bluuebunny/crossref_metadata_2025 huggingface dataset:
Synthesis of geochemical techniques to identify the origin and multistage evolution of saline water in a complex geothermal system
Authors: Hao Yinlei , Pang Zhonghe , Huang Tianming , Kong Yanlong , Tian Jiao , Wang Yingchun
Published: June 2019

Elucidating brine origin and evolution is a fundamental but not easy task especially for coastal geothermal systems with possible marine constituents and multistage evolution, as subsequently physical, chemical and biological alteration processes may mask the original and early-stage signatures. Here chemical and isotopic characteristics of water (D and 18O) and dissolved constituents (13C, 14C, 11B and 87Sr/86Sr) have been utilized to investigate the source and multistage evolution of the Jimo coastal geothermal system in eastern China, with dramatic differences of geochemical characteristics observed within a 0.2 km2 area. Results show that geothermal water is derived from paleo-meteoric water and has undergone a 3-stages evolution that involves: (1) Dissolution of marine halite and potash salts in the deep reservoir; (2) Water-rock reactions especially cation exchange produces a Cl-Na-Ca type water as deep geothermal water upwells along the fault zone; (3) A minor (<0.3%) addition of fossil seawater to the shallow aquifer that produces Cl-Na type waters in the west, whereas sulfide oxidation and dissolution of aluminosilicate and carbonates in the east produces Cl-Na-Ca type waters. The methodology utilized in this study offers a means of examining other similar complex geochemical systems having a multistage evolution.
13C-depleted moissanites in subduction-related rocks: tracers of slab fluids in the Earth’s mantle and a new diamond exploration tool
Authors: Kepezhinskas Pavel , Kepezhinskas Nikita
Published: June 2019

A magnesian low-Ti shoshonite dike intruding Archean Norwegian Terrane includes numerous grains of black, blue and gray moissanite (SiC). Moissanite contains inclusions of native Si, Fe and Al, Fe-Cr carbides, Fe-Cr-Mn alloys and diamond. The range of observed δ13C values (-24.8 to -29.6‰) is similar to ophiolite-hosted SiC, lower mantle diamonds and slab-derived biogenic carbon. Norwegian moissanites may have been formed during interaction of Earth’s mantle with carbonaceous slab fluids under extremely reducing conditions as suggested by native metal and carbide inclusions. 13C-depleted moissanite can be used as a new exploration tool for sub-lithospheric diamonds in Archean to Phanerozoic accreted terranes and mobile belts.
Comparison of Geochemical characteristics of geothermal fluids from Eastern and Western Syntaxes, Himalayan belt, China
Authors: Li Yiman , Pang Zhonghe , Fan Yifan
Published: June 2019

High temperature geothermal systems are widely present along the Himalayan belt in the NW China. Strong manifestations have been observed in the Eastern syntax, where geothermal fluids are alkaline with pH almost higher than 9.0 due to boiling and waters are mainly Cl·HCO3-Na and HCO3-Na types. Large amounts of CO2 from carbonate metamorphism are thought to be one of the reasons of calcite scaling and will be a problem for geothermal energy utilization. However, in the Western syntax, there are no obvious manifestations and geothermal fluid is Cl·SO4-Na type with pH about 7.0. High content of Mg, comparing with that of the Eastern syntax, is assumed to be sourced from dissolution of Mg containing minerals and longer water-rock interactions. Calcite is under-saturated indicating no scaling problem in the utilization. Both of the geothermal fluids are recharged by the local precipitation but with different water vapor sources based on water isotope analysis. Therefore, it’s concluded that geothermal fluids from these two syntaxes have gone through totally different geochemical processes and should be carefully monitored in utilization due to the calcite scaling problem.
Change of regional water chemistry driven by group coal mining in Changzhi basin, Shanxi, China
Authors: Han Zhantao , Zhang Fawang , Yao Hongchao
Published: June 2019

Coal mining can deplete groundwater resources and discharge polluted water into the surrounding water bodies, decreasing the quality of ground and surface water. Group coal mining can drastically deteriorate regional water quality due to the overlying effects of multiple mines. As a country, China produces the most coal per year in the world. Group coal mining in China has caused a decline in the quality of many watersheds. This study introduced the regional water quality in Changchi basin, Shanxi province, which contains coal seams covering over 60% of its area and has over 10 active collieries. Coal mine discharge caused the increase of TDS, especially NO3-, in the river receiving the mining discharge. Quality of groundwater was also affected by the contaminated surface water when it was recharged by surface water. Reuse and treatment of mining discharged should be promoted in the area with high intensity of group coal mining.
Distribution of As and As(V) adsorption potential onto topsoil in the Datong Basin, northern China
Authors: Chi Zeyong , Xie Xianjun , Wang Yanxin
Published: June 2019

Datong Basin is a typical groundwater irrigation area in northern China contaminated by As. In order to reveal the distribution of As and the adsorption behavior of As(V) onto topsoil in the Datong Basin, a total of 40 topsoil core samples were collected from a 2 m core. Samples were used for chemical composition analysis and an isothermal adsorption experiment. Results show that the collected samples are not contaminated. The adsorption capacity (Stot value) of topsoil for As(V) at the study site ranged from 257 to 1027 nmol/g with an average of 505.2 ± 200.2 nmol/g. Vertically, both Stot value and topsoil components exhibit a vertical zoning with a positive correlation between the Stot value and K2O, CaO, MgO, Al2O3, Fe2O3 contents while Na2O and SiO2 are inversely correlated. Fe2O3 content shows a strong positive correlation with As content while Al2O3 does not. Moreover, the correlation between Stot value and Al2O3 is much better than Fe2O3. These results indicate that Fe minerals are the main reservoirs of As in the natural topsoil at the study site. Furthermore, both Fe minerals and clay minerals can adsorb As(V), but clay minerals have a greater storage potential. This result indicate that a combined effect of Fe minerals and clay minerals for As(V) accumulation onto topsoil should be considered during geochemical modeling and further agricultural management.
Water defluoridation by carbonate calcined and uncalcined Layered Double Hydroxides (LDHs)
Authors: Dore Elisabetta , Frau Franco
Published: June 2019

Carbonate Layered Double Hydroxides (LDHs) with different cationic composition and M2+/M3+ molar ratio were tested, both untreated and calcined, for water defluoridation. The defluoridation capacity of calcined LDHs, produced by intercalation during the reconstruction of lamellar structure, is more effective than the anion exchange of untreated LDHs. Among the calcined LDHs tested, that with 3Mg/(0.5Al+0.5Fe) molar ratio yields a maximum fluoride sorption capacity up to 92.3 mg/g, in spite of the high pH value (up to 12) of the experiments. After four cycles of regeneration by calcination and subsequent water defluoridation, this LDH composition maintains more than 80% of its fluoride removal capacity.
Factors influencing the distribution of arsenic, fluorine and iodine in shallow groundwater in the oasis zone in the southern margin of the Tarim Basin in Xinjiang, P. R. China
Authors: Fan Wei , Zhou Jinlong , Zhou Yinzhu , Zeng Yanyan , Chen Yunfei
Published: June 2019

Test results on 302 shallow groundwater samples in an oasis zone in the southern margin of the Tarim Basin in Xinjiang were collected and analysed for a large number of chemical parameters and subjected to mathematical statistics methods. Results show that groundwater with high arsenic, fluorine and iodine concentrations were mainly distributed in the Minfeng County in the central part of the study area. Shallow groundwater in the southern margin of the entire Tarim Basin generally had high fluorine concentrations. The distribution of arsenic and fluorine in groundwater were similar. Intensive evaporation under the arid climatatic condition was an important factor for the enrichment of arsenic, fluorine and iodine in groundwater.
Hydrochemistry of pit lakes in the Portuguese sector of the Iberian Pyrite Belt
Authors: Gomes Patrícia , Valente Teresa , Cordeiro Mayara , Moreno Filipa
Published: June 2019

The Iberian Pyrite Belt (IPB) is a world-class metallogenic province with volcanogenic massive sulphide ore deposits. Most ore exploitation occurred since pre-Roman time, creating extensive galleries, wells, waste-dumps and pit lakes. These last structures are a concern for their potential environmental impact because they accumulate large volumes of mine water affected acid mine drainage. The present work classifies the pit lakes based on the surface water hydrochemistry. Using the Ficklin diagram for classification, pit lake waters vary from acid, high-metal to high-acid, extreme-metal and exhibit similarities with other pit lakes from the Spanish sector of the IPB.
Health Risk Assessment of Heavy Metals in Surface Water of the Lhasa River, China
Authors: He Liu , Gao Bai , Qin Huanhuan , Liu Junping , Ding Xiaoyan , Sanjay Dhawan , Dong Andre
Published: June 2019

This study was performed in Lhasa River Watershed to explore the contents of heavy metals in river water and assess the health risks. The test results show that the contents of arsenic (As), chromium (Cr) and uranium (U) are relatively low, reaching the Class III of the "Environmental quality standards for surface water (GB3838-2002)". Health risk assessment demonstrates that the average total risk value is 1.79×10-5 yr-1, which is lower than the maximum acceptable level of 5.05×10-5 yr-1 recommended by USEPA. Moreover, the average value of radionuclide risk (U) is calculated to be 9.47×10-9 yr-1, which is five orders of magnitude lower than the standard value of 5.0×10-4 yr-1. Therefore, the river water in the study area causes no significant health risk to people.
Redox hydrogeochemistry of organic rich floodplain exemplified by Ammer river
Authors: Kortunov Evgenii , Lu Chuanhe , Amos Richard , Grathwohl Peter
Published: June 2019

Diffusive groundwater pollution caused by agricultural and atmospheric inputs is a pressing issue in environmental management worldwide. Various researchers have studied nitrate contamination since the substantial increase of nitrogen fertilization in agriculture starting in the second half of the 20th century. This study addresses large scale reactive solute transport in typical landscapes and aquifers exemplified by geological analogues of southwestern Germany.. Fate of nitrate and other solutes (e.g. agricultural nitrate, ammonium, natural sulfate and dissolved organic carbon) was studied in a typical small river floodplain. Reactive transport model of Ammer river floodplain shows that agriculture nitrate is reduced rapidly in the Ammer floodplain sediments. However, there is a potential for geogenic production of ammonium in sediment layers high in organic carbon and peat, which might be a major source of nitrate in the drains. Part of the nitrate in drains and creeks in the Ammer valley thus could be of geogenic origin. Such findings are relevant for regional land and water quality management.
Role of unsaturated soil above a heavily contaminated aquifer in the natural attenuation of arsenic
Authors: Martínez-Villegas Nadia , Gómez Andrea del Pilar , Zamora-Morales Armando , Hernández Martínez Jejanny Lucero , Meza-Figueroa Diana , SenGupta Bhaskar
Published: June 2019

Large attenuation of arsenic is observed in a shallow aquifer in central Mexico, where the concentration decreases from 91.5 to 11.3 mg/L, over 1.3 km horizontal distance. To investigate possible mechanisms of attenuation of this pollutant, we dug a pit between the surface and the saturated zone. We then described the soil profile and determined arsenic and iron concentrations in soil samples as a function of depth. Next, we determined particle size distribution, bulk density, particle density, soil moisture and porosity of the soil material. We also analyzed arsenic concentration in the groundwater. The 2.2 m deep profile intersected 4 soil horizons (A, AB, B1 and B2). We found arsenic accumulation in the B2 horizon, directly overlying the saturated zone. These accumulations coincide with the thickness of the capillary fringe, revealing that arsenic is drawn up in the soil profile by capillary rise of arsenic-contaminated groundwater. Furthermore, arsenic accumulation showed a direct relationship with iron, likely due to arsenic adsorption on iron oxides. Results from this study contribute to understanding a capillary-driven natural attenuation mechanism that removes contaminants from groundwater by sinking them in the capillary fringe.
Residues of sulfonamides and their acetylated metabolites in the environment of aquaculture
Authors: Tong Lei , Qin Liting , Guan Chuan , Wilson Mesmire E.
Published: June 2019

This study documents the occurrence of sulfonamides (SA) and their acetylated metabolite compounds in an aquaculture environment in China. Five parent compounds of SAs and four metabolites were detected in fish water, surface water, and groundwater. The most relevant compounds in terms of frequency of detection and maximum concentrations detected were sulfamethoxazole and sulfachinoxalin in fish water and surface water, with average concentrations of 541 ng/L and 274 ng/L, respectively. The acetylated metabolites were detected at higher concentrations than their parent compounds in groundwater, although they were much lower than in the surface environment. The recent appearance of acetylated metabolites of SAs in groundwater samples demonstrates the environmental health concern and the need of further study.
Hydrochemistry and Environmental Isotope Characteristics of Groundwater in the Kashgar Delta Area, Xinjiang, China
Authors: Wei Xing , Zhou Jinlong , Zhou Yinzhu , Zeng Yanyan , Chen Yunfei
Published: June 2019

The hydrochemistry and isotope characteristics of groundwater of different geomorphological types in the Kashgar Delta in Xinjiang, China were studied using isotopic analyses. Results showed that groundwater in the study area was mainly recharged by precipitation, and that groundwater ions were mainly affected by weathering-leaching of rock salt and evaporite as well as cation exchange and adsorption. The hydrochemical type of phreatic water in the alluvial-proluvial gravel plain on the piedmont slope was mainly SO4·Cl-Na·Ca type, with an average age of 28a. The phreatic water in the fluvial plain had shallow buried depth and intense evaporation. The hydrochemical type of phreatic water varied from SO4·Cl-Na·Mg type to Cl-Na type, with an average age of 99a. Along the groundwater flow path eastward, confined water in the fluvial plain experienced gradually more intense cation exchange and stronger adsorption. The major hydrochemical type of which was the SO4·Cl-Na type, with an average age of 30405a.
Prediction of drainage water chemistry by experimental data (on the example of the Nakhodka porphyry copper prospect, Western Chukotka, Russia)
Authors: Yablonskaya Daria , Lubkova Tatiana , Shestakova Tatiana , Strilchuk Natalia
Published: June 2019

Prediction of drainage water chemistry is a critical part of mine planning; particularly water and mine waste management. This study investigates a potential composition of drainage water for various storage times of sulphide-bearing geological materials by experimental data. The paper presents the results of Short-Term Leach tests and Humidity Cell tests for geological materials of the Nakhodka porphyry copper ore field, the Baimka ore trend (Western Chukotka, Russia). The results obtained can be used to forecast of wastewater composition as well at the initial stage of storage of sulphide-bearing geological materials as over the long-term.
Reactive transport calculations to evaluate sulphide fluxes in the near-field of a SF/HLW repository
Authors: Pękala Marek , Wersin Paul , Cloet Veerle , Diomidis Nikitas
Published: June 2019

Radioactive waste is planned to be disposed in a deep geological repository in the Opalinus Clay (OPA) rock formation in Switzerland. Cu coating of the steel disposal canister is considered as potential a measure to ensure complete waste containment of spent nuclear fuel (SF) and vitrified high-level waste (HLW) or a period of 100,000 years. Sulphide is a potential corroding agent to Cu under reducing redox conditions. Background dissolved sulphide concentrations in pristine OPA are low, likely controlled by equilibrium with pyrite. At such concentrations, sulphide-assisted corrosion of Cu would be negligible. However, the possibility exists that sulphate reducing bacteria (SRB) might thrive at discrete locations of the repository’s near-field. The activity of SRB might then lead to significantly higher dissolved sulphide concentrations. The objective of this work is to employ reactive transport calculations to evaluate sulphide fluxes in the near-field of the SF/HLW repository in the OPA. Cu canister corrosion due to sulphide fluxes is also simplistically evaluated.
Mutual impact of reservoir sands and acidic liquid radioactive waste
Authors: Vlasova Irina , Romanchuk Anna , Gusev Ilya , Volkova Anna , Zakharova Elena , Rzhevskaia Alexandra , Kalmykov Stepan
Published: June 2019

The work is devoted to the study of the behavior of long-lived alpha-emitting radionuclides (Pu, Am, U, Np) under the conditions of injection of acidic liquid radioactive waste into a sandy rock reservoir bed. Different mineral phases of initial reservoir sands, secondary minerals formed when interacting with waste solutions, as well as phases precipitating from the waste solution are considered in terms of their retention properties towards to actinides.
Sources of atmospheric pollution in the rust belt, Ohio, USA
Authors: Kousehlar Masoomeh , Widom Elisabeth , Kuentz David
Published: June 2019

Industrial regions in the rust belt are sites of significant legacy atmospheric pollution, and despite recent regulations, continue to be sites of ongoing atmospheric emissions of toxic metals. Source attribution in such areas is complicated by the presence of multiple potential point sources, including steel production plants and coal fired power plants. This study focuses on a region in Ohio, USA, in the vicinity of a steel manufacturing plant and coke production facility. Scanning electron microscopy of atmospheric particulate matter, combined with major and trace element and Pb and Os isotopic analysis of dust, lichen and tree bark, have been employed to identify the primary contributors to the atmospheric metal load. Our results suggest that steel production, coal burning, and traffic emissions are all significant sources of metals to the local environment.
Isotopic fractionation associated to nitrate attenuation by ferrous iron containing minerals
Authors: Margalef-Marti Rosanna , Carrey Raul , Soler Albert , Otero Neus
Published: June 2019

Biotic and abiotic laboratory experiments of nitrate and nitrite reduction by Fe-containing minerals were performed and the isotopic fractionation of the different reactions was calculated in order to determine whether it is possible to distinguish biotic and abiotic reactions involving N compounds. Results of biotic experiments showed nitrate reduction up to 96 % with transient NO2- accumulation and no significant N2O production. No significant nitrate attenuation was observed in abiotic nitrate reduction experiments. Abiotic experiments of nitrite reduction showed a rapid decrease in nitrite concentrations in those experiments with added Fe2+ coupled with a significant N2O production. Preliminary results of the N and O isotopic fractionation of the biotic experiments of nitrate reduction show differences in the ε15NNO3 and ε18ONO3 when different minerals were added. The abiotic experiments of nitrite reduction contrarily, showed similar ε15NNO2 in all the experiments.
Chemical and isotopic characterization of nitrate retained and leached from soil after manure fertilization-by lysimeter experiments
Authors: Otero Neus , Llovet Alba , Carrey Raúl , Ribas Angela , Domene Xavier , Mattana Stefania , Chin-Pampillo Juan , Alcañiz Josep Mᵃ , Soler Albert
Published: June 2019

Increase of soil fertilization produces an increase of N exported to the hydrosphere. The amount of nitrate that reaches the aquifers is controlled by processes affecting N-species within the soils. The most relevant processes are nitrification, denitrification, assimilation, mineralization, and immobilization. This work studies the fate of N compounds in soil after manure application in a lysimeter study. To this end the isotopic composition of N and O of dissolved nitrate (δ15N-NO3- and δ18O-NO3-) was studied coupled with the evolution of N-compounds retained and leached from the soil. Results showed an increase in the δ15N-NO3- of the leached nitrate towards values similar to the δ15N-NH4+ from the applied manure. The highest δ15N-NO3- values were measured after 100 days of manure application, and thereafter, values decreased progressively towards the initial δ15N-NO3- of the soil before manure application.
Isotope ratio mass spectrometry application for environmental investigations
Authors: Simonova Galina , Kalashnikova Daria
Published: June 2019

Stable isotope ratio mass spectrometry (IRMS) is a useful technique with a broad range of applications. Stable isotopes are important indicators of environmental changes (including climate), and can be used to explain the functioning of biogeochemical cycles. Examples of IRMS aplications in the fields of environmental science, paleoecology/climatology, and analysis of atmospheric pollution are presented in this paper. These studies were carried out using an isotope mass spectrometry complex (Tomsk Regional Сore Facilities of Tomsk Scientific Center of Siberian Branch of the Russian Academy of Sciences (SB RAS)) at the Institute of Monitoring of Climatic and Ecological Systems of SB RAS.
Hydrogen, oxygen and carbon isotopes of carbonic waters in the Daurian hydromineral oblast’ (Transbaikalia, Russia)
Authors: Zamana Leonid V.
Published: June 2019

The results of isotopic analyzes of about 40 samples of carbonic waters are presented, dependencies of isotopic relationships between themselves and with bicarbonate were analyzed, the causes of these dependencies are explained, conclusions on the genesis of CO2 are made.
Selbst ist das Kind – Du+Ich-Konzept: Kinder übernehmen Eigenverantwortung
Authors: Hiebl Sarah , Junkers Anja
Published: June 2019

Lena hat mal wieder ihren Turnbeutel vergessen. Soll ihre Mutter ihn ihr hinterhertragen und zur Schule bringen? Wie Ergotherapeuten mithilfe des Du+Ich-Konzeptes Eltern dabei unterstützen können, ihren Kindern mehr Eigenverantwortung zu überlassen, zeigen Anja Junkers und Sara Hiebl.
Basis für die therapeutische Forschung am Menschen – Die Helsinki-Deklaration
Authors: Zalpour Christoff
Published: June 2019

Die Akademisierung der Ergo- und Physiotherapie und die damit verbundenen Forschungsvorhaben führen zu neuen berufs- und forschungsethischen Pflichten. Forschende Therapeuten sollten daher unbedingt die Deklaration von Helsinki kennen, die die ethischen Grundsätze für die Forschung am Menschen regelt.
CRYOPRECIPITATE COMPOSITION AS A FUNCTION OF PLASMA SOFTENING RATE
Authors: Winkelman L , Pinnell M
Published: June 2019

Plasma for fractionation is commonly stored at <−30°. Some warming is necessary to soften the frozen plasma in order that it may be crushed prior to thawing to 0° for cryo-precipitation. In an investigation of softening conditions, small-scale experiments were performed using frozen plasma warmed to various temperatures (−15° to −2.5°) and held for varying lengths of time (0-40h). Cryoprecipitate was prepared and recoveries of fVIII:C, fibrinogen and fibronectin assessed. Yields of fVIII:C and fibronectin were independent of softening conditions, but the yield of fibrinogen was directly related to the temperature profile of the plasma. If held for periods >1h at >−10°, the weight of fibrinogen in the cryoprecipitate increased. In the conditions used in these experiments (which were chosen to reflect those which might be possible on a large scale), fibrinogen recovery ranged from 600-1700 mg/kg plasma. These findings have important implications for the preparation of fVIII concentrate from cryoprecipitate where subsequent purification may be significantly easier and final yields improved if the initial burden of fibrinogen is reduced.
POSSIBLE ASSOCIATION OF HLA AND Gm WITH THE ALLOIMMUNE RESPONSE TO FVIII
Authors: Reisner H M , Reisner E A , Kostyu D D , Lubahn B C , McMillan C , White G C
Published: June 2019

Between 5 and 15% of individuals with severe hemophilia A are at risk of developing inhibitors (alloantibodies) to FVIII. Genetic factors are important in determining risk, but the nature of these factors is poorly defined. The human immune response to a wide variety of antigens has been associated with the HLA and/or Gm loci. Hence, we have investigated polymorphisms at these two loci in hemophilia A patients with and without inhibitors.

DATA SET: To date 127 hemophiliacs have been Gm or HLA typed. Forty-eight are inhibitor positive (I+) based on positive FVIII neutralization assays. This represents about 70% of all I+ hemophiliacs seen at UNC. To prevent familial bias, one member of each of 14 pairs of close relatives was randomly removed from the Data Set without regard to inhibitor status (Data Set 1). Twenty non-white individuals were also removed to constitute Data Set 2. GM TYPING: Samples were typed for Gm antigens 1, 3 and 5. Phenotype frequencies in Data Sets 1 and 2 did not deviate from expected values. I+ hemophiliacs showed an excess of Gm 1 in both Data Sets which was of possible significance (p = .13 and .21 respectively by chi square). Reanalysis of Data Set 2 to include only I- individuals without evidence of either circulating VIII:C or VIII:CAg (N=58) yields a p of .12. HLA TYPING: Analysis on 77 individuals in Data Set 2 (21 I+, 56 I-) has been done. In preliminary typing of HLA A, B, C, DR and DQ no significant differences in antigen frequency were found between the I+ and I- groups. INTERACTION BETWEEN HLA AND GM: A significant excess of Gm 1 I+ individuals was found among all HLA-A2 positive hemophiliacs (N=45, p=.034 by Fisher’s exact test. This was not significant after correction for multiple comparisons). This suggestion of an interaction between HLA-A2 and Gm 1 in determining alloreactivity to FVIII will require further prospective evaluation for confirmation.
LONG TERM, FREQUENT PLASMA EXCHANGE DONATION OF CRYOPRECIPITATE
Authors: McLeod B , Sassetti R , Cole E , Scott P
Published: June 2019

In plasma exchange donation (PED), several liters of fresh plasma are removed fran a donor with a pheresis instrument as a source of cryoprecipitate, and replaced with autologous cryoprecipitate-supernatant from the previous donation. Repetitive PED can produce large quantities of factor VIII from individual donors over time, with a favorable impact on donor exposure for factor VIII recipients. To clarify the implications for donor safety, we report our experience with several donors who have undergone multiple PEDs. Detailed observations are presented for one donor who has undergone PED 101 times between 5/83 and 1/87, and has provided all the factor VIII needed by his son (now age 14) with severe hemophilia A during this period. Exchange volume was gradually increased while donation frequency was gradually decreased. There were 23 exchanges of 2 L, 52 of 2.5 L, and 26 of 3 L for a total of 254 L plasma exchanged. Desmopressin (20 meg tV) was given before 45 more recent donations to augment factor VIII yield. A total of 343,274 IU factor VIII have been collected; the mean (±SD) yield from a 3 L, desmopressin- stimulated PED is 5598 ± 899 IU. The donor has remained in good health; he has noted no adverse effects fran any PED, and none have been found in laboratory monitoring. Prior to the 100th donation the following were within normal limits: CBC,platelet count, urinalysis, SMA-18, protein electrophoresis, IgG, IgA, IgM, hemolytic complement, C3, C4, fibronectin, prothrombin time, partial thromboplastin time, thranbin time, factor VIII:C (140%), factor VIII:Ag (134%), von Willebrand factor (86%) and fibrinogen (215 mg/dL). In another family, the father has donated 40 times since 1981 and the paternal grandmother has donated 31 times since 1984 with no untoward effects detected in clinical or laboratory monitoring. They have supported two moderately affected patients now ages 7 and 9. Extensive experience with these donors suggests that repeated PED is safe, and that a highly motivated donor can sometimes provide single donor support, even for a severe hemophiliac.
LIFE-EXPECTANCY OF DUTCH HEMOPHILIACS, 1972-1985
Authors: Rosendaal F , Smit C , Varekamp I , Bröcker-Vriends A , Suurmeijer T , Briít E
Published: June 2019

The 1ife-expectancy of hemophiliacs has risen during the last decades from (for severe hemophilia) a mere 16 yrs in 1940, 23 yrs in 1964 to almost normal after coagulation factor preparations became available. Still, many hemophiliacs encounter ratings or refusals when applying for life-insurance.

We carried out three mail surveys (1972, 1978, 1985) among Dutch hemophiliacs to establish excess mortality caused by hemophilia. Follow-up for non-respondents was carried out with help from the municipal authorities, while information on the deceased was obtained from the hemophilia treatment centers. To compare mortality with the general population, we constructed Kaplan-Meier reference curves for population groups with the same age and sex distribution, using national death tables. Pending completion of the follow-up for the 1972-1985 interval, this abstract supplies the results for the 1978-1985 period.

The 1978 cohort included 578 individuals, with a mean age of 26 yrs (general male population: 33 yrs). None were lost to follow-up, 22 (3.8%) had died during the 7i yrs of observation. In the reference group 1.9% mortality would have been expected. So, overall relative mortality is increased twofold compared to the general male population (95% confidence interval 1.3-3.0). Patients with an inhibitor excluded, excess mortality did not differ much for severe, moderate and mild hemophilia (relative mortality: 2.3, 1.8, 1.5). In 11 cases hemorrhage was the cause of death, but in 6 cases this was associated with an underlying lethal disorder. In 9 cases the cause of death was not related to hemophilia and in 2 it remained unknown. An inhibitor was present in 4 (18%) of these 22 patients, but in only 4% of the total group of 578. In the 1972 survey 43% of the respondents who had applied for a life-insurance (n=199) reported a rating or a refusal, in 1978 47% of 227 and in 1985 53% of 676.

The excess mortality caused by hemophilia is small and similar to that due to smoking cigarettes. Moreover, the excess may be caused in part by a carry-over effect from the time before modern treatment was introduced.
ISOLATION AND CHARACTERIZATION OF ACTIVATED FACTOR VIII
Authors: Van de pol H , Bihorean N , Sauger A , Ganne V
Published: June 2019

Recently a discussion has started in the scientific press about the molecular nature of activated human antihemophilic Factor. One group suggests that the generation of the 90KD/80KD complex and the subsequent cleavage on the light chain is sufficient to cause activation, another group insists that full activation is only achieved when also the heavy chain of Factor VIII is cleaved by thrombin. Most of the results reported, are obtained with very high pure preparations of Factor VIII. In order to test the significance of the reported molecular nature of Factor VIII activation, we activated the human antihemophilic Factor in commercial concentrates and analysed the activated plasma concentrates using biological assays or immunoprecipitation-techniques and characterized immunopurified activated Factor VIII by FPLC and PAGE analytical techniques. Using those methods we noticed that the activation of Factor VIII in plasma concentrates starts with a cleavage by thrombin of the light chain resulting in a three fold activation. The generated 70KD product is complexed with the heavy chain and its higher molecular weight parents (90KD, 110KD, 120KD, 130KD, 145DK, 155KD, 180KD and 210KD), an can be isolatedas those active complexes using immunopurification or Fast Pressure Liquid Chromatography. The isolated 70KD containing complexes can be activated another three fold without the expected proteolytic cleavage of the heavy chain. The generation of the 45KD and 55KD cleavage products of the heavy chain are first seen when an excess of thrombin is used for activation, and corresponds likely with the first inactivation of the molecule. Furthermore we report a protective role of region B against activation by thrombin: when partially activated Factor VIII was analysed by Fast Pressure Liquid Chromatography, we noticed a significant higher percentage uncleaved 80KD fragment in fractions containing region B.
EPITOPE MAPPING OF HUMAN COAGULATION FACTOR VIII WITH IN VITRO SYNTHESIZED FRAGMENTS OF THE PROTEIN
Authors: Leyte A , Evers R F , Verbeet M Ph
Published: June 2019

We have used recombinant DNA techniques to map the epitopes of the coagulation Factor VIII for several monoclonal antibodies, raised against this protein. For this purpose, we cloned full- length- and partial Factor VIII cDNA sequences in the vector pSP64. Corresponding RNA fragments were synthesized in vitro with SP6 RNA polymerase and translated in a rabbit reticulocyte lysate system. The resulting Factor VII. I polypeptide fragments were immunoprecipitated. We have located the binding sites of a panel of monoclonal anti-Factor VIII antibodies. Two examples are shown in the figure below. The epitopes for anti-Factor VIII CLB-9 and CLB-65 have been confined to areas of 29 (711-740) and 142 (1635-1777) amino acids, respectively. The results of these studies will be useful in determining structure-function relationships of Factor VIII
TWO FVIII GENE LESIONS DETECTED IN SEVERE AND MODERATE HAEMOPHILIA A
Authors: Bertagnolo V , Volinia S , Legnani C , Rodorigo G , De Rosa V De , Bernardi F
Published: June 2019

DNAs from 15 haemophilia A patients from different families have been hybridized to FVIII cDNA probes for the exons 14-26.

In a severely affected patient (FVIII:C 2 %) the TaqI site of exon 24 is absent originating an abnormal band of 4.2 Kb. A C toT transition in the CG dinucleotide of the TaqI site (TCGA) is the probable gene mutation. Since the transition in the sense strand should originate an additional Hind III site, which is not detected in our patient, we infer that the mutation occurred in the antisensestrand causing an aminoacid change (CGA →CAA, Arg → Gin). This isin accordance with the low activity of FVIII and with the absence of inhibitor. Infact Gitschier et Al reported in a patient with ahigh titre of anti-FVIII antibody and with <1% FVIII activity a C → T transition in the coding strand, originating a nonsense codon in the TaqI site of exon 24.

In the Hindlll pattern from a moderately affected patient (FVIII:C 4%) the fragment containing the exon 18 is 2.5 Kb in size (normal 2.6 Kb). Since the patterns with other restriction enzymes are indistinguishable from normal a small mutation originating a new Hind III site is likely. Both altered patterns have been detected in the patients' mothers.

Work supported by Ricerca Sanitaria Finalizzata Regione Emilia Romagna
VONWILLEBRAND FRAGMENTS IN CoMMERCIAL FVIII CONCENTRATES
Authors: Dayal V K , Hsieh A , Velasquez O , Arkel Y S
Published: June 2019

We have previously reported sane of the biochemical character-istics of varioias aomnercial FVIII concentrates. Two-dimensional gel analysis of these preparations indicated the presence of pep-tides in the 30-50 KDa basic region. We also noted a relative decrease of high molecular weight vonWillebrand factor (vWF) multimers on SDS-agarose gels. However, the proportion of low molecular weight vWF fragments seems to be increased as judged by the following studies. After reduction and electrophoresis in SDS- PAGE gels, the proteins in concentrates were electroblotted on to the nitrocellulose membranes. The vWF fragments were detected using sandwich of rabbit anti-human vWF antibody, followed by goat anti-rabbit antibody-horseradish peroxidase conjugate incubation and color development. These studies indicated the ore- senceof several vWF peptides in the 30-50 KDa region of one of the FVIII concentrates. These fragments were not detectable in normal plasma analyzed under identical conditions. In order to investigate any clinical significance of the 30-50 KDa peotides, the following experiments were carried out. A factor VIII concentrate preparation was subjected to electrophoresis in SDS-PAGE gels. After electrobiotting, the proteins nitrocellulose membrane were blocked and then reacted with hemoohilia a plasma. Subsequent reaction with 125I Protein A and autoradiograohy revealed the presence of a peptide in approximately 35 KDa region. A protein band of similar molecular weight also reacted with anti-vWF antibody in a separate identical run. Although the patient plasma did contain an antibody to a 35 KDa peotide, it is not certain whether this peptide is identical to the 35 KDa vWF fragment. The commercial FVIII concentrates that contain 30-59 KDa peptides also demonstrate a relative decrease in high molecular weight vWF mul timers. The possible role of proteolysis in the generation of vWF fragments is under study. The results reported here provide evidence that some of the conmercial FVIII concentrates contain peptides in 30-50 KDa region which are not detectable in normal plasma. Furthermore, sane of these low molecular weight peptides are attributable to vWF fragments.
THE ISOLATION OF COAGULATION FACTOR VIII FROM HUMAN BLOOD PLASMA BY AFFINITY CHROMATOGRAPHY
Authors: Riethorst W , te Booy M W P M , Beugeling T , Bantjes A , Over J , van Aken W G
Published: June 2019

The need for high quality concentrates of coagulation factor VIII (FVIII:C) for treatment of haemophilia A is increasing. As the purity of FVIII:C obtained with existing large scale methods is poor and yields are low, another method for the isolation of FVIII is being developed primarily to avoid losses incurred during cryoprecipitation.

Affinity gels were prepared by derivatizing Sepharose CL 4B with different positively charged ligand-spacer combinations. The adsorption of FVIII as well as the von Willebrand factor (VWF) from human blood plasma onto these gels was measured by a one-stage assay for FVIII:C, and enzyme immuno assays (ELISA) for FVIII:CAG and VWF:AG using monoclonal antibodies. The influences of pH, conductivity, ligand density, geltplasma ratio, and length and composition of the spacer as well as the adsorption kinetics were studied to obtain information about the types of interactions responsible for bonding of FVIII to the gels. A combination of at least electrostatic and hydrophobic interactions was concluded to play a role in most cases.

At optimal conditions more than 90 % of FVIII could be adsorbed batch-wise from plasma at room temperature in less than one hour with a gel:plasma ratio of 1:20 (i.e. 2.5 g dried gel/1 plasma). In different runs 65-75 % of the FVIII:C applied was recovered by column-wise elution with a salt gradient. The eluate contained less than 0.34 % of the protein applied, which implies that FVIII was purified 190 times. Using fresh-frozen plasma (0.8 IU FVIII/ml) as a starting material for this one-step procedure the final specific activity was 2.3 IU/mg, which is significantly better than that obtained for FVIII isolated by cryoprecipitation. Furthermore, the F. VIII:C to VWF ratio in the eluate was approximately 1:1. The isolated FVIII:C was stable at room temperature and the supernatant plasma appears suitable for further fractionation. It is concluded that this method is worth scaling up and its use for purification of FVIII from other sources is anticipated.
IMMUNOPURIFICATI0N OF FVIII/vWF FROM PLASMA
Authors: MEJAN O , DELEZAY M , FERT V , CHEBALLAH R , BOURGOIS A
Published: June 2019

At present, antihemophilic factor concentrates available for hemophilia A therapy raise some problems :

- weak recovery of FVIIIC (20 to 25%)

- specific activity generaly lower than 5IU/mg

- loss of a large part of the von Willebrand factor, carrier of the FVIIIC

- necessity to inactivate viral contaminants by treatments which induce a decrease in FVIII recovery. To solve these difficulties, an immunopurification process has been developed directly from plasma. A mouse monoclonal antibody (MAb) toward vWF, able to bind the FVIII/vWF complex under physiologic conditions and to release it in a suitable elution buffer preserving the FVIIIC activity has been selected. MAb coupling chemistry on gel beads has been optimised. The immunopurification process developed allows to treat 12 to 19 Liters of plasma with one gram of antibody in each cycle (i.e. 6h.). Immunoadsorbant can be used during 10 cycles with standard quality plasma and with an average recovery of 50% for FVIIIC. This process warrants an important elimination of contaminant virus (104 for HBs) an a release of the mouse MAb in purified FVIII lower than 10-9 M . These properties could justify an industrial application of this immunopurification process to produce therapeutic hemophilia A fractions.
LEWIS BLOOD TYPE HAS AN EFFECT ON THE PLASMA CONCENTRATION OF FACTOR VIII
Authors: Ørstavik K H , Kornstad L , Reisner H M
Published: June 2019

The plasma concentration of factor VIII is influenced by the ABO blood group. Individuals with blood group 0 have a lower concentration of both factor VIII coagulant activity, factor VIII coagulant antigen (VIIICAg) and factor VIII related antigen (VIHRAg) than individuals with group A, B and AB. Thirty percent of the genetic variance of VIIIRAg concentration is due to the ABO blood group. The Lewis substances Lea and Leb are closely related to the A, B and H substances. We therefore examined the effect of the presence of these antigens on the plasma concentration of VIIICAg and VIIIRAg. The material was 74 monozygotic and 84 dizygotic twin pairs and 58 blood donors with ABO blood group 0. VIIICAg was determined by a radioimmunoassay and VIIIRAg was determined by an electroimmunoassay. A higher mean concentration °f VIIICAg (147%) and VIIIRAg (81%) was found in individuals with the Le antigen on their red cell surface compared to the mean concentration of VIIICAg (101%) and VIIIRAg (66%) in individuals who lacked this antigen. The difference was found in individuals with ABO blood group 0 only. Individuals with red cell Lea antigen are non-secretors and individuals who lack this antigen but have the Leb antigen are secretors of the A, B and H substances. The lowest factor VIII concentration was found in group 0 secretors. The effect of the Lewis phenotype on factor VIII concentration is therefore most probably due to an effect of the secretor locus. This finding may have practical implications for the diagnosis of carriers of hemophilia A. it has been shown that information on the ABO blood group improves the discrimination between carriers and normals. We found that the effect of ABO blood group on the total variance of VIIIRAg was higher in secretors (21%) than in non-secretors (8%). Since the frequency of secretors varies widely, it is possible that the importance of the ABO locus in carrier detection is different in different populations. Lewis blood typing in materials of carriers and normals are necessary to determine the effect of the Lewis phenotype in carrier detection.
CLOTTING FACTOR DEPENDENCY OF PROTHROMBINASE ACTIVITY IN DICOUMAROL PLASMA. THE IMPORTANCE OF A FACTOR IX DEPENDENT PATHWAY IN EXTRINSIC COAGULATION
Authors: Xi Ma , Béguin S , Hemker H C , Devilée P P
Published: June 2019

We determined the generation of prothrombinase activity in plasma using a mathematical analysis of the thrombin generation curve (H. C. Hemker, G. M. Willems, S. Béguin. Thromb. Haemostas. 56, 9-17, 1986).

Addition of the purified factor VII, IX or X to plasma from deeply anticoagulated patients (<15% level >10%) did not influence the rate and amount of prothrombinase formed. Only the amount of prothrombin in the starting plasma determined the course of thrombin generation. Adding increasing amounts of purified human clotting factor preparations to deficient plasmas showed that the treshold concentration under which factor VII, and IX start to have an effect on prothrombinase activity are 5% or lower. For factor X it is lower than 10%.

From this it can be concluded that only the changes in prothrombin level must be held responsible for the anti thrombotic effect of oral anticoagulation. These conclusions are not modified if different types and concentrations of thromboplastin are used.

We were able to show that at dilutions of human brain thromboplastin higher than 1:50, a factor IX and factor VIII dependent pathway plays an increasingly important role. This directly demonstrates the Josso pathway. The concentration of factor IX necessary for full activity is again <5%.

We conclude that the antithrombotic effect of oral anticoagulant treatment, if it is mediated via the coagulation system, works via modification of the prothrombin level only.
ASIALO VON WILLEBRAND FACTOR ENHANCES PLATELET ADHESION TO VASCULAR SUBENDOTHELIUM
Authors: Ordinas A , Bastida E , Garrido M , Monteagudo J , de Marco L , Castillo R
Published: June 2019

Native Von Willebrand factor (NvWF) binds to platelets activated by thrombin, ADP or ristocetin, and also supports the adhesion of platelets to subendothelium at high shear rates. In contrast, asialo von Willebrand factor (AvWF) induces platelet aggregation in absence of platelet activators. We investigated the role of AvWF in supporting the adhesion of platelets to rabbit vessel subendothelium under flow conditions at a shear rate of 2000 sec-1 for 5 min using the Baumgartner perfusion system. We also studied the effects of blockage of platelet GPIb or GPIIb/IIIa on platelet adhesion using monoclonal antibodies (Mabs),and we measured the rate of binding of 111I-labeled NvWF and AvWF to subendothelium. Perfusates consisted of washed platelts and red cells resuspended in a 4% human albumin solution to which increasing concentrations of NvWF or AvWF had been added. Platelets interacting with the perfused vessels were evaluated morphometrically using a computerized system. At a concentration of 1.2 /ig/ml the percentage of total coverage surface was 21.3 ± 4.8% and 40.0±14.6%, for NvWF and AvWF, respectively (p<0.01). Addition of either Mab against GPIb (LJlbl) or against GPIIb/IIIa (CP8) to the perfusates, reduced platelet deposition (p <0.01). The rates of binding of 111I-labeled NvWF and AvWF to perfused vessel subendothelium were similar (0.83±0.1μg and 0.95±0.1 μg ,respectively).

Our results indicate that AvWF enhances the interaction of washed platelets with the vessel subendothelium under flow conditions. Furthermore, they suggest that this effect is related to the interaction of AvWF with platelets and not to an increased affinity of AvWF for subendothelium.