin, the on-bead digestion of protein corona by the immobilized trypsin could be accomplished within 1h, along with the significantly reduced self-digestion of trypsin and the improved reproducibility on the identification of proteins by the mass spectrometry-based proteomic approach. It showed that the number of identified bovine serum (BS) proteins on the commercial Fe3O4 nanoparticles was increased by 13% for the immobilized trypsin with 1h digestion as compared to that of using free trypsin with even overnight digestion. In addition, the on-bead digestion of using the immobilized trypsin was further applied on the identification of human plasma protein corona on the commercial Fe3O4 nanoparticles, which leads the efficient digestion of the human plasma proteins and the identification of 149 human plasma proteins corresponding to putative critical pathways and biological processes. Copyright © 2014 Elsevier B.V. All rights reserved.
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Kim, Nan-Sun; Yu, Hwa-Young; Chung, Nguyen-Duc; Kwon, Tae-Ho; Yang, Moon-Sik
2014-09-01
Productivity of recombinant bovine trypsin using a rice amylase 3D promoter has been studied in transgenic rice suspension culture. Alternative carbon sources were added to rice cell suspension cultures in order to improve the production of recombinant bovine trypsin. It was demonstrated that addition of alternative carbon sources such as succinic acid, fumaric acid and malic acid in the culture medium could increase the productivity of recombinant bovine trypsin 3.8-4.3-fold compared to those in the control medium without carbon sources. The highest accumulated trypsin reached 68.2 mg/L on day 5 in the culture medium with 40 mM fumaric acid. The feasibility of repeated use of the cells for recombinant trypsin production was tested in transgenic rice cell suspension culture with the culture medium containing the combination of variable sucrose concentration and 40 mM fumaric acid. Among the used combinations, the combination of 1% sucrose and 40 mM fumaric acid resulted in a yield of up to 53 mg/L five days after incubation. It also increased 31% (W/W) of dry cell weight and improved 43% of cell viability compared to that in control medium without sucrose. Based on these data, recycling of the trypsin production process with repeated 1% sucrose and 40 mM fumaric acid supplying-harvesting cycles was developed in flask scale culture. Recombinant bovine trypsin could be stably produced with a yield of up to 53-39 mg/L per cycle during five recycling cycles. Copyright © 2014 Elsevier Inc. All rights reserved.
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An overview of the Oyashio ecosystem
NASA Astrophysics Data System (ADS)
Sakurai, Yasunori
2007-11-01
The Oyashio shelf region and the seasonally ice-covered areas north of Hokkaido are highly productive, supporting a wide range of species including marine mammals, seabirds and commercially important species in the western subarctic Pacific. The fishes include gadids, such as walleye pollock and Pacific cod, and subarctic migratory pelagic fishes such as chum salmon and pink salmon. It is also an important summer feeding ground for subtropical migrants such as the Japanese sardine, Japanese anchovy, Pacific saury, mackerels, Japanese common squid, whales and seabirds. In recent decades, some components of the Oyashio ecosystem (i.e., phytoplankton, mesozooplankton, gadid fish, and subtropical migrants) have shown changes in species abundance or distribution that are correlated with environmental changes such as the 1976/1977 and 1988/1989 regime shifts. The First Oyashio Intrusion moved northward from the mid-1960s until the late 1970s, when it moved southward until the 1980s, after which it returned to the north again after the mid-1990s. The sea-surface temperature in spring decreased after the late 1970s, increased after the late 1980s, and remained high during the 1990s. The extent of ice cover in the Sea of Okhostk also decreased during the latest warming in the 1980-1990s but has increased again since the late 1990s. This and other variabilities affect the Oyashio ecosystem and the surrounding region.
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Code of Federal Regulations, 2011 CFR
2011-04-01
... Substances Affirmed as GRAS § 184.1914 Trypsin. (a) Trypsin (CAS Reg. No. 9002-07-7) is an enzyme preparation... characterizing enzyme activity is that of a peptide hydrolase (EC 3.4.21.4). (b) The ingredient meets the general requirements and additional requirements for enzyme preparations in the Food Chemicals Codex, 3d ed. (1981), p...
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USDA-ARS?s Scientific Manuscript database
Momordica charantia is a perennial plant with reported health benefits. BG-4, a novel peptide from Momordica charantia, was isolated, purified and characterized. The trypsin inhibitory activity of BG-4 is 8.6 times higher than purified soybean trypsin inhibitor. The high trypsin inhibitory activity ...
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Labò, G; Vezzadini, P; Gullo, L; Sternini, C; Bonora, G
1983-08-01
We studied the effect of bombesin (9 ng/kg X min for 30 min by intravenous infusion) on serum immunoreactive trypsin in healthy subjects and in chronic pancreatitis patients. Bombesin administration caused a marked and significant increase of serum immunoreactive trypsin concentration in healthy subjects. The increase occurred in the first 15 min after the beginning of bombesin infusion and persisted for the duration of the study (2 h). In patients with chronic pancreatitis, the increase was much less pronounced. In these patients, the integrated immunoreactive trypsin response to bombesin was significantly correlated with bicarbonate, lipase, and chymotrypsin outputs into the duodenum. The response of serum immunoreactive trypsin to bombesin stimulation seems to vary according to the degree of pancreatic exocrine dysfunction and to reflect the functional capacity of acinar cell mass.
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Han, Song; Lee, Constance W.; Trevino, Jose G.; Hughes, Steven J.; Sarosi, George A.
2013-01-01
Trypsin or Tumor associated trypsin (TAT) activation of Protease-activated receptor 2 (PAR-2) promotes tumor cell proliferation in gastrointestinal cancers. The role of the trypsin/PAR-2 network in esophageal adenocarcinoma (EA) development has not yet been investigated. The aim of this study is to investigate the role of trypsin/PAR-2 activation in EA tumorogenesis and therapy. We found that esophageal adenocarcinoma cells (EACs) and Barrett’s Metaplasia (BART) expressed high levels of type 3 extra-pancreatic trypsinogen (PRSS3), a novel type of TAT. Activity of secreted trypsin was detected in cultured media from EA OE19 and OE33 cultures but not from BART culture. Surface PAR-2 expression in BART and EACs was confirmed by both flow cytometry and immunofluorescence. Trypsin induced cell proliferation (∼ 2 fold; P<0.01) in all tested cell lines at a concentration of 10 nM. Inhibition of PAR-2 activity in EACs via the PAR-2 antagonist ENMD (500 µM), anti-PAR2 antibody SAM-11 (2 µg/ml), or siRNA PAR-2 knockdown, reduced cell proliferation and increased apoptosis by up to 4 fold (P<0.01). Trypsin stimulation led to phosphorylation of ERK1/2, suggesting involvement of MAPK pathway in PAR-2 signal transduction. Inhibition of PAR-2 activation or siRNA PAR-2 knockdown in EACs prior to treatment with 5 FU reduced cell viability of EACs by an additional 30% (P<0.01) compared to chemotherapy alone. Our data suggest that extra-pancreatic trypsinogen 3 is produced by EACs and activates PAR-2 in an autocrine manner. PAR-2 activation increases cancer cell proliferation, and promotes cancer cell survival. Targeting the trypsin activated PAR-2 pathway in conjunction with current chemotherapeutic agents may be a viable therapeutic strategy in EA. PMID:24146905
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Dias, Renata O; Via, Allegra; Brandão, Marcelo M; Tramontano, Anna; Silva-Filho, Marcio C
2015-03-01
Trypsins and chymotrypsins are well-studied serine peptidases that cleave peptide bonds at the carboxyl side of basic and hydrophobic L-amino acids, respectively. These enzymes are largely responsible for the digestion of proteins. Three primary processes regulate the activity of these peptidases: secretion, precursor (zymogen) activation and substrate-binding site recognition. Here, we present a detailed phylogenetic analysis of trypsins and chymotrypsins in three orders of holometabolous insects and reveal divergent characteristics of Lepidoptera enzymes in comparison with those of Coleoptera and Diptera. In particular, trypsin subsite S1 was more hydrophilic in Lepidoptera than in Coleoptera and Diptera, whereas subsites S2-S4 were more hydrophobic, suggesting different substrate preferences. Furthermore, Lepidoptera displayed a lineage-specific trypsin group belonging only to the Noctuidae family. Evidence for facilitated trypsin auto-activation events were also observed in all the insect orders studied, with the characteristic zymogen activation motif complementary to the trypsin active site. In contrast, insect chymotrypsins did not seem to have a peculiar evolutionary history with respect to their mammal counterparts. Overall, our findings suggest that the need for fast digestion allowed holometabolous insects to evolve divergent groups of peptidases with high auto-activation rates, and highlight that the evolution of trypsins led to a most diverse group of enzymes in Lepidoptera. Copyright © 2015 Elsevier Ltd. All rights reserved.
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Ding, Keke; Zhang, Huanxin; Wang, Haifei; Lv, Xuan; Pan, Liumeng; Zhang, Wenjing; Zhuang, Shulin
2015-12-15
Tetrabromobisphenol A (TBBPA) and its replacement alternative tetrabromobisphenol S (TBBPS) are used widely as brominated flame retardants (BFRs). However, the potential risk of their effects on bovine trypsin remains largely unknown. We investigated the effects of TBBPA and TBBPS to bovine trypsin by the fluorescence spectroscopy, circular dichroism and molecular dynamics (MD) simulations. They statically quenched the intrinsic fluorescence of bovine trypsin in a concentration-dependent mode and caused slight red-shifted fluorescence. The short and long fluorescence lifetime decay components of bovine trypsin were both affected, partly due to the disturbed microenvironmental changes of Trp215. The β-sheet content of bovine trypsin was significantly reduced from 82.4% to 75.7% and 76.6% by TBBPA and TBBPS, respectively, possibly impairing the physiological function of bovine trypsin. TBBPA and TBBPS bind at the 8-anilinonaphthalene-1-sulfonate (ANS) binding site with an association constant of 1.09×10(4) M(-1) and 2.41×10(4) M(-1) at 298 K, respectively. MD simulations revealed that van der Waals interactions and hydrogen bond interactions are dominant for TBBPA, whereas electrostatic interactions are critical for TBBPS. Our in vitro and in silico studies are beneficial to the understanding of risk assessment and future design of environmental benign BFRs. Copyright © 2015 Elsevier B.V. All rights reserved.
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NASA Astrophysics Data System (ADS)
Pečová, M.; Šebela, M.; Marková, Z.; Poláková, K.; Čuda, J.; Šafářová, K.; Zbořil, R.
2013-03-01
In this work, magnetosomes produced by microorganisms were chosen as a suitable magnetic carrier for covalent immobilization of thermostable trypsin conjugates with an expected applicability for efficient and rapid digestion of proteins at elevated temperatures. First, a biogenic magnetite was isolated from Magnetospirillum gryphiswaldense and its free surface was coated with the natural polysaccharide chitosan containing free amino and hydroxy groups. Prior to covalent immobilization, bovine trypsin was modified by conjugating with α-, β- and γ-cyclodextrin. Modified trypsin was bound to the magnetic carriers via amino groups using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide and N-hydroxysulfosuccinimide as coupling reagents. The magnetic biomaterial was characterized by magnetometric analysis and electron microscopy. With regard to their biochemical properties, the immobilized trypsin conjugates showed an increased resistance to elevated temperatures, eliminated autolysis, had an unchanged pH optimum and a significant storage stability and reusability. Considering these parameters, the presented enzymatic system exhibits properties that are superior to those of trypsin forms obtained by other frequently used approaches. The proteolytic performance was demonstrated during in-solution digestion of model proteins (horseradish peroxidase, bovine serum albumin and hen egg white lysozyme) followed by mass spectrometry. It is shown that both magnetic immobilization and chemical modification enhance the characteristics of trypsin making it a promising tool for protein digestion.
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21 CFR 524.2620 - Liquid crystalline trypsin, Peru balsam, castor oil.
Code of Federal Regulations, 2013 CFR
2013-04-01
...) Conditions of use. The drug is used as an aid in the treatment of external wounds and assists healing by... delivered to the wound site contains 0.12 milligram of crystalline trypsin, 87.0 milligrams of Peru balsam... gram delivered to the wound site contains 0.1 milligram of crystalline trypsin, 72.5 milligrams of Peru...
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21 CFR 524.2620 - Liquid crystalline trypsin, Peru balsam, castor oil.
Code of Federal Regulations, 2012 CFR
2012-04-01
...) Conditions of use. The drug is used as an aid in the treatment of external wounds and assists healing by... delivered to the wound site contains 0.12 milligram of crystalline trypsin, 87.0 milligrams of Peru balsam... gram delivered to the wound site contains 0.1 milligram of crystalline trypsin, 72.5 milligrams of Peru...
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21 CFR 524.2620 - Liquid crystalline trypsin, Peru balsam, castor oil.
Code of Federal Regulations, 2011 CFR
2011-04-01
... delivered to the wound site contains 0.12 milligram of crystalline trypsin, 87.0 milligrams of Peru balsam... gram delivered to the wound site contains 0.1 milligram of crystalline trypsin, 72.5 milligrams of Peru...) Conditions of use. The drug is used as an aid in the treatment of external wounds and assists healing by...
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21 CFR 524.2620 - Liquid crystalline trypsin, Peru balsam, castor oil.
Code of Federal Regulations, 2010 CFR
2010-04-01
...) Conditions of use. The drug is used as an aid in the treatment of external wounds and assists healing by... delivered to the wound site contains 0.12 milligram of crystalline trypsin, 87.0 milligrams of Peru balsam... gram delivered to the wound site contains 0.1 milligram of crystalline trypsin, 72.5 milligrams of Peru...
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Krishnamoorthy, Navaneethakrishnan; Gajendrarao, Poornima; Eom, Soo Hyun; Kwon, Yong Jung; Cheong, Gang-Won; Lee, Keun Woo
2008-08-01
In Bacillus subtilis, CodW peptidase and CodX ATPase function together as a distinctive ATP-dependent protease called CodWX, which participates in protein degradation and regulates cell division. The molecular structure of CodX and the assembly structure of CodW-CodX have not yet been resolved. Here we present the first three-dimensional structure of CodX N-terminal (N) and C-terminal (C) domain including possible structure of intermediate (I) domain based on the crystal structure of homologous Escherichia coli HslU ATPase. Moreover, the biologically relevant CodWX (W(6)W(6)X(6)) octadecamer complex structure was constructed using the recently identified CodW-HslU hybrid crystal structure. Molecular dynamics (MD) simulation shows a reasonably stable structure of modeled CodWX and explicit behavior of key segments in CodX N and C domain: nucleotide binding residues, GYVG pore motif and CodW-CodX interface. Predicted structure of the possible I domain is flexible in nature with highly coiled hydrophobic region (M153-M206) that could favor substrate binding and entry. Electrostatic surface potential observation unveiled charge complementarity based CodW-CodX interaction pattern could be a possible native interaction pattern in the interface of CodWX. CodX GYVG pore motif structural features, flexible nature of glycine (G92 and G95) residues and aromatic ring conformation preserved Y93 indicated that it may follow the similar mode during the proteolysis mechanism as in the HslU closed state. This molecular modeling study uncovers the significance of CodX N and C domain in CodWX complex and provides possible explanations which would be helpful to understand the CodWX-dependent proteolysis mechanism of B. subtilis.
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Trypsin activation pathway of rotavirus infectivity.
Arias, C F; Romero, P; Alvarez, V; López, S
1996-01-01
The infectivity of rotaviruses is increased by and most probably is dependent on trypsin treatment of the virus. This proteolytic treatment specifically cleaves VP4, the protein that forms the spikes on the surface of the virions, to polypeptides VP5 and VP8. This cleavage has been reported to occur in rotavirus SA114fM at two conserved, closely spaced arginine residues located at VP4 amino acids 241 and 247. In this work, we have characterized the VP4 cleavage products of rotavirus SA114S generated by in vitro treatment of the virus with increasing concentrations of trypsin and with proteases AspN and alpha-chymotrypsin. The VP8 and VP5 polypeptides were analyzed by gel electrophoresis and by Western blotting (immunoblotting) with antibodies raised to synthetic peptides that mimic the terminal regions of VP4 generated by the trypsin cleavage. It was shown that in addition to arginine residues 241 and 247, VP4 is cleaved at arginine residue 231. These three sites were found to have different susceptibilities to trypsin, Arg-241 > Arg-231 > Arg-247, with the enhancement of infectivity correlating with cleavage at Arg-247 rather than at Arg-231 or Arg-241. Proteases AspN and alpha-chymotrypsin cleaved VP4 at Asp-242 and Tyr-246, respectively, with no significant enhancement of infectivity, although this enhancement could be achieved by further treatment of the virus with trypsin. The VP4 end products of trypsin treatment were a homogeneous VP8 polypeptide comprising VP4 amino acids 1 to 231 and a heterogeneous VP5, which is formed by two polypeptide species (present at a ratio of approximately 1:5) as a result of cleavage at either Arg-241 or Arg-247. A pathway for the trypsin activation of rotavirus infectivity is proposed. PMID:8709201
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Fan, HuiYin; Dumont, Marie-Josée; Simpson, Benjamin K
2017-11-01
Gelatin from salmon ( Salmo salar ) skin with high molecular weight protein chains ( α -chains) was extracted using trypsin-aided process. Response surface methodology was used to optimise the extraction parameters. Yield, hydroxyproline content and protein electrophoretic profile via sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of gelatin were used as responses in the optimization study. The optimum conditions were determined as: trypsin concentration at 1.49 U/g; extraction temperature at 45 °C; and extraction time at 6 h 16 min. This response surface optimized model was significant and produced an experimental value (202.04 ± 8.64%) in good agreement with the predicted value (204.19%). Twofold higher yields of gelatin with high molecular weight protein chains were achieved in the optimized process with trypsin treatment when compared to the process without trypsin.
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Porphyrin Induced Laser Deactivation of Trypsinogen-Trypsin Conversion
NASA Astrophysics Data System (ADS)
Perido, Joanna; Brancaleon, Lorenzo
2015-03-01
Pancreatitis is caused by the inflammation of the pancreas, where the digestive enzyme trypsin is activated from the precursor enzyme trypsinogen while still in the pancreas. The presence of trypsin in the pancreas causes auto-activation of trypsinogen, resulting in greater inflammation and auto-digestion of the pancreas. In severe cases, this cascade effect can lead to organ failure, diabetes, and pancreatic cancer. Our hypothesis is that if trypsinogen is prevented from auto-activating into trypsin, then this cascade can be stopped. We propose to do this by inducing conformational changes in trypsinogen when bound to a photoactive porphyrin dye. Porphyrins are comprised of four linked heterocyclic groups forming a flat ring, and bind well with proteins such as trypsinogen. In this study we used spectroscopic techniques to probe the binding of meso-tetrakis (4-sulfonatephenyl) porphyrin (TSPP) to trypsinogen in vitro, as a preliminary step to then prompt and characterize conformational changes of trypsinogen through irradiation. If conformational changes are detected the trypsinogen will be tested for trypsin inactivation. This investigation may provide promising initial results to the possible use of porphyrins as an inhibitor of the self-activation of trypsinogen into trypsin, and a potential inhibitor of pancreatitis. MARC*U-STAR.
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Synthesis of Trypsin-Resistant Variants of the Listeria-Active Bacteriocin Salivaricin P▿
O'Shea, Eileen F.; O'Connor, Paula M.; Cotter, Paul D.; Ross, R. Paul; Hill, Colin
2010-01-01
Two-component salivaricin P-like bacteriocins have demonstrated potential as antimicrobials capable of controlling infections in the gastrointestinal tract (GIT). The anti-Listeria activity of salivaricin P is optimal when the individual peptides Sln1 and Sln2 are added in succession at a 1:1 ratio. However, as degradation by digestive proteases may compromise the functionality of these peptides within the GIT, we investigated the potential to create salivaricin variants with enhanced resistance to the intestinal protease trypsin. A total of 11 variants of the salivaricin P components, in which conservative modifications at the trypsin-specific cleavage sites were explored in order to protect the peptides from trypsin degradation while maintaining their potent antimicrobial activity, were generated. Analysis of these variants revealed that eight were resistant to trypsin digestion while retaining antimicrobial activity. Combining the complementary trypsin-resistant variants Sln1-5 and Sln2-3 resulted in a MIC50 of 300 nM against Listeria monocytogenes, a 3.75-fold reduction in activity compared to the level for wild-type salivaricin P. This study demonstrates the potential of engineering bacteriocin variants which are resistant to specific protease action but which retain significant antimicrobial activity. PMID:20581174
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Ahn, Hye-Kyung; Kim, Byoung Chan; Jun, Seung-Hyun
2010-12-15
An efficient protein digestion in proteomic analysis requires the stabilization of proteases such as trypsin. In the present work, trypsin was stabilized in the form of enzyme coating on electrospun polymer nanofibers (EC-TR), which crosslinks additional trypsin molecules onto covalently-attached trypsin (CA-TR). EC-TR showed better stability than CA-TR in rigorous conditions, such as at high temperatures of 40 °C and 50 °C, in the presence of organic co-solvents, and at various pH's. For example, the half-lives of CA-TR and EC-TR were 0.24 and 163.20 hours at 40 ºC, respectively. The improved stability of EC-TR can be explained by covalent-linkages onmore » the surface of trypsin molecules, which effectively inhibits the denaturation, autolysis, and leaching of trypsin. The protein digestion was performed at 40 °C by using both CA-TR and EC-TR in digesting a model protein, enolase. EC-TR showed better performance and stability than CA-TR by maintaining good performance of enolase digestion under recycled uses for a period of one week. In the same condition, CA-TR showed poor performance from the beginning, and could not be used for digestion at all after a few usages. The enzyme coating approach is anticipated to be successfully employed not only for protein digestion in proteomic analysis, but also for various other fields where the poor enzyme stability presently hampers the practical applications of enzymes.« less
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Molecular architecture of the ATP-dependent CodWX protease having an N-terminal serine active site
Kang, Min Suk; Kim, Soon Rae; Kwack, Pyeongsu; Lim, Byung Kook; Ahn, Sung Won; Rho, Young Min; Seong, Ihn Sik; Park, Seong-Chul; Eom, Soo Hyun; Cheong, Gang-Won; Chung, Chin Ha
2003-01-01
CodWX in Bacillus subtilis is an ATP-dependent, N-terminal serine protease, consisting of CodW peptidase and CodX ATPase. Here we show that CodWX is an alkaline protease and has a distinct molecular architecture. ATP hydrolysis is required for the formation of the CodWX complex and thus for its proteolytic function. Remarkably, CodX has a ‘spool-like’ structure that is formed by interaction of the intermediate domains of two hexameric or heptameric rings. In the CodWX complex, CodW consisting of two stacked hexameric rings (WW) binds to either or both ends of a CodX double ring (XX), forming asymmetric (WWXX) or symmetric cylindrical particles (WWXXWW). CodWX can also form an elongated particle, in which an additional CodX double ring is bound to the symmetric particle (WWXXWWXX). In addition, CodWX is capable of degrading EzrA, an inhibitor of FtsZ ring formation, implicating it in the regulation of cell division. Thus, CodWX appears to constitute a new type of protease that is distinct from other ATP-dependent proteases in its structure and proteolytic mechanism. PMID:12805205
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Mechanism of curcumin-induced trypsin inhibition: Computational and experimental studies
NASA Astrophysics Data System (ADS)
Wang, Yan-Qing; Zhang, Hong-Mei; Kang, Yi-Jun; Gu, Yun-Lan; Cao, Jian
2016-03-01
In the present study, the experimental and theoretical methods were used to analyze the binding interaction of food dye, curcumin with trypsin. The results of fluorescence spectroscopic measurements indicated that curcumin binding resulted in the obviously intrinsic fluorescence quenching with the increase concentration of curcumin. This binding interaction is a spontaneous process with the estimated enthalpy and entropy changes being -15.70 kJ mol-1 and 40.25 J mol-1 K-1, respectively. Hydrogen bonds and hydrophobic forces played an important role in the complex formation between curcumin and trypsin. Moreover, curcumin could enter into the primary substrate-binding pocket and makes the activity of trypsin decrease remarkably with the increasing concentration of curcumin.
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The involvement of the cysteine proteases of Clonorchis sinensis metacercariae in excystment.
Li, Shunyu; Chung, Young-Bae; Chung, Byung-Suk; Choi, Min-Ho; Yu, Jae-Ran; Hong, Sung-Tae
2004-05-01
The effects of trypsin, bile, trypsin-bile, pepsin, dithiothreitol (DTT) and metacercarial excretory-secretory product (ESP) on the in vitro excystment of Clonorchis sinensis metacercariae were investigated. The majority of metacercariae excysted immediately in trypsin-bile in PBS solution, a process which was complete after 30 min of incubation. When incubated in metacercarial ESP in PBS, excystment was potentiated in the presence of 5 mM DTT, but was inhibited dose-dependently by a cysteine protease inhibitor, iodoacetic acid. Two active protease bands of 28 and 40 kDa were identified in the ESP of metacercariae by gelatin substrate SDS-PAGE. Scanning electron microscopy demonstrated that the larvae in solutions of DTT and ESP migrated through a small hole on the metacercarial wall, whereas larvae were liberated by entire wall disruption in trypsin solution. These results suggest that trypsin is a major extrinsic factor of the rapid excystment of C. sinensis metacercariae, and that endogenous cysteine proteases are also involved in metacercarial excystment. Copyright 2004 Springer-Verlag
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Amino acid sequence of a trypsin inhibitor from a Spirometra (Spirometra erinaceieuropaei).
Sanda, A; Uchida, A; Itagaki, T; Kobayashi, H; Inokuchi, N; Koyama, T; Iwama, M; Ohgi, K; Irie, M
2001-12-01
A trypsin inhibitor that is highly homologous with bovine pancreatic trypsin inhibitor (BPTI) was co-purified along with RNase from Spirometra (Spirometra erinaceieuropaei). The amino acid sequence of this inhibitor (SETI) and the nucleotide sequence of the cDNA encoding this protein were determined by protein chemistry and gene technology. SETI contains 68 amino acid residues and has a molecular mass of 7,798 Da. SETI has 31 amino acid residues that are identical with BPTI's sequence, including 6 half-cystine and 5 aromatic amino acid residues. The active site Lys residue in BPTI is replaced by an Arg residue in SETI. SETI is an effective inhibitor of trypsin and moderately inhibits a-chymotrypsin, but less inhibits elastase or subtilisin. SETI was expressed by E. coli containing a PelB vector carrying the SETI encoding cDNA; an expression yield of 0.68 mg/l was obtained. The phylogenetic relationship of SETI and the other BPTI-like trypsin inhibitors was analyzed using most likelihood inference methods.
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You, Xiaoying; Li, Yinhuan; Li, Baoping; Ma, Jie
2016-01-15
A chemiluminescence resonance energy transfer (CRET) platform was developed for sensitive and label-free detection of protease by using trypsin as a model analyte. In this CRET platform, bis(2,4,6-trichlorophenyl)oxalate-hydrogen peroxide chemiluminescence (CL) reaction was utilized as an energy donor and bovine serum albumin (BSA)-stabilized gold nanoclusters (Au NCs) as an energy acceptor. The BSA-stabilized Au NCs triggered the CRET phenomenon by accepting the energy from TCPO-H2O2 CL reaction, thus producing intense CL. In the presence of trypsin, the protein template of BSA-stabilized Au NCs was digested, which frustrated the energy transfer efficiency between the CL donor and the BSA-stabilized Au NCs, leading to a significant decrease in the CL signal. The decreased CL signal was proportional to the logarithm of trypsin concentration in the range of 0.01-50.0µg mL(-1). The detection limit for trypsin was 9ng mL(-)(1) and the relative standard deviations were lesser than 3% (n=11). This Au NCs-based CRET platform was successfully applied to the determination of trypsin in human urine samples, demonstrating its potential application in clinical diagnosis. Copyright © 2015 Elsevier B.V. All rights reserved.
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Investigate the Binding of Catechins to Trypsin Using Docking and Molecular Dynamics Simulation
Cui, Fengchao; Yang, Kecheng; Li, Yunqi
2015-01-01
To explore the inhibitory mechanism of catechins for digestive enzymes, we investigated the binding mode of catechins to a typical digestive enzyme-trypsin and analyzed the structure-activity relationship of catechins, using an integration of molecular docking, molecular dynamics simulation and binding free energy calculation. We found that catechins with different structures bound to a conservative pocket S1 of trypsin, which is comprised of residues 189–195, 214–220 and 225–228. In the trypsin-catechin complexes, Asp189 by forming strong hydrogen bonding, and Gln192, Trp215 and Gly216 through hydrophobic interactions, all significantly contribute to the binding of catechins. The number and the position of hydroxyl and aromatic groups, the structure of stereoisomers, and the orientation of catechins in the binding pocket S1 of trypsin all affect the binding affinity. The binding affinity is in the order of Epigallocatechin gallate (EGCG) > Epicatechin gallate (ECG) > Epicatechin (EC) > Epigallocatechin (EGC), and 2R-3R EGCG shows the strongest binding affinity out of other stereoisomers. Meanwhile, the synergic conformational changes of residues and catechins were also analyzed. These findings will be helpful in understanding the knowledge of interactions between catechins and trypsin and referable for the design of novel polyphenol based functional food and nutriceutical formulas. PMID:25938485
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Cohen, A B; Gruenke, L D; Craig, J C; Geczy, D
1977-01-01
alpha-1-Antitrypsin is a serum protein that inhibits many proteolytic enzymes. Recently, it was suggested that the alpha-1-antitrypsin-trypsin complex is an acyl ester analogous to the acyl intermediate that forms between trypsin and its substrates. In previous work we showed that the alpha-1-antitrypsin-trypsin complex can be split at high pH, releasing a component of alpha-1-antitrypsin. This component had a new carboxyl-terminal lysine, and it had lost a peptide of about 4000 daltons. In order to determine whether the alpha-1-antitrypsin is bound to trypsin through the new carboxy-terminal lysine, as would be expected if the above hypothesis is correct, we split the complex in the presence of 18OH-. When the new carboxy-terminal lysine was cleaved with carboxypeptidase B, singly labeled, doubly labeled, and unlabeled lysine were recovered. These data support the hypothesis that the alpha-1-antitrypsin-trypsin complex is an acyl ester or a tetrahedral precursor that is transformed into the acyl ester form at high pH. If other enzymes are bound by a similar mechanism, the methods used may be useful in determining which amino acids on alpha-1-antitrypsin bind covalently to each enzyme. PMID:303770
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Effect of bombesin and cholecystokinin on plasma immunoreactive trypsin in humans.
de Jong, A J; Klamer, M; Jansen, J B; Hopman, W P; Lamers, C B
1987-01-01
Since bombesin is a potent stimulus of the release of cholecystokinin (CCK), it has been suggested that the stimulatory effect of bombesin on pancreatic enzyme secretion is mediated by CCK. The present study was undertaken to determine the role of CCK in the bombesin-induced stimulation of plasma immunoreactive trypsin. Plasma CCK was measured by radioimmunoassay using the antibody T204, which binds to all biologically active sulfated COOH-terminal CCK-peptides. Plasma trypsin was also measured by radioimmunoassay. Infusion of 5 ng/kg/min bombesin in 6 healthy volunteers increased plasma CCK from 1.2 +/- 0.2-8.9 +/- 0.7 pM (p less than 0.0001). The peak increment in plasma CCK during bombesin (9.3 +/- 0.6 pM) was accompanied by a significant rise in plasma trypsin from 206 +/- 21-334 +/- 44 ng/ml (p less than 0.01). However, when similar increases in plasma CCK were achieved by infusion of 0.018 CU/kg/min CCK-33 (9.9 +/- 0.8 pM) or by intraduodenal instillation of 250 ml 20% Intralipid (9.7 +/- 1.9 pM), no significant changes in plasma trypsin were observed. It is therefore concluded that the stimulatory effect of bombesin on plasma immunoreactive trypsin is not mediated by CCK.
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Structural stabilization of CNS synapses during postnatal development in rat cortex.
Khaing, Zin Z; Fidler, Lazar; Nandy, Nina; Phillips, Greg R
2006-07-01
CNS synapses are produced rapidly upon pre- and post-synaptic recruitment. However, their composition is known to change during development and we reasoned that this may be reflected in the gross biochemical properties of synapses. We found synaptic structure in adult cortical synaptosomes to be resistant to digestion with trypsin in the presence and absence of calcium ions, contrasting with previous observations. We evaluated the divalent cation dependence and trypsin sensitivities of synapses using synaptosomes from different developmental stages. In contrast to adult synapses, at postnatal day (P) 10 EDTA treatment eliminated approximately 60% of the synapses, and trypsin and EDTA, together, eliminated all junctions. Trypsinization in the presence of calcium eliminated approximately 60% of the junctions at P10. By P35, all synapses were calcium independent, whereas full trypsin resistance was not attained until P49. To compare the calcium dependence and trypsin sensitivity of synapses in another region of the adult brain, we examined synapses from adult (P50) hippocampus. Adult hippocampus maintained a population of synapses that resembled that of P35 cortex. Our results show that synapses are modified over a long time period in the developing cortex. We propose a model in which the addition of synergistic calcium-dependent and -independent adhesive systems stabilize synapses.
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Hu, Yong; Jing, Zhaoqian; Sudo, Yuta; Niu, Qigui; Du, Jingru; Wu, Jiang; Li, Yu-You
2015-07-01
The effect of the chemical oxygen demand/sulfate (COD/SO4(2-)) ratio on the anaerobic treatment of synthetic chemical wastewater containing acetate, ethanol, and sulfate, was investigated using a UASB reactor. The experimental results show that at a COD/SO4(2-) ratio of 20 and a COD loading rate of 25.2gCODL(-1)d(-1), a COD removal of as high as 87.8% was maintained. At a COD/SO4(2-) ratio of 0.5 (sulfate concentration 6000mgL(-1)), however, the COD removal was 79.2% and the methane yield was 0.20LCH4gCOD(-1). The conversion of influent COD to methane dropped from 80.5% to 54.4% as the COD/SO4(2-) ratio decreased from 20 to 0.5. At all the COD/SO4(2-) ratios applied, over 79.4% of the total electron flow was utilized by methane-producing archaea (MPA), indicating that methane fermentation was the predominant reaction. The majority of the methane was produced by acetoclastic MPA at high COD/SO4(2-) ratios and both acetoclastic and hydrogenthrophic MPA at low COD/SO4(2-) ratios. Only at low COD/SO4(2-) ratios were SRB species such as Desulfovibrio found to play a key role in ethanol degradation, whereas all the SRB species were found to be incomplete oxidizers at both high and low COD/SO4(2-) ratios. Copyright © 2015 Elsevier Ltd. All rights reserved.
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Improved COD Measurements for Organic Content in Flowback Water with High Chloride Concentrations.
Cardona, Isabel; Park, Ho Il; Lin, Lian-Shin
2016-03-01
An improved method was used to determine chemical oxygen demand (COD) as a measure of organic content in water samples containing high chloride content. A contour plot of COD percent error in the Cl(-)-Cl(-):COD domain showed that COD errors increased with Cl(-):COD. Substantial errors (>10%) could occur in low Cl(-):COD regions (<300) for samples with low (<10 g/L) and high chloride concentrations (>25 g/L). Applying the method to flowback water samples resulted in COD concentrations ranging in 130 to 1060 mg/L, which were substantially lower than the previously reported values for flowback water samples from Marcellus Shale (228 to 21 900 mg/L). It is likely that overestimations of COD in the previous studies occurred as result of chloride interferences. Pretreatment with mercuric sulfate, and use of a low-strength digestion solution, and the contour plot to correct COD measurements are feasible steps to significantly improve the accuracy of COD measurements.
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André, L; Pauss, A; Ribeiro, T
2017-03-01
The chemical oxygen demand (COD) is an essential parameter in waste management, particularly when monitoring wet anaerobic digestion processes. An adapted method to determine COD was developed for solid waste (total solids >15%). This method used commercial COD tubes and did not require sample dilution. A homemade plastic weighing support was used to transfer the solid sample into COD tubes. Potassium hydrogen phthalate and glucose used as standards showed an excellent repeatability. A small underestimation of the theoretical COD value (standard values around 5% lower than theoretical values) was also observed, mainly due to the intrinsic COD of the weighing support and to measurement uncertainties. The adapted COD method was tested using various solid wastes in the range of 1-8 mg COD , determining the COD of dried and ground cellulose, cattle manure, straw and a mixed-substrate sample. This new adapted method could be used to monitor and design dry anaerobic digestion processes.
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Suvilampi, J; Lehtomäki, A; Rintala, J
2003-07-01
A combined thermophilic-mesophilic wastewater treatment was studied using a laboratory-scale thermophilic activated sludge process (ASP) followed by mesophilic ASP or a thermophilic suspended carrier biofilm process (SCBP) followed by mesophilic ASP, both systems treating diluted molasses (dilution factor 1:500 corresponding GF/A-filtered COD (COD(filt)) of 1900+/-190 mgl(-1)). With hydraulic retention times (HRTs) of 12-18 h the thermophilic ASP and thermophilic SCBP removed 60+/-13% and 62+/-7% of COD(filt), respectively, with HRT of 8 h the removals were 48+/-1% and 69+/-4%. The sludge volume index (SVI) was notably lower in the thermophilic SCBP (measured from suspended sludge) than in the thermophilic ASP. Under the lowest HRT the mesophilic ASP gave better performance (as SVI, COD(filt), and COD(tot) removals) after the thermophilic SCBP than after the thermophilic ASP. Measured sludge yields were low (less than 0.1 kg suspended solids (SS) kg COD(filt removed)(-1)) in all processes. Both thermophilic treatments removed 80-85% of soluble COD (COD(sol)) whereas suspended COD (COD(susp)) and colloidal COD (COD(col)) were increased. Both mesophilic post-treatments removed all COD(col) and most of the COD(susp) from the thermophilic effluents. In conclusion, combined thermophilic-mesophilic treatment appeared to be easily operable and produced high effluent quality.
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Xie, Zong-Bo; Wang, Na; Wu, Wan-Xia; Le, Zhang-Gao; Yu, Xiao-Qi
2014-01-20
A simple, mild, one-pot tandem method catalyzed by trypsin was developed for the synthesis of 3,4-dihydropyrimidin-2(1H)-ones by the Biginelli reaction of urea, β-dicarbonyl compounds, and in situ-formed acetaldehyde. Trypsin was found to display dual promiscuous functions to catalyze transesterification and the Biginelli reaction in sequence. Copyright © 2013 Elsevier B.V. All rights reserved.
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Targeting Trypsin-Inflammation Axis for Pancreatitis Therapy in a Humanized Pancreatitis Model
2016-10-01
Award Number: W81XWH-15-1-0257 TITLE: Targeting Trypsin-Inflammation Axis for Pancreatitis Therapy in a Humanized Pancreatitis Model PRINCIPAL...AND SUBTITLE Targeting Trypsin-Inflammation Axis for Pancreatitis Therapy in a Humanized Pancreatitis Model 5a. CONTRACT NUMBER 5b. GRANT NUMBER...remains the same since it is covered under the institutional review. We set up monthly video conferences with our partnership PI to discuss any
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Inhibition of trypsin by condensed tannins and wine.
Gonçalves, Rui; Soares, Susana; Mateus, Nuno; de Freitas, Victor
2007-09-05
Phenolic compounds are abundant vegetable secondary metabolites in the human diet. The ability of procyanidin oligomers and wine polyphenols to inhibit trypsin activity was studied using a versatile and reliable in vitro method. The hydrolysis of the chromogenic substrate N-benzoyl-d,l-arginine-p-nitroanilide (BApNA) by trypsin was followed by spectrophotometry in the presence and absence of condensed tannins and wine. A clear relationship between the degree of polymerization of procyanidins and enzymatic inhibition was observed. Trypsin activity inhibition was also detected in several types of wine. In general, the inhibition increased with the concentration of phenolic compounds in wines. These results may be relevant when considering these compounds as antinutritional factors, thereby contributing to a reduced absorption of nutrients.
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Hui, Tabitha C. Y.; Gryba, Rowenna; Gregr, Edward J.; Trites, Andrew W.
2015-01-01
A leading hypothesis to explain the dramatic decline of Steller sea lions (Eumetopias jubatus) in western Alaska during the latter part of the 20th century is a change in prey availability due to commercial fisheries. We tested this hypothesis by exploring the relationships between sea lion population trends, fishery catches, and the prey biomass accessible to sea lions around 33 rookeries between 2000 and 2008. We focused on three commercially important species that have dominated the sea lion diet during the population decline: walleye pollock, Pacific cod and Atka mackerel. We estimated available prey biomass by removing fishery catches from predicted prey biomass distributions in the Aleutian Islands, Bering Sea and Gulf of Alaska; and modelled the likelihood of sea lions foraging at different distances from rookeries (accessibility) using satellite telemetry locations of tracked animals. We combined this accessibility model with the prey distributions to estimate the prey biomass accessible to sea lions by rookery. For each rookery, we compared sea lion population change to accessible prey biomass. Of 304 comparisons, we found 3 statistically significant relationships, all suggesting that sea lion populations increased with increasing prey accessibility. Given that the majority of comparisons showed no significant effect, it seems unlikely that the availability of pollock, cod or Atka mackerel was limiting sea lion populations in the 2000s. PMID:25950178
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Hui, Tabitha C Y; Gryba, Rowenna; Gregr, Edward J; Trites, Andrew W
2015-01-01
A leading hypothesis to explain the dramatic decline of Steller sea lions (Eumetopias jubatus) in western Alaska during the latter part of the 20th century is a change in prey availability due to commercial fisheries. We tested this hypothesis by exploring the relationships between sea lion population trends, fishery catches, and the prey biomass accessible to sea lions around 33 rookeries between 2000 and 2008. We focused on three commercially important species that have dominated the sea lion diet during the population decline: walleye pollock, Pacific cod and Atka mackerel. We estimated available prey biomass by removing fishery catches from predicted prey biomass distributions in the Aleutian Islands, Bering Sea and Gulf of Alaska; and modelled the likelihood of sea lions foraging at different distances from rookeries (accessibility) using satellite telemetry locations of tracked animals. We combined this accessibility model with the prey distributions to estimate the prey biomass accessible to sea lions by rookery. For each rookery, we compared sea lion population change to accessible prey biomass. Of 304 comparisons, we found 3 statistically significant relationships, all suggesting that sea lion populations increased with increasing prey accessibility. Given that the majority of comparisons showed no significant effect, it seems unlikely that the availability of pollock, cod or Atka mackerel was limiting sea lion populations in the 2000s.
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THE INACTIVATION OF DILUTE SOLUTIONS OF CRYSTALLINE TRYPSIN BY X-RADIATION
McDonald, Margaret R.
1955-01-01
The proteolytic activity of dilute solutions of clystalline trypsin is destroyed by x-rays, the amount of inactivation being an exponential function of the radiation dose. The reaction yield increases steadily with increasing concentration of trypsin, varying, as the concentration of enzyme is increased from 1 to 300 µM, from 0.068 to 0.958 micromole of trypsin per liter inactivated per 1000 r with 0.005 N hydrochloric acid as the solvent, from 0.273 to 0.866 with 0.005 N sulfuric acid as the solvent, and from 0.343 to 0.844 with 0.005 N nitric acid as the solvent. When the reaction yields are plotted as a function of the initial concentration of trypsin, they fall on a curve given by the expression Y α XK, in which Y is the reaction yield, X is the concentration of trypsin, and K is a constant equal to 0.46, 0.20, and 0.16, respectively, with 0.005 N hydrochloric, sulfuric, and nitric acids as solvents. The differences between the reaction yields found with chloride and sulfate ions in I to 10 µM trypsin solutions are significant only in the pH range from 2 to 4. The amount of inactivation obtained with a given dose of x-rays depends on the pH of the solution being irradiated and the nature of the solvent. The reaction yield-pH curve is a symmetrical one, with minimum yields at about pH 7. Buffers such as acetate, citrate, borate and barbiturate, and other organic molecules such as ethanol and glucose, in concentrations as low as 20 µM, inhibit the inactivation of trypsin by x-radiation. Sigmoid inactivation-dose curves instead of exponential ones are obtained in the presence of ethanol. The reaction yields for the inactivation of trypsin solutions by x-rays are approximately 1.5 times greater when the irradiation is done at 26°C. than when it is done at 5°C., when 0.005 N hydrochloric acid is the solvent. The dependence on temperature is less when 0.005 N sulfuric acid is used, and is negligible with 0.005 N nitric acid. The difficulties involved in interpreting radiation effects in aqueous systems, and in comparing the results obtained under different experimental conditions, are discussed. PMID:14367774
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Pallagi, Petra; Venglovecz, Viktória; Rakonczay, Zoltán; Borka, Katalin; Korompay, Anna; Ózsvári, Béla; Judák, Linda; Sahin-Tóth, Miklós; Geisz, Andrea; Schnúr, Andrea; Maléth, József; Takács, Tamás; Gray, Mike A.; Argent, Barry E.; Mayerle, Julia; Lerch, Markus M.; Wittmann, Tibor; Hegyi, Péter
2012-01-01
Background & Aims The effects of trypsin on pancreatic ductal epithelial cells (PDEC) vary among species and depend on localization of proteinase-activated receptor-2 (PAR-2). Bicarbonate secretion is similar in human and guinea pig PDEC; we compared its localization in these cell types and isolated guinea pig ducts to study the effects of trypsin and a PAR-2 agonist on this process. Methods PAR-2 localization was analyzed by immunohistochemistry in guinea pig and human pancreatic tissue samples (from 15 patients with chronic pancreatitis and 15 without pancreatic disease). Functions of guinea pig PDEC were studied by microperfusion of isolated ducts, measurements of intracellular pH (pHi) and Ca2+ concentration [Ca2+]i, and patch clamp analysis. The effect of pH on trypsinogen autoactivation was assessed using recombinant human cationic trypsinogen. Results PAR-2 localized to the apical membrane of human and guinea pig PDEC. Trypsin increased [Ca2+]i and pHi, and inhibited secretion of bicarbonate by the luminal anion exchanger and the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel. Autoactivation of human cationic trypsinogen accelerated when the pH was reduced from 8.5 to 6.0. PAR-2 expression was strongly down-regulated, at transcriptional and protein levels, in the ducts of patients with chronic pancreatitis, consistent with increased activity of intraductal trypsin. Importantly, in PAR-2 knockout mice, the effects of trypsin were PAR-2 dependent. Conclusions Trypsin reduces pancreatic ductal bicarbonate secretion via PAR-2–dependent inhibition of the apical anion exchanger and the CFTR Cl- channel. This could contribute to the development of chronic pancreatitis, decreasing luminal pH and promoting premature activation of trypsinogen in the pancreatic ducts. PMID:21893120
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Alloy, Alexandre P.; Kayode, Olumide; Wang, Ruiying
Human mesotrypsin is highly homologous to other mammalian trypsins, and yet it is functionally unique in possessing resistance to inhibition by canonical serine protease inhibitors and in cleaving these inhibitors as preferred substrates. Arg-193 and Ser-39 have been identified as contributors to the inhibitor resistance and cleavage capability of mesotrypsin, but it is not known whether these residues fully account for the unusual properties of mesotrypsin. Here, we use human cationic trypsin as a template for engineering a gain of catalytic function, assessing mutants containing mesotrypsin-like mutations for resistance to inhibition by bovine pancreatic trypsin inhibitor (BPTI) and amyloid precursormore » protein Kunitz protease inhibitor (APPI), and for the ability to hydrolyze these inhibitors as substrates. We find that Arg-193 and Ser-39 are sufficient to confer mesotrypsin-like resistance to inhibition; however, compared with mesotrypsin, the trypsin-Y39S/G193R double mutant remains 10-fold slower at hydrolyzing BPTI and 2.5-fold slower at hydrolyzing APPI. We identify two additional residues in mesotrypsin, Lys-74 and Asp-97, which in concert with Arg-193 and Ser-39 confer the full catalytic capability of mesotrypsin for proteolysis of BPTI and APPI. Novel crystal structures of trypsin mutants in complex with BPTI suggest that these four residues function cooperatively to favor conformational dynamics that assist in dissociation of cleaved inhibitors. Our results reveal that efficient inhibitor cleavage is a complex capability to which at least four spatially separated residues of mesotrypsin contribute. As a result, these findings suggest that inhibitor cleavage represents a functional adaptation of mesotrypsin that may have evolved in response to positive selection pressure.« less
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Efficacy of Trypsin in Treating Coral Snake Envenomation in the Porcine Model.
Parker-Cote, Jennifer L; O'Rourke, Dorcas P; Brewer, Kori L; Lertpiriyapong, Kvin; Punja, Mohan; Bush, Sean P; Miller, Susan N; Meggs, William J
2015-12-01
Antivenom is the definitive treatment for venomous snakebites. Alternative treatments warrant investigation because antivenom is sometimes unavailable, expensive, and can have deleterious side effects. This study assesses the efficacy of trypsin to treat coral snake envenomation in an in vivo porcine model. A randomized, blinded study was conducted. Subjects were 13 pigs injected subcutaneously with 1 mL of eastern coral snake venom (10 mg/mL) in the right distal hind limb. After 1 min, subjects were randomized to have the envenomation site injected with either 1 mL of saline or 1 mL of trypsin (100 mg/mL) by a blinded investigator. Clinical endpoint was survival for 72 h or respiratory depression defined as respiratory rate <15 breaths per minute, falling pulse oximetry, or agonal respirations. Fisher's exact t test was used for between group comparisons. Average time to toxicity for the saline control was 263 min (191-305 min). The development of respiratory depression occurred more frequently in control pigs than treated pigs (p = 0.009). Four of the six pigs that received trypsin survived to the end of the 3-day study. No control pigs survived. Two of the trypsin treatment pigs died with times to toxicity of 718 and 971 min. Survival to 12 and 24 h was significantly greater in the trypsin treatment group (p = 0.002, p = 0.009, respectively). Local injection of trypsin, a proteolytic enzyme, at the site of envenomation decreased the toxicity of eastern coral snake venom and increased survival significantly. Further investigation is required before these results can be extended to human snakebites.
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Xiaopeng, Bai; Tanaka, Yoshimasa; Ihara, Eikichi; Hirano, Katsuya; Nakano, Kayoko; Hirano, Mayumi; Oda, Yoshinao; Nakamura, Kazuhiko
2017-02-15
Duodenal reflux of fluids containing trypsin relates to refractory gastroesophageal reflux disease (GERD). Esophageal peristalsis and clearance are important factors in GERD pathogenesis. However, the function of trypsin in esophageal body contractility is not fully understood. In this study, effects of trypsin on circular smooth muscle (CSM) and longitudinal smooth muscle (LSM) of the porcine esophageal body were examined. Trypsin elicited a concentration dependent biphasic response, a major contraction and a subsequent relaxation only in CSM. In CSM, contraction occurred at trypsin concentrations of 100nM and relaxation at 1μM. A proteinase-activated receptor (PAR)2 activating peptide, SLIGKV-NH 2 (1mM), induced a monophasic contraction. Those responses were unaffected by tetrodotoxin though abolished by the gap junction uncouplers carbenoxolone and octanol. They were also partially inhibited by a transient receptor potential vanilloid type 1 (TRPV1) antagonist and abolished by combination of neurokinin receptor 1 (NK 1 ) and NK 2 antagonists, but not by an NK 3 antagonist, suggesting a PAR2-TRPV1-substance P pathway in sensory neurons. Substance P (100nM), an agonist for various NK receptors (NK 1 , NK 2 and NK 3 ) with differing affinities, induced significant contraction in CSM, but not in LSM. The contraction was also blocked by the combination of NK 1 and NK 2 antagonists, but not by the NK 3 antagonist. Moreover, substance P-induced contractions were unaffected by the TRPV1 antagonist, but inhibited by a gap junction uncoupler. In conclusion, trypsin induced a biphasic response only in CSM and this was mediated by PAR2, TRPV1 and NK 1/2 . Gap junctions were indispensable in this tachykinin-induced response. Copyright © 2017 Elsevier B.V. All rights reserved.
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Alloy, Alexandre P.; Kayode, Olumide; Wang, Ruiying; ...
2015-07-14
Human mesotrypsin is highly homologous to other mammalian trypsins, and yet it is functionally unique in possessing resistance to inhibition by canonical serine protease inhibitors and in cleaving these inhibitors as preferred substrates. Arg-193 and Ser-39 have been identified as contributors to the inhibitor resistance and cleavage capability of mesotrypsin, but it is not known whether these residues fully account for the unusual properties of mesotrypsin. Here, we use human cationic trypsin as a template for engineering a gain of catalytic function, assessing mutants containing mesotrypsin-like mutations for resistance to inhibition by bovine pancreatic trypsin inhibitor (BPTI) and amyloid precursormore » protein Kunitz protease inhibitor (APPI), and for the ability to hydrolyze these inhibitors as substrates. We find that Arg-193 and Ser-39 are sufficient to confer mesotrypsin-like resistance to inhibition; however, compared with mesotrypsin, the trypsin-Y39S/G193R double mutant remains 10-fold slower at hydrolyzing BPTI and 2.5-fold slower at hydrolyzing APPI. We identify two additional residues in mesotrypsin, Lys-74 and Asp-97, which in concert with Arg-193 and Ser-39 confer the full catalytic capability of mesotrypsin for proteolysis of BPTI and APPI. Novel crystal structures of trypsin mutants in complex with BPTI suggest that these four residues function cooperatively to favor conformational dynamics that assist in dissociation of cleaved inhibitors. Our results reveal that efficient inhibitor cleavage is a complex capability to which at least four spatially separated residues of mesotrypsin contribute. As a result, these findings suggest that inhibitor cleavage represents a functional adaptation of mesotrypsin that may have evolved in response to positive selection pressure.« less
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Infant motivation in dental health: attitude without constant reinforcement.
Teixeira Alves, Fabiana Bucholdz; Kuhn, Eunice; Bordin, Danielle; Kozlowski, Vitoldo Antonio; Raggio, Daniela Procida; Fadel, Cristina Berger
2014-01-01
Social factors determine the child's behavior and motivation is an important task in the teaching-learning process. This longitudinal and cross-sectional study aimed to analyze the effectiveness of a motivational activity program for oral hygiene habits formation after motivation and without constant reinforcement. The sample was constituted of 26 children (mean 6 years old) from a Public Kindergarten School in Ponta Grossa, PR, Brazil. Data were collected applying a test-chart, with figures reporting the process of dental health/illness. Some figures were considered positive to dental health (dentist/Cod 1, toothbrush/Cod 3, dentifrice/dental floss/Cod 6, fruits/vegetables/Cod 7 and tooth without caries lesion/Cod 8) and negative on dental health (sweets/Cod 2, bacteria/Cod 4, tooth with caries lesion/Cod 5). The figures presentation occurred in three different stages: First stage - figures were presented to children without previous knowledge; second stage - following the motivational presentation, and third stage - 30 days after the first contact. On the first stage, most children select good for the figures considered harmful to their teeth (Cod 2-88%; Cod 4-77% and Cod 5-65%). On the second stage, there was a lower percentage: 23% (P < 0.0001), 8% (P < 0.0001), and 23% (P = 0.0068) related to the Cod 2, 4, and 5. On the third stage, the results showed again an association with the good choice to these figures considered harmful (Cod 2-85%, Cod 4-65% and Cod 5-54%) similar the results obtained on the first stage. The motivational programs performed without constant reinforcement does not have a positive influence in changing the child's behavior related to a better dental care.
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Nishida, Ayumu; Naganuma, Tsuyoshi; Kanazawa, Takanori; Takashima, Yuuki; Yamada, Masaki; Okada, Hiroaki
2011-07-29
Aqueous preparations of silk protein (sericin) films were prepared to evaluate their biodegradation properties. In the absence of trypsin, sericin film swelled rapidly, kept its shape, and remained unaltered for 28 days or longer due to form β-sheet structures. In the presence of trypsin, sericin film gradually degraded; since the rate depended on the concentration of trypsin, the films likely underwent enzymatic hydrolysis. Sericin film incorporating the model protein drug fluorescein isothiocyanate-albumin (FA) also gradually degraded in the presence of trypsin and resulted in the sustained release of FA for 2 weeks or longer; in contrast, FA release was quite slow in the absence of trypsin. It is expected that sericin film has potential as a biodegradable and drug-releasing carrier. To evaluate the practical applicability of sericin film for the repair of defective tissues, fibroblast growth factor-2 (FGF-2) was incorporated into sericin films and the films were implanted on skull defects in rats. Whereas FGF-2 release was suppressed in the absence of trypsin in vitro, it appears that FGF-2, immobilized by ionic interactions between sericin and FGF-2, can be sustained-released in vivo from films incorporating 2500 or 250 ng of FGF-2 to support the growth of tissue around wounds. Copyright © 2011 Elsevier B.V. All rights reserved.
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The incomplete anti-Rh antibody agglutination mechanism of trypsinized ORh+ red cells.
Margni, R A; Leoni, J; Bazzurro, M
1977-01-01
The capacity for binding to trypsinized and non-trypsinized ORh+ red cells, of the IgG incomplete anti-Rh antibody and its F(ab')2 and Fc fragments has been investigated. An analysis has also been made of the capacity of non-specific human IgG, aggregated non-specific human IgG, human IgM (19S) and IgM (7S), and of fragments Fcgamma, Fcmu and Fc5mu to inhibit the agglutination of trypsinized ORh+ red cells by the IgG incomplete anti-Rh antibody. The results obtained indicate that these antibodies behave in a similar manner to that of nonprecipitating antibodies, and that the agglutination of trypsinized red cells seems to be a mixed reaction due to the interaction of an Fab fragment with its Rh antigenic determinant present in the surface of a red cell and the Fc of the same molecule with a receptor for Fc present in adjacent red cells. The trypsin treatment apparently results in the liberation of occult Fc receptors. It has also been demonstrated that in the agglutination of ORh+ red cells by IgG incomplete anti-Rh antibody in the presence of albumin, interaction must occur in some manner between the albumin and the Fc fragment since the F(ab')2 fragment does not give rise to agglutination under such conditions. Images Figure 1 PMID:415968
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Cario, Elke; Golenbock, Douglas T; Visintin, Alberto; Rünzi, Michael; Gerken, Guido; Podolsky, Daniel K
2006-04-01
Intestinal epithelial cells (IEC) are constantly exposed to both high concentrations of the bacterial ligand LPS and the serine protease trypsin. MD-2, which contains multiple trypsin cleavage sites, is an essential accessory glycoprotein required for LPS recognition and signaling through TLR4. The aim of this study was to characterize the expression and subcellular distribution of intestinal epithelial MD-2 and to delineate potential functional interactions with trypsin and then alteration in inflammatory bowel disease (IBD). Although MD-2 protein expression was minimal in primary IEC of normal colonic or ileal mucosa, expression was significantly increased in IEC from patients with active IBD colitis, but not in ileal areas from patients with severe Crohn's disease. Endogenous MD-2 was predominantly retained in the calnexin-calreticulin cycle of the endoplasmic reticulum; only a small fraction was exported to the Golgi. MD-2 expression correlated inversely with trypsin activity. Biochemical evidence and in vitro experiments demonstrated that trypsin exposure resulted in extensive proteolysis of endogenous and soluble MD-2 protein, but not of TLR4 in IEC, and was associated with desensitization of IEC to LPS. In conclusion, the present study suggests that endoplasmic reticulum-associated MD-2 expression in IBD may be altered by ileal protease in inflammation, leading to impaired LPS recognition and hyporesponsiveness through MD-2 proteolysis in IEC, thus implying a physiologic mechanism that helps maintain LPS tolerance in the intestine.
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Moulin, M M; Rodrigues, R; Ribeiro, S F F; Gonçalves, L S A; Bento, C S; Sudré, C P; Vasconcelos, I M; Gomes, V M
2014-11-07
Several plant organs contain proteinase inhibitors, which are produced during normal plant development or are induced upon pathogen attack to suppress the enzymatic activity of phytopathogenic microorganisms. In this study, we examined the presence of proteinase inhibitors, specifically trypsin inhibitors, in the leaf extract of Capsicum baccatum var. pendulum inoculated with PepYMV (Pepper yellow mosaic virus). Leaf extract from plants with the accession number UENF 1624, which is resistant to PepYMV, was collected at 7 different times (0, 24, 48, 72, 96, 120, and 144 h). Seedlings inoculated with PepYMV and control seedlings were grown in a growth chamber. Protein extract from leaf samples was partially purified by reversed-phase chromatography using a C2/C18 column. Residual trypsin activity was assayed to detect inhibitors followed by Tricine-SDS-PAGE analysis to determine the N-terminal peptide sequence. Based on trypsin inhibitor assays, trypsin inhibitors are likely constitutively synthesized in C. baccatum var. pendulum leaf tissue. These inhibitors are likely a defense mechanism for the C. baccatum var. pendulum- PepYMV pathosystem.
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Quantitative proteomics reveals the kinetics of trypsin-catalyzed protein digestion.
Pan, Yanbo; Cheng, Kai; Mao, Jiawei; Liu, Fangjie; Liu, Jing; Ye, Mingliang; Zou, Hanfa
2014-10-01
Trypsin is the popular protease to digest proteins into peptides in shotgun proteomics, but few studies have attempted to systematically investigate the kinetics of trypsin-catalyzed protein digestion in proteome samples. In this study, we applied quantitative proteomics via triplex stable isotope dimethyl labeling to investigate the kinetics of trypsin-catalyzed cleavage. It was found that trypsin cleaves the C-terminal to lysine (K) and arginine (R) residues with higher rates for R. And the cleavage sites surrounded by neutral residues could be quickly cut, while those with neighboring charged residues (D/E/K/R) or proline residue (P) could be slowly cut. In a proteome sample, a huge number of proteins with different physical chemical properties coexists. If any type of protein could be preferably digested, then limited digestion could be applied to reduce the sample complexity. However, we found that protein abundance and other physicochemical properties, such as molecular weight (Mw), grand average of hydropathicity (GRAVY), aliphatic index, and isoelectric point (pI) have no notable correlation with digestion priority of proteins.
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Wang, Yi; Zhou, Lele; Kang, Qi; Yu, Li
2018-06-01
Trypsin plays a central role in catalyzing the hydrolysis of peptide bonds, so a technique with simple operation is needed to monitor the activity of trypsin. Here a simple and label-free senor based on liquid crystals (LCs) was developed by employing bovine serum albumin (BSA) as the enzyme substrate and dodecyl trimethyl ammonium bromide (DTAB) as the controller for the alignment of LC. It was found that DTAB could form a self-assembled monolayer at the aqueous/LC interface to produce the dark optical images of LCs. And the addition of BSA could disturb the monolayer, so that the optical signal of LCs turned bright from dark. But the hydrolysis of BSA by trypsin resulted in the dark appearance. The sensing platform allows detection as low as 1 U/mL under the polarized light microscope based on at least three measurements. Moreover, this method was successfully applied in the detection of trypsin in human urines, suggesting its potential applications in clinic diagnosis. Copyright © 2018 Elsevier B.V. All rights reserved.
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Slack, Jeffrey M; Lawrence, Susan D; Krell, Peter J; Arif, Basil M
2008-10-01
Baculovirus occlusion-derived virions (ODVs) contain a number of infectivity factors essential for the initiation of infection in larval midgut cells. Deletion of any of these factors neutralizes infectivity by the per os route. We have observed that P74 of the group I alphabaculovirus Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is N-terminally cleaved when a soluble form of the protein was incubated with insect midgut tissues under alkaline conditions and that cleavage was prevented by soybean trypsin inhibitor (SBTI). Presently, biological assays were carried out that suggest SBTI inhibits and trypsin enhances baculovirus per os infectivity. We developed a method to rescue per os infectivity of a P74 null virus involving co-transfection of viral DNA with a plasmid that transiently expresses p74. We used this plasmid rescue method to functionally characterize P74. A series of site-directed mutants were generated at the N terminus to evaluate if trypsin cleavage sites were necessary for function. Mutagenesis of R195, R196 and R199 compromised per os infectivity and rendered P74 resistant to midgut trypsin.
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He, Ying-Ying; Liu, Shu-Bai; Lee, Wen-Hui; Qian, Jin-Qiao; Zhang, Yun
2008-10-01
Snake venom Kunitz/BPTI members are good tools for understanding of structure-functional relationship between serine proteases and their inhibitors. A novel dual Kunitz/BPTI serine proteinase inhibitor named OH-TCI (trypsin- and chymotrypsin-dual inhibitor from Ophiophagus hannah) was isolated from king cobra venom by three chromatographic steps of gel filtration, trypsin affinity and reverse phase HPLC. OH-TCI is composed of 58 amino acid residues with a molecular mass of 6339Da. Successful expression of OH-TCI was performed as the maltose-binding fusion protein in E. coli DH5alpha. Much different from Oh11-1, the purified native and recombinant OH-TCI both had strong inhibitory activities against trypsin and chymotrypsin although the sequence identity (74.1%) between them is very high. The inhibitor constants (K(i)) of recombinant OH-TCI were 3.91 x 10(-7) and 8.46 x10(-8)M for trypsin and chymotrypsin, respectively. To our knowledge, it was the first report of Kunitz/BPTI serine proteinase inhibitor from snake venom that had equivalent trypsin and chymotrypsin inhibitory activities.
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Isolation and characterization of the chicken trypsinogen gene family.
Wang, K; Gan, L; Lee, I; Hood, L
1995-01-01
Based on genomic Southern hybridizations and cDNA sequence analyses, the chicken trypsinogen gene family can be divided into two multi-member subfamilies, a six-member trypsinogen I subfamily which encodes the cationic trypsin isoenzymes and a three-member trypsinogen II subfamily which encodes the anionic trypsin isoenzymes. The chicken cDNA and genomic clones containing these two subfamilies were isolated and characterized by DNA sequence analysis. The results indicated that the chicken trypsinogen genes encoded a signal peptide of 15 to 16 amino acid residues, an activation peptide of 9 to 10 residues and a trypsin of 223 amino acid residues. The chicken trypsinogens contain all the common catalytic and structural features for trypsins, including the catalytic triad His, Asp and Ser and the six disulphide bonds. The trypsinogen I and II subfamilies share approximately 70% sequence identity at the nucleotide and amino acid level. The sequence comparison among chicken trypsinogen subfamily members and trypsin sequences from other species suggested that the chicken trypsinogen genes may have evolved in coincidental or concerted fashion. Images Figure 6 Figure 7 PMID:7733885
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NASA Astrophysics Data System (ADS)
Marquis, Jared Wayne
Passive longwave infrared radiometric satellite-based retrievals of sea surface temperature (SST) at instrument nadir are investigated for cold bias caused by unscreened optically-thin cirrus (OTC) clouds (cloud optical depth ≤ 0.3; COD). Level 2 split-window SST retrievals over tropical oceans (30° S - 30° N) from Moderate Resolution Imaging Spectroradiometer (MODIS) radiances collected aboard the NASA Aqua satellite (Aqua-MODIS) are collocated with cloud profiles from the Cloud-Aerosol Lidar with Orthogonal Polarization (CALIOP) instrument, mounted on the independent NASA CALIPSO satellite. OTC are present in approximately 25% of tropical quality-assured (QA) Aqua-MODIS Level-2 data, representing over 99% of all contaminating cirrus found. This results in cold-biased SST retrievals using either split- (MODIS, AVHRR and VIIRS) or triple-window (AVHRR and VIIRS only) retrieval methods. SST retrievals are modeled based on operational algorithms using radiative transfer model simulations conducted with a hypothetical 1.5 km thick OTC cloud placed incrementally from 10.0 - 18.0 km above mean sea level for cloud optical depths (COD) between 0.0 - 0.3. Corresponding cold bias estimates for each sensor are estimated using relative Aqua-MODIS cloud contamination frequencies as a function of cloud top height and COD (assuming them consistent across each platform) integrated within each corresponding modeled cold bias matrix. Split-window relative OTC cold biases, for any single observation, range from 0.40° - 0.49° C for the three sensors, with an absolute (bulk mean) bias between 0.10° - 0.13° C. Triple-window retrievals are more resilient, ranging from 0.03° - 0.04° C relative and 0.11° - 0.16° C absolute. Cold biases are constant across the Pacific and Indian Ocean domains. Absolute bias is smaller over the Atlantic, but relative bias is larger due to different cloud properties indicating that this issue persists globally.
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Banar, Maryam; Emaneini, Mohammad; Satarzadeh, Mhboubeh; Abdellahi, Nafiseh; Beigverdi, Reza; Leeuwen, Willem B van; Jabalameli, Fereshteh
2016-01-01
Biofilm is an important virulence factor in Pseudomonas aeruginosa and has a substantial role in antibiotic resistance and chronic burn wound infections. New therapeutic agents against P. aeruginosa, degrading biofilms in burn wounds and improving the efficacy of current antimicrobial agents, are required. In this study, the effects of α-mannosidase, β-mannosidase and trypsin enzymes on the degradation of P. aeruginosa biofilms and on the reduction of ceftazidime minimum biofilm eliminating concentrations (MBEC) were evaluated. All tested enzymes, destroyed the biofilms and reduced the ceftazidime MBECs. However, only trypsin had no cytotoxic effect on A-431 human epidermoid carcinoma cell lines. In conclusion, since trypsin had better features than mannosidase enzymes, it can be a promising agent in combatting P. aeruginosa burn wound infections.
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Effect of White Charcoal on COD Reduction in Wastewater Treatment
NASA Astrophysics Data System (ADS)
Pijarn, Nuchanaporn; Butsee, Manipa; Buakul, Kanokwan; Seng, Hasan; Sribuarai, Tinnphat; Phonprasert, Pongtep; Taneeto, Kla; Atthameth, Prasertsil
2017-06-01
The objective of this study is to compare the COD reduction in wastewater between using coconut shell and coconut spathe white charcoal from Khlong Wat NongPra-Ong, Krathumbaen, SamutSakhon province, Thailand. The waste water samples were collected using composite sampling method. The experimental section can be divided into 2 parts. The first part was study the optimum of COD adsorption time using both white charcoals. The second part was study the optimum amount of white charcoal for chemical oxygen demand (COD) reduction. The pre-treatment of wastewater was examined in parameters include temperature, alkalinity (pH), conductivity, turbidity, suspended solid (SS), total dissolved solid (TDS), and COD. The results show that both white charcoals can reduce COD of wastewater. The pH of pre-treatment wastewater had pH 9 but post-treatment wastewaters using both white charcoals have pH 8. The COD of pre-treatment wastewater had COD as 258 mg/L but post-treatment wastewater using coconut shell white charcoal had COD steady at 40 mg/L in 30 min and the amount of white charcoals 4 g. The COD of post-treatment wastewater using coconut spathe white charcoal had COD steady at 71 mg/L in 30 min and the amount of white charcoals 4 g. Therefore comparison of COD reduction between coconut shell white charcoal versus coconut spathe white charcoal found that the coconut shell white charcoal had efficiency for COD reduction better than coconut spathe white charcoal.
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STRANDEDNESS OF VICIA FABA CHROMOSOMES AS REVEALED BY ENZYME DIGESTION STUDIES
Trosko, James E.; Wolff, Sheldon
1965-01-01
Chromosomes and nuclei isolated from neutral formalin-fixed Vicia faba lateral roots were treated with trypsin, pepsin, RNase, or DNase. Only trypsin affected the morphology of the chromosomes and nuclei. The appearance of the chromosomes after trypsin digestion indicated that each chromatid contained four strands that could be seen with an ordinary light microscope. The experiments are interpreted as indicating that mitotic chromosomes of Vicia faba are multistranded and that the linear continuity of the chromosome is dependent on protein. PMID:5323605
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Zagatto, Alessandro M; Ardigò, Luca P; Barbieri, Fabio A; Milioni, Fabio; Dello Iacono, Antonio; Camargo, Bruno H F; Padulo, Johnny
2017-09-01
Zagatto, AM, Ardigò, LP, Barbieri, FA, Milioni, F, Dello Iacono, A, Camargo, BHF, and Padulo, J. Performance and metabolic demand of a new repeated-sprint ability test in basketball players: does the number of changes of direction matter? J Strength Cond Res 31(9): 2438-2446, 2017-This study compared 2 repeated-sprint ability (RSA) tests in basketball players. Both tests included 10 × 30-m sprints, with the difference that the previously validated test (RSA2COD) featured 2 changes of direction (COD) per sprint, whereas the experimental test (RSA5COD) featured 5 CODs per sprint. Test performances and metabolic demands were specifically assessed in 20 basketball players. First, RSA5COD test-retest reliability was investigated. Then, RSA2COD, RSA5COD sprint times, peak speeds, oxygen uptake (V[Combining Dot Above]O2) and posttest blood lactate concentration [La] were measured. The RSA5COD results showed to be reliable. RSA2COD performance resulted better than the RSA5COD version (p < 0.01), with shorter sprint times and higher peak speeds. Over sprints, the tests did not differ from each other in terms of V[Combining Dot Above]O2 (p > 0.05). Over whole bout, the RSA2COD was more demanding than the RSA5COD, considering overall metabolic power requirement (i.e., VO2-driven + [La]-driven components). Given that RSA5COD (a) mimics real game-play as sprint distance and action change frequency/direction and (b) has the same metabolic expenditure per task completion as metabolic cost, RSA5COD is a valuable option for players and coaches for training basketball-specific agility and assessing bioenergetic demands.
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The CodY regulator is essential for virulence in Streptococcus suis serotype 2
Feng, Liping; Zhu, Jiawen; Chang, Haitao; Gao, Xiaoping; Gao, Cheng; Wei, Xiaofeng; Yuan, Fangyan; Bei, Weicheng
2016-01-01
The main role of CodY, a global regulatory protein in most low G + C gram-positive bacteria, is in transcriptional repression. To study the functions of CodY in Streptococcus suis serotype 2 (S. suis 2), a mutant codY clone named ∆codY was constructed to explore the phenotypic variation between ∆codY and the wild-type strain. The result showed that the codY mutation significantly inhibited cell growth, adherence and invasion ability of S. suis 2 to HEp-2 cells. The codY mutation led to decreased binding of the pathogen to the host cells, easier clearance by RAW264.7 macrophages and decreased growth ability in fresh blood of Cavia porcellus. The codY mutation also attenuated the virulence of S. suis 2 in BALB/c mice. Morphological analysis revealed that the codY mutation decreased the thickness of the capsule of S. suis 2 and changed the surface structures analylized by SDS-PAGE. Finally, the codY mutation altered the expressions of many virulence related genes, including sialic acid synthesis genes, leading to a decreased sialic acid content in capsule. Overall, mutation of codY modulated bacterial virulence by affecting the growth and colonization of S. suis 2, and at least via regulating sialic acid synthesis and capsule thickness. PMID:26883762
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-04-16
... DEPARTMENT OF THE INTERIOR National Park Service [1730-SZM] Cape Cod National Seashore Advisory Commission; Cape Cod National Seashore, South Wellfleet, MA AGENCY: National Park Service, Interior. ACTION... Cod National Seashore, 99 Marconi Site Road, Wellfleet, MA 02667. Dated: April 9, 2012. George E...
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Anaerobic treatment of landfill leachate by sulfate reduction.
Henry, J G; Prasad, D
2000-01-01
The present study was conducted to investigate the effectiveness of the sulphate-reduction pathway in the anaerobic treatment of landfill leachate. The effects of several COD/SO4 ratios (keeping COD constant) and loadings on anaerobic filter performance were studied and compared with the results from anaerobic filters which followed the methanogenic pathway. Results indicated that the treatability of leachate by sulphate reducing bacteria (SRB) was dependent upon the leachate strength. With high strength leachate (COD = 15,000 mg/L) from the Keele Valley Landfill, it was found that at lower COD/SO4 ratios (< or = 1.6) toxic conditions developed in the system that were more inhibitory to the SRB than to the methane producing bacteria (MPB). As the COD/SO4 ratio increased, methanogenesis predominated. No predominance of SRB occurred at any COD/SO4 ratio with high strength leachate. The highest COD removal achieved was about 70% of which 20% was accomplished by the SRB at a COD/SO4 ratio of 1.6 and an organic loading rate (OLR) of 4 kg COD/m3.d. With low strength leachate (COD = 1500-3300 mg/L) from the Brock West Landfill, and a COD/SO4 ratio < or = 1, SRB became predominant. In these anaerobic filters in which SRB were predominant, the SRB reduced the COD as well as the MPB could. Sulphide inhibition did not take place at any loading in units treating low strength leachate. Consequently, both SRB and MPB should function at COD/SO4 ratios between 1 and 3. About 60% COD removal was achieved at a loading of 2.8 kg COD/m3.d and a COD/SO4 ratio of 1.0. However at a loading of 6 kg COD/m3.d only 27% COD removal was achieved, all of it through the sulphate-reduction pathway. These OLR values are comparable to those applied in systems where methanogenesis was dominant. It was also observed that once the methanogens were established in the units, it was not possible to displace them completely. However, where methanogenesis had not been previously established, it was found that sulphate-reduction could be the sole pathway for COD removal. From this study, it can be concluded that there is no advantage to the sulphate-reduction pathway in the anaerobic treatment of landfill leachate. The other options for increasing the loadings, i.e. the use of high surface/volume filter media (to achieve higher biomass concentrations) or high rate systems are likely to be more successful.
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COD::CIF::Parser: an error-correcting CIF parser for the Perl language.
Merkys, Andrius; Vaitkus, Antanas; Butkus, Justas; Okulič-Kazarinas, Mykolas; Kairys, Visvaldas; Gražulis, Saulius
2016-02-01
A syntax-correcting CIF parser, COD::CIF::Parser , is presented that can parse CIF 1.1 files and accurately report the position and the nature of the discovered syntactic problems. In addition, the parser is able to automatically fix the most common and the most obvious syntactic deficiencies of the input files. Bindings for Perl, C and Python programming environments are available. Based on COD::CIF::Parser , the cod-tools package for manipulating the CIFs in the Crystallography Open Database (COD) has been developed. The cod-tools package has been successfully used for continuous updates of the data in the automated COD data deposition pipeline, and to check the validity of COD data against the IUCr data validation guidelines. The performance, capabilities and applications of different parsers are compared.
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A bioactive film based on cashew gum polysaccharide for wound dressing applications.
Moreira, Bruna R; Batista, Karla A; Castro, Elisandra G; Lima, Eliana M; Fernandes, Kátia F
2015-05-20
This work presents the development of a new bioactive material for wound therapeutics which may play a dual role of modulate metallo proteinases activity while prevents infection blocking out pathogenic microorganisms and foreign materials. A CGP/PVA film was activated by covalent immobilization of trypsin. Results from biocompatibility test revealed that PDL fibroblasts grown on the surface of CGP/PVA and the high amount of viable cells proved absence of cytotoxicity. Trypsin immobilized onto CGP/PVA film remained 100% active after 28 days stored dried at room temperature. In addition, CGP/PVA-trypsin film could be used for 9 cycles of storage/use without loss of activity. After immobilization, trypsin retained its collagenolytic activity, indicating this material as a promising material for wound dressing applications. Copyright © 2015 Elsevier Ltd. All rights reserved.
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Jennings, Robert Charles; Forti, Giorgio; Gerola, Paolo Domenico; Garlaschi, Flavio Massimo
1978-01-01
Trypsin digestion of photosynthetic membranes isolated from spinach (Spinacia oleracea L.) leaves eliminates the cation stimulation of chlorophyll fluorescence. High concentrations of cations protect the fluorescence yield against trypsin digestion, and the cation specificity for this protection closely resembles that required for the stimulation of fluorescence by cations. Trypsin digestion reverses cation-induced thylakoid stacking, and the time course of this effect seems to parallel that of the reversal of cation fluorescence. High concentrations of cations protect thylakoid stacking and cation-stimulated fluorescence alike. The cation stimulation of photosytem II photochemistry remains intact after trypsinization has reversed both cation-induced thylakoid stacking and fluorescence yield. It is concluded that cation-stimulated fluorescence yield, and not the cation stimulation of photosystem II photochemistry, is associated with thylakoid membrane stacking. ImagesFig. 2Fig. 3 PMID:16660630
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Rai, R; Katzenellenbogen, J A
1992-11-13
Previously, we have reported that some guanidino-substituted alpha- and beta-aryl enol lactones I and II behaved as selective, mechanism-based inhibitors of some trypsin-like proteases (Rai, R.; Katzenellenbogen, J.A. J. Med. Chem., submitted). In this study, we describe the synthesis and kinetic evaluation of some related, guanidino-substituted enol lactones having greater conformational mobility and affording additional hydrogen-bonding sites at the active site. The alpha-aryl-substituted lactones 1 and 2, which have greater conformational mobility in the guanidinoaryl linkage than I, selectively inhibited the trypsin-like enzymes, and they were relatively poor inactivators of alpha-chymotrypsin and human neutrophil elastase (HNE). The iodo enol lactone 2 permanently inactivated trypsin, urokinase, tissue plasminogen activator, and plasmin, showing exceptionally high specificity in its interaction with trypsin and urokinase. The selectivity pattern exhibited by the closely related, conformationally less mobile alpha-aryl-substituted iodo lactone Ib, which was previously shown to be a selective suicide substrate of urokinase and plasmin, provides an interesting comparison. The alpha-benzamido-substituted lactones 3 and 4, which afford an additional site for active-site hydrogen bonding, were found to be very potent alternate substrate inhibitors of trypsin and urokinase. In addition, the iodo lactone 4 permanently inactivated alpha-chymotrypsin. The importance of secondary interactions in increasing the specificities in the case of alpha-chymotrypsin is discussed.
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Pandey, Prabhash K; Jamal, Farrukh
2014-11-01
A trypsin inhibitor purified from the seeds of Tamarindus indica by Sephadex G-75, DEAE-Sepharose and Trypsin-Sepharose CL-4B columns was studied for its antifeedant, larvicidal, pupicidal and growth inhibitory activities against Helicoverpa armigera larvae. Tamarindus trypsin inhibitor (TTI) exhibited inhibitory activity towards total gut proteolytic enzymes of H. armigera (~87%) and bovine trypsin (~84%). Lethal doses which caused mortality and weight reduction by 50% were 1% w/w and 0.50% w/w, respectively. IC50 of TTI against Helicoverpa midgut proteases and bovine trypsin were ~2.10 µg/ml and 1.68 µg/ml respectively. In larval feeding studies the 21 kDa Kunitz-type protein was found to retard growth and development, prolonged the larval-pupal development durations along with adversely affecting the fertility and fecundity of H. armigera. In artificial diet at 0.5% w/w TTI, the efficiency of conversion of ingested food as well as of digested food, relative growth rate, growth index declined whereas approximate digestibility, metabolic cost, relative consumption rate, consumption index and total developmental period enhanced for H. armigera larvae. These results suggest that TTI has toxic and adverse effect on the developmental physiology of H. armigera and could be useful in controlling the pest H. armigera. Copyright © 2014 Elsevier Inc. All rights reserved.
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Expanding proteome coverage with orthogonal-specificity α-Lytic proteases
DOE Office of Scientific and Technical Information (OSTI.GOV)
Meyer, Jesse G.; Kim, Sangtae; Maltby, David A.
2014-03-01
Bottom-up proteomics studies traditionally involve proteome digestion with a single protease, trypsin. However, trypsin alone does not generate peptides that encompass the entire proteome. Alternative proteases have been explored, but most have specificity for charged amino acid side chains. Therefore, additional proteases that improve proteome coverage by cleavage at sequences complimentary to trypsin may increase proteome coverage. We demonstrate the novel application of two proteases for bottom-up proteomics: wild type alpha-lytic protease (WaLP), and an active site mutant of WaLP, M190A alpha-lytic protease (MaLP). We assess several relevant factors including MS/MS fragmentation, peptide length, peptide yield, and protease specificity. Bymore » combining data from separate digestions with trypsin, LysC, WaLP, and MaLP, proteome coverage was increased 101% compared to trypsin digestion alone. To demonstrate how the gained sequence coverage can access additional PTM information, we show identification of a number of novel phosphorylation sites in the S. pombe proteome and include an illustrative example from the protein MPD2, wherein two novel sites are identified, one in a tryptic peptide too short to identify and the other in a sequence devoid of tryptic sites. The specificity of WaLP and MaLP for aliphatic amino acid side chains was particularly valuable for coverage of membrane protein sequences, which increased 350% when the data from trypsin, LysC, WaLP, and MaLP were combined.« less
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CodY-Dependent Regulation of Sporulation in Clostridium difficile.
Nawrocki, Kathryn L; Edwards, Adrianne N; Daou, Nadine; Bouillaut, Laurent; McBride, Shonna M
2016-08-01
Clostridium difficile must form a spore to survive outside the gastrointestinal tract. The factors that trigger sporulation in C. difficile remain poorly understood. Previous studies have suggested that a link exists between nutritional status and sporulation initiation in C. difficile In this study, we investigated the impact of the global nutritional regulator CodY on sporulation in C. difficile strains from the historical 012 ribotype and the current epidemic 027 ribotype. Sporulation frequencies were increased in both backgrounds, demonstrating that CodY represses sporulation in C. difficile The 027 codY mutant exhibited a greater increase in spore formation than the 012 codY mutant. To determine the role of CodY in the observed sporulation phenotypes, we examined several factors that are known to influence sporulation in C. difficile Using transcriptional reporter fusions and quantitative reverse transcription-PCR (qRT-PCR) analysis, we found that two loci associated with the initiation of sporulation, opp and sinR, are regulated by CodY. The data demonstrate that CodY is a repressor of sporulation in C. difficile and that the impact of CodY on sporulation and expression of specific genes is significantly influenced by the strain background. These results suggest that the variability of CodY-dependent regulation is an important contributor to virulence and sporulation in current epidemic isolates. This report provides further evidence that nutritional state, virulence, and sporulation are linked in C. difficile This study sought to examine the relationship between nutrition and sporulation in C. difficile by examining the global nutritional regulator CodY. CodY is a known virulence and nutritional regulator of C. difficile, but its role in sporulation was unknown. Here, we demonstrate that CodY is a negative regulator of sporulation in two different ribotypes of C. difficile We also demonstrate that CodY regulates known effectors of sporulation, Opp and SinR. These results support the idea that nutrient limitation is a trigger for sporulation in C. difficile and that the response to nutrient limitation is coordinated by CodY. Additionally, we demonstrate that CodY has an altered role in sporulation regulation for some strains. Copyright © 2016, American Society for Microbiology. All Rights Reserved.
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CodY-Dependent Regulation of Sporulation in Clostridium difficile
Nawrocki, Kathryn L.; Edwards, Adrianne N.; Daou, Nadine; Bouillaut, Laurent
2016-01-01
ABSTRACT Clostridium difficile must form a spore to survive outside the gastrointestinal tract. The factors that trigger sporulation in C. difficile remain poorly understood. Previous studies have suggested that a link exists between nutritional status and sporulation initiation in C. difficile. In this study, we investigated the impact of the global nutritional regulator CodY on sporulation in C. difficile strains from the historical 012 ribotype and the current epidemic 027 ribotype. Sporulation frequencies were increased in both backgrounds, demonstrating that CodY represses sporulation in C. difficile. The 027 codY mutant exhibited a greater increase in spore formation than the 012 codY mutant. To determine the role of CodY in the observed sporulation phenotypes, we examined several factors that are known to influence sporulation in C. difficile. Using transcriptional reporter fusions and quantitative reverse transcription-PCR (qRT-PCR) analysis, we found that two loci associated with the initiation of sporulation, opp and sinR, are regulated by CodY. The data demonstrate that CodY is a repressor of sporulation in C. difficile and that the impact of CodY on sporulation and expression of specific genes is significantly influenced by the strain background. These results suggest that the variability of CodY-dependent regulation is an important contributor to virulence and sporulation in current epidemic isolates. This report provides further evidence that nutritional state, virulence, and sporulation are linked in C. difficile. IMPORTANCE This study sought to examine the relationship between nutrition and sporulation in C. difficile by examining the global nutritional regulator CodY. CodY is a known virulence and nutritional regulator of C. difficile, but its role in sporulation was unknown. Here, we demonstrate that CodY is a negative regulator of sporulation in two different ribotypes of C. difficile. We also demonstrate that CodY regulates known effectors of sporulation, Opp and SinR. These results support the idea that nutrient limitation is a trigger for sporulation in C. difficile and that the response to nutrient limitation is coordinated by CodY. Additionally, we demonstrate that CodY has an altered role in sporulation regulation for some strains. PMID:27246573
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Dubber, Donata; Gray, Nicholas F
2010-10-01
Chemical oxygen demand (COD) is widely used for wastewater monitoring, design, modeling and plant operational analysis. However this method results in the production of hazardous wastes including mercury and hexavalent chromium. The study examined the replacement of COD with total organic carbon (TOC) for general performance monitoring by comparing their relationship with influent and effluent samples from 11 wastewater treatment plants. Biochemical oxygen demand (BOD5) was also included in the comparison as a control. The results show significant linear relationships between TOC, COD and BOD5 in settled (influent) domestic and municipal wastewaters, but only between COD and TOC in treated effluents. The study concludes that TOC can be reliably used for the generic replacement of both COD (COD=49.2+3.00*TOC) and BOD5 (BOD5=23.7+1.68*TOC) in influent wastewaters but only for COD (COD=7.25+2.99*TOC) in final effluents.
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Sai, Wenbo; Tian, Hong; Yang, Kangmin; Tang, Daoqi; Bao, Jinxiao; Ge, Yang; Song, Xiaoda; Zhang, Yu; Luo, Cheng; Gao, Xiangdong; Yao, Wenbing
2017-03-08
Exendin-4 is a strong therapeutic candidate for the treatment of metabolic syndrome. Related receptor agonist drugs have been on the market since 2005. However, technical limitations and the pain caused by subcutaneous injection have severely limited patient compliance. The goal of the study is to investigate a biologically active exendin-4 analog could be administered orally. Using intraperitoneal glucose tolerance tests, we discovered that exendin4-cysteine administered by oral gavage had a distinct hypoglycemic effect in C57BL/6J mice. Using Rosetta Design and Amber, we designed and screened a series of exendin4-cysteine analogs to identify those that retained biological activity while resisting trypsin digestion. Trypsin Cleavage Site Mutated Exendin4-cysteine 1 (TSME-1), an analog whose bioactivity was similar to exendin-4 and was almost completely resistant to trypsin, was screened out. In addition, TSME-1 significantly normalized the blood glucose levels and the availability of TSME-1 was significantly higher than that of exendin-4 and exendin4-cysteine. Collectively orally administered TSME-1, a trypsin-resistant exendin-4 analog obtained by the system, is a strong candidate for future treatments of type 2 diabetes.
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Pazhang, Mohammad; Mehrnejad, Faramarz; Pazhang, Yaghub; Falahati, Hanieh; Chaparzadeh, Nader
2016-01-01
The effect of glycerol and sorbitol on the stability of porcine pancreas trypsin was investigated in this work. Molecular dynamics simulation and thermostability results showed that trypsin has two flexible regions, and polyols (sorbitol and glycerol) stabilize the enzyme by decreasing the flexibility of these regions. Radial distribution function results exhibited that sorbitol and glycerol were excluded from the first water layer of the enzyme, therefore decrease the flexibility of the regions by preferential exclusion. Also, results showed that the stabilization effect of sorbitol is more than glycerol. This observation could be because of the larger decrease in the fluctuations of trypsin in the presence of sorbitol. We also examined the role of solvent's hydrophobicity in enzyme stabilization by sorbitol and glycerol. To do so, the thermostability of trypsin was evaluated in the presence of solvents with different hydrophobicity (methanol, ethanol, isopropanol and n-propanol) in addition to the polyols. Our results depicted that glycerol is a better stabilizer than sorbitol in the presence of hydrophobic solvents (n-propanol), whereas sorbitol is a better stabilizer than glycerol in the presence of hydrophilic solvents (methanol). © 2015 International Union of Biochemistry and Molecular Biology, Inc.
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Pawlak, J; Nadolna-Ałtyn, K; Szostakowska, B; Pachur, M; Podolska, M
2017-10-12
The parasite fauna of cod (Gadus morhus) is well described, but the life cycles of Baltic cod parasites are known only in general terms. Invertebrates commonly found in the stomach of cod are recognized as intermediate hosts in the life cycles of nematodes or acanthocephalans. The aim of this study was to determine the source of infection of Baltic cod with parasites found in situ in invertebrates present in the cod stomach. Our results indicate that Saduria entomon is both a source of infection of Baltic cod with parasites and an intermediate host in the life cycle of Hysterothylacium aduncum in the Baltic Sea.
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Application of microbial electrolysis cells to treat spent yeast from an alcoholic fermentation.
Sosa-Hernández, Ornella; Popat, Sudeep C; Parameswaran, Prathap; Alemán-Nava, Gibrán Sidney; Torres, César I; Buitrón, Germán; Parra-Saldívar, Roberto
2016-01-01
Spent yeast (SY), a major challenge for the brewing industry, was treated using a microbial electrolysis cell to recover energy. Concentrations of SY from bench alcoholic fermentation and ethanol were tested, ranging from 750 to 1500mgCOD/L and 0 to 2400mgCOD/L respectively. COD removal efficiency (RE), coulombic efficiency (CE), coulombic recovery (CR), hydrogen production and current density were evaluated. The best treatment condition was 750mgCOD/LSY+1200mgCOD/L ethanol giving higher COD RE, CE, CR (90±1%, 90±2% and 81±1% respectively), as compared with 1500mgCOD/LSY (76±2%, 63±7% and 48±4% respectively); ethanol addition was significantly favorable (p value=0.011), possibly due to electron availability and SY autolysis. 1500mgCOD/LSY+1200mgCOD/L ethanol achieved higher current density (222.0±31.3A/m(3)) and hydrogen production (2.18±0.66 [Formula: see text] ) but with lower efficiencies (87±2% COD RE, 71.0±.4% CE). Future work should focus on electron sinks, acclimation and optimizing SY breakdown. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.
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Fox, C J; Taylor, M I; Pereyra, R; Villasana, M I; Rico, C
2005-03-01
Recent substantial declines in northeastern Atlantic cod stocks necessitate improved biological knowledge and the development of techniques to complement standard stock assessment methods (which largely depend on accurate commercial catch data). In 2003, an ichthyoplankton survey was undertaken in the Irish Sea and subsamples of 'cod-like' eggs were analysed using a TaqMan multiplex, PCR (polymerase chain reaction) assay (with specific probes for cod, haddock and whiting). The TaqMan method was readily applied to the large number of samples (n = 2770) generated during the survey and when combined with a manual DNA extraction protocol had a low failure rate of 6%. Of the early stage 'cod-like' eggs (1.2-1.75 mm diameter) positively identified: 34% were cod, 8% haddock and 58% whiting. As previous stock estimates based on egg surveys for Irish Sea cod assumed that the majority of 'cod-like' eggs were from cod, the TaqMan results confirm that there was probably substantial contamination by eggs of whiting and haddock that would have inflated estimates of the stock biomass.
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Who’s your mama? Riverine hybridisation of threatened freshwater Trout Cod and Murray Cod
Unmack, Peter J.; Dyer, Fiona J.; Lintermans, Mark
2016-01-01
Rates of hybridization and introgression are increasing dramatically worldwide because of translocations, restocking of organisms and habitat modifications; thus, determining whether hybridization is occuring after reintroducing extirpated congeneric species is commensurately important for conservation. Restocking programs are sometimes criticized because of the genetic consequences of hatchery-bred fish breeding with wild populations. These concerns are important to conservation restocking programs, including those from the Australian freshwater fish family, Percichthyidae. Two of the better known Australian Percichthyidae are the Murray Cod, Maccullochella peelii and Trout Cod, Maccullochella macquariensis which were formerly widespread over the Murray Darling Basin. In much of the Murrumbidgee River, Trout Cod and Murray Cod were sympatric until the late 1970s when Trout Cod were extirpated. Here we use genetic single nucleotide polymorphism (SNP) data together with mitochondrial sequences to examine hybridization and introgression between Murray Cod and Trout Cod in the upper Murrumbidgee River and consider implications for restocking programs. We have confirmed restocked riverine Trout Cod reproducing, but only as inter-specific matings, in the wild. We detected hybrid Trout Cod–Murray Cod in the Upper Murrumbidgee, recording the first hybrid larvae in the wild. Although hybrid larvae, juveniles and adults have been recorded in hatcheries and impoundments, and hybrid adults have been recorded in rivers previously, this is the first time fertile F1 have been recorded in a wild riverine population. The F1 backcrosses with Murray cod have also been found to be fertile. All backcrosses noted were with pure Murray Cod. Such introgression has not been recorded previously in these two species, and the imbalance in hybridization direction may have important implications for restocking programs. PMID:27812407
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Vildmyren, Iselin; Cao, Huy John Vu; Haug, Lina Bowitz; Valand, Ida Ulrikke; Eng, Øyvin; Oterhals, Åge; Austgulen, Maren Hoff; Halstensen, Alfred; Mellgren, Gunnar; Gudbrandsen, Oddrun A
2018-06-05
Improved process technologies have allowed fishing vessels to utilize residuals from cod fillet production (head, backbone, skin, cuttings, and entrails) and convert this to high-quality protein powders for human consumption. In this double-blind pilot study, 42 healthy overweight or obese adults were randomized to three experimental groups consuming tablets corresponding to 6 g/day of proteins from cod residuals as presscake meal (Cod-PC), presscake and stickwater meal (Cod-PCW), or placebo tablets (control) for eight weeks. The primary outcome of this study was changes in metabolites related to glucose regulation in overweight or obese healthy adults after intake of proteins from cod residuals. Cod-PC supplementation decreased postprandial serum nonesterified fatty acids (NEFA) concentration and increased gene expressions of diglyceride acyltransferase 1 and 2 in subcutaneous adipose tissue compared with controls. Fasting insulin increased while fasting NEFA and 120-min postprandial glucose decreased within the Cod-PC group, but these changes did not differ from the other groups. In conclusion, supplementation with Cod-PC beneficially affected postprandial serum NEFA concentration compared with the other groups in overweight or obese adults. Supplementation with Cod-PCW, which contains a higher fraction of water-soluble protein compared to Cod-PC, did not affect serum markers of glucose regulation.
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Machado, Richele J. A.; Monteiro, Norberto K. V.; Migliolo, Ludovico; Silva, Osmar N.; Pinto, Michele F. S.; Oliveira, Adeliana S.; Franco, Octávio L.; Kiyota, Sumika; Bemquerer, Marcelo P.; Uchoa, Adriana F.; Morais, Ana H. A.; Santos, Elizeu A.
2013-01-01
Inhibitors of peptidases isolated from leguminous seeds have been studied for their pharmacological properties. The present study focused on purification, biochemical characterization and anti-inflammatory and anticoagulant evaluation of a novel Kunitz trypsin inhibitor from Erythrina velutina seeds (EvTI). Trypsin inhibitors were purified by ammonium sulfate (30–60%), fractionation followed by Trypsin-Sepharose affinity chromatography and reversed-phase high performance liquid chromatography. The purified inhibitor showed molecular mass of 19,210.48 Da. Furthermore, a second isoform with 19,228.16 Da was also observed. The inhibitor that showed highest trypsin specificity and enhanced recovery yield was named EvTI (P2) and was selected for further analysis. The EvTI peptide fragments, generated by trypsin and pepsin digestion, were further analyzed by MALDI-ToF-ToF mass spectrometry, allowing a partial primary structure elucidation. EvTI exhibited inhibitory activity against trypsin with IC50 of 2.2×10−8 mol.L−1 and constant inhibition (Ki) of 1.0×10−8 mol.L−1, by a non-competitive mechanism. In addition to inhibit the activity of trypsin, EvTI also inhibited factor Xa and neutrophil elastase, but do not inhibit thrombin, chymotrypsin or peptidase 3. EvTI was investigated for its anti-inflammatory and anti-coagulant properties. Firstly, EvTI showed no cytotoxic effect on human peripheral blood cells. Nevertheless, the inhibitor was able to prolong the clotting time in a dose-dependent manner by using in vitro and in vivo models. Due to anti-inflammatory and anticoagulant EvTI properties, two sepsis models were here challenged. EvTI inhibited leukocyte migration and specifically acted by inhibiting TNF-α release and stimulating IFN-α and IL-12 synthesis. The data presented clearly contribute to a better understanding of the use of Kunitz inhibitors in sepsis as a bioactive agent capable of interfering in blood coagulation and inflammation. PMID:23737945
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Siriangkanakun, Siriphon; Li-Chan, Eunice C Y; Yongsawadigul, Jirawat
2014-07-01
Sarcoplasmic proteins from 3 fish species were fractionated by 50% to 70% ammonium sulfate precipitation. Lyophilized fractionated sarcoplasmic proteins of threadfin bream (TB-SP), bigeye snapper (BS-SP), and yellow croaker (YC-SP) showed 80% to 92% trypsin inhibitory activity. Trypsin inhibitory activity staining gel electrophoresis revealed bands at 32, 33, 37, 45, 48, and 50 kDa for the 3 species, and a band at 95 kDa was observed for TB-SP and YC-SP. Alpha-1-antitrypsin with molecular mass of 45 to 50 kDa was identified in YC-SP by gel-based liquid chromatography-tandem mass spectrometry (GeLC-MS/MS). Other major protein bands appeared on trypsin activity staining included phosphorylase, glyceraldehyde-3-phosphate dehydrogenase, and creatine kinase with molecular mass of 95 and 35 to 40 kDa, respectively. But, these 3 proteins did not show true trypsin inhibitory activity. Trypsin inhibitory activity of fractionated sarcoplasmic proteins showed good stability, with >80% activity retained at 60 °C and up to 0.6 M NaCl. TB-SP showed the highest inhibitory activity against autolysis of washed threadfin bream mince at 65 °C. Addition of 0.5% or 1% TB-SP improved textural properties of threadfin bream surimi gels preincubated at 37 or 65 °C followed by heating at 90 °C. Therefore, TB-SP could be a promising protein ingredient for enhancing surimi gel texture. Threadfin bream, bigeye snapper, and yellow croaker are the main species used as raw material for tropical surimi production. Sarcoplasmic proteins from 3 species contain trypsin inhibitor(s) that can minimize proteolytic activity and improve gel texture of proteinase-laden fish muscle. Therefore, sarcoplasmic proteins that are byproducts from surimi processing of these species could be recovered, fractionated, and utilized as a functional protein ingredient. © 2014 Institute of Food Technologists®
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Xu, Kaijia; Wang, Yuzhi; Li, Yixue; Lin, Yunxuan; Zhang, Haibao; Zhou, Yigang
2016-11-23
Novel poly(deep eutectic solvent) grafted silica-coated magnetic microspheres (Fe 3 O 4 @SiO 2 -MPS@PDES) were prepared by polymerization of choline chloride-itaconic acid (ChCl-IA) and γ-MPS-modified magnetic silica composites, and were characterized by vibrating sample magnetometer (VSM), Fourier transform infrared spectrometry (FT-IR), X-ray photoelectron spectra (XPS), thermal gravimetric analysis (TGA) and transmission electron microscope (TEM). Then the synthetic Fe 3 O 4 @SiO 2 -MPS@PDES microspheres were applied for the magnetic solid-phase extraction (MSPE) of trypsin for the first time. After extraction, the concentration of trypsin in the supernatant was determined by a UV-vis spectrophotometer. Single factor experiments were carried out to investigate the effects of the extraction process, including the concentration of trypsin, the ionic strength, the pH value, the extraction time and the temperature. Experimental results showed the extraction capacity could reach up to 287.5 mg/g under optimized conditions. In comparison with Fe 3 O 4 @SiO 2 -MPS, Fe 3 O 4 @SiO 2 -MPS@PDES displayed higher extraction capacity and selectivity for trypsin. According to the regeneration studies, Fe 3 O 4 @SiO 2 -MPS@PDES microspheres can be recycled six times without significant loss of its extraction capacity, and retained a high extraction capacity of 233 mg/g after eight cycles. Besides, the activity studies also demonstrated that the activity of the extracted trypsin was well retained. Furthermore, the analysis of real sample revealed that the prepared magnetic microspheres can be used to purify trypsin in crude bovine pancreas extract. These results highlight the potential of the proposed Fe 3 O 4 @SiO 2 -MPS@PDES-MSPE method in separation of biomolecules. Copyright © 2016 Elsevier B.V. All rights reserved.
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Shimoda, Masayuki; Noguchi, Hirofumi; Fujita, Yasutaka; Takita, Morihito; Ikemoto, Tetsuya; Chujo, Daisuke; Naziruddin, Bashoo; Levy, Marlon F; Kobayashi, Naoya; Grayburn, Paul A; Matsumoto, Shinichi
2012-01-01
Porcine islets are considered to be a promising resource for xenotransplantation. However, it is difficult to isolate porcine islets because of the marked fragility and rapid dissociation. Endogenous trypsin is one of the main factors to damage islets during the isolation procedure. Recent studies have suggested that trypsin inhibitors during the preservation of pancreas or the collagenase digestion can improve the result of islet isolation. In this study, we examined whether α1-antitrypsin (Aralast™), which inhibits several endogenous proteases and has immunomodulatory properties, can protect islets from the proteases and improve the results of porcine islet isolation. Twelve porcine pancreata were divided into three groups: without Aralast group (standard, n = 5), preserved with Aralast using the ductal injection (DI) method (DI, n = 3), and with Aralast using the DI method and in the collagenase solution (DI+C, n = 4). Efficacy of islet isolation was assessed by islet yields, purity, and viability. The trypsin activity of the preservation and the digestion solution during the isolation procedure was measured. During islet isolation, the trypsin activity in DI+C group was significantly inhibited compared to the standard group, whereas DI group showed less effect than DI+C group. The average of postpurification islet equivalents (IEQ) per pancreas weight in the DI+C group was significantly higher than the standard group (standard: 3516 ± 497 IEQ/g, DI: 4607 ± 1090 IEQ/g, DI+C: 7097 ± 995 IEQ/g; p = 0.017 between standard and DI+C). In the DI+C group, stimulation index was higher than in other groups, although there was no significant difference. The presence of Aralast in both DI solution and collagenase solution markedly inhibited trypsin activity during pancreas digestion procedure and improved the porcine islet isolation. Inhibition of trypsin activity by Aralast could improve porcine islet isolation.
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Increased (/sup 125/I)trypsin-binding in serum from cystic fibrosis patients
DOE Office of Scientific and Technical Information (OSTI.GOV)
Cox, K.L.; Frates, R.C. Jr.; Sheikholislam, B.M.
1982-01-01
The capacities of normal and cystic fibrosis (CF) sera to bind to exogenous human (/sup 125/I)trypsin were compared. Sera from eight older CF patients bound significantly more exogenous human (/sup 125/I)trypsin than did sera from eight normal subjects (p less than 0.001). Disregarding the increased trypsin-binding (TB) of CF sera, serum immunoreactive trypsinogen (SIRT) levels were not detectable in these eight older CF patients. However, when SIRT levels were corrected for TB, four CF patients had normal SIRT concentrations and four had low but detectable SIRT levels. As compared to five normal newborns' sera, serum from a newborn with CFmore » had normal TB and the SIRT levels were very high. In conclusion, increased TB in CF serum lowers results of SIRT assays. Therefore, unless SIRT levels are corrected for TB, results obtained from currently available SIRT kits may be invalid.« less
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Effects of Mucuna pruriens protease inhibitors on Echis carinatus venom.
Hope-Onyekwere, Nnadozie Stanley; Ogueli, Godwin Ifeanyi; Cortelazzo, Alessio; Cerutti, Helena; Cito, Annarita; Aguiyi, John C; Guerranti, Roberto
2012-12-01
The medicinal plant Mucuna pruriens, with reputed anti-snake venom properties has been reported to contain a kunitz-type trypsin inhibitor. This study was undertaken to further evaluate the protease inhibitory potential of gpMuc, a multiform glycoprotein, and other protein fractions from M. pruriens seeds against trypsin, chymotrypsin, Echis carinatus snake venom, ecarin and thrombin. The results showed that gpMuc inhibited both trypsin and chymotrypsin activities and was thermally stable, maintaining its trypsin inhibitory activity at temperatures of up to 50°C. Its structural conformation was also maintained at pH ranges of 4-7. Immunoreactivity study confirms that it contains protease-recognizing epitope on one of its isoforms. The whole protein extract of M. pruriens seeds inhibited prothrombin activation by ecarin and whole E. carinatus venom, and also thrombin-like activity using chromogenic assay. Copyright © 2012 John Wiley & Sons, Ltd.
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Coulter-Mackie, M B; Lian, Q
2008-07-01
Alanine:glyoxylate aminotransferase (AGT) is a liver peroxisomal enzyme whose deficiency results in primary hyperoxaluria type 1 (PH1). More than 75 PH1 mutations are now documented in the AGT gene (AGXT), of which about 50% are missense. We have previously demonstrated that many such mutants expressed by transcription/translation are subject to generalized degradation by the proteasome and a specific limited trimming by an endogenous ATP-independent protease activity. Here, we report the results of partial digestion using trypsin as a mimic for the endogenous non-proteasomal protease and the use of N-terminal protein sequencing to determine the sensitive site. Partial trypsin digestion also provided an indicator of proper folding of the mutant enzyme. For selected mutations the sensitivity to trypsin could be ameliorated by addition of pyridoxal phosphate or aminooxy acetic acid as specific pharmacological chaperones.
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Yamaoka, Masaoki; Makino, Akiko; Sasahara, Kenji; Nastri, Aldise Mareta; Krisna, Luh Ade Wilan; Purhito, Edith Frederika; Poetranto, Emmanuel Djoko; Wulandari, Laksmi; Yudhawati, Resti; Setiawati, Landia; Setyoningrum, Retno Asih; Shinya, Kyoko
2013-01-01
The human influenza A virus (H3N2) has been the predominant influenza strain since 1992, and one property of this virus is non-agglutination of chicken erythrocytes [Ch(-) virus]. The Ch(-) virus in our study was able to acquire chicken hemagglutination [Ch(+)] by trypsin passage but not by chymotrypsin passage. Moreover, the trypsin-passaged Ch(+) viruses reacquired the Ch(-) property after a further chymotrypsin passage. In particular, genetic analysis showed no evidence of mutations in the hemagglutinin (HA) gene during either trypsin or chymotrypsin passages: the only differences found were in the HA cleavage sites between the trypsin-passaged virus and the chymotrypsin-passaged virus as determined by the N-terminal amino acid sequence. These results suggested that protease-dependent differences at the viral HA cleavage site, rather than genetic mutations, are likely to have a significant effect on the viral ability to produce chicken hemagglutination.
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DOE Office of Scientific and Technical Information (OSTI.GOV)
Jung, D.W.; Laties, G.G.
Potato mitochondria (Solanum tuberosum var. Russet Burbank), which readily phosphorylate ADP in oxidative phosphorylation, show low levels of ATPase activity which is stimulated neither by Mg/sup 2 +/, 2,4-dinitrophenol, incubation with respiratory substrates, nor disruption by sonication or treatment with Triton X-100, individually or in concert. Treatment of disrupted potato mitochondria with trypsin stimulates Mg/sup 2 +/-dependent, oligomycin-sensitive ATPase activity 10- to 15-fold, suggesting the presence of an ATPase inhibitor protein. Trypsin-induced ATPase activity was unaffected by uncoupler. Oligomycin-sensitive ATPase activity decreases as exposure to trypsin is increased. Incubation at alkaline pH or heating at 60/sup 0/C for 2 minutesmore » also activates ATPase of sonicated potato mitochondria. Disruption of cauliflower (Brassica oleracea), red sweet potato (Ipomoea batatas), and carrot (Daucus carota) mitochondria increases ATPase activity, which is further enhanced by treatment with trypsin. The significance of the tight association of the inhibitor protein and ATPase in potato mitochondria is not clear.« less
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Zhang, Yun-lin; Yang, Long-yuan; Qin, Bo-qiang; Gao, Guang; Luo, Lian-cong; Zhu, Guang-wei; Liu, Ming-liang
2008-06-01
Spatial variation of chemical oxygen demand (COD) concentration was documented and significant correlations between COD concentration and chromophoric dissolved organic matter (CDOM) absorption, fluorescence, DOC concentration were found based on a cruise sampling in the northern region of Lake Taihu in summer including 42 samplings. The possible source of COD was also discussed using every two cruise samplings in summer and winter, respectively. The COD concentration ranged from 3.77 to 7.96 mg x L(-1) with a mean value of (5.90 +/- 1.54) mg x L(-1). The mean COD concentrations in Meiliang Bay and the central lake basin were (6.93 +/- 0.89) mg x L(-1) and (4.21 +/- 0.49) mg x L(-1) respectively. A significant spatial difference was found between Meiliang Bay and the central lake basin in COD concentration, CDOM absorption coefficient, fluorescence, DOC and phytoplankton pigment concentrations, decreasing from the river mouth to inner bay, outer bay and the central lake basin. Significant correlations between COD concentration and CDOM absorption, fluorescence, DOC concentration, suggested that COD concentration could be estimated and organic pollution could be assessed using CDOM absorption retrieved from remote sensing images. Significant and positive correlation was found between COD concentration and chlorophyll a concentration in summer. However, the correlation was weak or no correlation was found in winter. Furthermore, a significant higher COD concentration was found in summer than in winter (p < 0.001). Our results indicated that degradation of phytoplankton blooms was the main source of COD in summer, except for river terrestrial input.
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Cape Cod Transit Task Force : Five-Year Public Transportation Plan
DOT National Transportation Integrated Search
2002-06-30
The U.S. Department of Transportation's Volpe National Transportation Systems : Center has been working in cooperation with the Cape Cod Regional Transit : Authority, the Cape Cod Commission, and other organizations participating on : the Cape Cod Tr...
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Lin, Kun; Gao, Feng; Chen, Qingquan; Liu, Qicai; Chen, Shu
2015-01-01
Early intracellular premature trypsinogen activation was interpreted as the key initiator of pancreatitis. When the balance in the homeostasis of trypsin and antitrypsin system is disequilibrated, elevated aggressive enzymes directly attack the pancreatic tissue, which leads to pancreatic destruction and inflammation. However, trypsin alone is not enough to cause complications in pancreatitis, which may play a crucial role in modulating signaling events in the initial phase of the disease. NFκB activation is the major inflammatory pathway involved in the occurrence and development of pancreatitis and it can be induced by intrapancreatic activation of trypsinogen. Synthesis of trypsinogen occurs in endoplasmic reticulum (ER), and ER stress is an important early acinar cell event. Components of ER stress response are known to be able to trigger cell death as well as NFκB signaling cascade. The strongest evidence supporting the trypsin-centered theory is that gene mutations, which lead to the generation of more trypsin, or reduce the activity of trypsin inhibitors or trypsin degradation, are associated with pancreatitis. Thus, trypsin–antitrypsin imbalance may be the first step leading to pancreatic autodigestion and inducing other pathways. Continued experimental studies are necessary to determine the specific relationships between trypsin–antitrypsin imbalance and genetic heterogeneity in pancreatitis. In this article, we review the latest advances that contributed to the understanding of the basic mechanisms behind the occurrence and development of pancreatitis with a focus on the interpretation of trypsin–antitrypsin imbalance and their relationships with other inflammation pathways. We additionally highlight genetic predispositions to pancreatitis and possible mechanisms associated with them. PMID:26228362
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Treatment of low-strength soluble wastewater using an anaerobic baffled reactor (ABR).
Gopala Krishna, G V T; Kumar, Pramod; Kumar, Pradeep
2009-01-01
Treatment of low-strength soluble wastewater (COD approximately 500 mg/L) was studied using an eight chambered anaerobic baffled reactor (ABR). At pseudo steady-state (PSS), the average total and soluble COD values (COD(T) and COD(S)) at 8h hydraulic retention time (HRT) were found to be around 50 and 40 mg/L, respectively, while at 10h HRT average COD(T) and COD(S) values were of the order of 47 and 37 mg/L, respectively. COD and BOD (3 day, 27 degrees C) removal averaged more than 90%. Effluent conformed to Indian standards laid down for BOD (less than 30 mg/L). Reactor effluent characteristics exhibited very low values of standard deviation indicating excellent reactor stability at PSS in terms of effluent characteristics. Based on mass balance calculations, more than 60% of raw wastewater COD was estimated to be recovered as CH(4) in the gas phase. Compartment-wise profiles indicated that most of the BOD and COD got reduced in the initial compartments only. Sudden drop in pH (7.8-6.7) and formation of volatile fatty acids (VFA) (53-85 mg/L) were observed in the first compartment due to acidogenesis and acetogenesis. The pH increased and VFA concentration decreased longitudinally down the reactor. Residence time distribution (RTD) studies revealed that the flow pattern in the ABR was neither completely plug-flow nor perfectly mixed. Observations from scanning electron micrographs (SEM) suggest that distinct phase separation takes place in an ABR.
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Petersen, Maria Faldborg; Steffensen, John Fleng
2003-01-01
Atlantic cod Gadus morhua has polymorphic haemoglobin, which can be separated into two homozygous types, HbI-1 and HbI-2, and one heterozygous type HbI-1/2. The geographical distribution of Atlantic cod with the different haemoglobin types varies, with the HbI(2) allele occurring at high frequency in northern regions, and the HbI(1) allele dominant in warmer areas. To determine if temperature is a selective parameter in the distribution of the haemoglobin types, the preferred temperature of the homozygous genotypes HbI-1 and HbI-2 was measured. We found that HbI-2 cod preferred a temperature of 8.2+/-1.5 degrees C while HbI-1 cod preferred 15.4+/-1.1 degrees C, and this preference was significant. The effect of hypoxia (35% oxygen saturation) on the preferred temperature was also measured. Previous studies showed that the preferred temperature of fish decreases during hypoxia, and this was the case for HbI-1 cod, which preferred 9.8+/-1.8 degrees C during hypoxia, whereas HbI-2 cod did not show this effect. The results indicate that environmental temperature changes will lead to a distributional change in the different haemoglobin types of Atlantic cod, global warming providing an advantage for HbI-1 cod. However, since HbI-1 cod prefer a low temperature under hypoxic conditions, a combination of increased water temperature and hypoxia could be unfavourable for Atlantic cod stocks.
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Booman, Marije; Borza, Tudor; Feng, Charles Y; Hori, Tiago S; Higgins, Brent; Culf, Adrian; Léger, Daniel; Chute, Ian C; Belkaid, Anissa; Rise, Marlies; Gamperl, A Kurt; Hubert, Sophie; Kimball, Jennifer; Ouellette, Rodney J; Johnson, Stewart C; Bowman, Sharen; Rise, Matthew L
2011-08-01
The collapse of Atlantic cod (Gadus morhua) wild populations strongly impacted the Atlantic cod fishery and led to the development of cod aquaculture. In order to improve aquaculture and broodstock quality, we need to gain knowledge of genes and pathways involved in Atlantic cod responses to pathogens and other stressors. The Atlantic Cod Genomics and Broodstock Development Project has generated over 150,000 expressed sequence tags from 42 cDNA libraries representing various tissues, developmental stages, and stimuli. We used this resource to develop an Atlantic cod oligonucleotide microarray containing 20,000 unique probes. Selection of sequences from the full range of cDNA libraries enables application of the microarray for a broad spectrum of Atlantic cod functional genomics studies. We included sequences that were highly abundant in suppression subtractive hybridization (SSH) libraries, which were enriched for transcripts responsive to pathogens or other stressors. These sequences represent genes that potentially play an important role in stress and/or immune responses, making the microarray particularly useful for studies of Atlantic cod gene expression responses to immune stimuli and other stressors. To demonstrate its value, we used the microarray to analyze the Atlantic cod spleen response to stimulation with formalin-killed, atypical Aeromonas salmonicida, resulting in a gene expression profile that indicates a strong innate immune response. These results were further validated by quantitative PCR analysis and comparison to results from previous analysis of an SSH library. This study shows that the Atlantic cod 20K oligonucleotide microarray is a valuable new tool for Atlantic cod functional genomics research.
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Levdikov, Vladimir M; Blagova, Elena; Young, Vicki L; Belitsky, Boris R; Lebedev, Andrey; Sonenshein, Abraham L; Wilkinson, Anthony J
2017-02-17
CodY is a branched-chain amino acid (BCAA) and GTP sensor and a global regulator of transcription in low G + C Gram-positive bacteria. It controls the expression of over 100 genes and operons, principally by repressing during growth genes whose products are required for adaptations to nutrient limitation. However, the mechanism by which BCAA binding regulates transcriptional changes is not clear. It is known that CodY consists of a GAF (c G MP-stimulated phosphodiesterases, a denylate cyclases, F hlA) domain that binds BCAAs and a winged helix-turn-helix (wHTH) domain that binds to DNA, but the way in which these domains interact and the structural basis of the BCAA dependence of this interaction are unknown. To gain new insights, we determined the crystal structure of unliganded CodY from Bacillus subtilis revealing a 10-turn α-helix linking otherwise discrete GAF and wHTH domains. The structure of CodY in complex with isoleucine revealed a reorganized GAF domain. In both complexes CodY was tetrameric. Size exclusion chromatography with multiangle laser light scattering (SEC-MALLS) experiments showed that CodY is a dimer at concentrations found in bacterial cells. Comparison of structures of dimers of unliganded CodY and CodY-Ile derived from the tetramers showed a splaying of the wHTH domains when Ile was bound; splaying is likely to account for the increased affinity of Ile-bound CodY for DNA. Electrophoretic mobility shift and SEC-MALLS analyses of CodY binding to 19-36-bp operator fragments are consistent with isoleucine-dependent binding of two CodY dimers per duplex. The implications of these observations for effector control of CodY activity are discussed. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.
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Li, Dong; Lv, Yufeng; Zeng, Huiping; Zhang, Jie
2016-09-01
In this study, a continuous-flow system with enhanced biological phosphorus removal (EBPR) granules was operated at different COD concentrations (200, 300 and 400mgL(-)(1)) to investigate the effect of COD loading on this system. The results showed that when the COD concentration in influent was increased to 400mgL(-)(1), the anaerobic COD removal efficiency and total phosphorus removal efficiency reduced obviously and the settling ability of granules deteriorated due to the proliferation of filamentous bacteria. Moreover, high COD loading inhibited the EPS secretion and destroyed the stability of granules. Results of high-through pyrosequencing indicated that filamentous bacteria had a competitive advantage over polyphosphate-accumulating organisms (PAOs) at high COD loading. The performance of system, settling ability of granules and proportion of PAOs gradually recovered to the initial level after the COD concentration was reduced to 200mgL(-)(1) on day 81. Copyright © 2016 Elsevier Ltd. All rights reserved.
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Li, Na; Kakarla, Ramesh; Moon, Jung Mi; Min, Booki
2015-07-01
Microbial fuel cells (MFCs) have gathered attention as a novel bioenergy technology to simultaneously treat wastewater with less sludge production than the conventional activated sludge system. In two different operations of the MFC and aerobic process, microbial growth was determined by the protein assay method and their biomass yields using real wastewater were compared. The biomass yield on the anode electrode of the MFC was 0.02 g-COD-cell/g- COD-substrate and the anolyte planktonic biomass was 0.14 g-COD-cell/g-COD-substrate. An MFC without anode electrode resulted in the biomass yield of 0.07 ± 0.03 g-COD-cell/g-COD-substrate, suggesting that oxygen diffusion from the cathode possibly supported the microbial growth. In a comparative test, the biomass yield under aerobic environment was 0.46 ± 0.07 g-COD-cell/g-COD-substrate, which was about 3 times higher than the total biomass value in the MFC operation.
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Carrier free immobilization and characterization of trypsin.
Menfaatli, Esra; Zihnioglu, Figen
2015-04-01
Pancreatic trypsin was immobilized by cross-linked enzyme aggregates (CLEA) which is a carrier free immobilization method. Ammonium sulfate was chosen for enzyme precipitation which was followed by cross linking of formed aggregates via glutaraldehyde. Concentrations of precipitant and cross linker were respectively optimized as 60% ammonium sulfate and 1% glutaraldehyde. Optimum pH and temperature for CLEA was increased compared to free enzyme. Furthermore, pH, thermal and storage stability were improved. Presence of additives had no effects on enzyme activity. Prepared cross-linked trypsin aggregates are convenient for in situ protein fragmentation and can be used for protein identification.
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High-Yield Method for Dispersing Simian Kidneys for Cell Cultures
de Oca, H. Montes; Probst, P.; Grubbs, R.
1971-01-01
A technique for dispersion of animal tissue cells is described. The proposed technique is based on the concomitant use of trypsin and disodium ethylenediamine tetraacetate (EDTA). The use of the two dispersing agents (trypsin and disodium EDTA) markedly enhances cell yield as compared with the standard cell dispersion methods. Moreover, significant reduction in the amount of time required for complete tissue dispersal, presence of a very low number of nonviable cells, less cell clumping, and more uniform monolayer formation upon cultivation compare favorably with the results usually obtained with the standard trypsinization technique. Images PMID:4993235
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Biochemical Characterization of An Arginine-Specific Alkaline Trypsin from Bacillus licheniformis.
Gong, Jin-Song; Li, Wei; Zhang, Dan-Dan; Xie, Min-Feng; Yang, Biao; Zhang, Rong-Xian; Li, Heng; Lu, Zhen-Ming; Xu, Zheng-Hong; Shi, Jin-Song
2015-12-17
In the present study, we isolated a trypsin-producing strain DMN6 from the leather waste and identified it as Bacillus licheniformis through a two-step screening strategy. The trypsin activity was increased up to 140 from 20 U/mL through culture optimization. The enzyme was purified to electrophoretic homogeneity with a molecular mass of 44 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and the specific activity of purified enzyme is 350 U/mg with Nα-Benzoyl-L-arginine ethylester as the substrate. The optimum temperature and pH for the trypsin are 65 °C and pH 9.0, respectively. Also, the enzyme can be significantly activated by Ba(2+). This enzyme is relatively stable in alkaline environment and displays excellent activity at low temperatures. It could retain over 95% of enzyme activity after 180 min of incubation at 45 °C. The distinguished activity under low temperature and prominent stability enhance its catalytic potential. In the current work, the open reading frame was obtained with a length of 1371 nucleotides that encoded a protein of 456 amino acids. These data would warrant the B. licheniformis trypsin as a promising candidate for catalytic application in collagen preparation and leather bating through further protein engineering.
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Miersch, Claudia; Stange, Katja; Röntgen, Monika
2018-06-01
Muscle stem cells, termed satellite cells (SC), and SC-derived myogenic progenitor cells (MPC) are involved in postnatal muscle growth, regeneration, and muscle adaptability. They can be released from their natural environment by mechanical disruption and tissue digestion. The literature contains several isolation protocols for porcine SC/MPC including various digestion procedures, but comparative studies are missing. In this report, classic trypsinization and a more complex trypsin, collagenase, and DNase (TCD) digestion were performed with skeletal muscle tissue from 4- to 5-d-old piglets. The two digestion procedures were compared regarding cell yield, viability, myogenic purity, and in vitro cell function. The TCD digestion tended to result in higher cell yields than digestion with solely trypsin (statistical trend p = 0.096), whereas cell size and viability did not differ. Isolated myogenic cells from both digestion procedures showed comparable proliferation rates, expressed the myogenic marker Desmin, and initiated myogenic differentiation in vitro at similar levels. Thus, TCD digestion tended to liberate slightly more cells without changes in the tested in vitro properties of the isolated cells. Both procedures are adequate for the isolation of SC/MPC from juvenile porcine muscles but the developmental state of the animal should always be considered.
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Rueda, Analiz; Sifuentes, Cecilia; Gilman, Robert H; Gutiérrez, Andrés H; Piña, Ruby; Chile, Nancy; Carrasco, Sebastián; Larson, Sandra; Mayta, Holger; Verástegui, Manuela; Rodriguez, Silvia; Gutiérrez-Correa, Marcel; García, Héctor H; Sheen, Patricia; Zimic, Mirko
2011-12-01
Neurocysticercosis is an endemic parasitic disease caused by Taenia solium larva. Although the mechanism of infection is not completely understood, it is likely driven by proteolytic activity that degrades the intestinal wall to facilitate oncosphere penetration and further infection. We analyzed the publicly available T. solium EST/DNA library and identified two contigs comprising a full-length cDNA fragment very similar to Echinococcus granulosus Ag5 protein. The T. solium cDNA sequence included a proteolytic trypsin-like-domain in the C-terminal region, and a thrombospondin type-1 adherence-domain in the N-terminal region. Both the trypsin-like and adherence domains were expressed independently as recombinant proteins in bacterial systems. TsAg5 showed marginal trypsin-like activity and high sequence similarity to Ag5. The purified antigens were tested in a Western immunoblot assay to diagnose human neurocysticercosis. The sensitivity of the trypsin-like-domain was 96.36% in patients infected with extraparenchymal cysts, 75.44% in patients infected with multiple cysts, and 39.62% in patients with a single cyst. Specificity was 76.70%. The thrombospondin type-1 adherence-domain was not specific for neurocysticercosis. Copyright © 2011 Elsevier B.V. All rights reserved.
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Blankenship, Elise; Vukoti, Krishna; Miyagi, Masaru; Lodowski, David T.
2014-01-01
With more than 500 crystal structures determined, serine proteases make up greater than one-third of all proteases structurally examined to date, making them among the best biochemically and structurally characterized enzymes. Despite the numerous crystallographic and biochemical studies of trypsin and related serine proteases, there are still considerable shortcomings in the understanding of their catalytic mechanism. Streptomyces erythraeus trypsin (SET) does not exhibit autolysis and crystallizes readily at physiological pH; hence, it is well suited for structural studies aimed at extending the understanding of the catalytic mechanism of serine proteases. While X-ray crystallographic structures of this enzyme have been reported, no coordinates have ever been made available in the Protein Data Bank. Based on this, and observations on the extreme stability and unique properties of this particular trypsin, it was decided to crystallize it and determine its structure. Here, the first sub-angstrom resolution structure of an unmodified, unliganded trypsin crystallized at physiological pH is reported. Detailed structural analysis reveals the geometry and structural rigidity of the catalytic triad in the unoccupied active site and comparison to related serine proteases provides a context for interpretation of biochemical studies of catalytic mechanism and activity. PMID:24598752
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A crystalline protein-proteinase inhibitor from pinto bean seeds.
Wang, D
1975-06-26
A crystalline protein-proteinase inhibitor has been isolated from seeds of Pinto bean (Phaseolus vulgaris cultvar. Pinto). It has an average molecular weight of 19 000 as estimated by gel filtration. This crystalline inhibitor is highly active against both bovine pancreatic trypsin and alpha-chymotrypsin. Complexes of both trypsin-inhibitor and alpha-chymotrypsin-inhibitor have been isolated. The inhibitor which was derived from the dissociated trypsin-inhibitor complex was only 62% as effective as the original compound against either enzyme. In contrast, the inhibitor obtained from alpha-chymotrypsin-inhibitor complex retained its full original inhibitory activity for trypsin, but only 25% of its original activity against alpha-chymotrypsin. The dissociated inhibitor from alpha-chymotrypsin-inhibitor compex, despite its full inhibitory activity, had been modified to such an extent that it could no longer form any precipitable complex with trypsin. The crystalline protein-proteinase inhibitor is not homogeneous and has been resolved into two distinct inhibitors in terms of their physical and chemical properties. These two inhibitors are designated as Pinto bean proteinase inhibitor I and II and their respective minimum molecular weights are 9100 and 10 000. They differ most strikingly in their amino acid composition in that inhibitor II is void of both valine and methionine.
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Trypsin inhibitory activity and gel-enhancing effect of sarcoplasmic proteins from common carp.
Siriangkanakun, Siriphon; Yongsawatdigul, Jirawat
2012-10-01
Proteinase inhibitory activity of sarcoplasmic protein (SP) extracted from common carp (Cyprinus carpio) muscle and its gel-improving ability were investigated. SPs displayed 89% and 54% inhibitory activity toward trypsin at 40 and 65 °C, respectively. Protein bands with molecular mass of 69, 50, 44, 41, and 35 kDa appeared on trypsin inhibitory activity staining under nonreducing condition when incubated at 40 °C, while 2 protein bands at 54 and 35 kDa were observed at 65 °C. Addition of SP at 0.18 g protein/100 g increased textural properties of threadfin bream surimi gel. However, when SP was added in combination with various CaCl(2) concentrations (0.1% to 0.5%) it did not further improve textural properties as compared to the addition of SP alone. Retention of myosin heavy chain of threadfin bream surimi was greater with the addition of SP. These results indicated that the gel-enhancing effect of common carp SP was due to the inhibitory activity toward endogenous trypsin-like proteinases in threadfin bream surimi. Sarcoplasmic protein from common carp muscle could be used as a functional protein ingredient that minimizes muscle proteolysis and improves textural properties of surimi containing trypsin-like endogenous proteinases. © 2012 Institute of Food Technologists®
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Blankenship, Elise; Vukoti, Krishna; Miyagi, Masaru; Lodowski, David T
2014-03-01
With more than 500 crystal structures determined, serine proteases make up greater than one-third of all proteases structurally examined to date, making them among the best biochemically and structurally characterized enzymes. Despite the numerous crystallographic and biochemical studies of trypsin and related serine proteases, there are still considerable shortcomings in the understanding of their catalytic mechanism. Streptomyces erythraeus trypsin (SET) does not exhibit autolysis and crystallizes readily at physiological pH; hence, it is well suited for structural studies aimed at extending the understanding of the catalytic mechanism of serine proteases. While X-ray crystallographic structures of this enzyme have been reported, no coordinates have ever been made available in the Protein Data Bank. Based on this, and observations on the extreme stability and unique properties of this particular trypsin, it was decided to crystallize it and determine its structure. Here, the first sub-angstrom resolution structure of an unmodified, unliganded trypsin crystallized at physiological pH is reported. Detailed structural analysis reveals the geometry and structural rigidity of the catalytic triad in the unoccupied active site and comparison to related serine proteases provides a context for interpretation of biochemical studies of catalytic mechanism and activity.
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Co-Inactivation of GlnR and CodY Regulators Impacts Pneumococcal Cell Wall Physiology.
Johnston, Calum; Bootsma, Hester J; Aldridge, Christine; Manuse, Sylvie; Gisch, Nicolas; Schwudke, Dominik; Hermans, Peter W M; Grangeasse, Christophe; Polard, Patrice; Vollmer, Waldemar; Claverys, Jean-Pierre
2015-01-01
CodY, a nutritional regulator highly conserved in low G+C Gram-positive bacteria, is essential in Streptococcus pneumoniae (the pneumococcus). A published codY mutant possessed suppressing mutations inactivating the fatC and amiC genes, respectively belonging to iron (Fat/Fec) and oligopeptide (Ami) ABC permease operons, which are directly repressed by CodY. Here we analyzed two additional published codY mutants to further explore the essentiality of CodY. We show that one, in which the regulator of glutamine/glutamate metabolism glnR had been inactivated by design, had only a suppressor in fecE (a gene in the fat/fec operon), while the other possessed both fecE and amiC mutations. Independent isolation of three different fat/fec suppressors thus establishes that reduction of iron import is crucial for survival without CodY. We refer to these as primary suppressors, while inactivation of ami, which is not essential for survival of codY mutants and acquired after initial fat/fec inactivation, can be regarded as a secondary suppressor. The availability of codY- ami+ cells allowed us to establish that CodY activates competence for genetic transformation indirectly, presumably by repressing ami which is known to antagonize competence. The glnR codY fecE mutant was then found to be only partially viable on solid medium and hypersensitive to peptidoglycan (PG) targeting agents such as the antibiotic cefotaxime and the muramidase lysozyme. While analysis of PG and teichoic acid composition uncovered no alteration in the glnR codY fecE mutant compared to wildtype, electron microscopy revealed altered ultrastructure of the cell wall in the mutant, establishing that co-inactivation of GlnR and CodY regulators impacts pneumococcal cell wall physiology. In light of rising levels of resistance to PG-targeting antibiotics of natural pneumococcal isolates, GlnR and CodY constitute potential alternative therapeutic targets to combat this debilitating pathogen, as co-inactivation of these regulators renders pneumococci sensitive to iron and PG-targeting agents.
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Zuo, Shaozhi; Huwer, Bastian; Bahlool, Qusay; Al-Jubury, Azmi; Daugbjerg Christensen, Nanna; Korbut, Rozalia; Kania, Per; Buchmann, Kurt
2016-06-15
A significant increase in the infection level of Baltic cod Gadus morhua with the anisakid nematode larvae Contracaecum osculatum and Pseudoterranova decipiens has been recorded during recent years due to the expanding local population of grey seals Halichoerus grypus, which act as final hosts for these parasites. Here, we report from an investigation of 368 cod (total length [TL] 6-49 cm; caught in ICES Subdivision 25) that the infection level of juvenile cod (TL 6-30 cm) with larvae of C. osculatum and P. decipiens is absent or very low, whereas it increases drastically in larger cod (TL 31-48 cm). A third nematode Hysterothylacium aduncum was rarely found. The study indicates that the prey animals for large cod act as transport hosts for the parasite larvae. Analyses of stomach contents of cod caught in the same area (2007-2014) showed that small benthic organisms (including polychaetes Harmothoë sarsi) are preferred food items by small cod, the isopod Saduria entomon is taken by all size classes, and sprat Sprattus sprattus are common prey items for cod larger than 30 cm. Parasitological investigations (microscopic and molecular analyses) of H. sarsi (100 specimens) and S. entomon (40 specimens) did not reveal infection in these invertebrates, but 11.6% of sprat (265 specimens examined) was shown to be infected with 1-8 C. osculatum third stage larvae per fish. Analyses of sprat stomach contents confirmed that copepods and cladocerans are the main food items of sprat. These observations suggest that the C. osculatum life cycle in the Baltic Sea includes grey seals as final hosts, sprat as the first transport host and cod as second transport host. It may be speculated that sprat obtain infection by feeding on copepods and/or cladocerans, which could serve as the first intermediate hosts. One cannot exclude the possibility that the size-dependent C. osculatum infection of cod may contribute (indirectly or directly) to the differential mortality of larger cod (>38 cm) compared to smaller cod (<30 cm) recently recorded in the Baltic cod population.
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Yu, Zheng-zhe; Sun, De-zhi; Li, Chang-hai; Shi, Peng-fei; Duan, Xiao-dong; Sun, Guo-rong; Liu, Jun-xin
2004-01-01
The performance of UV/H2O2, UV/O3 and UV/H2O2/O3 oxidation systems for treating spent caustic from an ethylene plant was investigated. In UV/H2O2 system, with the increase of H2O2 dosage, removal efficiencies of COD and the ratio of biochemical oxygen demand (BOD) to chemical oxygen demand (COD) of the effluent were increased and a better performance was obtained than the H2O2 system alone. In UV/H2O2 system, removal efficiency of COD reach 68% under the optimum condition, and BOD/COD ratio was significantly increased from 0.22 to 0.52. In UV/O3 system, with the increase of O3 dosage, removal efficiency of COD and BOD/COD ratio were increased, and a better performance was obtained than the O3 system alone. Under the optimum condition, removal efficiency of COD was 54%, and BOD/COD ratio was significantly increased from 0.22 to 0.48. In UV/H2O2/O3 system, COD removal efficiency was found to be 22.0% higher than UV/O3 system.
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Orton, David C.; Makowiecki, Daniel; de Roo, Tessa; Johnstone, Cluny; Harland, Jennifer; Jonsson, Leif; Heinrich, Dirk; Enghoff, Inge Bødker; Lõugas, Lembi; Van Neer, Wim; Ervynck, Anton; Hufthammer, Anne Karin; Amundsen, Colin; Jones, Andrew K. G.; Locker, Alison; Hamilton-Dyer, Sheila; Pope, Peter; MacKenzie, Brian R.; Richards, Michael; O'Connell, Tamsin C.; Barrett, James H.
2011-01-01
Although recent historical ecology studies have extended quantitative knowledge of eastern Baltic cod (Gadus morhua) exploitation back as far as the 16th century, the historical origin of the modern fishery remains obscure. Widespread archaeological evidence for cod consumption around the eastern Baltic littoral emerges around the 13th century, three centuries before systematic documentation, but it is not clear whether this represents (1) development of a substantial eastern Baltic cod fishery, or (2) large-scale importation of preserved cod from elsewhere. To distinguish between these hypotheses we use stable carbon and nitrogen isotope analysis to determine likely catch regions of 74 cod vertebrae and cleithra from 19 Baltic archaeological sites dated from the 8th to the 16th centuries. δ13C and δ15N signatures for six possible catch regions were established using a larger sample of archaeological cod cranial bones (n = 249). The data strongly support the second hypothesis, revealing widespread importation of cod during the 13th to 14th centuries, most of it probably from Arctic Norway. By the 15th century, however, eastern Baltic cod dominate within our sample, indicating the development of a substantial late medieval fishery. Potential human impact on cod stocks in the eastern Baltic must thus be taken into account for at least the last 600 years. PMID:22110675
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Cemento-Osseous Dysplasias: Imaging Features Based on Cone Beam Computed Tomography Scans.
Cavalcanti, Paulo Henrique Pereira; Nascimento, Eduarda Helena Leandro; Pontual, Maria Luiza Dos Anjos; Pontual, Andréa Dos Anjos; Marcelos, Priscylla Gonçalves Correia Leite de; Perez, Danyel Elias da Cruz; Ramos-Perez, Flávia Maria de Moraes
2018-01-01
Imaging exams have important role in diagnosis of cemento-osseous dysplasia (COD). Cone beam computed tomography (CBCT) stands out for allowing three-dimensional image evaluation. This study aimed to assess the prevalence of cases diagnosed as COD on CBCT scans, as well identify the main imaging features related to these lesions. An analysis was performed in a database containing 22,400 radiological reports, in which all cases showing some type of COD were initially selected. These CBCT exams were reevaluated to confirm the radiographic diagnosis and determine the prevalence and distribution of the types of COD with regard to gender, age and preferred location, while describing its most common imaging aspects. Data were presented using descriptive analyses. There were 82 cases diagnosed as COD in the CBCT images (prevalence of 0.4%). The distribution of patients was 11 (13.4%) male and 71 (86.6%) female, with a mean age of 49.8 years (age-range 17-85 years). There were 47 (57.3%) cases of periapical COD, 23 (28%) of focal COD and 12 (14.6%) of florid COD. The mandible was more affected than the maxilla. In most cases, the lesions were mixed or hyperdense. All COD had well-defined limits and there were no cases of tooth displacement. In conclusion, periapical COD was the most common type and the most affected bone was the mandible. Imaging evaluation is critical for diagnosis and dentists should bear in mind all possible radiographic presentations of COD in order to prevent misleading diagnoses and consequently, inadequate treatments.
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ERIC Educational Resources Information Center
Russo, Salvatore F.; Holzman, Tom
1977-01-01
Describes an experiment that illustrates the following points concerning the experimental determination of trypsin activity: (1) there is a difference in basing enzyme concentration on weight, absorbance, or active sites; and (2) the method of expressing enzyme concentration determines the value of specific, molecular, and catalytic center…
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Mesotrypsin Has Evolved Four Unique Residues to Cleave Trypsin Inhibitors as Substrates.
Alloy, Alexandre P; Kayode, Olumide; Wang, Ruiying; Hockla, Alexandra; Soares, Alexei S; Radisky, Evette S
2015-08-28
Human mesotrypsin is highly homologous to other mammalian trypsins, and yet it is functionally unique in possessing resistance to inhibition by canonical serine protease inhibitors and in cleaving these inhibitors as preferred substrates. Arg-193 and Ser-39 have been identified as contributors to the inhibitor resistance and cleavage capability of mesotrypsin, but it is not known whether these residues fully account for the unusual properties of mesotrypsin. Here, we use human cationic trypsin as a template for engineering a gain of catalytic function, assessing mutants containing mesotrypsin-like mutations for resistance to inhibition by bovine pancreatic trypsin inhibitor (BPTI) and amyloid precursor protein Kunitz protease inhibitor (APPI), and for the ability to hydrolyze these inhibitors as substrates. We find that Arg-193 and Ser-39 are sufficient to confer mesotrypsin-like resistance to inhibition; however, compared with mesotrypsin, the trypsin-Y39S/G193R double mutant remains 10-fold slower at hydrolyzing BPTI and 2.5-fold slower at hydrolyzing APPI. We identify two additional residues in mesotrypsin, Lys-74 and Asp-97, which in concert with Arg-193 and Ser-39 confer the full catalytic capability of mesotrypsin for proteolysis of BPTI and APPI. Novel crystal structures of trypsin mutants in complex with BPTI suggest that these four residues function cooperatively to favor conformational dynamics that assist in dissociation of cleaved inhibitors. Our results reveal that efficient inhibitor cleavage is a complex capability to which at least four spatially separated residues of mesotrypsin contribute. These findings suggest that inhibitor cleavage represents a functional adaptation of mesotrypsin that may have evolved in response to positive selection pressure. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.
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T-cell epitope-containing hypoallergenic β-lactoglobulin for oral immunotherapy in milk allergy.
Ueno, Hiroshi M; Kato, Teruhiko; Ohnishi, Hidenori; Kawamoto, Norio; Kato, Zenichiro; Kaneko, Hideo; Kondo, Naomi; Nakano, Taku
2016-12-01
Optimally hydrolyzed β-Lactoglobulin (βLg) is a promising milk oral immunotherapy (OIT) candidate with respect to showing reduced B-cell reactivity but retaining the T-cell epitope. To demonstrate that an edible hypoallergenic βLg hydrolysate containing the T-cell epitope is suitable for OIT. We tested how chymotrypsin affected the retention of the T-cell epitope of βLg when preparing βLg hydrolysates using food-grade trypsin. We investigated the effect of chymotrypsin activity on the formation of the T-cell epitope-containing peptide of βLg (βLg 102-124 ) and prepared an edible βLg hydrolysate containing βLg 102-124 using screened food-grade trypsins. B-cell reactivity was determined using immunoassays in which ELISA was performed with anti-βLg rabbit IgG and Western blotting was performed with a milk-specific IgE antiserum. In βLg hydrolysis performed by varying the activity of trypsin and chymotrypsin, chymotrypsin activity inhibited the formation of βLg 102-124 with an increase in hydrolysis time in a dose-dependent manner. βLg 102-124 was generated by two of five food-grade trypsins used at a ratio of 1:50 (w/w, enzyme/substrate) for 20 h at 40°C. The edible βLg hydrolysate retained βLg 102-124 and showed a reduction in molecular weight distribution and antigenicity against IgG and IgE. Chymotrypsin activity inhibited the formation of βLg 102-124 in the trypsin hydrolysate of βLg. This βLg trypsin hydrolysate is a novel candidate for peptide-based OIT in cow's milk allergy for safely inducing desensitization. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
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Hu, Qiong-Zheng; Jang, Chang-Hyun
2012-03-01
In this study, we developed a new type of liquid crystal (LC)-based sensor for the real-time and label-free monitoring of enzymatic activity through changes in the orientation of LCs coupled to the interactions between polyelectrolyte and phospholipid. The LCs changed from dark to bright after an aqueous solution of poly-l-lysine (PLL) was transferred onto a self-assembled monolayer of the phospholipid, dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), at the aqueous/LC interface. Interactions between the positively charged PLL and the negatively charged DOPG drove the reorganization of the phospholipid membrane, which induced an orientational transition in the LCs from a homeotropic to planar state. Since the serine endopeptidase trypsin can enzymatically catalyze the hydrolysis of PLL, the dark-to-bright shift in the optical response was not observed after transferring a mixed solution of PLL and trypsin onto the DOPG-decorated LC interface, indicating that no orientational transitions in the LCs occurred. However, the optical response from dark to bright was observed when the mixture in the optical cell was replaced by an aqueous solution of PLL. Control experiments with trypsin or an aqueous mixture of PLL and deactivated trypsin further confirmed the feasibility of this approach. The detection limit of trypsin was determined to be ~1 μg/mL. This approach holds great promise for use in the development of LC-based sensors for the detection of enzymatic reactions in cases where the biological polyelectrolyte substrates of enzymes could disrupt the organization of the membrane and induce orientational transitions of LCs at the aqueous/LC interface. © 2012 American Chemical Society
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A novel helper phage enabling construction of genome-scale ORF-enriched phage display libraries.
Gupta, Amita; Shrivastava, Nimisha; Grover, Payal; Singh, Ajay; Mathur, Kapil; Verma, Vaishali; Kaur, Charanpreet; Chaudhary, Vijay K
2013-01-01
Phagemid-based expression of cloned genes fused to the gIIIP coding sequence and rescue using helper phages, such as VCSM13, has been used extensively for constructing large antibody phage display libraries. However, for randomly primed cDNA and gene fragment libraries, this system encounters reading frame problems wherein only one of 18 phages display the translated foreign peptide/protein fused to phagemid-encoded gIIIP. The elimination of phages carrying out-of-frame inserts is vital in order to improve the quality of phage display libraries. In this study, we designed a novel helper phage, AGM13, which carries trypsin-sensitive sites within the linker regions of gIIIP. This renders the phage highly sensitive to trypsin digestion, which abolishes its infectivity. For open reading frame (ORF) selection, the phagemid-borne phages are rescued using AGM13, so that clones with in-frame inserts express fusion proteins with phagemid-encoded trypsin-resistant gIIIP, which becomes incorporated into the phages along with a few copies of AGM13-encoded trypsin-sensitive gIIIP. In contrast, clones with out-of-frame inserts produce phages carrying only AGM13-encoded trypsin-sensitive gIIIP. Trypsin treatment of the phage population renders the phages with out-of-frame inserts non-infectious, whereas phages carrying in-frame inserts remain fully infectious and can hence be enriched by infection. This strategy was applied efficiently at a genome scale to generate an ORF-enriched whole genome fragment library from Mycobacterium tuberculosis, in which nearly 100% of the clones carried in-frame inserts after selection. The ORF-enriched libraries were successfully used for identification of linear and conformational epitopes for monoclonal antibodies specific to mycobacterial proteins.
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Khangembam, Bronson Kumar; Chakrabarti, Rina
2015-05-15
Trypsin was purified 35.64-fold with 4.97% recovery from the viscera of carp Cirrhinus mrigala (mrigal) by ammonium sulfate precipitation, ion exchange and affinity chromatography. The purified enzyme was active at a wide range of pH (7.0-9.2) and temperature (10-50°C). The purified enzyme exhibited high thermal stability up to 50°C for 1h. The enzyme activity was stabilized by Ca(+2) (2mM) up to 7h at 40°C. The Km and kcat values of purified enzyme were 0.0672 mM and 92.09/s/mM, respectively. Soybean trypsin inhibitor and phenylmethylsulphonylflouride completely inhibited the enzyme activity. The specific inhibitor of trypsin, N-α-p-tosyl-L-lysine chloromethyl ketone inhibited 99.67% activity. Na(+), K(+) and Li(+) inhibited 20.99 ± 5.25%, 16.53 ± 4.80% and 18.99 ± 1.42% of enzyme activity, respectively. Divalent ions Mg(+2), Zn(+2), Co(+2), Hg(+2) and Cd(+2) inhibited 21.61 ± 2.22%, 31.62 ± 1.78%, 31.62 ± 1.96%, 85.68 ± 1.51% and 47.95 ± 2.13% enzyme activity, respectively. SDS-PAGE showed that the molecular mass of purified enzyme was 21.7 kDa. MALDI-TOF study showed a peptide sequence of AFCGGSLVNENKMHSAGHCYKSRIQV at the N-Terminal. This sequence recorded 76-84% identity with trypsin from Thunnus thynnus and other fish species. This confirmed that the purified protein was trypsin. The purified enzyme has potential applications in detergent and food industry because of its thermal stability and alkaline nature. Copyright © 2014 Elsevier Ltd. All rights reserved.
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Yang, Qin; Wang, Yuzhi; Zhang, Hongmei; Xu, Kaijia; Wei, Xiaoxiao; Xu, Panli; Zhou, Yigang
2017-11-01
A novel magnetic extractant, PEG 4000 modified Fe 3 O 4 nanomaterial that coated with dianionic amino acid ionic liquid (Fe 3 O 4 @PEG@DAAAIL), was successfully synthesized and characterized. X-ray diffraction (XRD), transmission electron microscope (TEM), vibrating sample magnetometer (VSM), fourier transform infrared spectrometry (FT-IR), thermal gravimetric analysis (TGA) and zeta potentials were used to confirm that the novel nanocomposite was successfully synthesized. Subsequently, the prepared Fe 3 O 4 @PEG@DAAAIL nanocomposite was used as the extractant for trypsin coupled with magnetic solid-phase extraction (MSPE). The concentrations of trypsin in the supernatant were detected by UV-vis spectrophotometer at 278nm. The extraction ability turned out to be better than the other four kinds of extractants prepared in this work. Furthermore, the influence of a series of factors, such as extraction time and temperature, initial trypsin concentration, the value of pH and ionic strength, was systematically investigated. Under the optimal extraction condition, the extraction capacity for trypsin could reach up to 718.73mg/g, absolutely higher than that of other adsorbents reported. This satisfactory extraction capacity could be maintained unchangeable after at least eight days, and kept over 90% of initial extraction capacity after eight recycles. What's more, the activity of trypsin after extraction retained 92.29% of initial activity, verifying the biocompatibility of the prepared extractant. Finally, the developed Fe 3 O 4 @PEG@DAAAIL-MSPE method was successfully applied to the real sample analysis with satisfactory results. All of above proves the potential value of Fe 3 O 4 @PEG@DAAAIL-MSPE in the analysis of biomass. Copyright © 2017 Elsevier B.V. All rights reserved.
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Swathi, Marri; Mishra, Prashant K.; Lokya, Vadthya; Swaroop, Vanka; Mallikarjuna, Nalini; Dutta-Gupta, Aparna; Padmasree, Kollipara
2016-01-01
Proteinase inhibitors (PIs) are natural defense proteins of plants found to be active against gut proteases of various insects. A pigeonpea wild relative Cajanus platycarpus was identified as a source of resistance against Helicoverpa armigera, a most devastating pest of several crops including pigeonpea. In the light of earlier studies, trypsin-specific PIs (CpPI 63) were purified from mature dry seeds of C. platycarpus (ICPW-63) and characterized their biochemical properties in contributing to H. armigera resistance. CpPI 63 possessed significant H. armigera gut trypsin-like proteinase inhibitor (HGPI) activity than trypsin inhibitor (TI) activity. Analysis of CpPI 63 using two-dimensional (2-D) electrophoresis and matrix assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry revealed that it contained several isoinhibitors and small oligomers with masses ranging between 6 and 58 kDa. The gelatin activity staining studies suggest that these isoinhibitors and oligomers possessed strong inhibitory activity against H. armigera gut trypsin-like proteases (HGPs). The N-terminal sequence of the isoinhibitors (pI 6.6 and pI 5.6) of CpPI 63 exhibited 80% homology with several Kunitz trypsin inhibitors (KTIs) as well as miraculin-like proteins (MLPs). Further, modification of lysine residue(s) lead to 80% loss in both TI and HGPI activities of CpPI 63. In contrast, the TI and HGPI activities of CpPI 63 were stable over a wide range of temperature and pH conditions. The reported results provide a biochemical basis for pod borer resistance in C. platycarpus. PMID:27656149
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Methylselenium and Prostate Cancer Apoptosis
2007-02-01
adherent cells were collected by gentle trypsinization and were combined with the floaters for pelleting by centrifugation. After gentle lysis of the...Cancer Ther 2006;5(7). July 2006 trypsinization and were combined with the floaters for pelleting by centrifugation. After gentle lysis of the cells with
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Okamura, Masashi; Yokoyama, Naoaki; Takabatake, Noriyuki; Okubo, Kazuhiro; Ikehara, Yuzuru; Igarashi, Ikuo
2007-02-01
In the present study, we investigated the effects of protease pretreatments of host erythrocytes (RBC) on the in vitro growth of bovine Babesia parasites (Babesia bovis and B. bigemina) and equine Babesia parasites (B. equi and B. caballi). The selected proteases, trypsin and chymotrypsin, clearly modified several membrane proteins of both bovine and equine RBC, as demonstrated by SDS-PAGE analysis; however, the protease treatments also modified the sialic acid content exclusively in bovine RBC, as demonstrated by lectin blot analysis. An in vitro growth assay using the protease-treated RBC showed that the trypsin-treated bovine RBC, but not the chymotrypsin-treated ones, significantly reduced the growth of B. bovis and B. bigemina as compared to the control. In contrast, the growth of B. equi and B. caballi was not affected by any of these proteases. Thus, the bovine, but not the equine, Babesia parasites require the trypsin-sensitive membrane (sialoglyco) proteins to infect the RBC.
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Zdarta, Jakub; Antecka, Katarzyna; Jędrzak, Artur; Synoradzki, Karol; Łuczak, Magdalena; Jesionowski, Teofil
2018-05-08
In the presented study synthesized magnetic nanoparticles were used as an inorganic precursor for the preparation of novel magnetite-lignin and magnetite-chitin hybrid supports for enzyme immobilization. Effective synthesis of the hybrids was confirmed by Fourier transform infrared spectroscopy and powder X-ray diffraction analysis. The materials exhibited good thermal stability and surface areas of 4.3 and 5.6 m 2 /g respectively. The magnetite-lignin + trypsin and magnetite-chitin + trypsin systems were found to have good storage stability and reusability. After 20 days they retained over 75% and 90% respectively of their initial activity, and after 10 consecutive biocatalytic cycles retained over 60% and 80% respectively of their initial activity. The kinetic parameters of the free and immobilized enzyme were also comprehensively examined and compared. The results of peptide digestion tests confirmed the high proteolytic activity of the produced trypsin-based magnetic biocatalytic systems. Copyright © 2018 Elsevier B.V. All rights reserved.
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Perreault, Véronique; Hénaux, Loïc; Bazinet, Laurent; Doyen, Alain
2017-04-15
The effect of high hydrostatic pressure (HHP) on flaxseed protein structure and peptide profiles, obtained after protein hydrolysis, was investigated. Isolated flaxseed protein (1%, m/v) was subjected to HHP (600MPa, 5min or 20min at 20°C) prior to hydrolysis with trypsin only and trypsin-pronase. The results demonstrated that HHP treatment induced dissociation of flaxseed proteins and generated higher molecular weight aggregates as a function of processing duration. Fluorescence spectroscopy showed that HHP treatment, as well as processing duration, had an impact on flaxseed protein structure since exposition of hydrophobic amino acid tyrosine was modified. Except for some specific peptides, the concentrations of which were modified, similar peptide profiles were obtained after hydrolysis of pressure-treated proteins using trypsin. Finally, hydrolysates obtained using trypsin-pronase had a greater antioxidant capacity (ORAC) than control samples; these results confirmed that HHP enhanced the generation of antioxidant peptides. Copyright © 2016 Elsevier Ltd. All rights reserved.
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Shazly, Ahmed Behdal; He, Zhiyong; El-Aziz, Mahmoud Abd; Zeng, Maomao; Zhang, Shuang; Qin, Fang; Chen, Jie
2017-10-01
Buffalo and bovine caseins were hydrolysed by alcalase and trypsin to produce novel antioxidant peptides. The casein hydrolysates were purified using ultrafiltration (UF) and further characterized by RP-HPLC. The fractions produced higher antioxidant activities were identified for their peptides using LC MS/MS. All UF-VI (MW<1kDa) fractions showed higher antioxidant activity. Hydrolysate produced by alcalase for buffalo casein (UF-VI with 54.84-fold purification) showed higher antioxidant activity than that obtained by trypsin. Trypsin hydrolysate contained high amount of hydrophobic amino acids while alcalase hydrolysate consisted mainly of Ser, Arg, Ala and Leu. The antioxidant peptides identified by LC MS/MS were RELEE, MEDNKQ and TVA, EQL in buffalo casein hydrolysates produced by trypsin and alcalase, respectively. Mechanism and reaction pathways of selected antioxidant peptides with ABTS were proposed. Conclusively, buffalo casein provided antioxidant peptides similar to bovine, suggesting that buffalo casein is a novel source of antioxidant. Copyright © 2017 Elsevier Ltd. All rights reserved.
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Ramare, F; Nicoli, J; Dabard, J; Corring, T; Ladire, M; Gueugneau, A M; Raibaud, P
1993-09-01
An antibacterial substance appeared within 1 day in feces of gnotobiotic rats harboring a human intestinal Peptostreptococcus strain. It disappeared when the rat bile-pancreatic duct was ligatured or when the rats ingested a trypsin inhibitor. Anaerobic cultures of the Peptostreptococcus strain in a medium supplemented with trypsin also exhibited an antibacterial activity, which was also inhibited by the trypsin inhibitor. In vitro the antibacterial substance from both feces and culture medium was active against several gram-positive bacteria, including other Peptostreptococcus spp., potentially pathogenic Clostridium spp. such as C. perfringens, C. difficile, C. butyricum, C. septicum, and C. sordellii, Eubacterium spp., Bifidobacterium spp., and Bacillus spp. Whatever the order of inoculation of the strains, a sensitive strain of C. perfringens was eliminated within 1 day from the intestine of rats monoassociated with the Peptostreptococcus strain. These findings demonstrate for the first time that very potent antibacterial substances can be produced through a mechanism involving intestinal bacteria and exocrine pancreatic secretions.
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Comparison of polymer induced and solvent induced trypsin denaturation: the role of hydrophobicity.
Jasti, Lakshmi S; Fadnavis, Nitin W; Addepally, Uma; Daniels, Siona; Deokar, Sarika; Ponrathnam, Surendra
2014-04-01
Trypsin adsorption from aqueous buffer by various copolymers of allyl glycidyl ether-ethylene glycol dimethacrylate (AGE-EGDM) copolymer with varying crosslink density increases with increasing crosslink density and the effect slowly wears off after reaching a plateau at 50% crosslink density. The copolymer with 25% crosslink density was reacted with different amines with alkyl/aryl side chains to obtain a series of copolymers with 1,2-amino alcohol functional groups and varying hydrophobicity. Trypsin binding capacity again increases with hydrophobicity of the reacting amine and a good correlation between logPoctanol of the amine and protein binding is observed. The bound trypsin is denatured to the extent of 90% in spite of the presence of hydrophilic hydroxyl and amino groups. The behavior was comparable to that in mixtures of aqueous buffer and water-miscible organic co-solvents where the solvent concentration required to deactivate 50% of the enzyme (C50) is dependent on logPoctanol of the co-solvent. Copyright © 2014 Elsevier B.V. All rights reserved.
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Fresh and aged human lymphocyte metaphase slides are equally usable for GTG banding.
Sajjad, Naheed; Haque, Sayedul; SBurney, Syed Intesar; Shahid, Syed Muhammad; Zehra, Sitwat; Azhar, Abid
2014-09-01
The identification of chromosomes for routine cytogenetic analysis is based on quality of metaphases and good banding pattern. Fresh slides of human lymphocytes have been shown to produce good bands for the identification of chromosomes morphology. G-bands by Trypsin using Giemsa (GTG) banding of aged slides is generally considered hard to get desired band pattern of chromosomes persistently. The current study is focused on GTG banding of aged slides. A total of 340 subjects including 290 primary infertile and 50 fertile were selected. The blood samples were drawn aseptically for cytogenetic analysis. Lymphocytes were cultured and GTG banding was done on 1440 glass slides. Giemsa trypsin banding of aged slides were done by adjusting average trypsin time for each month according to the slide age and metaphase concentration. Correlation analyses showed a significant and positive correlation between slide ageing and trypsin pre-treatment time. The results of this study suggest that, the fresh and aged human lymphocyte metaphases are equally usable for GTG banding.
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Birkenhäger, J C; Upton, G V; Seldenrath, H J; Krieger, D T; Tashjian, A H
1976-10-01
A 45-year-old women had medullary tyroid carcinoma associated with Cushing's syndrome and galactorrhoea. Elevated plasma immunoreactive ACTH and cortisol were partially suppressed by intravenous dexamethasone, appreciably raised by lysine vasopressin, and urinary excretion of 17-oxogenic steroids slightly elevated by metyrapone. A large arterio-venous increase in plasma corticotrophin releasing factor-like activity across the thyroid gland was observed and tumour tissue contained corticotrophin releasing factor-like activity. Biologically active ACTH was not detected in tumour extracts before incubation with trypsin, but after trypsinization a value of 3.2 mU per gram was obtained. Arterial plasma contained biologically active ACTH (1.5 mU/100 ml) prior to trypsinization. Venous effluent from the thyroid gland contained biologically active (9.6 mU/100 ml) and immunoreactive ACTH (970 pg/ml) before trypsinization. Tumour extracts also contained prolactin production-stimulating activity. These findings can explain the Cushing's syndrome and the galactorrhoea both of which disappeared completely after thyroidectomy.
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A Murray Cod Assemblage: Re/Considering Riverscape Pedagogy
ERIC Educational Resources Information Center
Stewart, Alistair James
2018-01-01
This article enacts Deleuze and Guattari's (1987) concept "assemblage" to craft a riverScape pedagogy that is informed by, and responsive to, the Murray Cod, the river, and its circumstances. The Murray Cod, the largest fish species in Australia's Murray-Darling Basin, has diverse cultural meanings. Cod are at once a creation being of…
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Federal Register 2010, 2011, 2012, 2013, 2014
2012-02-17
... DEPARTMENT OF THE INTERIOR National Park Service [1730-SZM] Cape Cod National Seashore Advisory Commission; Cape Cod National Seashore, South Wellfleet, MA AGENCY: National Park Service, Interior. ACTION... National Seashore, 99 Marconi Site Road, Wellfleet, MA 02667. Date: February 7, 2012. George E. Price, Jr...
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Ghunmi, Lina Abu; Zeeman, Grietje; Fayyad, Manar; van Lier, Jules B
2011-02-01
Knowing the biodegradability characteristics of grey water constituents is imperative for a proper design and operation of a biological treatment system of grey water. This study characterizes the different COD fractions of dormitory grey water and investigates the effect of applying different conditions in the biodegradation test. The maximum aerobic and anaerobic biodegradability and conversion rate for the different COD fractions is determined. The results show that, on average, dormitory grey water COD fractions are 28% suspended, 32% colloidal and 40% dissolved. The studied factors incubation time, inoculum addition and temperature are influencing the determined biodegradability. The maximum biodegradability and biodegradation rate differ between different COD fractions, viz. COD(ss), COD(col) and COD(diss). The dissolved COD fraction is characterised by the lowest degradation rate, both for anaerobic and aerobic conditions. The maximum biodegradability for aerobic and anaerobic conditions is 86 and 70% respectively, whereas the first order conversion rate constant, k₂₀, is 0.119 and 0.005 day⁻¹, respectively. The anaerobic and aerobic conversion rates in relation to temperature can be described by the Arrhenius relation, with temperature coefficients of 1.069 and 1.099, respectively.
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NASA Astrophysics Data System (ADS)
Castelló Serrano, Iván; Stoica, Georgiana; Matas Adams, Alba; Palomares, Emilio
2014-10-01
We present herein two colour encoded silica nanospheres (2nanoSi) for the fluorescence quantitative ratiometric determination of trypsin in humans. Current detection methods for cystic fibrosis diagnosis are slow, costly and suffer from false positives. The 2nanoSi proved to be a highly sensitive, fast (minutes), and single-step approach nanosensor for the screening and diagnosis of cystic fibrosis, allowing the quantification of trypsin concentrations in a wide range relevant for clinical applications (25-350 μg L-1). Furthermore, as trypsin is directly related to the development of cystic fibrosis (CF), different human genotypes, i.e. CF homozygotic, CF heterozygotic, and unaffected, respectively, can be determined using our 2nanoSi nanospheres. We anticipate the 2nanoSi system to be a starting point for non-invasive, easy-to-use and cost effective ratiometric fluorescent biomarkers for recessive genetic diseases like human cystic fibrosis. In a screening program in which the goal is to detect disease and also the carrier status, early diagnosis could be of great help.We present herein two colour encoded silica nanospheres (2nanoSi) for the fluorescence quantitative ratiometric determination of trypsin in humans. Current detection methods for cystic fibrosis diagnosis are slow, costly and suffer from false positives. The 2nanoSi proved to be a highly sensitive, fast (minutes), and single-step approach nanosensor for the screening and diagnosis of cystic fibrosis, allowing the quantification of trypsin concentrations in a wide range relevant for clinical applications (25-350 μg L-1). Furthermore, as trypsin is directly related to the development of cystic fibrosis (CF), different human genotypes, i.e. CF homozygotic, CF heterozygotic, and unaffected, respectively, can be determined using our 2nanoSi nanospheres. We anticipate the 2nanoSi system to be a starting point for non-invasive, easy-to-use and cost effective ratiometric fluorescent biomarkers for recessive genetic diseases like human cystic fibrosis. In a screening program in which the goal is to detect disease and also the carrier status, early diagnosis could be of great help. Electronic supplementary information (ESI) available. See DOI: 10.1039/c4nr03952a
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Guldager, H S; Bøknaes, N; Osterberg, C; Nielsen, J; Dalgaard, P
1998-09-01
The effect of two months of frozen storage at -20 degrees C on the spoilage characteristics and shelf life of thawed and modified atmosphere packed (MAP) cod fillets stored at 2 degrees C was studied. Thawed MAP cod fillets were compared with fresh cod fillets stored in CO2-containing modified atmospheres with and without added oxygen. The shelf life of 11 to 12 days in the fresh MAP cod was extended to more than 20 days in the thawed MAP cod at 2 degrees C. This shelf life extension was most likely due to the inactivation of the spoilage bacterium Photobacterium phosphoreum during frozen storage as reflected both in chemical analyses and sensory evaluation. In contrast to fresh MAP cod fillets no significant production of trimethylamine occurred and almost no amine odor and taste were detected during 20 days of chill storage of thawed MAP cod fillets. The use of frozen fillets as raw material not only provides a more stable product in MAP but also allows much greater flexibility for production and distribution. However, a slightly increased concentration of dimethylamine, a larger drip loss, and detection of weak frozen storage flavor were observed in the thawed MAP cod fillets.
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NASA Astrophysics Data System (ADS)
Samudro, Ganjar; Syafrudin; Nugraha, Winardi Dwi; Sutrisno, Endro; Priyambada, Ika Bagus; Muthi'ah, Hilma; Sinaga, Glory Natalia; Hakiem, Rahmat Tubagus
2018-02-01
This research is conducted to analyze and determine the optimum of COD concentration containing leaves litter, canteen and composite waste to power density and COD removal efficiency as the indicator of SMFC performance. COD as the one of organic matter parameters perform as substrate, nutrient and dominating the whole process of SMFC. Leaves litter and canteen based food waste were obtained from TPST UNDIP in Semarang and treated in SMFC reactor. Its reactor was designed 2 liter volume and equipped by homemade graphene electrodes that were utilized at the surface of organic waste as cathode and in a half of reactor height as anode. COD concentration was initially characterized and became variations of initial COD concentration. Waste volume was maintained 2/3 of volume of reactor. Bacteria sources as the important process factor in SMFC were obtained from river sediment which contain bacteroides and exoelectrogenic bacteria. Temperature and pH were not maintained while power density and COD concentration were periodically observed and measured during 44 days. The results showed that power density up to 4 mW/m2 and COD removal efficiency performance up to 70% were reached by leaves litter, canteen and composite waste at days 11 up to days 44 days. Leaves litter contain 16,567 mg COD/l providing higher COD removal efficiency reached approximately 87.67%, more stable power density reached approximately 4.71 mW/m2, and faster optimum time in the third day than canteen based food waste and composite waste. High COD removal efficiency has not yet resulted in high power density.
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Diet of Norwegian coastal cod (Gadus morhua) studied by using citizen science
NASA Astrophysics Data System (ADS)
Enoksen, Siri Elise; Reiss, Henning
2018-04-01
The Norwegian coastal cod (Gadus morhua) is a keystone species in the food web of northern Norwegian fjords. Their relatively stationary populations might specifically depend on local food resources, but the diet of cod has rarely been studied in fjord systems. Using a citizen science approach, where recreational anglers and tourists participated in the sampling, we studied small-scale differences in the diet composition of cod in a fjord system in northern Norway. We compared the cod diet from the MPA Saltstraumen, characterised by strong tidal currents and a highly diverse and abundant fauna, with the inner fjord area of Skjerstadfjord. The diet composition of cod significantly differed between both areas within the fjord. Although fish was the dominant prey in both areas, cod consumed > 40% invertebrates in terms of weight, even in the cod size class of 70-99 cm. The invertebrate prey also caused the observed spatial differences. In Saltstraumen, brittle stars (Ophiuroidea), crabs (Brachyura) and sea cucumbers (Holothuroidea) were important food sources for cod, while sea urchins (Echinoidea), clams (Bivalvia), shrimps (Caridea) and krill (Euphausiacea) dominated the diet in the inner Skjerstadfjord. The high densities of sessile fauna in the dynamic environment of Saltstraumen, was only partly reflected in the diet of cod, with only Holothuroidea found in 17% of the stomachs. High rates of empty stomachs (24%), cannibalism as well as a higher proportion of low-energy prey in the diet of large cod, may indicate a shortage of high quality food in Skjerstadfjord. The samples for this study were collected through a citizen science campaign. This approach might provide opportunities to be used for coastal ecological monitoring with potential applications in local ecosystem management strategies through public involvement.
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Elutriated acid fermentation of municipal primary sludge.
Ahn, Young Ho; Speece, Richard E
2006-06-01
The performance of a novel fermentation process, adopting a sludge blanket type configuration, for higher hydrolysis/acidogenesis of the municipal primary sludge was investigated under batch and semi-continuous conditions with varying pH and temperature. This acid elutriation slurry reactor provided higher system performance with a short HRT (5d) and higher acidogenic effluent quality under pH 9 and thermophilic (55 degrees C) conditions. The hydrolysis of the sludge was revealed to be significantly dependent on seasonal effects for sludge characteristics but with little impact on acidogenesis. Based on the rainy season at the optimal conditions, VFA production and recovery fraction (VFA(COD)/COD) were 0.18 g VFA(COD)/g VSS(COD) and 63%. As byproducts, nitrogen and phosphorus release were measured at 0.006 g N/g VSS(COD) and 0.003 g P/g VSS(COD), respectively. For the mass balance in a full-scale plant (Q=158,880 m(3)/d) based on the rainy season, the VFA and non-VFA (as COD) production were 3110 kg VFA(COD)/d and 1800 kg COD/d, resulting in an increase of organics of 31 mg COD/L and 20mg VFA(COD)/L and nutrients of 0.7 mg N/L and 0.3 mg P/L in the influent sewage. The economical benefit from this process application was estimated to be about 67 dollars per 1000 m(3) of sewage except for energy requirements and also, better benefits can be expected during the dry season. Moreover, the results revealed that the process has various additional advantages such as pathogen-free stabilized solids production, excellent solids control and economical benefits.
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Leo, Elettra; Kunz, Kristina L; Schmidt, Matthias; Storch, Daniela; Pörtner, Hans-O; Mark, Felix C
2017-01-01
Ocean acidification and warming are happening fast in the Arctic but little is known about the effects of ocean acidification and warming on the physiological performance and survival of Arctic fish. In this study we investigated the metabolic background of performance through analyses of cardiac mitochondrial function in response to control and elevated water temperatures and P CO 2 of two gadoid fish species, Polar cod ( Boreogadus saida ), an endemic Arctic species, and Atlantic cod ( Gadus morhua ), which is a temperate to cold eurytherm and currently expanding into Arctic waters in the wake of ocean warming. We studied their responses to the above-mentioned drivers and their acclimation potential through analysing the cardiac mitochondrial function in permeabilised cardiac muscle fibres after 4 months of incubation at different temperatures (Polar cod: 0, 3, 6, 8 °C and Atlantic cod: 3, 8, 12, 16 °C), combined with exposure to present (400μatm) and year 2100 (1170μatm) levels of CO 2 . OXPHOS, proton leak and ATP production efficiency in Polar cod were similar in the groups acclimated at 400μatm and 1170μatm of CO 2 , while incubation at 8 °C evoked increased proton leak resulting in decreased ATP production efficiency and decreased Complex IV capacity. In contrast, OXPHOS of Atlantic cod increased with temperature without compromising the ATP production efficiency, whereas the combination of high temperature and high P CO 2 depressed OXPHOS and ATP production efficiency. Polar cod mitochondrial efficiency decreased at 8 °C while Atlantic cod mitochondria were more resilient to elevated temperature; however, this resilience was constrained by high P CO 2 . In line with its lower habitat temperature and higher degree of stenothermy, Polar cod has a lower acclimation potential to warming than Atlantic cod.
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Lobel, Lior; Herskovits, Anat A.
2016-01-01
Bacteria sense and respond to many environmental cues, rewiring their regulatory network to facilitate adaptation to new conditions/niches. Global transcription factors that co-regulate multiple pathways simultaneously are essential to this regulatory rewiring. CodY is one such global regulator, controlling expression of both metabolic and virulence genes in Gram-positive bacteria. Branch chained amino acids (BCAAs) serve as a ligand for CodY and modulate its activity. Classically, CodY was considered to function primarily as a repressor under rich growth conditions. However, our previous studies of the bacterial pathogen Listeria monocytogenes revealed that CodY is active also when the bacteria are starved for BCAAs. Under these conditions, CodY loses the ability to repress genes (e.g., metabolic genes) and functions as a direct activator of the master virulence regulator gene, prfA. This observation raised the possibility that CodY possesses multiple functions that allow it to coordinate gene expression across a wide spectrum of metabolic growth conditions, and thus better adapt bacteria to the mammalian niche. To gain a deeper understanding of CodY’s regulatory repertoire and identify direct target genes, we performed a genome wide analysis of the CodY regulon and DNA binding under both rich and minimal growth conditions, using RNA-Seq and ChIP-Seq techniques. We demonstrate here that CodY is indeed active (i.e., binds DNA) under both conditions, serving as a repressor and activator of different genes. Further, we identified new genes and pathways that are directly regulated by CodY (e.g., sigB, arg, his, actA, glpF, gadG, gdhA, poxB, glnR and fla genes), integrating metabolism, stress responses, motility and virulence in L. monocytogenes. This study establishes CodY as a multifaceted factor regulating L. monocytogenes physiology in a highly versatile manner. PMID:26895237
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NASA Astrophysics Data System (ADS)
Vanyushin, George
2015-04-01
Continuous long-term database (1998-2014) on the sea surface temperature (SST) comprising results of regional satellite monitoring (the Norwegian and the Barents seas) is used to resolve several applied problems. Authors have analyzed indirect influence the SST (the NOAA satellite data) on modern cod total stock biomass (abundance of the Northeast Arctic cod at age 3+). In this study, we went on the consideration of the relationship between the SST anomalies for March-April in the main spawning ground of the cod off the Lofoten islands in the Norwegian Shelf Waters and forecasting assessment of future cod generation success and its future abundance of 3 year old. Mean monthly SST and SST anomalies are computed for the selected area on the basis of the weekly SST maps which made by using the NOAA satellites data for the period 1998-2014. Comparison of the SST anomalies in the main spawning ground with abundance of the cod year class at age 3+ shows that survival of the cod generations was inhibited on the whole as negative (below -0,1C) well as positive SST anomalies (above +1,3C) during March and April. Finally, the results indicate that poor and low middle generations of cod at age 3+ (2002, 2004, 2010) occurred in years with negative or extremely high positive the SST anomalies in the spawning area. The SST anomalies in years which were close to normal significances provide conditions for appearance middle or strong generations of cod (2001-2003, 2005-2009, 2011-2013). So, the SST and SST anomalies (by the NOAA satellite data) characterize of increase in input of warm Atlantic waters which form numerous eddies along the main stream thus creating favorable conditions for spawning and development of the cod larvae and fry and provide them with food stock, finally direct influence on forming total stock biomass of cod and helping its population forecast. Key words: satellite monitoring of SST, the Northeast Arctic cod, spawning ground, forecast of the cod year class strength at age 3+.
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Hypotrypsinaemia in diabetes mellitus.
Adrian, T E; Barnes, A J; Bloom, S R
1979-10-01
Plasma trypsin concentrations were measured in 403 fasting diabetics and 106 healthy controls. Basal trypsin concentrations in the normal subjects were 88 +/- 6 ng/ml (mean +/- S.E.M.). Mean plasma trypsin concentrations in diabetics treated with diet alone (n = 74) were 45 +/- 2 ng/ml, while in a group of young (less than 35 years, n = 88) insulin-dependent diabetics, they were very low at 29 +/- 2 ng/ml and these levels were inversely related to insulin dosage. The findings may help in the understanding of the pathophysiological changes in the exocrine pancreas in the diabetic state and may also shed some light on the physiological interrelationship between the endocrine and exocrine pancreas.
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Li, Jihong; Freedman, John C; McClane, Bruce A
2015-10-01
Clostridium perfringens type D strains are usually associated with diseases of livestock, and their virulence requires the production of epsilon toxin (ETX). We previously showed (J. Li, S. Sayeed, S. Robertson, J. Chen, and B. A. McClane, PLoS Pathog 7:e1002429, 2011, http://dx.doi.org/10.1371/journal.ppat.1002429) that BMC202, a nanI null mutant of type D strain CN3718, produces less ETX than wild-type CN3718 does. The current study proved that the lower ETX production by strain BMC202 is due to nanI gene disruption, since both genetic and physical (NanI or sialic acid) complementation increased ETX production by BMC202. Furthermore, a sialidase inhibitor that interfered with NanI activity also reduced ETX production by wild-type CN3718. The NanI effect on ETX production was shown to involve reductions in codY and ccpA gene transcription levels in BMC202 versus wild-type CN3718. Similar to CodY, CcpA was found to positively control ETX production. A double codY ccpA null mutant produced even less ETX than a codY or ccpA single null mutant. CcpA bound directly to sequences upstream of the etx or codY start codon, and bioinformatics identified putative CcpA-binding cre sites immediately upstream of both the codY and etx start codons, suggesting possible direct CcpA regulatory effects. A ccpA mutation also decreased codY transcription, suggesting that CcpA effects on ETX production can be both direct and indirect, including effects on codY transcription. Collectively, these results suggest that NanI, CcpA, and CodY work together to regulate ETX production, with NanI-generated sialic acid from the intestines possibly signaling type D strains to upregulate their ETX production and induce disease. Clostridium perfringens NanI was previously shown to increase ETX binding to, and cytotoxicity for, MDCK host cells. The current study demonstrates that NanI also regulates ETX production via increased transcription of genes encoding the CodY and CcpA global regulators. Results obtained using single ccpA or codY null mutants and a ccpA codY double null mutant showed that codY and ccpA regulate ETX production independently of one another but that ccpA also affects codY transcription. Electrophoretic mobility shift assays and bioinformatic analyses suggest that both CodY and CcpA may directly regulate etx transcription. Collectively, results of this study suggest that sialic acid generated by NanI from intestinal sources signals ETX-producing C. perfringens strains, via CcpA and CodY, to upregulate ETX production and cause disease. Copyright © 2015, American Society for Microbiology. All Rights Reserved.
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Trypsin level in gallbladder bile and ductitis and width of the cystic duct.
Vracko, J; Wiechel, K L
2000-01-01
The change from laparotomy to laparoscopy for cholecystectomy has raised the question of how to manage concomitant bile duct stones. The present-day interest--and controversy--has focused on a transcystic approach reported to be feasible in 66-96% of cases, but without explaining the necessary prerequisite: the widening of the cystic duct. The cystic duct, wide mainly in patients with bile duct stones, has been reported to be highly variable: from strictured to very wide. The present study aims at comparing the trypsin level in the gallbladder bile and the cystic duct morphology and width in patients with and without bile duct stones. A prospective series of 63 gallstone patients, 30 with and 33 without bile duct stones (controls), underwent cholecystectomy and bile duct clearance. The study includes the trypsin level in the gallbladder bile, the width and morphology of the cystic duct, and the size of the gallstones. The patients with bile duct stones had, in contrast to the controls, higher trypsin levels in the gallbladder bile (P < 0.001) and wider cystic ducts (P < 0.001) with more pronounced signs of chronic ductitis. The obtained results strongly suggest that the increased trypsin level, a sign of reflux of pancreatic juice, caused changes in the cystic duct that facilitate gallstone migration, which also ought to render a transcystic stone extraction feasible.
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Rahman, Shafiq; Griffin, Michelle; Naik, Anish; Szarko, Matthew; Butler, Peter E M
2018-02-15
Decellularized scaffolds can induce chondrogenic differentiation of stem cells. This study compares different methods to optimise the decellularization of auricular cartilage. The process consisted of an initial 12 hour dry freeze thaw which froze the cartilage specimens in an empty tube at -20 °C. Samples were allowed to thaw at room temperature followed by submersion in phosphate buffer solution in which they were frozen at -20 °C for a 12 hour period. They were then allowed to thaw at room temperature as before. Protocol A subsequently involved subjecting specimens to both deoxyribonuclease and sodium deoxycholate. Protocol B and C were adaptations of this using 0.25% trypsin (7 cycles) and a 0.5 molar solution of ethylenediaminetetraacetic acid (3 hours for each cycle) respectively as additional steps. Trypsin accelerated the decellularization process with a reduction in DNA content from 55.4 ng/μL (native) to 17.3 ng/μL (P-value < 0.05) after 14 days. Protocol B showed a faster reduction in DNA content when compared with protocol A. In comparison to protocol C after 14 days, trypsin also showed greater decellularization with a mean difference of 11.7 ng/μL (P-value < 0.05). Histological analysis with H&E and DAPI confirmed depletion of cells at 14 days with trypsin.
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Proteases induce secretion of collagenase and plasminogen activator by fibroblasts
DOE Office of Scientific and Technical Information (OSTI.GOV)
Werb, Z.; Aggeler, J.
1978-04-01
We have observed that treatment of rabbit synovial fibroblasts with proteolytic enzymes can induce secretion of collagenase (EC 3.4.24.7) and plasminogen activator (EC 3.4.21.-). Cells treated for 2 to 24 hr with plasmin, trypsin, chymotrypsin, pancreatic elastase, papain, bromelain, thermolysin, or ..cap alpha..-protease but not with thrombin or neuraminidase secreted detectable amounts of collagenase within 16 to 48 hr. Treatment of fibroblasts with trypsin also induced secretion of plasminogen activator. Proteases initiated secretion of collagenase (up to 20 units per 10/sup 6/ cells per 24 hr) only when treatment produced decreased cell adhesion. Collagenase production did not depend on continuedmore » presence of proteolytic activity or on subsequent cell adhesion, spreading, or proliferation. Routine subculturing with crude trypsin also induced collagenase secretion by cells. Secretion of collagenase was prevented and normal spreading was obtained if the trypsinized cells were placed into medium containing fetal calf serum. Soybean trypsin inhibitor, ..cap alpha../sub 1/-antitrypsin, bovine serum albumin, collagen, and fibronectin did not inhibit collagenase production. Although proteases that induced collagenase secretion also removed surface glycoprotein, the kinetics of induction of cell protease secretion were different from those for removal of fibronectin. Physiological inducers of secretion of collagenase and plasminogen activator by cells have not been identified. These results suggest that extracellular proteases in conjunction with plasma proteins may govern protease secretion by cells.« less
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Zhou, Tingting; Wang, Xichang; Yan, Juan; Li, Yan
2018-06-01
The ability of Antarctic krill, Euphausia superba (Dana, 1852), to thrive in a cold environment comes from its capacity to synthesize cold-adapted enzymes. Its trypsin, as a main substance in the metabolic reactions, plays a key role in the adaption to low temperatures. However, the progress of research on its cold-adaption mechanism is being influenced due to the limited information on its gene and spatial structure. We studied the gene of E. superba trypsin with transcriptome sequencing first, and then discussed its cold-adaption mechanism with the full gene and predicted structure basing on bioinformatics. The results showed the proportion of certain residues played important roles in the cold-adaptation behavior for trypsin. Furthermore, a higher proportion of random coils and reduced steric hindrance might also be key factors promoting its cold adaption. This research aimed to reveal the cold-adaption mechanism of E. superba trypsin and provide support for basic research on molecular modification by site-directed mutagenesis of complementary DNA used to produce new and improved recombinant variants with cold adaption. Furthermore, it may broaden its commercial application on minimizing undesirable changes elevated at higher temperature in food processing and in treatment of trauma and inflammation in medicine. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.
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USDA-ARS?s Scientific Manuscript database
Trypsin modulating oostatic factor (TMOF), a decapaptide isolated from the ovaries of Aedes aegypti, is the physiological factor that terminates the trypsin biosynthesis after the blood meal. Earlier results obtained from feeding mosquito larvae and injecting female mosquitoes with TMOF show that tr...
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Removal of slowly biodegradable COD in combined thermophilic UASB and MBBR systems.
Ji, M; Yu, J; Chen, H; Yue, P L
2001-09-01
Starch, cellulose and polyvinyl alcohol (PVA) are common substrates of the slowly biodegradable COD (SBCOD) in industrial wastewaters. Removal of the individual and mixed SbCOD substrates was investigated in a combined system of thermophilic upflow anaerobic sludge blanket (TUASB) reactor (55 degrees C) and aerobic moving bed biofilm reactor (MBBR). The removal mechanisms of the three SBCOD substrates were quite different. Starch-COD was almost equally utilized and removed in the two reactors. Cellulose-COD was completely (97-98%) removed from water in the TUASB reactor by microbial entrapment and sedimentation of the cellulose fibers. PVA alone was hardly biodegraded and removed by the combined reactors. However, PVA-COD could be removed to some extent in a binary solution of starch (77%) plus PVA (23%). The PVA macromolecules in the binary solution actually affected the microbial activity in the TUASB reactor resulting accumulation of volatile fatty acids, which shifted the overall COD removal from the TUASB to the MBBR reactor where SBCOD including PVA-COD was removed. Since the three SBCOD substrates were removed by different mechanisms, the combined reactors showed a better and more stable performance than individual reactors.
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Combined thermo-chemo-sonic disintegration of waste activated sludge for biogas production.
Kavitha, S; Yukesh Kannah, R; Yeom, Ick Tae; Do, Khac-Uan; Banu, J Rajesh
2015-12-01
In the present study, there was an investigation about the impact of a new combined thermo-chemo-sonic disintegration of waste activated sludge (WAS) on biodegradability. The outcome of sludge disintegration reveals that maximum Suspended Solids (SS) reduction and Chemical Oxygen Demand (COD) solubilization effectuated at a specific energy input of 5290.5kJ/kgTS, and was found to be 20%, 16.4%, 15% and 27%, 22%, and 20%, respectively for the three alkalis (NaOH, KOH, and Ca(OH)2). The conversion coefficient of the Volatile Suspended Solids (VSS) to product Soluble COD (SCOD), calculated by nonlinear regression modeling, was found to be 0.5530gSCOD/gVSS, 0.4587gSCOD/gVSS, and 0.4195gSCOD/gVSS for NaOH, KOH, and Ca(OH)2, respectively. In the biodegradability studies, the parameter evaluation provides an estimate of parameter uncertainty and correlation, and elucidates that there is no significant difference in biodegradability (0.413gCOD/gCOD, 0.367gCOD/gCOD, and 0.342gCOD/gCOD) for three alkalis (NaOH, KOH, and Ca(OH)2). Copyright © 2015 Elsevier Ltd. All rights reserved.
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Lin, Richen; Cheng, Jun; Yang, Zongbo; Ding, Lingkan; Zhang, Jiabei; Zhou, Junhu; Cen, Kefa
2016-08-01
Cassava ethanol wastewater (CEW) was subjected to sequential dark H2, photo H2 and CH4 fermentation to maximize H2 production and energy yield. A relatively low H2 yield of 23.6mL/g soluble chemical oxygen demand (CODs) was obtained in dark fermentation. To eliminate the inhibition of excessive NH4(+) on sequential photo fermentation, zeolite was used to remove NH4(+) in residual dark solution (86.5% removal efficiency). The treated solution from 5gCODs/L of CEW achieved the highest photo H2 yield of 369.7mL/gCODs, while the solution from 20gCODs/L gave the lowest yield of 259.6mL/gCODs. This can be explained that photo H2 yield was correlated to soluble metabolic products (SMPs) yield in dark fermentation, and specific SMPs yield decreased from 38.0 to 18.1mM/g CODs. The total energy yield significantly increased to 8.39kJ/gCODs by combining methanogenesis with a CH4 yield of 117.9mL/gCODs. Copyright © 2016 Elsevier Ltd. All rights reserved.
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Earth observations taken from shuttle orbiter Atlantis during STS-84 mission
1997-07-09
STS084-703-003 (15-24 May 1997) --- Cape Cod extends 105 kilometers (65 miles) into the Atlantic Ocean. To the south of Cape Cod are the islands of Martha's Vineyard (west) and Nantucket (east). The city of Boston can be seen surrounding the bay above the "hook" on Cape Cod. To the south are the cities of New Bedford on Buzzards Bay, and Providence, Rhode Island. The Cape Cod Canal is an artificial waterway that connects Buzzards Bay and Cape Cod Bay. The waterway is 28 kilometers (17.5 miles) and does not contains locks. The canal was built to shorten the distance over water between New York City and Boston. In 1620 the Pilgrims landed at Provincetown, on the upper tip of Cape Cod, before they proceeded to Plymouth.
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Tawfik, Ahmed; Wahab, Rifaat Abdel; Al-Asmer, Azza; Matary, Fatma
2011-08-01
Grey wastewater (GW) treatment via down-flow hanging sponge (DHS) system was the subject of the study. The reactor was operated at different hydraulic retention times (HRTs) of 11.7, 5.8 and 2.9 h, corresponding to organic loading rates (OLRs) of 1.9, 3.6 and 6.8 kgCOD/m3 day, respectively. The results obtained revealed that decreasing the HRT from 11.7 to 2.9 h negatively affected on the performance of the DHS system. COD(total), COD(soluble), COD(particulate) and detergent removal efficiency were reduced from 96 ± 2.4 to 90 ± 2.3%, from 83 ± 10 to 69 ± 8%, from 98 ± 2 to 94 ± 3% and from 96 ± 12 to 88 ± 6.9%, respectively. However, the removal efficiency of the distinguished COD fractions and detergent remained unaffected when decreasing the HRT from 11.7 to 5.8 h. The DHS system provided a removal efficiency of 95 ± 1% for COD(total), 79 ± 8% for COD(soluble), 98 ± 2 for COD(particulate) and 94.7% for detergent at an HRT of 5.8 h. Based on these results, it is recommended to operate such a system at an HRT of 5.8 h and OLR not exceeding 3.6 kgCOD/m3 day for producing an effluent quality complying for reuse in unrestricted irrigation purposes. The removal of TKj-N and nitrification efficiency in the DHS system was significantly affected by increasing the OLR from 1.9 to 3.6 kgCOD/m3 day and from 3.6 to 6.8 kgCOD/m3 day. At an OLR of 1.9 kgCOD/m3 day, the DHS system removed 80 ± 12% of TKj-N and 91 ± 22% of ammonia which is significantly higher than that at an OLR of 3.6 (58.5 ± 13%) and 6.8 kgCOD/m3 day (26.8 ± 16%). Similar results were recorded for the removal of total coliform (TC), viz., the efficiencies dropped for TC from 99.8 ± 0.2 to 99.4 ± 0.8% and from 99.4 ± 0.8 to 90.0 ± 7.6%, respectively. DHS profile results showed that the major part of COD was removed in the upper portion of the system while the nitrification process was taken place in the lower part of the DHS system at OLR of 1.9 kgCOD/m3 day and HRT of 11.7 h.
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NASA Astrophysics Data System (ADS)
Drinkwater, Ken
2009-10-01
Concern about future anthropogenic warming has lead to demands for information on what might happen to fish and fisheries under various climate-change scenarios. One suggestion has been to use past events as a proxy for what will happen in the future. In this paper a comparison between the responses of Atlantic cod ( Gadus morhua) to two major warm periods in the North Atlantic during the 20th century is carried out to determine how reliable the past might be as a predictor of the future. The first warm period began during the 1920s, remained relatively warm through the 1960s, and was limited primarily to the northern regions (>60°N). The second warm period, which again covered the northern regions but also extended farther south (30°N), began in the 1990s and has continued into the present century. During the earlier warm period, the most northern of the cod stocks (West Greenland, Icelandic, and Northeast Arctic cod in the Barents Sea) increased in abundance, individual growth was high, recruitment was strong, and their distribution spread northward. Available plankton data suggest that these cod responses were driven by bottom-up processes. Fishing pressure increased during this period of high cod abundance and the northern cod stocks began to decline, as early as the 1950s in the Barents Sea but during the 1960s elsewhere. Individual growth declined as temperatures cooled and the cod distributions retracted southward. During the warming in the 1990s, the spawning stock biomass of cod in the Barents Sea again increased, recruitment rose, and the stock spread northward, but the individual growth did not improve significantly. Cod off West Greenland also have shown signs of improving recruitment and increasing biomass, albeit they are still very low in comparison to the earlier warming period. The abundance of Icelandic cod, on the other hand, has remained low through the recent warm period and spawning stock biomass and total biomass are at levels near the lowest on record. The different responses of cod to the two warm events, in particular the reduced cod production during the recent warm period, are attributed to the effects of intense fishing pressure and possibly related ecosystem changes. The implications of the results of the comparisons on the development of cod scenarios under future climate change are addressed.
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Zhang, Ming; Sun, Bo; Zhang, Qi; Gao, Rong; Liu, Qiao; Dong, Fangting; Fang, Haiqin; Peng, Shuangqing; Li, Famei; Yan, Xianzhong
2017-01-15
A quenching, harvesting, and extraction protocol was optimized for cardiomyocytes NMR metabonomics analysis in this study. Trypsin treatment and direct scraping cells in acetonitrile were compared for sample harvesting. The results showed trypsin treatment cause normalized concentration increasing of phosphocholine and metabolites leakage, since the trypsin-induced membrane broken and long term harvesting procedures. Then the intracellular metabolite extraction efficiency of methanol and acetonitrile were compared. As a result, washing twice with phosphate buffer, direct scraping cells and extracting with acetonitrile were chosen to prepare cardiomyocytes extracts samples for metabonomics studies. This optimized protocol is rapid, effective, and exhibits greater metabolite retention. Copyright © 2016 Elsevier Inc. All rights reserved.
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Tejel, Cristina; Villarroya, B. Eva; Ciriano, Miguel A.; Oro, Luis A.; Lanfranchi, Maurizio; Tiripicchio, Antonio; Tiripicchio-Camellini, Marisa
1996-07-17
Reactions of [M(2)(&mgr;-Cl)(2)(cod)(2)] (cod = 1,5-cyclooctadiene, M = Rh, Ir) with benzimidazole-2-thiol (H(2)Bzimt) afford the mononuclear complexes [MCl(H(2)Bzimt)(cod)] (M = Rh (1), Ir (2)) for which a S-coordination of the ligand is proposed based on their spectroscopic data. The dinuclear complexes [M(2)(&mgr;-HBzimt)(2)(cod)(2)] (M = Rh (3), Ir (4)) are isolated from the reaction of [M(acac)(cod)] and benzimidazole-2-thiol. They contain the monodeprotonated ligand (HBzimt(-)) bridging the two metals in a &mgr;(2)-(1kappaN,2kappaS) coordination mode and in a relative cis,cis-HT arrangement. Complexes 3 and 4 react with the appropriate species [M(cod)(Me(2)CO)(2)](+) to afford the trinuclear cationic aggregates [M(3)(&mgr;-HBzimt)(2)(cod)(3)](+) (M = Rh (5), Ir (6)) and with the [M'(2)(&mgr;-OMe)(2)(cod)(2)] compounds to give the homo- and heterotetranuclear complexes [MM'(&mgr;-Bzimt)(cod)(2)](2) (M = M' = Rh (7), Ir (8); M = Ir, M' = Rh (9)) containing the dideprotonated ligand (Bzimt(2)(-)). The trinuclear neutral complexes [M(3)(&mgr;-Bzimt)(&mgr;-HBzimt)(cod)(3)] are intermediates detected in the synthesis of the tetranuclear complexes. Protonation of 9 with HBF(4) gives the unsymmetrical complex [Ir(2)Rh(&mgr;-HBzimt)(2)(cod)(3)]BF(4) (10). This reaction involves the protonation of the bridging ligands followed by the removal of one "Rh(cod)" moiety to give a single isomer. The molecular structure of [Rh(2)(&mgr;-Bzimt)(cod)(2)](2) (7) has been determined by X-ray diffraction methods. Crystals are monoclinic, space group P2(1)/n, a = 20.173(5) Å, b = 42.076(8) Å, c = 10.983(3) Å, beta = 93.32(2) degrees, Z = 8, 7145 reflections, R = 0.0622, and R(w) = 0.0779. The complete assignment of the resonances of the (1)H NMR spectra of the complexes 3, 4, and 7-9 was carried out by selective decoupling, NOE, and H,H-COSY experiments. The differences in the chemical shifts of the olefinic protons are discussed on the basis of steric and magnetic anisotropy effects.
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Niwas, Ram; Osama, Khwaja; Khan, Saif; Haque, Shafiul; Tripathi, C. K. M.; Mishra, B. N.
2015-01-01
Cholesterol oxidase (COD) is a bi-functional FAD-containing oxidoreductase which catalyzes the oxidation of cholesterol into 4-cholesten-3-one. The wider biological functions and clinical applications of COD have urged the screening, isolation and characterization of newer microbes from diverse habitats as a source of COD and optimization and over-production of COD for various uses. The practicability of statistical/ artificial intelligence techniques, such as response surface methodology (RSM), artificial neural network (ANN) and genetic algorithm (GA) have been tested to optimize the medium composition for the production of COD from novel strain Streptomyces sp. NCIM 5500. All experiments were performed according to the five factor central composite design (CCD) and the generated data was analysed using RSM and ANN. GA was employed to optimize the models generated by RSM and ANN. Based upon the predicted COD concentration, the model developed with ANN was found to be superior to the model developed with RSM. The RSM-GA approach predicted maximum of 6.283 U/mL COD production, whereas the ANN-GA approach predicted a maximum of 9.93 U/mL COD concentration. The optimum concentrations of the medium variables predicted through ANN-GA approach were: 1.431 g/50 mL soybean, 1.389 g/50 mL maltose, 0.029 g/50 mL MgSO4, 0.45 g/50 mL NaCl and 2.235 ml/50 mL glycerol. The experimental COD concentration was concurrent with the GA predicted yield and led to 9.75 U/mL COD production, which was nearly two times higher than the yield (4.2 U/mL) obtained with the un-optimized medium. This is the very first time we are reporting the statistical versus artificial intelligence based modeling and optimization of COD production by Streptomyces sp. NCIM 5500. PMID:26368924
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Pathak, Lakshmi; Singh, Vineeta; Niwas, Ram; Osama, Khwaja; Khan, Saif; Haque, Shafiul; Tripathi, C K M; Mishra, B N
2015-01-01
Cholesterol oxidase (COD) is a bi-functional FAD-containing oxidoreductase which catalyzes the oxidation of cholesterol into 4-cholesten-3-one. The wider biological functions and clinical applications of COD have urged the screening, isolation and characterization of newer microbes from diverse habitats as a source of COD and optimization and over-production of COD for various uses. The practicability of statistical/ artificial intelligence techniques, such as response surface methodology (RSM), artificial neural network (ANN) and genetic algorithm (GA) have been tested to optimize the medium composition for the production of COD from novel strain Streptomyces sp. NCIM 5500. All experiments were performed according to the five factor central composite design (CCD) and the generated data was analysed using RSM and ANN. GA was employed to optimize the models generated by RSM and ANN. Based upon the predicted COD concentration, the model developed with ANN was found to be superior to the model developed with RSM. The RSM-GA approach predicted maximum of 6.283 U/mL COD production, whereas the ANN-GA approach predicted a maximum of 9.93 U/mL COD concentration. The optimum concentrations of the medium variables predicted through ANN-GA approach were: 1.431 g/50 mL soybean, 1.389 g/50 mL maltose, 0.029 g/50 mL MgSO4, 0.45 g/50 mL NaCl and 2.235 ml/50 mL glycerol. The experimental COD concentration was concurrent with the GA predicted yield and led to 9.75 U/mL COD production, which was nearly two times higher than the yield (4.2 U/mL) obtained with the un-optimized medium. This is the very first time we are reporting the statistical versus artificial intelligence based modeling and optimization of COD production by Streptomyces sp. NCIM 5500.
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NASA Astrophysics Data System (ADS)
Kohlbach, Doreen; Schaafsma, Fokje L.; Graeve, Martin; Lebreton, Benoit; Lange, Benjamin Allen; David, Carmen; Vortkamp, Martina; Flores, Hauke
2017-03-01
The polar cod (Boreogadus saida) is considered an ecological key species, because it reaches high stock biomasses and constitutes an important carbon source for seabirds and marine mammals in high-Arctic ecosystems. Young polar cod (1-2 years) are often associated with the underside of sea ice. To evaluate the impact of changing Arctic sea ice habitats on polar cod, we examined the diet composition and quantified the contribution of ice algae-produced carbon (αIce) to the carbon budget of polar cod. Young polar cod were sampled in the ice-water interface layer in the central Arctic Ocean during late summer 2012. Diets and carbon sources of these fish were examined using 4 approaches: (1) stomach content analysis, (2) fatty acid (FA) analysis, (3) bulk nitrogen and carbon stable isotope analysis (BSIA) and (4) compound-specific stable isotope analysis (CSIA) of FAs. The ice-associated (sympagic) amphipod Apherusa glacialis dominated the stomach contents by mass, indicating a high importance of sympagic fauna in young polar cod diets. The biomass of food measured in stomachs implied constant feeding at daily rates of ∼1.2% body mass per fish, indicating the potential for positive growth. FA profiles of polar cod indicated that diatoms were the primary carbon source, indirectly obtained via amphipods and copepods. The αIce using bulk isotope data from muscle was estimated to be >90%. In comparison, αIce based on CSIA ranged from 34 to 65%, with the highest estimates from muscle and the lowest from liver tissue. Overall, our results indicate a strong dependency of polar cod on ice-algae produced carbon. This suggests that young polar cod may be particularly vulnerable to changes in the distribution and structure of sea ice habitats. Due to the ecological key role of polar cod, changes at the base of the sea ice-associated food web are likely to affect the higher trophic levels of high-Arctic ecosystems.
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Beato, Marco; Bianchi, Mattia; Coratella, Giuseppe; Merlini, Michele; Drust, Barry
2018-02-01
Beato, M, Bianchi, M, Coratella, G, Merlini, M, and Drust, B. Effects of plyometric and directional training on speed and jump performance in elite youth soccer players. J Strength Cond Res 32(2): 289-296, 2018-Soccer players perform approximately 1,350 activities (every 4-6 seconds), such as accelerations/decelerations and changes of direction (CODs) during matches. It is well established that COD and plyometric training have a positive impact on fitness parameters in football players. This study analyzed the effect of a complex COD and plyometric protocol (CODJ-G) compared with an isolated COD protocol (COD-G) training on elite football players. A randomized pre-post parallel group trial was used in this study. Twenty-one youth players were enrolled in this study (mean ± SD; age 17 ± 0.8 years, mass 70.1 ± 6.4 kg, and height 177.4 ± 6.2 cm). Players were randomized into 2 different groups: CODJ-G (n = 11) and COD-G (n = 10), training frequency of 2 times a week more than 6 weeks. Sprint 10, 30, and 40 m, long jump, triple hop jump, and 505 COD test were considered. Exercise-induced within-group changes in performance for both CODJ-G and COD-G: long jump (effect size [ES] = 0.32 and ES = 0.26, respectively) and sprint 10 m (ES = -0.51 and ES = -0.22, respectively), after 6 weeks of training. Moreover, CODJ-G reported substantially better results (between-group changes) in long jump test (ES = 0.32). In conclusion, this study showed that short-term protocols (CODJ-G and COD-G) are important and able to give meaningful improvements on power and speed parameters in a specific soccer population. CODJ-G showed a larger effect in sprint and jump parameters compared with COD-G after the training protocol. This study offers important implications for designing COD and jumps training in elite soccer.
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NASA Astrophysics Data System (ADS)
Ma, Zhen; Wan, Rong; Song, Xiefa; Gao, Lei
2013-09-01
Different culture methods may affect the intensive culture system of Pacific white shrimp ( Litopenaeus vannamei) regarding water quality and growth and economic performance. This study evaluated the potential effects of three culture methods through cultivation of juvenile shrimps under consistent tank management conditions for 84 d. The three methods involved shrimp cultivation in different tanks, i.e., outdoor tanks with cement bottom (mode-C), greenhouse tanks with cement bottom (mode-G) and outdoor tanks with mud-substrate (mode-M). Results showed that water temperature was significantly higher in mode-G than that in mode-C ( P < 0.05). In contrast to the other two treatments, mode-M had stable pH after 50 d cultivation of shrimps. In the mid-late period, the average concentrations of TAN, NO2-N, DIP and COD were significantly lower in mode-M and mode-G compared with those in mode-C ( P < 0.05). Despite lack of differences in the final shrimp weight among different treatments ( P > 0.05), mode-M had significantly higher shrimp yield, survival rate and feed conversion rate ( P < 0.05) than other modes. There were significant differences in revenue and net return among different treatments ( P < 0.05). These demonstrated that the treatments of mode-G and mode-M were conductive to the intensive culture system of L. vannamei.
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USDA-ARS?s Scientific Manuscript database
Soybean seeds possess anti-nutritional compounds which inactivate digestive proteases, principally corresponding to two families: Kunitz Trypsin Inhibitors (KTi) and Bowman-Birk Inhibitors (BBI). High levels of raw soybeans/soybean meal in feed mixtures can cause poor weight gain and pancreatic abno...
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USDA-ARS?s Scientific Manuscript database
Soybean contains several biologically active components and one of this belongs to the bioactive peptide group. The objectives of this study were to produce different lunasin-enriched preparations (LEP) and determine the effect of Bowman-Birk inhibitor and Kunitz trypsin concentrations on the stabil...
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Peters, Roger H; Young, M Scott; Rojas, Elizabeth C; Gorey, Claire M
2017-07-01
Over seven million persons in the United States are supervised by the criminal justice system, including many who have co-occurring mental and substance use disorders (CODs). This population is at high risk for recidivism and presents numerous challenges to those working in the justice system. To provide a contemporary review of the existing research and examine key issues and evidence-based treatment and supervision practices related to CODs in the justice system. We reviewed COD research involving offenders that has been conducted over the past 20 years and provide an analysis of key findings. Several empirically supported frameworks are available to guide services for offenders who have CODs, including Integrated Dual Disorders Treatment (IDDT), the Risk-Need-Responsivity (RNR) model, and Cognitive-Behavioral Therapy (CBT). Evidence-based services include integrated assessment that addresses both sets of disorders and the risk for criminal recidivism. Although several evidence-based COD interventions have been implemented at different points in the justice system, there remains a significant gap in services for offenders who have CODs. Existing program models include Crisis Intervention Teams (CIT), day reporting centers, specialized community supervision teams, pre- and post-booking diversion programs, and treatment-based courts (e.g., drug courts, mental health courts, COD dockets). Jail-based COD treatment programs provide stabilization of acute symptoms, medication consultation, and triage to community services, while longer-term prison COD programs feature Modified Therapeutic Communities (MTCs). Despite the availability of multiple evidence-based interventions that have been implemented across diverse justice system settings, these services are not sufficiently used to address the scope of treatment and supervision needs among offenders with CODs.
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Farghaly, Ahmed; Tawfik, Ahmed
2017-01-01
Multi-phase anaerobic reactor for H 2 and CH 4 production from paperboard mill wastewater was studied. The reactor was operated at hydraulic retention times (HRTs) of 12, 18, 24, and 36 h, and organic loading rates (OLRs) of 2.2, 1.5, 1.1, and 0.75 kg chemical oxygen demand (COD)/m 3 day, respectively. HRT of 12 h and OLR of 2.2 kg COD/m 3 day provided maximum hydrogen yield of 42.76 ± 14.5 ml/g COD removed and volumetric substrate uptake rate (-rS) of 16.51 ± 4.43 mg COD/L h. This corresponded to the highest soluble COD/total COD (SCOD/TCOD) ratio of 56.25 ± 3.3 % and the maximum volatile fatty acid (VFA) yield (Y VFA ) of 0.21 ± 0.03 g VFA/g COD, confirming that H 2 was mainly produced through SCOD conversion. The highest methane yield (18.78 ± 3.8 ml/g COD removed ) and -rS of 21.74 ± 1.34 mgCOD/L h were achieved at an HRT of 36 h and OLR of 0.75 kg COD/m 3 day. The maximum hydrogen production rate (HPR) and methane production rate (MPR) were achieved at carbon to nitrogen (C/N) ratio of 47.9 and 14.3, respectively. This implies the important effect of C/N ratio on the distinction between the dominant microorganism bioactivities responsible for H 2 and CH 4 production.
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[Pollution load and the first flush effect of BOD5 and COD in urban runoff of Wenzhou City].
Wang, Jun; Bi, Chun-juan; Chen, Zhen-lou; Zhou, Dong
2013-05-01
Four typical rainfalls were monitored in two different research areas of Wenzhou Municipality. Concentrations of BOD5 and COD in six different urban runoffs were measured. In addition the event mean concentration (EMC), M (V) curve and BOD5/COD of pollutant were calculated. The results showed that concentrations of BOD5 and COD in different urban runoffs of Wenzhou ranged from ND to 69.21 mg x L(-1) and ND to 636 mg x L(-1). Concentrations of BOD5 and COD in different urban runoffs were decreasing over time, so it is greatly significant to manage the initial runoff for reducing organic pollution. Judged by EMC of BOD5 and COD in these five rainfalls, concentrations of pollutant in some urban runoffs were out of the integrated wastewater discharge standard. If these runoffs flowed into river, it would cause environmental pressure to the next level receiving water bodies. According to the M (V) curve, the first flush effect of COD in most urban runoffs was common; while the first flush effect of BOD5 was same as that of COD. The result also showed that organic pollution was serious at the beginning of runoff. The underlying surface type could affect the concentration of BOD5 and COD in urban runoff. While the results of BOD5/COD also suggested that biodegradation was considered as one of the effective ways to decrease the pollution load of organics in urban runoff, and the best management plans (BMPs) should be selected for various urban runoff types for the treatment of organic pollution.
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Jimenez, Jose; Miller, Mark; Bott, Charles; Murthy, Sudhir; De Clippeleir, Haydee; Wett, Bernhard
2015-12-15
The high-rate activated sludge (HRAS) process is a technology suitable for the removal and redirection of organics from wastewater to energy generating processes in an efficient manner. A HRAS pilot plant was operated under controlled conditions resulting in concentrating the influent particulate, colloidal, and soluble COD to a waste solids stream with minimal energy input by maximizing sludge production, bacterial storage, and bioflocculation. The impact of important process parameters such as solids retention time (SRT), hydraulic residence time (HRT) and dissolved oxygen (DO) levels on the performance of a HRAS system was demonstrated in a pilot study. The results showed that maximum removal efficiencies of soluble COD were reached at a DO > 0.3 mg O2/L, SRT > 0.5 days and HRT > 15 min which indicates that minimizing the oxidation of the soluble COD in the high-rate activated sludge process is difficult. The study of DO, SRT and HRT exhibited high degree of impact on the colloidal and particulate COD removal. Thus, more attention should be focused on controlling the removal of these COD fractions. Colloidal COD removal plateaued at a DO > 0.7 mg O2/L, SRT > 1.5 days and HRT > 30 min, similar to particulate COD removal. Concurrent increase in extracellular polymers (EPS) production in the reactor and the association of particulate and colloidal material into sludge flocs (bioflocculation) indicated carbon capture by biomass. The SRT impacted the overall mass and energy balance of the high-rate process indicating that at low SRT conditions, lower COD mineralization or loss of COD content occurred. In addition, the lower SRT conditions resulted in higher sludge yields and higher COD content in the WAS. Copyright © 2015 Elsevier Ltd. All rights reserved.
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Barzideh, Zoha; Latiff, Aishah Abd; Gan, Chee-Yuen; Abedin, Md Zainul; Alias, Abd Karim
2014-12-01
Collagen isolated from the ribbon jellyfish ( Chrysaora sp.) was hydrolysed using three different proteases ( i.e. trypsin, alcalase and Protamex) to obtain bioactive peptides. Angiotensin-I-converting enzyme (ACE) inhibitory activity and antioxidant activities ( i.e. ferric reducing antioxidant power (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity) of the peptides were measured and compared, and the effect of the duration of hydrolysis on the bioactivity (ACE inhibitory and antioxidant activities) of peptides was also evaluated. FRAP activity was the highest in Protamex-induced (25-27 mM) and trypsin-induced hydrolysates (24-26 mM) at 7 and 9 h, respectively. Conversely, hydrolysates produced by trypsin for 1 and 3 h showed the highest DPPH radical scavenging activities (94 and 92%, respectively). Trypsin-induced hydrolysates (at 3 h) also showed the highest ACE inhibitory activity (89%). The peptide sequences with the highest activities were identified using tandem mass spectrometry, and the results show that the hydrolysates had a high content of hydrophobic amino acids as well as unique amino acid sequences, which likely contribute to their biological activities.
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NASA Astrophysics Data System (ADS)
Sumiyati, Sri; Purwanto; Sutrisno, Endro; Sudarno; Arthawidya, Jalu; Izzudin, Humam
2018-02-01
Household wastewater contains contaminants that harm the environment. One of the pollutants found COD. If being discharged into the environment directly, COD concentrations exceeding the existing quality standard will disrupt the ecosystem in the receiving water body. One of the technologies that can degrade COD is biofilm technology with honeycomb tube media. This research aims to analyze the decrease of COD concentration present in wastewater household with biofilm technology of honeycomb media. The reactor used in was made of glass with a thickness of 4 mm, a volume of 18 litres and operated continuously. The media used is made of a PVC pipe, cut to 3 cm in size and then glued to one another, forming a honey comb. The results showed that there has been a decrease in COD concentration of household wastewater.
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Walsh, Philip; Favaro, Brett
2017-01-01
Pots (also known as traps) are baited fishing gears widely used in commercial fisheries, and are being considered as a tool for harvesting Atlantic cod (Gadus morhua) in Newfoundland and Labrador, Canada. Pots produce lower environmental impacts than many other fishing gears, but they will only be a viable fishing strategy if they are efficient and selective at catching their target species. To study the behaviour of cod in and around pots, and how those behaviours affect pot efficiency, we used long-duration underwater video cameras to assess two models of cod pot deployed in the nearshore waters of Fogo Island, NL. We examined the number of cod that approached the pot, the number and proportion that successfully completed entries into the pot openings, and the number that exited, and related these factors to the direction of water movement. We observed very few entry attempts relative to the number of approaches by cod, and only 22% of all entry attempts were successful. We observed that 50% of approaches, 70% of entry attempts, and 73% of successful entrances occurred against the current, and 25% of cod were able to exit the pot following capture. Based on our observations, we suggest that future cod pots should have a greater number of entrances, or a mechanism to ensure that entrances rotate in line with the current, in order to maximize their catch efficiency for cod. PMID:28194312
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Meintzer, Phillip; Walsh, Philip; Favaro, Brett
2017-01-01
Pots (also known as traps) are baited fishing gears widely used in commercial fisheries, and are being considered as a tool for harvesting Atlantic cod ( Gadus morhua ) in Newfoundland and Labrador, Canada. Pots produce lower environmental impacts than many other fishing gears, but they will only be a viable fishing strategy if they are efficient and selective at catching their target species. To study the behaviour of cod in and around pots, and how those behaviours affect pot efficiency, we used long-duration underwater video cameras to assess two models of cod pot deployed in the nearshore waters of Fogo Island, NL. We examined the number of cod that approached the pot, the number and proportion that successfully completed entries into the pot openings, and the number that exited, and related these factors to the direction of water movement. We observed very few entry attempts relative to the number of approaches by cod, and only 22% of all entry attempts were successful. We observed that 50% of approaches, 70% of entry attempts, and 73% of successful entrances occurred against the current, and 25% of cod were able to exit the pot following capture. Based on our observations, we suggest that future cod pots should have a greater number of entrances, or a mechanism to ensure that entrances rotate in line with the current, in order to maximize their catch efficiency for cod.
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Anaerobic biodegradation of aircraft deicing fluid in UASB reactors.
Tham, P T Pham thi; Kennedy, K J Kevin J
2004-05-01
A central composite design was employed to methodically investigate anaerobic treatment of aircraft deicing fluid (ADF) in bench-scale Upflow Anaerobic Sludge Blanket (UASB) reactors. A total of 23 runs at 17 different operating conditions (0.8% 1.6% ADF (6000-12,000mg/L COD), 12-56h HRT, and 18-36gVSS/L) were conducted in continuous mode. The development of four empirical models describing process responses (i.e. COD removal efficiency, biomass-specific acetoclastic activity, methane production rate, and methane production potential) as functions of ADF concentration, hydraulic retention time, and biomass concentration is presented. Model verification indicated that predicted responses (COD removal efficiencies, biomass-specific acetoclastic activity, and methane production rates and potential) were in good agreement with experimental results. Biomass-specific acetoclastic activity was improved two-fold from 0.23gCOD/gVSS/d for inoculum to a maximum of 0.55gCOD/gVSS/d during ADF treatment in UASB reactors. For the design window, COD removal efficiencies were higher than 90%. The predicted methane production potentials were close to theoretical values, and methane production rates increased as the organic loading rate is increased. ADF toxicity effects were evident for 1.6% ADF at medium organic loadings (SOLR above 0.5gCOD/gVSS/d). In contrast, good reactor stability and excellent COD removal efficiencies were achieved at 1.2% ADF for reactor loadings approaching that of highly loaded systems (0.73gCOD/gVSS/d).
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Could Seals Prevent Cod Recovery in the Baltic Sea?
MacKenzie, Brian R.; Eero, Margit; Ojaveer, Henn
2011-01-01
Fish populations are increasingly affected by multiple human and natural impacts including exploitation, eutrophication, habitat alteration and climate change. As a result many collapsed populations may have to recover in ecosystems whose structure and functioning differ from those in which they were formerly productive and supported sustainable fisheries. Here we investigate how a cod (Gadus morhua) population in the Baltic Sea whose biomass was reduced due to a combination of high exploitation and deteriorating environmental conditions might recover and develop in the 21st century in an ecosystem that likely will change due to both the already started recovery of a cod predator, the grey seal Halichoerus grypus, and projected climate impacts. Simulation modelling, assuming increased seal predation, fishing levels consistent with management plan targets and stable salinity, shows that the cod population could reach high levels well above the long-term average. Scenarios with similar seal and fishing levels but with 15% lower salinity suggest that the Baltic will still be able to support a cod population which can sustain a fishery, but biomass and yields will be lower. At present knowledge of cod and seal interactions, seal predation was found to have much lower impact on cod recovery, compared to the effects of exploitation and salinity. These results suggest that dual management objectives (recovery of both seal and cod populations) are realistic but success in achieving these goals will also depend on how climate change affects cod recruitment. PMID:21573062
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Oliveira-Neto, Osmundo B; Batista, João A N; Rigden, Daniel J; Fragoso, Rodrigo R; Silva, Rodrigo O; Gomes, Eliane A; Franco, Octávio L; Dias, Simoni C; Cordeiro, Célia M T; Monnerat, Rose G; Grossi-De-Sá, Maria F
2004-09-01
Fourteen different cDNA fragments encoding serine proteinases were isolated by reverse transcription-PCR from cotton boll weevil (Anthonomus grandis) larvae. A large diversity between the sequences was observed, with a mean pairwise identity of 22% in the amino acid sequence. The cDNAs encompassed 11 trypsin-like sequences classifiable into three families and three chymotrypsin-like sequences belonging to a single family. Using a combination of 5' and 3' RACE, the full-length sequence was obtained for five of the cDNAs, named Agser2, Agser5, Agser6, Agser10 and Agser21. The encoded proteins included amino acid sequence motifs of serine proteinase active sites, conserved cysteine residues, and both zymogen activation and signal peptides. Southern blotting analysis suggested that one or two copies of these serine proteinase genes exist in the A. grandis genome. Northern blotting analysis of Agser2 and Agser5 showed that for both genes, expression is induced upon feeding and is concentrated in the gut of larvae and adult insects. Reverse northern analysis of the 14 cDNA fragments showed that only two trypsin-like and two chymotrypsin-like were expressed at detectable levels. Under the effect of the serine proteinase inhibitors soybean Kunitz trypsin inhibitor and black-eyed pea trypsin/chymotrypsin inhibitor, expression of one of the trypsin-like sequences was upregulated while expression of the two chymotrypsin-like sequences was downregulated. Copyright 2004 Elsevier Ltd.
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Evaluation of hyaluronic acid-protein conjugates for polymer masked-unmasked protein therapy.
Ferguson, Elaine L; Alshame, Alshame M J; Thomas, David W
2010-12-15
Bioresponsive polymers may effectively be utilized to enhance the circulation time and stability of biologically active proteins and peptides, while reducing their immunogenicity and toxicity. Recently, dextrin-epidermal growth factor (EGF) conjugates, which make use of the Polymer-masked UnMasked Protein Therapy (PUMPT) concept, have been developed and shown potential as modulators of impaired wound healing. This study investigated the potential of PUMPT using hyaluronic acid (HA) conjugates to mask activity and enhance protein stability, while allowing restoration of biological activity following triggered degradation. HA fragments (Mw ∼90,000g/mol), obtained by acid hydrolysis of Rooster comb HA, were conjugated to trypsin as a model enzyme or to EGF as a model growth factor. Conjugates contained 2.45 and 0.98% (w/w) trypsin or EGF, respectively, and contained <5% free protein. HA conjugation did not significantly alter trypsin's activity. However, incubation of the conjugate with physiological concentrations of HAase increased its activity to ∼145% (p<0.001) that of the free enzyme. In contrast, when HA-EGF conjugates were tested in vitro, no effect on cell proliferation was seen, even in the presence of HAase. HA conjugates did not display typical masking/unmasking behavior, HA-trypsin conjugates exhibited ∼52% greater stability in the presence of elastase, compared to free trypsin, demonstrating the potential of HA conjugates for further development as modulators of tissue repair. Copyright © 2010 Elsevier B.V. All rights reserved.
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Alternative Enzymes Lead to Improvements in Sequence Coverage and PTM Analysis
Hooper, Kyle; Rosenblatt, Michael; Urh, Marjeta; Saveliev, Sergei; Hosfield, Chris; Kobs, Gary; Ford, Michael; Jones, Richard; Amunugama, Ravi; Allen, David; Brazas, Robert
2013-01-01
The profiling of proteins using biological mass spectrometry (bottom up proteomics) most commonly requires trypsin. Trypsin is advantageous in that it produces peptides of optimal charge and size. However, for applications in which the proteins under investigation are part of a complex mixture or not isolated at high levels (i.e. low ng from an immunoprecipitation), sequence coverage is rarely complete. In addition, we have found that in several cases, like phosphorylation, acetylation, and methylation, alternative proteases are required to prepare peptides suitable for MS detection. This poster will provide specific examples which demonstrate this observation. For example, the application of a combined Trypsin/ Lys-C mixture reduces the number of missed cleavages by more than 3-fold producing samples with lower CV's (for biological replicates). The mixture is also well-suited for the complete proteolysis of hydrophobic, compact proteins. The addition of chymotrypsin and elastase has been found to be useful for identifying phosphorylation sites on proteins, especially on sequences where the site of phosphorylation inhibits trypsin (i.e. proximal to K or R). Many epigenetic applications have focused on histone modifications, like lysine acetylation and arginine methylation. Alternative proteases like Asp-N, Glu-C, and chymotrypsin have been especially useful given the fact that the modified K and R residues are resistant to c-terminal cleavage by trypsin. Finally, in the case of serum profiling, the addition of the endoglycosidase, PNGase F has been found to improve sequence coverage due to the removal of N-linked glycans.
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Magnetic nanoparticles coated with polyaniline to stabilize immobilized trypsin
NASA Astrophysics Data System (ADS)
Maciel, J. C.; D. Mercês, A. A.; Cabrera, M.; Shigeyosi, W. T.; de Souza, S. D.; Olzon-Dionysio, M.; Fabris, J. D.; Cardoso, C. A.; Neri, D. F. M.; C. Silva, M. P.; Carvalho, L. B.
2016-12-01
It is reported the synthesis of magnetic nanoparticles via the chemical co-precipitation of Fe 3+ ions and their preparation by coating them with polyaniline. The electronic micrograph analysis showed that the mean diameter for the nanoparticles is ˜15 nm. FTIR, powder X-ray diffraction and Mössbauer spectroscopy were used to understand the chemical, crystallographic and 57Fe hyperfine structures for the two samples. The nanoparticles, which exhibited magnetic behavior with relatively high spontaneous magnetization at room temperature, were identified as being mainly formed by maghemite ( γFe2O3). The coated magnetic nanoparticles (sample labeled "mPANI") presented a real ability to bind biological molecules such as trypsin, forming the magnetic enzyme derivative (sample "mPANIG-Trypsin"). The amount of protein and specific activity of the immobilized trypsin were found to be 13±5 μg of protein/mg of mPANI (49.3 % of immobilized protein) and 24.1±0.7 U/mg of immobilized protein, respectively. After 48 days of storage at 4 ∘C, the activity of the immobilized trypsin was found to be 89 % of its initial activity. This simple, fast and low-cost procedure was revealed to be a promising way to prepare mPANI nanoparticles if technological applications addressed to covalently link biomolecules are envisaged. This route yields chemically stable derivatives, which can be easily recovered from the reaction mixture with a magnetic field and recyclable reused.
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Liao, Yi-Shan; Zhuo, Mu-Ning; Li, Ding-Qiang; Guo, Tai-Long
2013-08-01
In the Pearl Delta region, urban rivers have been seriously polluted, and the input of non-point source pollution materials, such as chemical oxygen demand (COD), into rivers cannot be neglected. During 2009-2010, the water qualities at eight different catchments in the Fenjiang River of Foshan city were monitored, and the COD loads for eight rivulet sewages were calculated in respect of different rainfall conditions. Interesting results were concluded in our paper. The rainfall and landuse type played important roles in the COD loading, with greater influence of rainfall than landuse type. Consequently, a COD loading formula was constructed that was defined as a function of runoff and landuse type that were derived SCS model and land use map. Loading of COD could be evaluated and predicted with the constructed formula. The mean simulation accuracy for single rainfall event was 75.51%. Long-term simulation accuracy was better than that of single rainfall. In 2009, the estimated COD loading and its loading intensity were 8 053 t and 339 kg x (hm2 x a)(-1), and the industrial land was regarded as the main source of COD pollution area. The severe non-point source pollution such as COD in Fenjiang River must be paid more attention in the future.
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Kalyuzhnyi, S; Gladchenko, M; Epov, A
2003-01-01
As a first step of treatment of landfill leachates (total COD--1,430-3,810 mg/l, total nitrogen 90-162 mg/l), a performance of laboratory UASB reactors has been investigated under mesophilic (30 degrees C), sub-mesophilic (20 degrees C) and psychrophilic (10 degrees C) conditions. Under hydraulic retention times (HRT) of around 7 h, when the average organic loading rates (OLR) were around 5 g COD/l/day, the total COD removal accounted for 81% (on the average) with the effluent concentrations close to anaerobic biodegradability limit (0.25 g COD/l) for mesophilic and sub-mesophilic regimes. The psychrophilic treatment conducted under the average HRT of 8 h and the average OLR of 4.22 g COD/l/day showed a total COD removal of 47% producing the effluents (0.75 g COD/l) more suitable for subsequent biological nitrogen removal. All three anaerobic regimes used for leachate treatment were quite efficient for elimination of heavy metals (Fe, Zn, Cu, Pb, Cd) by concomitant precipitation in the form of insoluble sulphides inside the sludge bed. The application of aerobic/anoxic biofilter as a sole polishing step for psychrophilic anaerobic effluents was acceptable for elimination of biodegradable COD and nitrogen approaching the current standards for direct discharge of treated wastewater.
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Zhao, F; Stein, D J; Paborji, M; Cash, P W; Root, B J; Wei, Z; Knupp, C J
2001-01-01
BMS-196843 (Oncostatin M) is a therapeutic recombinant protein in development. Scale-up process changes led to unexpected instability of the bulk drug substance solution during storage. A product with an apparent higher MW than the parent protein was observed by the size-exclusion chromatography (SEC). This study was aimed to fully characterize the product and to identify a solution to stabilize the protein. SEC, SDS-PAGE, tryptic mapping, and N-terminal sequencing were performed to characterize the unknown product. The effect of pH, temperature, bulk concentration, and immobilized trypsin inhibitor on the degradation rate was studied to elucidate the mechanism and to identify stabilization strategies. Despite the apparent high MW indicated initially by SEC, the unknown was characterized to be a degradation product resulted from a backbone cleavage between residues Arg145-Gly146. The resulting fragments from the backbone cleavage were, however, still linked through an intramolecular disulfide bond. Thus, the final product had a more open structure with an increased hydrodynamic radius compared to the parent protein, which explains the initial SEC results. The site-specific backbone cleavage was suspected to be catalyzed by trypsin-like protease impurities in the bulk solution. The bulk drug substance solution was subsequently treated with immobilized soybean trypsin inhibitor, and the degradation rate was significantly reduced. Furthermore, increasing the solution pH from 5 to 8 led to an increase in the degradation rate, which was consistent with the expected pH dependency of trypsin activity. In addition, the effect of bulk concentration also supported the involvement of protease impurities rather than a spontaneous peptide bond hydrolysis reaction. Trace trypsin-like protease impurities led to an unusual site-specific backbone cleavage of BMS-196854. The proteolytic degradation can be minimized by treating the bulk solution with immobilized soybean trypsin inhibitor and/or controlling the solution pH and storage temperature.
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NASA Astrophysics Data System (ADS)
Crawford, Justin A.; Quakenbush, Lori T.; Citta, John J.
2015-08-01
Reductions in summer sea ice in the Chukchi and Beaufort seas are expected to affect what has been an ice-adapted marine food web in the Pacific Arctic. To determine whether recent decreases in sea ice have affected ice-associated marine predators (i.e., ringed, Pusa hispida, and bearded seals, Erignathus barbatus) in the Bering and Chukchi seas we compared diet, body condition, growth, productivity, and the proportion of pups harvested (an index of weaning success) for seals of each species harvested by 11 Alaskan villages during two periods; a historical (1975-1984) and a recent period (2003-2012). We also examined how changes in indices of seal health may be correlated with the reduction of sea ice characteristic of the recent period. For ringed seals ⩾1 year of age, the % frequency of occurrence (%FO) of Arctic cod (Boreogadus saida), walleye pollock (Gadus chalcogramma), rainbow smelt (Osmerus mordax), and Pacific herring (Clupea pallasi) increased from the historic to the recent period, while the %FO of invertebrates decreased for both pups and seals ⩾1 year of age. For bearded seals ⩾1 year of age, the %FO of Arctic cod, pricklebacks, and flatfish increased during the recent period, while the %FO of saffron cod (Eleginus gracilis) decreased for pups. Although invertebrates did not change overall for either age class, decreases occurred in 10 of 24 specific prey categories, for bearded seals ⩾1 year of age; only echiurids increased. The %FO of gastropods and bivalves increased for pups while isopods and one species of shrimp and crab decreased in occurrence. During the recent period ringed seals grew faster, had thicker blubber, had no change in pregnancy rate, matured 2 years earlier, and a larger proportion of pups was harvested than during the historical period. Correlations with spring ice concentration showed that the growth and blubber thickness of seals ⩾1 year of age, blubber thickness of pups, and the proportion of pups in the harvest all declined for ringed seals when ice concentrations were higher in the historic period. However, only the correlations between high ice concentrations and growth of ringed seals ⩾1 year of age and the proportion of ringed seal pups in the harvest were statistically significant. Although growth of bearded seals ⩾1 year of age was slower during the recent period, it was similar to the average over the entire time series, and blubber thickness increased. Pup growth and blubber thickness did not change between periods. There was no change in pregnancy rate, but females matured 1.6 years earlier, and a larger proportion of pups were harvested. Correlations with spring ice concentration showed that the growth of seals ⩾1 year of age, the growth of pups, blubber thickness of pups, and proportion of pups in the harvest also declined for bearded seals when sea ice concentrations were higher. However, no relationships between bearded seals and sea ice were statistically significant. Overall, our results suggest that ringed seals in the Alaskan Bering and Chukchi seas have adjusted to changes in diet, are growing faster and possibly weaning more pups in the recent compared to the historic period. These patterns are less evident for bearded seals. Although the ringed and bearded seals we examined have not exhibited the declines in body condition, growth, or reproduction observed in other populations, continued monitoring and comparison among seal populations is vital to understanding the effects of changing environmental conditions in the Pacific Arctic region.
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[Effect of ERK/AP-1 signaling pathway on proliferation of hepatoma cells induced by PAR-2 agonists].
Zheng, Yan-min; Xie, Li-qun; Li, Xuan; Zhao, Jun-yan; Chen, Xiao-yi; Chen, Li; Zhou, Jing; Li, Fei
2009-12-01
To investigate the expression of protease activated receptor-2 (PAR-2) in human HepG2 hepatoma cells and elucidate the effects of trypsin and PAR-2 agonist peptide SLIGKV-NH(2) upon the proliferation of hepatoma cells and its intracellular signaling mechanism. PAR-2 protein and mRNA expression were detected by immunofluorescence and RT-PCR. The cells were treated with SLIGKV-NH(2), trypsin, reverse PAR-2 agonist peptide VKGILS-NH(2) or PD98059. The changes of cell cycle distribution were evaluated by flow cytometry. The proliferative potential of HepG2 cells was estimated by MTT. The changes of PAR-2, c-fos and PCNA mRNA expression were detected by RT-PCR. The changes of c-fos and PCNA protein expression were detected by Western blotting. PAR-2 protein and mRNA were expressed in HepG2 cells. PAR-2 mRNA expression (PAR-2/beta-actin) were 0.70 +/- 0.04 and 0.99 +/- 0.05 respectively in cells treated with trypsin and SLIGKV-NH(2). They were both significantly higher than that in the control group (0.35 +/- 0.05, F = 135.534, P < 0.01). Percent G(0)/G(1) phase of HepG2 cells treated with trypsin or SLIGKV-NH(2) were significantly lower than those in the control group [(56.11 +/- 0.85)%, (57.85 +/- 0.46)% vs (79.12 +/- 0.67)%, both P < 0.01] Percent S phase, G(2)/M phase and proliferation index (PI) of HepG2 cells treated with trypsin or SLIGKV-NH(2) were significantly elevated (P < 0.01). The proliferation-enhancing effects and the up-regulation of mRNA and protein of c-fos and PCNA induced by trypsin or SLIGKV-NH(2) were significantly blocked by pretreatment with PD98059 (P < 0.01). There was no statistical significance in proliferation of HepG2 cells between the reverse PAR-2 agonist peptide VKGILS-NH(2) and control group (P > 0.05). PAR-2 is expressed in HepG2 hepatoma cells. PAR-2 activation induced by trypsin or SLIGKV-NH(2) promotes the proliferation of HepG2 cells partially via the ERK/AP-1 pathway.
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Dostálová, Anna; Votýpka, Jan; Favreau, Amanda J; Barbian, Kent D; Volf, Petr; Valenzuela, Jesus G; Jochim, Ryan C
2011-05-10
Parasite-vector interactions are fundamental in the transmission of vector-borne diseases such as leishmaniasis. Leishmania development in the vector sand fly is confined to the digestive tract, where sand fly midgut molecules interact with the parasites. In this work we sequenced and analyzed two midgut-specific cDNA libraries from sugar fed and blood fed female Phlebotomus perniciosus and compared the transcript expression profiles. A total of 4111 high quality sequences were obtained from the two libraries and assembled into 370 contigs and 1085 singletons. Molecules with putative roles in blood meal digestion, peritrophic matrix formation, immunity and response to oxidative stress were identified, including proteins that were not previously reported in sand flies. These molecules were evaluated relative to other published sand fly transcripts. Comparative analysis of the two libraries revealed transcripts differentially expressed in response to blood feeding. Molecules up regulated by blood feeding include a putative peritrophin (PperPer1), two chymotrypsin-like proteins (PperChym1 and PperChym2), a putative trypsin (PperTryp3) and four putative microvillar proteins (PperMVP1, 2, 4 and 5). Additionally, several transcripts were more abundant in the sugar fed midgut, such as two putative trypsins (PperTryp1 and PperTryp2), a chymotrypsin (PperChym3) and a microvillar protein (PperMVP3). We performed a detailed temporal expression profile analysis of the putative trypsin transcripts using qPCR and confirmed the expression of blood-induced and blood-repressed trypsins. Trypsin expression was measured in Leishmania infantum-infected and uninfected sand flies, which identified the L. infantum-induced down regulation of PperTryp3 at 24 hours post-blood meal. This midgut tissue-specific transcriptome provides insight into the molecules expressed in the midgut of P. perniciosus, an important vector of visceral leishmaniasis in the Old World. Through the comparative analysis of the libraries we identified molecules differentially expressed during blood meal digestion. Additionally, this study provides a detailed comparison to transcripts of other sand flies. Moreover, our analysis of putative trypsins demonstrated that L. infantum infection can reduce the transcript abundance of trypsin PperTryp3 in the midgut of P. perniciosus.
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Karaseva, E M
2011-01-01
Proceeding from long-term data on the numbers of eggs of cod in ichthyoplankton, the total annual production of cod eggs at four main spawning grounds of the Baltic Sea was calculated. It was shown that the long-term fluctuations of cod egg production were positively related to the dynamics of the volume of waters coming to the Baltic Sea in years of the North Sea advections. It is suggested that this dependence was determined by a set of adaptations providing the extension of cod reproduction upon the improvement of the environment.
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Cod Fractions In Mechanical-Biological Wastewater Treatment Plant
NASA Astrophysics Data System (ADS)
Płuciennik-Koropczuk, Ewelina; Jakubaszek, Anita; Myszograj, Sylwia; Uszakiewicz, Sylwia
2017-03-01
The paper presents results of studies concerning the designation of COD fraction in the raw, mechanically treated and biologically treated wastewater. The test object was a wastewater treatment plant with the output of over 20,000 PE. The results were compared with data received in the ASM models. During investigation following fractions of COD were determined: dissolved non-biodegradable SI, dissolved easily biodegradable SS, in organic suspension slowly degradable XS and in organic suspension non-biodegradable XI. Methodology for determining the COD fraction was based on the guidelines ATV-A 131. The real percentage of each fraction in total COD in raw wastewater are different from data received in ASM models.
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Hermon, S; Grange, D; Pellet, Y; Lloret, G; Oyonarte, S; Bosch, F; Coste, M
2008-01-01
Degreasing waste effluents issued from a surface treatment plant were treated by electrochemical techniques in an attempt to reduce COD so that clean water can be returned to the rinse bath. Electrocoagulation, both with iron and aluminium anodes, and anodic oxidation with boron doped diamond (BDD) anodes were tested. In the electrocoagulation tests, the nature of the anodes did not impact significantly the reduction of COD. Electrocoagulation showed good COD removal rates, superior to 80%, but it was not able to reduce COD down to low levels. Anodic oxidation was able to reduce COD down to discharge limits; the oxidation efficiency was superior to 50%. Economical calculations show that anodic oxidation is best used as a polishing step after electrocoagulation. The bulk of the COD would be reduced by electrocoagulation and, then, anodic oxidation would reduce COD below discharge limits. The maximum treatable flow is somewhat hindered by the small sizes of current BDD installation but it would reach 600 m(3)/year if anodic oxidation is coupled with electrocoagulation, the operational cost being 2.90 Euros /m(3). (c) IWA Publishing 2008.
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Kiyuna, Luma Sayuri Mazine; Fuess, Lucas Tadeu; Zaiat, Marcelo
2017-05-01
Throughout the sugarcane harvest, it is common for sulfate to accumulate in the vinasse of sugar and ethanol plants. However, little is known regarding the influence of sulfate on the anaerobic digestion (AD) of vinasse, which may lead to severe performance losses. This study assessed the influence of various COD/sulfate ratios (12.0, 10.0 and 7.5) on both COD removal and methane (CH 4 ) production from sugarcane vinasse AD. Batch assays were conducted in thermophilic conditions. At a COD/sulfate ratio of 7.5, CH 4 production was 35% lower compared with a ratio of 12.0, considering a diversion of approximately 13.6% of the electron flow to sulfidogenesis. The diversion of electrons to sulfidogenesis was negligible at COD/sulfate ratios higher than 25, considering the exponential increase in CH 4 production. Organic matter degradation was not greatly affected by sulfidogenesis, with COD removal levels higher than 80%, regardless of the initial COD/sulfate ratio. Copyright © 2017 Elsevier Ltd. All rights reserved.
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Xu, Lei; Zhao, Yaqian; Fan, Chuang; Fan, Zhiren; Zhao, Fangchao
2017-11-01
Chemical oxygen demand (COD) is one of the major targets to remove in constructed wetlands (CWs) system. Traditional method for COD measurement is a complex, time-consuming and highly toxic reagents participated procedure. In this study, microbial fuel cell (MFC) was successfully integrated into CW for indicating COD concentration. Results showed that there are two linear correlations between bioelectrical signals (output voltage from MFC) and COD concentration (acetate), which are COD from 0 to 500mg/L (101.99±7.42 to 631.74±7.41mV, R 2 =0.9710) and then from 500 to 1000mg/L (631.74±7.41 to 668.46±0.01mV, R 2 =0.9245). Furthermore, results also revealed the specificity of the system in terms of different types of carbon source. Overall, this work presented the feasibility of using CW-MFC for in-situ sensing COD during the wastewater treatment process, which will be a promising technique for water quality monitoring within CWs. Copyright © 2017 Elsevier Ltd. All rights reserved.
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Food web dynamics affect Northeast Arctic cod recruitment.
Hjermann, Dag Ø; Bogstad, Bjarte; Eikeset, Anne Maria; Ottersen, Geir; Gjøsaeter, Harald; Stenseth, Nils Chr
2007-03-07
Proper management of ecosystems requires an understanding of both the species interactions as well as the effect of climate variation. However, a common problem is that the available time-series are of different lengths. Here, we present a general approach for studying the dynamic structure of such interactions. Specifically, we analyse the recruitment of the world's largest cod stock, the Northeast Arctic cod. Studies based on data starting in the 1970-1980s indicate that this stock is affected by temperature through a variety of pathways. However, the value of such studies is somewhat limited by the fact that they are based on a quite specific ecological and climatic situation. Recently, this stock has consisted of fairly young fish and the spawning stock has consisted of relatively few age groups. In this study, we develop a model for the effect of capelin (the cod's main prey) and herring on cod recruitment since 1973. Based on this model, we analyse data on cod, herring and temperature going back to 1921 and find that food-web effects explain a significant part of the cod recruitment variation back to around 1950.
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NASA Astrophysics Data System (ADS)
Shiller, A. M.; Grissom, K.
2014-12-01
Interest in the oceanic geochemistry of barium (Ba) stems from a variety of reasons including its use as a paleo-productivity indicator, its chemical similarity to Ra, and its utility as a water source tracer. To better constrain these uses of Ba, we have obtained trace element clean samples from both the North Atlantic and Eastern Pacific US GEOTRACES cruises. Analytical work on the Pacific samples is proceeding while work on the Atlantic samples is complete. For the Pacific, 36 stations were occupied from Peru to Tahiti. For the Atlantic, dissolved Ba was determined at 32 stations across the North Atlantic during US cruises GT10 and GT11 along the meridional transect from Lisbon to the Cape Verde Islands and the zonal transect from Cape Cod to the Mauritanian coast. In the Atlantic, the general distribution of dissolved Ba exhibits a vertical bifurcation at approximately 500 m into shallow versus deep water. The greatest variation is found on the eastern side of the basin with concentrations ranging from 35 nmol/kg at the near surface (100 m) to over 83 nmol/kg at depth. A reduction of Ba in excess of 20% compared to the average of mesopelagic depths less than 500 m is observed within the Canary Current upwelling zone east of the Cape Verde Islands and accompanied to some extent by a subsequent regeneration at depth. Below 500 m, dissolved Ba correlates well with dissolved Si, whereas the correlation with alkalinity is poor at depth and shows a decoupling above 500 m. There is evidence of hydrothermal Ba input at the TAG vent system of the Mid-Atlantic Ridge along transect GT11 as indicated by the rapid increase in the dissolved Ba below 2500 m in conjunction with increases in Fe and Mn. In addition to the hydrothermal source, a near surface (~40 m) maximum of 51 nmol/kg is found along the continental slope of North America in correspondence with a minimum surface salinity (34.75) and increased dissolved manganese indicating either fluvial or sediment input; however, a similar input is not evident along the North African continental margin. We will contrast the Atlantic distribution with the Pacific as data become available.
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Uncertainties in stormwater runoff data collection from a small urban catchment, Southeast China.
Huang, Jinliang; Tu, Zhenshun; Du, Pengfei; Lin, Jie; Li, Qingsheng
2010-01-01
Monitoring data are often used to identify stormwater runoff characteristics and in stormwater runoff modelling without consideration of their inherent uncertainties. Integrated with discrete sample analysis and error propagation analysis, this study attempted to quantify the uncertainties of discrete chemical oxygen demand (COD), total suspended solids (TSS) concentration, stormwater flowrate, stormwater event volumes, COD event mean concentration (EMC), and COD event loads in terms of flow measurement, sample collection, storage and laboratory analysis. The results showed that the uncertainties due to sample collection, storage and laboratory analysis of COD from stormwater runoff are 13.99%, 19.48% and 12.28%. Meanwhile, flow measurement uncertainty was 12.82%, and the sample collection uncertainty of TSS from stormwater runoff was 31.63%. Based on the law of propagation of uncertainties, the uncertainties regarding event flow volume, COD EMC and COD event loads were quantified as 7.03%, 10.26% and 18.47%.
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Effect of organic matter to nitrogen ratio on membrane bioreactor performance.
Hao, L; Liao, B Q
2015-01-01
Effect of chemical oxygen demand (COD) to nitrogen (COD:N) ratio in feed on the performance of aerobic membrane bioreactor (MBR) for treating a synthetic high-strength industrial waste water containing glucose was studied for over 370 days. The widely recommended nutrients ratio (COD:N:P = 100:5:1) is not necessary for aerobic biological industrial waste water treatment. An increased COD:N ratio from 100:5 to 100:2.5 and 100:1.8 had a limited impact on COD removal efficiency and further led to a significant improvement in membrane performance, a reduced sludge yield, and improved effluent quality in terms of residual nutrients. An increased COD:N ratio will benefit the industrial waste water treatment using MBRs by reducing membrane fouling and sludge yield, saving chemical costs, and reducing secondary pollution by nutrients addition. Optimization of nutrients usage should be conducted for specific industrial waste water streams.
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Treatment of hospital waste water by ozone technology
NASA Astrophysics Data System (ADS)
Indah Dianawati, Rina; Endah Wahyuningsih, Nur; Nur, Muhammad
2018-05-01
Conventional treatment hospital wastewater need high cost, large area, long time and the final result leaves a new waste known as sludge. Alternative to more efficient and new technologies for treated hospital wastewaters was ozonation. Ozonation is able to oxidized pollutant materials in wastewater. This research is to know the decrease of COD and TDS levels with ozone. Waste water samples used by dr. Adhyatma, MPH Hospitals Semarang. Kruskal-Wallis test for COD and TDS with variation of concentration p-value = 0,029 and 0,001 (p≤0,05) or there is significantly difference between COD and TDS with level of concentration but there were no different between levels of COD, and TDS with reactions time variations p-value = 0,735, and 0,870 (p≥0.05). Ozone efficiently reduction of COD and TDS at a concentration of 100 mg/liter, the lowest mean value at COD 17.47 mg/liter and TDS 409.75 mg/liter.
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Zhang, Chuansong; Wang, Xiulin; Shi, Xiaoyong; Han, Xiurong; Sun, Xia; Zhu, Chenjian; Lu, Rong
2003-07-01
Based on the data of COD and petroleum hydrocarbons collected in the cruise from April 25 to May 2, 2002 in intensive red tide occurrence areas in East China Sea, the distribution of COD, and petroleum hydrocarbons and the eutrophication index(EI) were analyzed. The results showed that the EI and COD value were both high in coastal water, and decreased gradually away from shore. After the preliminary study on the relationships between correlative factors and occurrence of red tide, it was found that high EI and COD were necessary. There would be great chances for the red tide to break out under conditions that the EI was between 2.5 and 15 and COD concentration was between 0.8 to 1.4 mg.L-1 in seawater, along with the favorable temperature and salinity.
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Bayesian estimation of the discrete coefficient of determination.
Chen, Ting; Braga-Neto, Ulisses M
2016-12-01
The discrete coefficient of determination (CoD) measures the nonlinear interaction between discrete predictor and target variables and has had far-reaching applications in Genomic Signal Processing. Previous work has addressed the inference of the discrete CoD using classical parametric and nonparametric approaches. In this paper, we introduce a Bayesian framework for the inference of the discrete CoD. We derive analytically the optimal minimum mean-square error (MMSE) CoD estimator, as well as a CoD estimator based on the Optimal Bayesian Predictor (OBP). For the latter estimator, exact expressions for its bias, variance, and root-mean-square (RMS) are given. The accuracy of both Bayesian CoD estimators with non-informative and informative priors, under fixed or random parameters, is studied via analytical and numerical approaches. We also demonstrate the application of the proposed Bayesian approach in the inference of gene regulatory networks, using gene-expression data from a previously published study on metastatic melanoma.
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Long-term effects of the transient COD concentration on the performance of microbial fuel cells.
Mateo, S; Gonzalez Del Campo, A; Lobato, J; Rodrigo, M; Cañizares, P; Fernandez-Morales, F J
2016-07-08
In this work, the long-term effects of transient chemical oxygen demands (COD) concentrations over the performance of a microbial fuel cell were studied. From the obtained results, it was observed that the repetitive change in the COD loading rate during 12 h conditioned the behavior of the system during periods of up to 7 days. The main modifications were the enhancement of the COD consumption rate and the exerted current. These enhancements yielded increasing Coulombic efficiencies (CEs) when working with COD concentrations of 300 mg/L, but constant CEs when working with COD concentrations from 900 to 1800 mg/L. This effect could be explained by the higher affinity for the substrate of Geobacter than that of the nonelectrogenic organisms such as Clostridia. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:883-890, 2016. © 2016 American Institute of Chemical Engineers.
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Removing organic matter from sulfate-rich wastewater via sulfidogenic and methanogenic pathways.
Vilela, Rogerio Silveira; Damianovic, Márcia Helena Rissato Zamariolli; Foresti, Eugenio
2014-01-01
The simultaneous organic matter removal and sulfate reduction in synthetic sulfate-rich wastewater was evaluated for various chemical oxygen demand (COD)/sulfate ratios applied in a horizontal-flow anaerobic immobilized sludge (HAIS) reactor. At higher COD/sulfate ratios (12.5 and 7.5), the removal of organic matter was stable, likely due to methanogenesis. A combination of sulfate reduction and methanogenesis was clearly established at COD/sulfate ratios of 3.0 and 1.9. At a COD/sulfate ratio of 1.0, the organic matter removal was likely influenced by methanogenesis inhibition. The quantity of sulfate removed at a COD/sulfate ratio of 1.0 was identical to that obtained at a ratio of 1.9, indicating a lack of available electron donors for sulfidogenesis. The sulfate reduction and organic matter removal were not maximized at the same COD/sulfate ratio; therefore, competitive inhibition must be the predominant mechanism in establishing an electron flow.
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USDA-ARS?s Scientific Manuscript database
Serine proteases, such as trypsin and chymotrypsin, are the primary digestive enzymes in lepidopteran larvae, and are also involved in Bacillus thuringiensis (Bt) protoxin activation and protoxin/toxin degradation. We isolated and sequenced 34 cDNAs putatively encoding trypsins, chymotrypsins and th...
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Influence of surface-imprinted nanoparticles on trypsin activity.
Guerreiro, António; Poma, Alessandro; Karim, Kal; Moczko, Ewa; Takarada, Jessica; de Vargas-Sansalvador, Isabel Perez; Turner, Nicholas; Piletska, Elena; de Magalhães, Cristiana Schmidt; Glazova, Natalia; Serkova, Anastasia; Omelianova, Aleksandra; Piletsky, Sergey
2014-09-01
Here, the modulation of enzyme activity is presented by protein-imprinted nanoparticles produced using a solid-phase approach. Using trypsin as target, binding of the nanoparticles to the enzyme results in its inhibition or in stabilization, depending on the orientation of the immobilized enzyme used during imprinting. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
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Code of Federal Regulations, 2012 CFR
2012-04-01
... defined in § 170.3(o)(9) of this chapter to hydrolyze proteins or polypeptides. (2) The ingredient is used... 21 Food and Drugs 3 2012-04-01 2012-04-01 false Trypsin. 184.1914 Section 184.1914 Food and Drugs... characterizing enzyme activity is that of a peptide hydrolase (EC 3.4.21.4). (b) The ingredient meets the general...
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Code of Federal Regulations, 2014 CFR
2014-04-01
...: (1) The ingredient is used as an enzyme as defined in § 170.3(o)(9) of this chapter to hydrolyze... 21 Food and Drugs 3 2014-04-01 2014-04-01 false Trypsin. 184.1914 Section 184.1914 Food and Drugs... activity is that of a peptide hydrolase (EC 3.4.21.4). (b) The ingredient meets the general requirements...
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Code of Federal Regulations, 2010 CFR
2010-04-01
... defined in § 170.3(o)(9) of this chapter to hydrolyze proteins or polypeptides. (2) The ingredient is used... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Trypsin. 184.1914 Section 184.1914 Food and Drugs... characterizing enzyme activity is that of a peptide hydrolase (EC 3.4.21.4). (b) The ingredient meets the general...
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Use of microwaves to improve nutritional value of soybeans for future space inhabitants
NASA Technical Reports Server (NTRS)
Singh, G.
1983-01-01
Whole soybeans from four different varieties at different moisture contents were microwaved for varying times to determine the conditions for maximum destruction of trypsin inhibitor and lipoxygenase activities, and optimal growth of chicks. Microwaving 150 gm samples of soybeans (at 14 to 28% moisture) for 1.5 min was found optimal for reduction of trypsin inhibitor and lipoxygenase activities. Microwaving 1 kgm samples of soybeans for 9 minutes destroyed 82% of the trypsin inhibitor activity and gave optimal chick growth. It should be pointed out that the microwaving time would vary according to the weight of the sample and the power of the microwave oven. The microwave oven used in the above experiments was rated at 650 watts 2450 MHz.
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Methacrylate gels with epoxide groups as supports for immobilization of enzymes in pH range 3-12.
Turková, J; Bláha, K; Malaníková, M; Vancurová, D; Svec, F; Kálal, J
1978-05-11
Glycidyl methacrylate gels are carriers suitable for attachment of enzymes and for use in affinity chromatography. Experiments on the coupling of glycyl-L-leucine and acetyl-L-leucine to these gels have shown a high pH-dependence of the bond formation between the support and the alpha-amino group (pH optimum 9.7); the coupling reaction between the epoxide group and the carboxyl group is practically pH-independent. Serum albumin and trypsin were attached to a greater extent in acidic than in alkaline media. The effects of time and temperature were also studied. The catalytic action of immobilized trypsin, as well as its use for affinity chromatography of trypsin inhibitor, were studied.
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Comparing growth rates of Arctic Cod Boreogadus saida across the Chukchi and Beaufort Seas
NASA Astrophysics Data System (ADS)
Frothingham, A. M.; Norcross, B.
2016-02-01
Dramatic changes to the Arctic have highlighted the need for a greater understanding of the present ecosystem. Arctic Cod, Boreogadus saida, commonly dominate fish assemblages in the Arctic region and inhabit two geographically unique seas in the U.S. Due to the importance of Arctic Cod in the Arctic food web, establishing current benchmark information such as growth rates, will provide a better understanding as to how the species will adapt to the effects of climate change. To investigate differences in Arctic Cod life history across nearly 1500 km of vital habitat, growth rates were examined using a von Bertalanffy growth equation. Arctic Cod were collected from 2009 to 2014 from the Chukchi and Beaufort seas. Arctic Cod collected from the Chukchi Sea had an overall smaller maximum achievable length (210 mm) compared to the Beaufort Sea (253 mm) despite a larger sample size in the Chukchi Sea (n=1569) than the Beaufort Sea (n=1140). Growth rates indicated faster growth in the Chukchi Sea (K =0.33) than in the Beaufort Sea (K= 0.29). Arctic Cod collected from the Chukchi Sea had similar achievable maximum lengths throughout, but those collected from the southern Chukchi Sea grew at faster rates (K=0.45).Arctic Cod in the eastern Beaufort Sea region had a higher overall maximum achievable length (243 mm) than in the western Beaufort Sea region (186 mm). Knowledge about contemporary growth rates of Arctic Cod in the Chukchi and Beaufort Seas can be used in future comparisons to evaluate potential effects of increasing climate change and anthropogenic influences.
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Ruglass, Lesia M; Shevorykin, Alina; Brezing, Christina; Hu, Mei-Chen; Hien, Denise A
2017-09-01
While the detrimental effects of concurrent substance use disorders (SUDs) are now being well documented, very few studies have examined this comorbidity among women with posttraumatic stress disorder (PTSD). Data for these analyses were derived from the "Women and Trauma" study conducted within the National Drug Abuse Treatment Clinical Trials Network. Women with full or subthreshold PTSD and co-occurring cannabis use disorder (CUD) and cocaine use disorder (COD; N=99) were compared to their counterparts with co-occurring CUD only (N=26) and co-occurring COD only (N=161) on rates of trauma exposure, psychiatric disorders, psychosocial problems, and other substance use utilizing a set of multivariate logistic regressions. In models adjusted for age and race/ethnicity, women with PTSD and COD only were significantly older than their counterparts with CUD only and concurrent CUD+COD. Relative to those with CUD only, women with concurrent CUD+COD had higher odds of adult sexual assault. Relative to those with COD only, women with concurrent CUD+COD had higher odds of alcohol use disorder in the past 12months. Finally, relative to those with CUD only, women with COD only had higher odds of ever being arrested/convicted and adult sexual assault. The higher rates of adult sexual assault and alcohol use disorder among those with concurrent CUD+COD suggest the need for trauma-informed approaches that can respond to the needs of this dually-diagnosed population. Moreover, the causal link between repeated traumatic stress exposure and polysubstance use requires further examination. Copyright © 2017 Elsevier Inc. All rights reserved.
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Svedäng, Henrik; Hornborg, Sara
2017-08-01
Understanding how combinations of fishing effort and selectivity affect productivity is central to fisheries research. We investigate the roles of fishing regulation in comparison with ecosystem status for Baltic Sea cod stock productivity, growth performance, and population stability. This case study is interesting because three cod populations with different exploitation patterns and stock status are located in three adjacent but partially, ecologically different areas. In assessing stock status, growth, and productivity, we use survey information and rather basic stock parameters without relying on age readings. Because there is an urgent interest of better understanding of the current development of the Eastern Baltic cod stock, we argue that our approach represents partly a novel way of interpreting monitoring information together with catch data in a simplified yet more informative way. Our study reports how the Eastern and Western Baltic cod have gone toward more truncated size structures between 1991 and 2016, in particular for the Eastern Baltic cod, whereas the Öresund cod show no trend. We suggest that selective fishing may disrupt fish population dynamic stability and that lower natural productivity might amplify the effects of selective fishing. In support of earlier findings on a density-dependent growth of Eastern Baltic cod, management is advised to acknowledge that sustainable exploitation levels for Eastern Baltic cod are much more limited than perceived in regular assessments. Of more general importance, our results emphasize the need to embrace a more realistic view on what ecosystems can produce regarding tractable fish biomass to facilitate a more ecosystem-based fisheries management.
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Bejankiwar, Rajesh S; Lokesh, K S; Gowda, T P Halappa
2003-05-01
The treatment of biologically treated wastewater of coffee-curing industry by the electrochemical oxidation using steel anode was investigated. Bench-scale experiments were conducted for activated sludge process on raw wastewater and the treated effluents were further treated by electrochemical oxidation method for its colour and organic content removal. The efficiency of the process was determined in terms of removal percentage of COD, BOD and colour during the course of reaction. Several operating parameters like time, pH and current density were examined to ascertain their effects on the treatment efficiency. Steel anode was found to be effective for the COD and colour removal with anode efficiency of 0.118 kgCOD x h(-1) x A(-1) x m(-2) and energy consumption 20.61 kWh x kg(-1) of COD at pH 9. The decrease in pH from 9 to 3 found to increase the anode efficiency from 0.118 kgCOD x h(-1) x A(-1) x m(-2) to 0.144 kWh x kg(-1) of COD while decrease the energy consumption from 20.61 kWh x kg(-1) of COD to 12.86 kWh x kg(-1) of COD. The pH of 5 was considered an ideal from the present treatment process as it avoids the addition of chemicals for neutralization of treated effluents and also economical with respect to energy consumption. An empirical relation developed for relationship between applied current density and COD removal efficiency showed strong predictive capability with coefficient of determination of 96.5%.
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Piña-Salazar, E Z; Cervantes, F J; Meraz, M; Celis, L B
2011-01-01
In sulfate-reducing reactors, it has been reported that the sulfate removal efficiency increases when the COD/SO4(2-) ratio is increased. The start-up of a down-flow fluidized bed reactor constitutes an important step to establish a microbial community in the biofilm able to survive under the operational bioreactor conditions in order to achieve effective removal of both sulfate and organic matter. In this work the influence of COD/SO4(2-) ratio and HRT in the development of a biofilm during reactor start-up (35 days) was studied. The reactor was inoculated with 1.6 g VSS/L of granular sludge, ground low density polyethylene was used as support material; the feed consisted of mineral medium at pH 5.5 containing 1 g COD/L (acetate:lactate, 70:30) and sodium sulfate. Four experiments were conducted at HRT of 1 or 2 days and COD/SO4(2-) ratio of 0.67 or 2.5. The results obtained indicated that a COD/SO4(2-) ratio of 2.5 and HRT 2 days allowed high sulfate and COD removal (66.1 and 69.8%, respectively), whereas maximum amount of attached biomass (1.9 g SVI/L support) and highest sulfate reducing biofilm activity (10.1 g COD-H2S/g VSS-d) was achieved at HRT of 1 day and at COD/sulfate ratios of 0.67 and 2.5, respectively, which suggests that suspended biomass also played a key role in the performance of the reactors.
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Cape Cod National Seashore intelligent transportation systems implementation plan: final report
DOT National Transportation Integrated Search
2011-03-18
The Volpe National Transportation Systems Center (Volpe Center) is providing technical support to the Cape Cod National Seashore in the planning of an Intelligent Transportation System (ITS). In collaboration with the Cape Cod Commission, National Se...
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Mendez-Villanueva, Alberto; Palazzi, Dino; Ahmaidi, Saïd
2016-01-01
Purpose The aims of this study were to 1) compare the metabolic power demand of straight-line and change of direction (COD) sprints including 45° or 90°-turns, and 2) examine the relation between estimated metabolic demands and muscular activity throughout the 3 phases of COD-sprints. Methods Twelve highly-trained soccer players performed one 25-m and three 20-m sprints, either in straight-line or with one 45°- or 90°-COD. Sprints were monitored with 2 synchronized 100-Hz laser guns to assess players’ velocities before, during and after the COD. Acceleration and deceleration were derived from changes in speed over time. Metabolic power was estimated based on di Prampero’s approach (2005). Electromyography amplitude (RMS) of 2 lower limb muscles was measured. The expected energy expenditure during time-adjusted straight-line sprints (matching COD sprints time) was also calculated. Results Locomotor-dependant metabolic demand was largely lower with COD (90°, 142.1±13.5 J.kg-1) compared with time-adjusted (effect size, ES = -3.0; 193.2±18.6 J.kg-1) and non-adjusted straight-line sprints (ES = -1.7; 168.4±15.3 J.kg-1). Metabolic power requirement was angle-dependent, moderately lower for 90°-COD vs. 45°-COD sprint (ES = -1.0; 149.5±10.4 J.kg-1). Conversely, the RMS was slightly- (45°, ES = +0.5; +2.1%, 90% confidence limits (±3.6) for vastus lateralis muscle (VL)) to-largely (90°, ES = +1.6; +6.1 (3.3%) for VL) greater for COD-sprints. Metabolic power/RMS ratio was 2 to 4 times lower during deceleration than acceleration phases. Conclusion Present results show that COD-sprints are largely less metabolically demanding than linear sprints. This may be related to the very low metabolic demand associated with the deceleration phase during COD-sprints that may not be compensated by the increased requirement of the reacceleration phase. These results also highlight the dissociation between metabolic and muscle activity demands during COD-sprints, which questions the use of metabolic power as a single measure of running load in soccer. PMID:26930649
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Seasonal Temperature Estimates From Late Holocene Barents Sea cod Otoliths: Problems and Potential
NASA Astrophysics Data System (ADS)
Andersson, C.; Folkvord, A.; Geffen, A.; Høie, H.
2008-12-01
Realistic models of climate change require accurate data of past seasonal temperature regimes. In many marine settings this is difficult to achieve using the marine sedimentary record. This study tests the feasibility to use archaeological cod otoliths to reconstruct changes in the seasonal temperature cycle in the Barents Sea. Potentially, stable oxygen isotope records from cod otoliths could be used to produce records of Holocene temperature change on a seasonal time scale. Human settlements along the coast of northern Norway have exploited the cod (Gadus morhua) population of the Barents Sea from the earliest occupation, through the middle ages, to the present day. A pilot project has been carried out in Bergen to exploit the availability of a collection of cod otoliths from archeological excavations that is housed by the Bergen Museum. Cod otoliths from archaeological excavations in northern Norway, 26 specimens from from Måsøy (Finnmark) and 17 specimens from Vanna (Troms), were selected for this study. These specimens were AMS radiocarbon dated and the ages are spanning approximately 1400 to 1780 AD. Most of the otoliths are from cod that were captured during the Little Ice Age. Seasonal growth patterns were identifiable in the archeological otoliths, comparable to those in modern otoliths. Micromilling was used to sample for stable oxygen isotopes over a 2-year growth period in each of a total of 43 fossil cod otoliths. There are large differences in the temperature ranges experienced by the fish over the 2-year period analyzed. For the Måsøy specimens the temperature range experienced by the fish is between 2.7 to 9.9 °C. The Vanna cod experienced temperature ranges between 1.7 to 7.2 °C. The maximum temperature ranges for both Vanna and Måsøy specimens are higher than the seasonal instrumental measurements for the 0-200 m depth interval in the Kola section in the Barents Sea. The cod otoliths in our study have not yet been divided into different age groups, which is necessary in order to be able to interpret the calculated temperature ranges because the annual temperature cycles experienced by fish at different ages are different. Another problem that needs to address is the annual migration of cod of northern Norway. It is known that both mature and immature cod undertake long seasonal migrations.
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Research progress of on-line automatic monitoring of chemical oxygen demand (COD) of water
NASA Astrophysics Data System (ADS)
Cai, Youfa; Fu, Xing; Gao, Xiaolu; Li, Lianyin
2018-02-01
With the increasingly stricter control of pollutant emission in China, the on-line automatic monitoring of water quality is particularly urgent. The chemical oxygen demand (COD) is a comprehensive index to measure the contamination caused by organic matters, and thus it is taken as one important index of energy-saving and emission reduction in China’s “Twelve-Five” program. So far, the COD on-line automatic monitoring instrument has played an important role in the field of sewage monitoring. This paper reviews the existing methods to achieve on-line automatic monitoring of COD, and on the basis, points out the future trend of the COD on-line automatic monitoring instruments.
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Thomas, Christopher; Comfort, Paul; Jones, Paul A; Dos'Santos, Thomas
2017-08-01
To investigate the relationships between maximal isometric strength, vertical jump (VJ), sprint speed, and change-of-direction speed (CoDS) in academy netball players and determine whether players who have high performance in isometric strength testing would demonstrate superior performance in VJ, sprint speed, and CoDS measures. Twenty-six young female netball players (age 16.1 ± 1.2 y, height 173.9 ± 5.7 cm, body mass 66.0 ± 7.2 kg) from a regional netball academy performed isometric midthigh pull (IMTP), squat jumps (SJs), countermovement jumps (CMJs), 10-m sprints, and CoDS (505). IMTP measures displayed moderate to strong correlations with sprint and CoDS performance (r = -.41 to -.66). The VJs, which included SJs and CMJs, demonstrated strong correlations with 10-m sprint times (r = -.60 to -.65; P < .01) and CoDS (r = -.60 to -.71; P = .01). Stronger players displayed significantly faster sprint (ES = 1.1-1.2) and CoDS times (ES = 1.2-1.7) and greater VJ height (ES = 0.9-1.0) than weaker players. The results of this study illustrate the importance of developing high levels of lower-body strength to enhance VJ, sprint, and CoDS performance in youth netball players, with stronger athletes demonstrating superior VJ, sprint, and CoDS performances.
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Dalgaard, P; Garcia Munoz, L; Mejlholm, O
1998-09-01
Inhibition of the specific spoilage organism, Photobacterium phosphoreum, was studied in model substrates and in modified-atmosphere-packed cod fillets. The objective was to determine how inhibition of this organism influenced spoilage. The spoilage reactions limiting shelf life were studied rather than the development of a new product. In naturally contaminated modified-atmosphere-packed cod fillets, 500 ppm Na2CaEDTA reduced the growth rate of P. phosphoreum by 40% and shelf life was increased proportionally by 40%, from 15 to 17 days to 21 to 23 days at 0 degree C. In aerobically stored cod fillets other microorganisms were responsible for spoilage and Na2CaEDTA had no effect on shelf life. The extension of the shelf life of modified-atmosphere-packed cod therefore was a result of the reduced growth of P. phosphoreum and no other microbial or nonmicrobial spoilage reactions limited shelf life. These results confirmed P. phosphoreum as the specific spoilage organism in modified-atmosphere-packed cod and showed the organism to have an extensive spoilage domain. Consequently, any preservation procedure able to reduce growth of P. phosphoreum is likely to extend shelf life of packed cod. However, the effect of different inhibitory compounds in model systems as well as results from packed cod indicated the need to include product studies in the screening of antimicrobials and in the development of preservation procedures.
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Raghu, S; Ahmed Basha, C
2007-10-22
This paper examines the use of chemical or electrocoagulation treatment process followed by ion-exchange process of the textile dye effluent. The dye effluent was treated using polymeric coagulant (cationic dye-fixing agent) or electrocoagulation (iron and aluminum electrode) process under various conditions such as various current densities and effect of pH. Efficiencies of COD reduction, colour removal and power consumption were studied for each process. The chemical or electrochemical treatment are indented primarily to remove colour and COD of wastewater while ion exchange is used to further improve the removal efficiency of the colour, COD, Fe concentration, conductivity, alkalinity and total dissolved solids (TDS). From the results chemical coagulation, maximum COD reduction of about 81.3% was obtained at 300 mg/l of coagulant whereas in electrocoagulation process, maximum COD removal of about 92.31% (0.25 A/dm2) was achieved with energy consumption of about 19.29 k Wh/kg of COD and 80% (1A/dm(2)) COD removal was obtained with energy consumption of about 130.095 k Wh/kg of COD at iron and aluminum electrodes, respectively. All the experimental results, throughout the present study, have indicated that chemical or electrocoagulation treatment followed by ion-exchange methods were very effective and were capable of elevating quality of the treated wastewater effluent to the reuse standard of the textile industry.
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Chadwick, J W; Alsufyani, N A; Lam, E W N
2011-05-01
The simple bone cyst (SBC) is a pseudocyst that can occur as a solitary entity in the jaws or may occur in association with cemento-osseous dysplasia (COD). The purpose of this study was to review the clinical and radiographic features of solitary and COD-associated SBCs. Archived imaging reports from the Special Procedures Clinic in Oral and Maxillofacial Radiology at the Faculty of Dentistry at the University of Toronto between 1 January 1989 and 31 December 2009 revealed 23 COD-associated SBCs and 68 solitary SBCs. Almost all solitary and COD-associated SBCs were found in the mandible. Furthermore, 87.0% of COD-associated SBCs were found in females in their fifth decade of life (P < 0.001) while solitary SBCs were found in equal numbers in both sexes in their second decade of life (P < 0.005). COD-associated SBCs were also more likely to cause thinning of the endosteal cortex, bone expansion and scalloping of the superior border between teeth (all P < 0.001) than solitary SBCs that are classically described as having these characteristics. Finally, COD-associated SBC demonstrated a loss of lamina dura more often (P < 0.05) than solitary SBCs. Knowledge of the sporadic association between COD and SBC and their potential radiographic appearances should prevent inappropriate treatment and management of these patients.
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Behavioral responses of Atlantic cod to sea temperature changes.
Freitas, Carla; Olsen, Esben Moland; Moland, Even; Ciannelli, Lorenzo; Knutsen, Halvor
2015-05-01
Understanding responses of marine species to temperature variability is essential to predict impacts of future climate change in the oceans. Most ectotherms are expected to adjust their behavior to avoid extreme temperatures and minimize acute changes in body temperature. However, measuring such behavioral plasticity in the wild is challenging. Combining 4 years of telemetry-derived behavioral data on juvenile and adult (30-80 cm) Atlantic cod (Gadus morhua), and in situ ocean temperature measurements, we found a significant effect of sea temperature on cod depth use and activity level in coastal Skagerrak. During summer, cod were found in deeper waters when sea surface temperature increased. Further, this effect of temperature was stronger on larger cod. Diel vertical migration, which consists in a nighttime rise to shallow feeding habitats, was stronger among smaller cod. As surface temperature increased beyond ∼15°C, their vertical migration was limited to deeper waters. In addition to larger diel vertical migrations, smaller cod were more active and travelled larger distances compared to larger specimens. Cold temperatures during winter tended, however, to reduce the magnitude of diel vertical migrations, as well as the activity level and distance moved by those smaller individuals. Our findings suggest that future and ongoing rises in sea surface temperature may increasingly deprive cod in this region from shallow feeding areas during summer, which may be detrimental for local populations of the species.
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Kalyuzhnyi, Sergey; Gladchenko, Marina; Epov, Andrey; Appanna, Vasu
2003-01-01
As a first step of treatment of landfill leachates (total chemical oxygen demand [COD]: 1.43-3.81 g/L; total nitrogen: 90-162 mg/L), performance of laboratory upflow anaerobic sludge bed reactors was investigated under mesophilic (30 degrees C), submesophilic (20 degrees C), and psychrophilic (10 degrees C) conditions. Under hydraulic retention times (HRTs) of about 0.3 d, when the average organic loading rates (OLRs) were about 5 g of COD/(L.d), the total COD removal accounted for 81% (on average) with the effluent concentrations close to the anaerobic biodegradability limit (0.25 g of COD/L) for mesophilic and submesophilic regimes. The psychrophilic treatment conducted under an average HRT of 0.34 d and an average OLR of 4.22 g of COD/(L.d) showed a total COD removal of 47%, giving effluents (0.75 g of COD/L) more suitable for subsequent biologic nitrogen removal. All three anaerobic regimes used for leachate treatment were quite efficient for elimination of heavy metals (Fe, Zn, Cu, Pb, Cd) by concomitant precipitation in the form of insoluble sulfides inside the sludge bed. The application of aerobic/ anoxic biofilter as a sole polishing step for psychrophilic anaerobic effluents was acceptable for elimination of biodegradable COD and nitrogen approaching the current standards for direct discharge of treated wastewater.
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Behavioral responses of Atlantic cod to sea temperature changes
Freitas, Carla; Olsen, Esben Moland; Moland, Even; Ciannelli, Lorenzo; Knutsen, Halvor
2015-01-01
Understanding responses of marine species to temperature variability is essential to predict impacts of future climate change in the oceans. Most ectotherms are expected to adjust their behavior to avoid extreme temperatures and minimize acute changes in body temperature. However, measuring such behavioral plasticity in the wild is challenging. Combining 4 years of telemetry-derived behavioral data on juvenile and adult (30–80 cm) Atlantic cod (Gadus morhua), and in situ ocean temperature measurements, we found a significant effect of sea temperature on cod depth use and activity level in coastal Skagerrak. During summer, cod were found in deeper waters when sea surface temperature increased. Further, this effect of temperature was stronger on larger cod. Diel vertical migration, which consists in a nighttime rise to shallow feeding habitats, was stronger among smaller cod. As surface temperature increased beyond ∼15°C, their vertical migration was limited to deeper waters. In addition to larger diel vertical migrations, smaller cod were more active and travelled larger distances compared to larger specimens. Cold temperatures during winter tended, however, to reduce the magnitude of diel vertical migrations, as well as the activity level and distance moved by those smaller individuals. Our findings suggest that future and ongoing rises in sea surface temperature may increasingly deprive cod in this region from shallow feeding areas during summer, which may be detrimental for local populations of the species. PMID:26045957
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Shuler, Monique; Suzuki, Sumihiro; Podesta, Arwen; Qualls-Hampton, Raquel; Wallington, Sherrie Flynt
2017-01-01
There is increasing literature supporting the adverse effects of disasters on substance use and psychiatric disorders. The co-occurrence of psychiatric disorders with substance use intensifies the challenge of treatment delivery. Thus the aim of this study was to examine the prevalence of substance use, treatment characteristics, and demographics of discharges from substance abuse treatment in New Orleans, post-Hurricane Katrina. Trends associated with discharges that have a co-occurring psychiatric and substance use disorder (COD) were also assessed. The secondary aim of this study was to examine the association of successful substance abuse treatment completion among those with a COD post-Hurricane Katrina. Substance abuse treatment discharge data (N = 16,507) from New Orleans, Louisiana, for years 2006 through 2011 were obtained from the Treatment Episode Data Set-Discharge. Multiple logistic regression analysis was employed to examine the association of discharges with a COD and completion of substance abuse treatment. Demographic, psychiatric, and treatment characteristics of discharges in 2006 were compared to characteristics in 2011. Trends of characteristics were also assessed through the study period. Roughly a third (35.2%) of all discharges in New Orleans from 2006 to 2011 had a COD. After controlling for race, employment, treatment service setting at discharge, primary substance problem, and the discharge's principal source of referral, discharges with a COD were 29% less likely to complete treatment as compared to those with no COD (AOR = 0.71, 95% CI [0.56, 0.90], p = .004). Treatment completion among discharges with a COD has significantly declined from 36.8% in 2006 to 18.7% in 2011 (p < .0001). Notable significant trends in homelessness, criminality, and heroin use were identified among discharges with a COD. Substance abuse treatment undergoes various changes in the event of a natural disaster. These changes may increase challenges for successful treatment completion for vulnerable populations such as those with a COD. Results of this study demonstrate that discharges with a COD are less likely to complete treatment as compared to those with no COD disorder. Unmet treatment needs may also increase odds of criminalization and homelessness.
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The Relationship between Running Velocity and the Energy Cost of Turning during Running
Hatamoto, Yoichi; Yamada, Yosuke; Sagayama, Hiroyuki; Higaki, Yasuki; Kiyonaga, Akira; Tanaka, Hiroaki
2014-01-01
Ball game players frequently perform changes of direction (CODs) while running; however, there has been little research on the physiological impact of CODs. In particular, the effect of running velocity on the physiological and energy demands of CODs while running has not been clearly determined. The purpose of this study was to examine the relationship between running velocity and the energy cost of a 180°COD and to quantify the energy cost of a 180°COD. Nine male university students (aged 18–22 years) participated in the study. Five shuttle trials were performed in which the subjects were required to run at different velocities (3, 4, 5, 6, 7, and 8 km/h). Each trial consisted of four stages with different turn frequencies (13, 18, 24 and 30 per minute), and each stage lasted 3 minutes. Oxygen consumption was measured during the trial. The energy cost of a COD significantly increased with running velocity (except between 7 and 8 km/h, p = 0.110). The relationship between running velocity and the energy cost of a 180°COD is best represented by a quadratic function (y = −0.012+0.066x +0.008x2, [r = 0.994, p = 0.001]), but is also well represented by a linear (y = −0.228+0.152x, [r = 0.991, p<0.001]). These data suggest that even low running velocities have relatively high physiological demands if the COD frequency increases, and that running velocities affect the physiological demands of CODs. These results also showed that the energy expenditure of COD can be evaluated using only two data points. These results may be useful for estimating the energy expenditure of players during a match and designing shuttle exercise training programs. PMID:24497913
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CodY Promotes Sporulation and Enterotoxin Production by Clostridium perfringens Type A Strain SM101.
Li, Jihong; Freedman, John C; Evans, Daniel R; McClane, Bruce A
2017-03-01
Clostridium perfringens type D strains cause enterotoxemia and enteritis in livestock via epsilon toxin production. In type D strain CN3718, CodY was previously shown to increase the level of epsilon toxin production and repress sporulation. C. perfringens type A strains producing C. perfringens enterotoxin (CPE) cause human food poisoning and antibiotic-associated diarrhea. Sporulation is critical for C. perfringens type A food poisoning since spores contribute to transmission and resistance in the harsh food environment and sporulation is essential for CPE production. Therefore, the current study asked whether CodY also regulates sporulation and CPE production in SM101, a derivative of C. perfringens type A food-poisoning strain NCTC8798. An isogenic codY -null mutant of SM101 showed decreased levels of spore formation, along with lower levels of CPE production. A complemented strain recovered wild-type levels of both sporulation and CPE production. When this result was coupled with the earlier results obtained with CN3718, it became apparent that CodY regulation of sporulation varies among different C. perfringens strains. Results from quantitative reverse transcriptase PCR analysis clearly demonstrated that, during sporulation, codY transcript levels remained high in SM101 but rapidly declined in CN3718. In addition, abrB gene expression patterns varied significantly between codY -null mutants of SM101 and CN3718. Compared to the levels in their wild-type parents, the level of abrB gene expression decreased in the CN3718 codY -null mutant strain but significantly increased in the SM101 codY -null mutant strain, demonstrating CodY-dependent regulation differences in abrB expression between these two strains. This difference appears to be important since overexpression of the abrB gene in SM101 reduced the levels of sporulation and enterotoxin production, supporting the involvement of AbrB repression in regulating C. perfringens sporulation. Copyright © 2017 American Society for Microbiology.
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CodY Promotes Sporulation and Enterotoxin Production by Clostridium perfringens Type A Strain SM101
Li, Jihong; Freedman, John C.; Evans, Daniel R.
2017-01-01
ABSTRACT Clostridium perfringens type D strains cause enterotoxemia and enteritis in livestock via epsilon toxin production. In type D strain CN3718, CodY was previously shown to increase the level of epsilon toxin production and repress sporulation. C. perfringens type A strains producing C. perfringens enterotoxin (CPE) cause human food poisoning and antibiotic-associated diarrhea. Sporulation is critical for C. perfringens type A food poisoning since spores contribute to transmission and resistance in the harsh food environment and sporulation is essential for CPE production. Therefore, the current study asked whether CodY also regulates sporulation and CPE production in SM101, a derivative of C. perfringens type A food-poisoning strain NCTC8798. An isogenic codY-null mutant of SM101 showed decreased levels of spore formation, along with lower levels of CPE production. A complemented strain recovered wild-type levels of both sporulation and CPE production. When this result was coupled with the earlier results obtained with CN3718, it became apparent that CodY regulation of sporulation varies among different C. perfringens strains. Results from quantitative reverse transcriptase PCR analysis clearly demonstrated that, during sporulation, codY transcript levels remained high in SM101 but rapidly declined in CN3718. In addition, abrB gene expression patterns varied significantly between codY-null mutants of SM101 and CN3718. Compared to the levels in their wild-type parents, the level of abrB gene expression decreased in the CN3718 codY-null mutant strain but significantly increased in the SM101 codY-null mutant strain, demonstrating CodY-dependent regulation differences in abrB expression between these two strains. This difference appears to be important since overexpression of the abrB gene in SM101 reduced the levels of sporulation and enterotoxin production, supporting the involvement of AbrB repression in regulating C. perfringens sporulation. PMID:28052992
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Muhamad, Mohd Hafizuddin; Sheikh Abdullah, Siti Rozaimah; Mohamad, Abu Bakar; Rahman, Rakmi Abdul; Kadhum, Abdul Amir Hasan
2012-01-01
A pilot scale granular activated carbon-sequencing batch biofilm reactor with a capacity of 2.2 m3 was operated for over three months to evaluate its performance treating real recycled paper industry wastewater under different operational conditions. In this study, dissolved air floatation (DAF) and clarifier effluents were used as influent sources of the pilot plant. During the course of the study, the reactor was able to biodegrade the contaminants in the incoming recycled paper mill wastewater in terms of chemical oxygen demand (COD), adsorbable organic halides (AOX; specifically 2,4-dichlorophenol (2,4-DCP)) and ammoniacal nitrogen (NH3-N) removal efficiencies at varying hydraulic retention times (HRTs) of 1-3 days, aeration rates (ARs) of 2.1-3.4 m3/min and influent feed concentration of 40-950 mg COD/l. Percentages of COD, 2,4-DCP and NH3-N removals increased with increasing HRT, resulting in more than 90% COD, 2,4-DCP and NH3-N removals at HRT values above two days. Degradation of COD, 2,4-DCP and NH3-N were seriously affected by variation of ARs, which resulted in significant decrease of COD, 2,4-DCP and NH3-N removals by decreasing ARs from 3.4 m3/min to 2.1 m3/min, varying in the ranges of 24-80%, 6-96% and 5-42%, respectively. In comparison to the clarifier effluent, the treatment performance of DAF effluent, containing high COD concentration, resulted in a higher COD removal of 82%. The use of diluted DAF effluent did not improve significantly the COD removal. Higher NH3-N removal efficiency of almost 100% was observed during operation after maintenance shutdown compared to normal operation, even at the same HRT of one day due to the higher dissolved oxygen concentrations (1-7 mg/l), while no significant difference in COD removal efficiency was observed.
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Causes of death among commercially insured multiple sclerosis patients in the United States.
Goodin, Douglas S; Corwin, Michael; Kaufman, David; Golub, Howard; Reshef, Shoshana; Rametta, Mark J; Knappertz, Volker; Cutter, Gary; Pleimes, Dirk
2014-01-01
Information on causes of death (CODs) for patients with multiple sclerosis (MS) in the United States is sparse and limited by standard categorizations of underlying and immediate CODs on death certificates. Prior research indicated that excess mortality among MS patients was largely due to greater mortality from infectious, cardiovascular, or pulmonary causes. To analyze disease categories in order to gain insight to pathways, which lead directly to death in MS patients. Commercially insured MS patients enrolled in the OptumInsight Research database between 1996 and 2009 were matched to non-MS comparators on age/residence at index year and sex. The cause most-directly leading to death from the death certificate, referred to as the "principal" COD, was determined using an algorithm to minimize the selection of either MS or cardiac/pulmonary arrest as the COD. Principal CODs were categorized into MS, cancer, cardiovascular, infectious, suicide, accidental, pulmonary, other, or unknown. Infectious, cardiovascular, and pulmonary CODs were further subcategorized. 30,402 MS patients were matched to 89,818 controls, with mortality rates of 899 and 446 deaths/100,000 person-years, respectively. Excluding MS, differences in mortality rate between MS patients and non-MS comparators were largely attributable to infections, cardiovascular causes, and pulmonary problems. Of the 95 excessive deaths (per 100,000 person-years) related to infectious causes, 41 (43.2%) were due to pulmonary infections and 45 (47.4%) were attributed to sepsis. Of the 46 excessive deaths (per 100,000 person-years) related to pulmonary causes, 27 (58.7%) were due to aspiration. No single diagnostic entity predominated for the 60 excessive deaths (per 100,000 person-years) attributable to cardiac CODs. The principal COD algorithm improved on other methods of determining COD in MS patients from death certificates. A greater awareness of the common CODs in MS patients will allow physicians to anticipate potential problems and, thereby, improve the care that they provide.
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Llanos, Ricardo J; Barrera, Daniel; Valz-Gianinet, Jorge N; Miceli, Dora C
2006-10-01
We describe the morphological and biochemical changes in Bufo arenarum coelomic egg envelopes (CE) following passage through the oviduct. In this species, the transformation of the CE into the vitelline envelope (VE) leads to the acquisition of fertilizability and involves the cleavage of a glycoprotein component. Electrophoretic patterns indicate that a pars recta oviductal protease selectively hydrolyzes in vitro the 84 and the 55 kDa glycoproteins of the CE. During the CE to VE transformation, the relative concentrations of gp48, 42 and 39 kDa also change. In in vitro tests, sperm binding to envelope glycoprotein occurs when they are exposed to VE but not when treated with CE, and VE labeled glycoproteins bind to the head and mid piece of the sperm. The gp39 VE component has 100% identity with internal domains of the sequence deduced from ovarian cDNA for the homologous zona pellucida glycoprotein type C (ZPC) protein precursor in B. arenarum. The effects of trypsin as a substitute for oviductal protease were also examined. Trypsin selectively attacks the 84 and the 55 kDa glycoproteins without hydrolyzing other components and renders coelomic eggs fertilizable in a jelly water preparation. Therefore, trypsin can mimic in vitro the biological action of the oviductal protease. However, it does not wholly mimic the biological action of the oviduct which, in B. arenarum at least, exceeds a mere proteolytic effect. This fact was verified by the lower fertility rates and the abnormal embryo development found when trypsin-treated coelomic eggs were fertilized in vitro. 2006 Wiley-Liss, Inc.
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Clearance of bile and trypsin in rat lungs following aspiration of human gastric fluid
Leung, Jason H.; Chang, Jui-Chih; Foltz, Emily; Bell, Sadé M.; Pi, Cinthia; Azad, Sassan; Everett, Mary Lou; Holzknecht, Zoie E.; Sanders, Nathan L.; Parker, William; Davis, R. Duane; Keshavjee, Shaf; Lin, Shu S.
2016-01-01
ABSTRACT Purpose: In the clinical setting, there is no reliable tool for diagnosing gastric aspiration. A potential way of diagnosing gastric fluid aspiration entails bronchoalveolar lavage (BAL) with subsequent examination of the BAL fluid for gastric fluid components that are exogenous to the lungs. The objective of this study was to determine the longevity of the gastric fluid components bile and trypsin in the lung, in order to provide an estimate of the time frame in which assessment of these components in the BAL might effectively be used as a measure of aspiration. Materials and Methods: Human gastric fluid (0.5 mg/kg) was infused in the right lung of intubated male Fischer 344 rats (n = 30). Animals were sacrificed at specified times following the experimentally induced aspiration, and bronchoalveolar lavage fluid (BALF) was collected. Bile concentrations were analyzed by an enzyme-linked chromatogenic method, and the concentration of trypsin was quantified using an ELISA. Data were analyzed using non-linear regression and a one-phase decay equation. Results: In this experimental model, the half-life of bile was 9.3 hours (r 2 = 0.81), and the half-life of trypsin was 9.0 hours (r 2 = 0.68). Conclusions: The half-lives of bile and trypsin in the rodent aspiration model suggest that the ability to detect aspiration may be limited to a few days post-aspiration. If studies using rats are any indication, it may be most effective to collect BAL samples within the first 24 hours of suspected aspiration events in order to detect aspiration. PMID:26873328
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Clearance of bile and trypsin in rat lungs following aspiration of human gastric fluid.
Leung, Jason H; Chang, Jui-Chih; Foltz, Emily; Bell, Sadé M; Pi, Cinthia; Azad, Sassan; Everett, Mary Lou; Holzknecht, Zoie E; Sanders, Nathan L; Parker, William; Davis, R Duane; Keshavjee, Shaf; Lin, Shu S
2016-01-01
In the clinical setting, there is no reliable tool for diagnosing gastric aspiration. A potential way of diagnosing gastric fluid aspiration entails bronchoalveolar lavage (BAL) with subsequent examination of the BAL fluid for gastric fluid components that are exogenous to the lungs. The objective of this study was to determine the longevity of the gastric fluid components bile and trypsin in the lung, in order to provide an estimate of the time frame in which assessment of these components in the BAL might effectively be used as a measure of aspiration. Human gastric fluid (0.5 mg/kg) was infused in the right lung of intubated male Fischer 344 rats (n = 30). Animals were sacrificed at specified times following the experimentally induced aspiration, and bronchoalveolar lavage fluid (BALF) was collected. Bile concentrations were analyzed by an enzyme-linked chromatogenic method, and the concentration of trypsin was quantified using an ELISA. Data were analyzed using non-linear regression and a one-phase decay equation. In this experimental model, the half-life of bile was 9.3 hours (r(2) = 0.81), and the half-life of trypsin was 9.0 hours (r(2) = 0.68). The half-lives of bile and trypsin in the rodent aspiration model suggest that the ability to detect aspiration may be limited to a few days post-aspiration. If studies using rats are any indication, it may be most effective to collect BAL samples within the first 24 hours of suspected aspiration events in order to detect aspiration.
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Rivera-Ciprian, José Pedro; Aceituno-Medina, Marysol; Guillen, Karina
2017-01-01
Abstract In this study, we examined the activity of two serine proteases (chymotrypsin and trypsin) and two metalloproteases (carboxypeptidases A and B) during larval development in Anastrepha obliqua fed natural (mango fruit) and artificial (formulation used in mass-rearing) diets. Proteolytic activity of chymotrypsin, trypsin, carboxypeptidase A, and carboxypeptidase B was detected in the midgut of different instars of A. obliqua and was strongly affected by the pH and diet type. The protein content of the natural and artificial diets was similar. Enzymatic activity was higher in the midgut of the larvae fed the natural diet than in larvae fed the artificial diet. The activity of the endopeptidases (chymotrypsin and trypsin) was lower than those of the exopeptidases (carboxypeptidases A and B). The pH of the midgut varied from acidic to neutral. The results indicate that in the midgut of the larvae reared on both types of diet, the level of carboxypeptidase activity was approximately 100-fold greater than the level of chymotrypsin activity and 10,000-fold greater than the level of trypsin. In conclusion, carboxypeptidase A and B are the main proteases involved in the digestion of proteins in the larvae of A. obliqua. The natural diet showed a high bioaccessibility. A clear tendency to express high activities of chymotrypsin and trypsin was observed by the third instar. Our research contributes to the planning and development of novel bioaccessibility assays to understand the nutrition processing of A. obliqua larvae under mass-rearing conditions for sterile insect technique.
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Cell-free released components of Streptococcus sanguis inhibit human platelet aggregation.
Herzberg, M C; Brintzenhofe, K L; Clawson, C C
1983-01-01
To study the role of surface components in the selective binding and aggregation of platelet-rich plasma (PRP) by strains of viridans streptococci, we treated the binding, aggregation strain Streptococcus sanguis I 2017-78 by sonication or trypsinization. Morphologically identifiable electron-dense fibrils were released from the cell wall, apparently from an inner electron-dense layer, under conditions that left cells intact. These controlled conditions were determined to cause submaximal loss in adhesion to platelet ghosts and PRP aggregation by treated, washed S. sanguis. Soluble components were recovered from the controlled sonic or L-(tosylamido 2-phenyl)ethyl chloromethyl ketone-trypsin treatments. Each showed dose-response inhibition of aggregation when preincubated with PRP before challenge with fresh, untreated S. sanguis. The time to onset of PRP aggregation was inhibited by 50% with 0.2 mg of TPCK-trypsin peptides or 1.0 mg of the sonicate per ml per 2 X 10(8) platelets. Components of both preparations were immunologically cross-reactive, but lipoteichoic acid was not a major antigen of either. By weight, the TPCK-trypsin peptides were virtually all protein; the sonicate residues identified were about 50% protein and 7% hexose. Each was a complex mixture of components as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. More than 8 TPCK-trypsin peptides and 16 sonicate components were so identified. In contrast, at least four or five components from either preparation were recognized as surface determinants by a rabbit antiserum to whole homologous microbes. Platelet-binding ligands of S. sanguis could be among these determinants. Images PMID:6618669
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Oliva, M L V; Andrade, S A; Juliano, M A; Sallai, R C; Torquato, R J; Sampaio, M U; Pott, V J; Sampaio, C A M
2003-07-01
The specific Kunitz Bauhinia ungulata factor Xa inhibitor (BuXI) and the Bauhinia variegata trypsin inhibitor (BvTI) blocked the activity of trypsin, chymotrypsin, plasmin, plasma kallikrein and factor XIIa, and factor Xa inhibition was achieved only by BuXI (K(i) 14 nM). BuXI and BvTI are highly homologous (70%). The major differences are the methionine residues at BuXI reactive site, which are involved in the inhibition, since the oxidized protein no longer inhibits factor Xa but maintains the trypsin inhibition. Quenched fluorescent substrates based on the reactive site sequence of the inhibitors were synthesized and the kinetic parameters of the hydrolysis were determined using factor Xa and trypsin. The catalytic efficiency k(cat)/K(m) 4.3 x 10(7) M(-1)sec(>-1) for Abz-VMIAALPRTMFIQ-EDDnp (lead peptide) hydrolysis by factor Xa was 10(4)-fold higher than that of Boc-Ile-Glu-Gly-Arg-AMC, widely used as factor Xa substrate. Lengthening of the substrate changed its susceptibility to factor Xa hydrolysis. Both methionine residues in the substrate influence the binding to factor Xa. Serine replacement of threonine (P(1)') decreases the catalytic efficiency by four orders of magnitude. Factor Xa did not hydrolyze the substrate containing the reactive site sequence of BvTI, that inhibits trypsin inhibitor but not factor Xa. Abz-VMIAALPRTMFIQ-EDDnp prolonged both the prothrombin time and the activated partial thromboplastin time, and the other modified substrates used in this experiment altered blood-clotting assays.
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Umoh, J. U.; Blenden, D. C.
1981-01-01
Formalin-fixed central nervous system tissue from clinically rabid animals was treated with 0.25% trypsin and tested for the presence of rabies virus antigen by direct immunofluorescent (IF) staining. The results were comparable with those obtained from direct IF staining of acetone-fixed standard smears or fresh frozen-cut sections. Experiments were conducted using coded brain specimens (classified as IF-negative, weakly positive, or strongly positive) and showed a specificity of 100% for sections and 92% for smears; the latter figure was subsequently improved by modifying the preparation technique. The specificity of the technique was checked by standard virus neutralization of the conjugate, and by known antibody neutralization of the virus antigen in the specimens. The optimal duration for the trypsin digestion was found to be a minimum of 60 minutes at 37 °C or 120 minutes at 4 °C. The tissues could be held in buffered formalin for between 3 days and 7 weeks with no apparent difference in the results. Satisfactory concentrations of formalin were 0.125% or 0.25%. Trypsin was found to have no effect on non-formalinized tissues, with the exception that softening occurred making tissues harder to cut and process. The results suggest that trypsinization of formalin-fixed tissue is a valid procedure for the preparation of tissues for IF examination, which would be useful in cases where the current standard techniques cannot be used. However, further evaluation of the method is still required. ImagesFig. 3Fig. 1Fig. 2 PMID:6172212
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Tae, G S; Black, M T; Cramer, W A; Vallon, O; Bogorad, L
1988-12-27
Protease accessibility and antibody to a COOH-terminal peptide were used as probes for the in situ topography of the Mr 10,000 psbE gene product (alpha subunit) of the chloroplast cytochrome b-559. Exposure of thylakoid membranes to trypsin or Staphylococcus aureus V8 protease cleaved the alpha subunit to a slightly smaller polypeptide (delta Mr approximately -1000) as detected on Western blots, without loss of reactivity to COOH-terminal antibody. The disappearance of the parent Mr 10,000 polypeptide from thylakoids in the presence of trypsin correlated with the appearance of the smaller polypeptide with delta Mr = -750, the conversion having a half-time of approximately 15 min. Exposure of inside-out vesicles to trypsin resulted in almost complete loss of reactivity to the antibody, showing that the COOH terminus is exposed on the lumenal side of the membrane. Removal of the extrinsic polypeptides of the oxygen-evolving complex resulted in an increase of the accessibility of the alpha subunit to trypsin. These data establish that the alpha subunit of cytochrome b-559 crosses the membrane once, as predicted from its single, 26-residue, hydrophobic domain. The NH2 terminus of the alpha polypeptide is on the stromal side of the membrane, where it is accessible, most likely at Arg-7 or Glu-6/Asp-11, to trypsin or V8 protease, respectively. As a consequence of this orientation, the single histidine residue in the alpha subunit is located on the stromal side of the hydrophobic domain.(ABSTRACT TRUNCATED AT 250 WORDS)
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Lei, Ge; Ren, Hongqiang; Ding, Lili; Wang, Feifei; Zhang, Xingsong
2010-08-01
A full-scale combined biological system is used for the treatment of treated wastewater discharged from a pharmaceutical industrial park. This treated water is rich in NH(4)(+)-N (average in 86.4 mg/L), low in COD/NH(4)(+)-N (average in 3.4) and low in BOD(5)/COD ratio (average in 0.24) with pH varying from 7.16 to 7.78. The final effluent of the combined treatment process was stably below 100mg/L COD and 20mg/L NH(4)(+)-N, separately, with organic loading rate of 4954 kg COD/d and 92.5 kg NH(4)(+)-N/d. It is found that the BOD(5)/COD ratio could be raised from 0.24 to 0.35, and the production of total VFAs account for 9.57% of the total COD via the treatment of hydrolysis/acidification. MBBR and oxidation ditch represent 35.4% and 60.7% of NH(4)(+)-N removal, 30.2% and 61.5% of COD removal, separately, of the total treatment process. PCR-DGGE is used for microbial community analysis of MBBR and oxidation ditch. (c) 2010. Published by Elsevier Ltd. All rights reserved.
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NASA Astrophysics Data System (ADS)
Zhu, Tong; Zhu, Yufang; Fienko, Udo; Yuanhua, Xie; Kuo, Zhang
2017-01-01
A multilevel contact oxidation system was applied in a pilot-scale experiment to treat the automobile painting wastewater, which had poor biodegradability and contained high concentration of Chemical Oxygen Demand (COD). The wastewater used for this experiment study was the actual painting wastewater which had been pre-treated by the physic-chemical process, and its Biological Oxygen Demand (BOD5)/COD was less than 0.1,COD concentration was 800∼1500mg/L. The results showed that the multilevel contact oxidation system could efficiently degrade the COD of the painting wastewater. When the experimental system kept stable operation, the total removal rate of COD and suspended solid (SS) were 84% and 82.5% respectively with the Hydraulic Retention Time (HRT) of 8 hours. Meanwhile, this system had a strong ability to resist the impact of COD concentration change. The COD concentration of final treated wastewater was less than 500 mg/L, which could reach the factory discharge requirement for the paint shop. Besides, this system with simple structure was able to reduce the excess sludge production greatly, which would reduce much cost for the treatment of painting wastewater.
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Kinetics of heterotrophic biomass and storage mechanism in wetland cores measured by respirometry.
Ortigara, A R C; Foladori, P; Andreottola, G
2011-01-01
Although oxygen uptake rate has been widely used in activated sludge for measuring kinetic and stoichiometric parameters or for wastewater characterization, its application in constructed wetlands (CWs) cores has been recently proposed. The aim of this research is to estimate the kinetic and stoichiometric parameters of the heterotrophic biomass in CW cores. Respirometric tests were carried out with pure carbonaceous substrate and real wastewater. Endogenous respiration was about 2 gO2 m(-3) h(-1) (per unit of bed volume), while the kinetic parameters obtained for COD oxidation were very high (maximum rate per unit of bed volume of 10.7-26.8 gCOD m(-3) h(-1)) which indicates high biodegradation potential in fully aerobic environment. Regarding to stoichiometric parameter, the maximum growth yield, Y(H), was 0.56-0.59 mgCOD/mgCOD, while the storage yield, Y(STO), was 0.75-0.77 mgCOD/mgCOD. The storage mechanism was observed in CW cores during COD oxidation, which leads to the transformation of the external soluble substrate in internal storage products, probably as response to intermittent loads applied in CW systems, transient concentrations of readily biodegradable substrate and alternance of feast/famine periods.
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Community onsite treatment of cold strong sewage in a UASB-septic tank.
Al-Jamal, Wafa; Mahmoud, Nidal
2009-02-01
Two community onsite UASB-septic tanks namely R1 and R2 were operated under two different HRT (2 days for R1 and 4 days for R2) in parallel over a year and monitored over the cold half of the year. During the monitoring period, the sewage was characterised by a high COD(tot) of 905mg/l with a high fraction of COD(ss), viz. about 43.7%, and rather low temperature of 17.3 degrees C. The achieved removal efficiencies in R1 and R2 for COD(tot), COD(sus), COD(col), COD(dis), BOD(5) and TSS were "51%, 83%, 20%, 24%, 45% and 74%" and "54%, 87%, 10%, 28%, 49% and 78%", respectively. The difference in the removal efficiencies of those parameters in R1 and R2 is marginal and was only significant (p<0.05) for COD(sus). The sludge filling period of the reactors is expected to be 4-7 years. In view of that, the UASB-septic tank system is a robust and compact system as it can be adequately designed in Palestine at 2 days HRT.
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Meng, Jia; Li, Jiuling; Li, Jianzheng; Astals, Sergi; Nan, Jun; Deng, Kaiwen; Antwi, Philip; Xu, Pianpian
2018-07-01
This study investigated the role of COD/N ratio on the start-up and performance of an upflow microaerobic sludge reactor (UMSR) treating piggery wastewater at 0.5 mgO 2 /L. At high COD/N ratio (6.24 and 4.52), results showed that the competition for oxygen between ammonia-oxidizing bacteria, nitrite-oxidizing bacteria and heterotrophic bacteria limited the removal of nitrogen. Nitrogen removal efficiency was below 40% in both scenarios. Decreasing the influent COD/N ratio to 0.88 allowed achieving high removal efficiencies for COD (∼75%) and nitrogen (∼85%) due to the lower oxygen consumption for COD mineralization. Molecular biology techniques showed that nitrogen conversion at a COD/N ratio 0.88 was dominated by the anammox pathway and that Candidatus Brocadia sp. was the most important anammox bacteria in the reactor with a relative abundance of 58.5% among the anammox bacteria. Molecular techniques also showed that Nitrosomonas spp. was the major ammonia-oxidiser bacteria (relative abundance of 86.3%) and that denitrification via NO 3 - and NO 2 - also contributed to remove nitrogen from the system. Copyright © 2018 Elsevier Ltd. All rights reserved.
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Treatment of oilfield produced water by anaerobic process coupled with micro-electrolysis.
Li, Gang; Guo, Shuhai; Li, Fengmei
2010-01-01
Treatment of oilfield produced water was investigated using an anaerobic process coupled with micro-electrolysis (ME), focusing on changes in chemical oxygen demand (COD) and biodegradability. Results showed that COD exhibited an abnormal change in the single anaerobic system in which it increased within the first 168 hr, but then decreased to 222 mg/L after 360 hr. The biological oxygen demand (five-day) (BODs)/COD ratio of the water increased from 0.05 to 0.15. Hydrocarbons in the wastewater, such as pectin, degraded to small molecules during the hydrolytic acidification process. Comparatively, the effect of ME was also investigated. The COD underwent a slight decrease and the BOD5/COD ratio of the water improved from 0.05 to 0.17 after ME. Removal of COD was 38.3% under the idealized ME conditions (pH 6.0), using iron and active carbon (80 and 40 g/L, respectively). Coupling the anaerobic process with ME accelerated the COD removal ratio (average removal was 53.3%). Gas chromatography/mass spectrometry was used to analyze organic species conversion. This integrated system appeared to be a useful option for the treatment of water produced in oilfields.
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EPA Cape Cod 208 Plan 2015 Update Approval Letter
EPA approval letter re: certification by the Governor of MA that the Cape Cod WQM Plan Update is consistent with CWA section 208(b)(3) & accepted the Commonwealth’s reaffirmation of the existing designations of Cape Cod Towns as waste management agencies.
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43. CAPE COD AIR STATION PAVE PAWS FACILITY WITH ...
43. CAPE COD AIR STATION PAVE PAWS FACILITY - WITH BUILDING METAL SIDING BEING APPLIED ON "C" FACE (RIGHT) AND "B" FACE BEING PREPARED FOR INSTALLATION. - Cape Cod Air Station, Technical Facility-Scanner Building & Power Plant, Massachusetts Military Reservation, Sandwich, Barnstable County, MA
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Code of Federal Regulations, 2013 CFR
2013-04-01
... practice conditions of use: (1) The ingredient is used as an enzyme as defined in § 170.3(o)(9) of this... 21 Food and Drugs 3 2013-04-01 2013-04-01 false Trypsin. 184.1914 Section 184.1914 Food and Drugs... characterizing enzyme activity is that of a peptide hydrolase (EC 3.4.21.4). (b) The ingredient meets the general...
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"XANSONS for COD": a new small BOINC project in crystallography
NASA Astrophysics Data System (ADS)
Neverov, Vladislav S.; Khrapov, Nikolay P.
2018-04-01
"XANSONS for COD" (http://xansons4cod.com) is a new BOINC project aimed at creating the open-access database of simulated x-ray and neutron powder diffraction patterns for nanocrystalline phase of materials from the collection of the Crystallography Open Database (COD). The project uses original open-source software XaNSoNS to simulate diffraction patterns on CPU and GPU. This paper describes the scientific problem this project solves, the project's internal structure, its operation principles and organization of the final database.
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[Ni(cod) 2][Al(OR F) 4], a Source for Naked Nickel(I) Chemistry
DOE Office of Scientific and Technical Information (OSTI.GOV)
Schwab, Miriam M.; Himmel, Daniel; Kacprzak, Sylwia
The straightforward synthesis of the cationic, purely organometallic Ni I salt [Ni(cod) 2] +[Al(OR F) 4] - was realized through a reaction between [Ni(cod) 2] and Ag[Al(OR F) 4] (cod=1,5-cyclooctadiene). Crystal-structure analysis and EPR, XANES, and cyclic voltammetry studies confirmed the presence of a homoleptic NiI olefin complex. Weak interactions between the metal center, the ligands, and the anion provide a good starting material for further cationic NiI complexes.
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Enhanced proliferation of human hepatoma cells by PAR-2 agonists via the ERK/AP-1 pathway.
Xie, Liqun; Zheng, Yanmin; Li, Xuan; Zhao, Junyan; Chen, Xiaoyi; Chen, Li; Zhou, Jing; Hai, Ou; Li, Fei
2012-11-01
To investigate the expression and role of PAR-2 in the proliferation of the human hepatoma cell line HepG2, PAR-2 protein and mRNA expression were evaluated by immuno-histochemistry, immunofluorescence and RT-PCR analysis. The signaling pathways downstream of PAR-2 activation that lead to hepatoma cell proliferation were analyzed. The results showed that PAR-2 is expressed in human hepatoma cells and PAR-2 mRNA expression was found to be upregulated in cells treated with trypsin or SLIGKV-NH2 (P<0.001). The proliferation rate of HepG2 cells treated with trypsin or SLIGKV-NH2 was significantly increased (P<0.001). The percentage of S phase, G2/M phase and the proliferation index (PI) of HepG2 cells treated with trypsin or SLIGKV-NH2 were significantly elevated (P<0.001). The proliferative responses of HepG2 to trypsin and SLIGKV-NH2 were associated with the upregulation of c-fos and PCNA, which were significantly blocked by PD98059 pretreatment. In conclusion, our results indicate that PAR-2 enhances proliferation of human hepatoma cells possibly via the ERK/AP-1 pathway.