A critical assessment of widely used techniques for nitrate source apportionment in arid and semi-arid regions.

The assessment of nitrate pollution origin using stable isotope techniques is a fundamental prerequisite for the application of sustainable groundwater management plans. Although nitrate pollution is a worldwide groundwater quality problem, existing knowledge on the origin of nitrate pollution in arid and semi-arid regions is still scarce. Using the example of the Grombalia aquifer (NE Tunisia), this work summarizes the main strengths and constraints of multi-isotope techniques targeting at nitrate source identification and apportionment The results highlighted that, even in the case of well-established methodologies, like those of isotope hydrogeochemistry (δ15NNO3, δ 18ONO3 and δ 11B) and mixing modelling for source apportionment, it is fundamental to take into account regional and local end-members to avoid biased data interpretation and to fully exploit the potential of such accurate tools.

Groundwater-dependent ecosystems as transfer vectors of nitrogen from the aquifer to surface waters in agricultural basins: The fontanili of the Po Plain (Italy).

The most spread groundwater-dependent ecosystems in the River Po valley are semi-natural lowland springs called "fontanili". They provide specific habitats and support high biodiversity, but are often strongly impaired by agricultural pollution. In the present study we seasonally monitored the discharge and nitrogen concentration of 48 fontanili from the Adda and the Ticino river basins. We observed a wide spatial variability of both NO3-N concentrations and flows. The annual NO3-N loads ranged from <1 to 75 t y-1 and < 1 to 29 t y-1 in the Adda and Ticino basins respectively. In the springs characterized by variable discharge the N loads were exported mostly during the summer season when water table level was elevated mainly due to irrigation. Upscaling the mean NO3-N load to each river catchment based on the total number of springs, we obtained an aerial export of 33.2 ± 6.0 and 12.5 ± 3.2 kg y-1 ha-1. Such loads accounted for the 30.4 and 21.5% of the N surplus estimated for the Adda and Ticino basins respectively. Random Forest analysis was performed to identify the most important environmental variables influencing the nitrate contamination in the spring waters. A total of 22 explanatory variables related to N sources, land uses, intrinsic hydrogeologic and soil proprieties, in "situ" and remotely sensed variables were considered. The percent of soil cultivated with maize in a 500 m radius buffer area surrounding the sampling site, the N from manure and the distance of each spring from the main river were the most effective factors in controlling the NO3-N concentration in the fontanili water. The outcomes of this work open up to achievable management prospects for the protection and recovery of fontanili waters, and can be particularly useful for water managers in identifying areas and sites where restoration plans should be a priority.

Integrated socio-hydrogeological approach to tackle nitrate contamination in groundwater resources. The case of Grombalia Basin (Tunisia).

Nitrate contamination still remains one of the main groundwater quality issues in several aquifers worldwide, despite the perduring efforts of the international scientific community to effectively tackle this problem. The classical hydrogeological and isotopic investigations are obviously of paramount importance for the characterization of contaminant sources, but are clearly not sufficient for the correct and long-term protection of groundwater resources. This paper aims at demonstrating the effectiveness of the socio-hydrogeological approach as the best tool to tackle groundwater quality issues, while contributing bridging the gap between science and society. An integrated survey, including land use, hydrochemical (physicochemical parameters and major ions) and isotopic (δ15NNO3 and δ18ONO3) analyses, coupled to capacity building and participatory activities was carried out to correctly attribute the nitrate origin in groundwater from the Grombalia Basin (North Tunisia), a region where only synthetic fertilizers have been generally identified as the main source of such pollution. Results demonstrates that the basin is characterized by high nitrate concentrations, often exceeding the statutory limits for drinking water, in both the shallow and deep aquifers, whereas sources are associated to both agricultural and urban activities. The public participation of local actors proved to be a fundamental element for the development of the hydrogeological investigation, as it permitted to obtain relevant information to support data interpretation, and eventually guaranteed the correct assessment of contaminant sources in the studied area. In addition, such activity, if adequately transferred to regulators, will ensure the effective adoption of management practices based on the research outcomes and tailored on the real needs of the local population, proving the added value to include it in any integrated investigation.

Tackling the salinity-pollution nexus in coastal aquifers from arid regions using nitrate and boron isotopes.

Salinization and nitrate pollution are generally ascertained as the main issues affecting coastal aquifers worldwide. In arid zones, where agricultural activities also result in soil salinization, both phenomena tend to co-exist and synergically contribute to alter groundwater quality, with severe negative impacts on human populations and natural ecosystems' wellbeing. It becomes therefore necessary to understand if and to what extent integrated hydrogeochemical tools can help in distinguishing among possible different salinization and nitrate contamination origins, in order to provide adequate science-based support to local development and environmental protection. The alluvial plain of Bou-Areg (North Morocco) extends over about 190 km2 and is separated from the Mediterranean Sea by the coastal Lagoon of Nador. Its surface is covered for more than 60% by agricultural activities, although the region has been recently concerned by urban population increase and tourism expansion. All these activities mainly rely on groundwater exploitation and at the same time are the main causes of both aquifer and lagoon water quality degradation. For this reason, it was chosen as a case study representative of the typical situation of coastal aquifers in arid zones worldwide, where a clear identification of salinization and pollution sources is fundamental for the implementation of locally oriented remedies and long-term management strategies. Results of a hydrogeochemical investigation performed between 2009 and 2011 show that the Bou-Areg aquifer presents high salinity (often exceeding 100 mg/L in TDS) due to both natural and anthropogenic processes. The area is also impacted by nitrate contamination, with concentrations generally exceeding the WHO statutory limits for drinking water (50 mg/L) and reaching up to about 300 mg/L, in both the rural and urban/peri-urban areas. The isotopic composition of dissolved nitrates (δ15NNO3 and δ18ONO) was used to constrain pollution drivers. The results indicate two main origins for human-induced pollution: (i) manure and septic effluents, especially in urban areas, and (ii) synthetic fertilizers in agricultural areas. In the latter, δ15N-enriched values highlight a mixture of those sources, possibly related to unbalanced fertilization and agricultural return flow. Boron isotopes (δ11B) were hence studied to further distinguish the nitrate origin in the presence of multiple sources and mixing processes. The results indicate that in the study area, the high geochemical background for B and Cl, associated to the complex water-rock interaction processes, limit the application of the coupled δ11B and δ15N isotopic systematics to the detection of sources of groundwater pollution. In fact, despite the exceedingly high nitrate contents, the depleted δ11B values that characterize synthetic fertilizers and sewage leakages could not be detected. Therefore, even if in saline groundwater the anthropogenic contribution has a negligible effect in terms of salinity input, with both sewage and irrigation water not very charged, the associated nitrate content fuels up water-rock interaction processes, eventually leading to a mineralization increase.

Assessment of groundwater pollution from ash ponds using stable and unstable isotopes around the Koradi and Khaperkheda thermal power plants (Maharashtra, India).

The impact on local water resources due to fly ash produced in the Koradi and Khaperkheda thermal power plants (district of Nagpur, Maharashtra - India) and disposed in large ponds at the surface was assessed through the study of environmental variation of ratios of stable and unstable isotopes. Analyses of oxygen and hydrogen isotopes suggest scarce interaction between the water temporarily stored in the ponds and the groundwater in the study area. Data also highlight that the high salinity of groundwater measured in the polluted wells is not due to evaporation, but to subsequent infiltration of stream waters draining from the ponds to the local aquifer. (87)Sr/(86)Sr values, when associated with Sr/Ca ratios, demonstrate the dominant role of waste waters coming from tens of brick kilns surrounding the pond sulfate pollution. Uranium isotopic analyses clearly show evidence of the interaction between groundwater and aquifer rocks, and confirm again the low influence of ash ponds. A new conceptual model based on the study of the isotopes of radium is also proposed and used to estimate residence times of groundwater in the area. This model highlights that high salinity cannot be in any case attributed to a prolonged water-rock interaction, but is due to the influence of untreated waste water of domestic or brick kiln origin on the shallow and vulnerable aquifers.

Identifying the effects of human pressure on groundwater quality to support water management strategies in coastal regions: a multi-tracer and statistical approach (Bou-Areg region, Morocco).

Groundwater pollution from anthropogenic sources is a serious concern affecting several coastal aquifers worldwide. Increasing groundwater exploitation, coupled with point and non-point pollution sources, are the main anthropogenic impacts on coastal environments and are responsible for severe health and food security issues. Adequate management strategies to protect groundwater from contamination and overexploitation are of paramount importance, especially in arid prone regions, where coastal aquifers often represent the main freshwater resource to sustain human needs. The Bou-Areg Aquifer (Morocco) is a perfect example of a coastal aquifer constantly exposed to all the negative externalities associated with groundwater use for agricultural purposes, which lead to a general increase in aquifer salinization. In this study data on 61 water samples, collected in June and November 2010, were used to: (i) track groundwater composition changes related to the use of irrigation water from different sources, (ii) highlight seasonal variations to assess aquifer vulnerability, and (iii) present a reproducible example of multi-tracer approach for groundwater management in rural coastal areas. Hydrogeochemical results show that Bou-Areg groundwater is characterized by - high salinity, associated with a remarkable increase in bicarbonate content in the crop growing season, due to more intense biological activity in irrigated soils. The coupled multi-tracer and statistical analysis confirms the strong dependency on irrigation activities as well as a clear identification of the processes governing the aquifer's hydrochemistry in the different seasons. Water Rock Interaction (WRI) dominates the composition of most of groundwater samples in the Low Irrigation season (L-IR) and Agricultural Return Flow (ARF) mainly affects groundwater salinization in the High Irrigation season (H-IR) in the same areas naturally affected by WRI. In the central part of the plain River Recharge (RR) from the Selouane River is responsible for the high groundwater salinity whilst Mixing Processes (MIX) occur in absence of irrigation activities.

Impact of the disposal and re-use of fly ash on water quality: the case of the Koradi and Khaperkheda thermal power plants (Maharashtra, India).

An increasing amount of fly ash from thermal power plants is produced in India every year. Its disposal is generally done in ponds after it is mixed together in suitable proportion of water to form a slurry. Fly ash from Koradi and Khaperkheda thermal power plants (Nagpur, Maharashtra) is commonly disposed in an area characterized by the presence of many small villages where the population uses the groundwater for drinking and domestic purposes. Here, the groundwater locally exceeds the concentration limits recommended by the Bureau of Indian Standards (BIS, 2005) and by the World Health Organization (WHO, 2008) for Mg(2+), Ca(2+), NO3(-), SO4(2-), Total Dissolved Solids (TDS) and for some minor elements like As, Mo, V and U. A new geological map of the study area has been prepared to understand the possible water-rock interactions. An extensive geochemical survey of groundwater, stream water and fly ash was also carried out to clarify the possible origin of the pollutants by discriminating between geogenic and anthropogenic sources and to assess the influence of the ash ponds on water quality. The analytical results suggest that a large part of the sulfates in the groundwater of the villages of Masada, Khairi and Kawatha originate from the infiltration of industrial water from tens of factories that mix fly ash with relatively high quantities of gypsum and lime for the production of bricks. In addition, the interaction with the relatively U-rich Gondwana units, like Talchir formation, is probably the cause of the high concentration of this element. Results showed how the relatively high concentrations of Mo, As, B and F in circulating waters are linked to the leaching from fly ash, also pointing out a direct spatial correlation between the concentration of fluorides in the groundwater and their closeness to the ash ponds.

Nitrogen inputs to a river course in a heavily impacted watershed: a combined hydrochemical and isotopic evaluation (Oglio River Basin, N Italy).

This study aims at evaluating sources and processes affecting NO3(-) concentrations in the Oglio River. Five sampling campaigns considered the main watercourse, tributaries, point pollution sources, springs, and groundwater. Physico-chemical parameters, N forms, B, Sr(2+), stable isotopes (δ(2)HH2O, δ(18)OH2O, δ(15)NNO3, δ(18)ONO3, δ(11)B) and discharge were measured. Hydrological modelling was performed using mass balance and End Member Mixing Analysis equations. During the irrigation period, in the upstream reach, up to 90% of the natural river flow is diverted for irrigation and industrial purposes; excess water drained from agricultural fields is returned to river in the downstream reach. Results evidenced, in the middle reach, a large input of NO3(-)-rich groundwater which could be quantified using hydrological modelling. Groundwater inputs are responsible for the sharp, tenfold increase in NO3(-) in the river water, from 2.2-4.4 up to 33.5 mgL(-1), and are more evident in summer, when discharge is lower. Nevertheless, river water preserves its natural B isotopic composition, indicating that the two tracers do not have a common origin and are not co-migrant. In the lower plain, surface-groundwater interconnections and human disturbances in the water cycle favour the recycling of the compounds in the environment, and lead to a similarity in composition of the different water bodies (Oglio River, tributaries and groundwater). The long lasting agronomical practices have profoundly modified the surface-groundwater equilibrium and chemical characteristics, resulting in a highly buffered system. Infiltrating irrigation water leaches down NO3(-) which is subsequently denitrified; when returned to the Oglio River, groundwater modifies the river water composition by dilution, in the case of NO3(-), or by addition, for other constituents (e.g. Cl(-), B). The results of this study indicate that, in order to reduce the NO3(-) transport towards the Adriatic Sea, groundwater contamination should be addressed first, with expected long recovery times.

C-reactive protein is directly related to plasminogen activator inhibitor type 1 (PAI-1) levels in diabetic subjects with the 4G allele at position -675 of the PAI-1 gene.

BACKGROUND AND AIMS: C-reactive protein (CRP) has been identified as a possible factor able to promote atherosclerosis. "In vitro" studies have demonstrated that CRP induces plasminogen activator inhibitor type 1 (PAI-1) expression, suggesting a hypofibrinolytic role for CRP. As CRP and PAI-1 levels increase in type 2 diabetic subjects, we decided to study the relationship between CRP and PAI-1, and the role of the 4G/5G polymorphism of the PAI-1 gene on this relationship in a diabetic population without complications. METHODS AND RESULTS: Two hundred and ninety-five type 2 diabetic patients (age 60.9+/-10.5 years) and 290 healthy controls (age 59.2+/-11.5 years) were enrolled. A significant correlation between PAI-1 and CRP in diabetic subjects was found (r=0.45, p<0.001), whereas no relationship was evident in the control subjects between these inflammatory markers. Multiple regression analysis highlighted that CRP is the only one significant variable of PAI-1 antigen in diabetic subjects (partial r=0.31, p<0.01). Stratifying by genotype, a positive correlation between PAI-1 and CRP in 4G/4G (partial r=0.64 p<0.001) and 4G/5G (partial r=0.47, p<0.001) subjects was found, whereas no correlation in 5G/5G was present. Multiple regression analysis confirmed the presence of this correlation in 4G/4G (partial r=0.45, p<0.001) and in 4G/5G (partial r=0.34, p=0.007) diabetic patients. CONCLUSIONS: These findings demonstrate that CRP plays an important role in the complex mechanism regulating PAI-1 antigen in 4G diabetic carriers.

Effect of 4G/5G PAI-1 polymorphism on the response of PAI-1 activity to vitamin E supplementation in Type 2 diabetic patients.

Plasminogen activator inhibitor type 1 (PAI-1) is an independent cardiovascular risk factor and increases in patients with Type 2 diabetes mellitus. The 4G/5G polymorphism of PAI-1 has been reported to be involved in the incidence of cardiovascular disease by regulation of PAI-1 levels, but this relation is still under debate. The aim of the study was to test the effect of 4G/5G polymorphism on the lowering of PAI-1 levels in Type 2 diabetic patients during vitamin E supplementation. Ninety-three Type 2 diabetic subjects (age +/- SD, 62.1 +/- 6.1 yr) were enrolled and treated with vitamin E (500 IU/die) for 10 weeks. We determined the 4G/5G polymorphism and PAI-1 activity at baseline, during (5th and 10th week) and after (30th week) vitamin E supplementation. No significant differences were found in PAI-1 and its determinants among the three genotypic groups at baseline. Decrements were detected in the whole group in PAI-1 at the 5th and the 10th week from baseline followed by an increase at the 30th week (p<0.001). Patients with 4G/4G and 4G/5G genotypes showed a different trend with respect to those with 5G/5G in PAI-1. In particular, there was a decrease in 4G/4G and 4G/5G PAI-1 levels from the 10th week, while a decrease in 5G/5G PAI-1 was observed from the 5th week (p<0.01). The delayed decrease, found in patients with at least one 4G allele with respect to those with 5G/5G genotype, demonstrates that 4G/5G polymorphism mainly influences the rate of decrease of PAI-1 after supplementation with vitamin E in Type 2 diabetic subjects.

Gamma dose rate calculation and mapping of piemonte (North-West Italy) from gamma spectrometry soil data.

In this study the air gamma dose rate map of Piemonte, a region in the North-West of Italy, was produced from gamma spectrometry soil data. Soil samples collected in 110 different sites of Piemonte were analysed with Hyperpure Germanium (HPGe) detectors (30% relative efficiency), which allow the evaluation of the activity concentrations of natural radionuclides and (137)Cs. Then, using the available mathematical models, the gamma absorbed dose rate in air due to radionuclides was calculated. The contribution of the cosmic radiation to the total absorbed dose rate, which depend on the site altitude was also evaluated and added to the soil contribution. Finally, the map of the whole region was obtained by fitting the dose rate values of the different sites with kriging algorithms.

Multivariate statistical and GIS-based approach to identify heavy metal sources in soils.

The knowledge of the regional variability, the background values and the anthropic vs. natural origin for potentially harmful elements in soils is of critical importance to assess human impact and to fix guide values and quality standards. The present study was undertaken as a preliminary survey on soil contamination on a regional scale in Piemonte (NW Italy). The aims of the study were: (1) to determine average regional concentrations of some heavy metals (Cr, Co, Ni, Cu, Zn, Pb); (2) to find out their large-scale variability; (3) to define their natural or artificial origin; and (4) to identify possible non-point sources of contamination. Multivariate statistic approaches (Principal Component Analysis and Cluster Analysis) were adopted for data treatment, allowing the identification of three main factors controlling the heavy metal variability in cultivated soils. Geostatistics were used to construct regional distribution maps, to be compared with the geographical, geologic and land use regional database using GIS software. This approach, evidencing spatial relationships, proved very useful to the confirmation and refinement of geochemical interpretations of the statistical output. Cr, Co and Ni were associated with and controlled by parent rocks, whereas Cu together with Zn, and Pb alone were controlled by anthropic activities. The study indicates that background values and realistic mandatory guidelines are impossible to fix without an extensive data collection and without a correct geochemical interpretation of the data.

HELLP syndrome and factor V Leiden.

The association of thrombophilia and obstetrical complications is documented and well consistent with the hypothesis of an insufficient placental perfusion due to fibrin deposition as a major underlying pathophysiological mechanism. Factor V Leiden is one of the most frequent thrombophilic mutations. A high prevalence of this mutation has recently been reported in a group of 21 German women with haemolysis, elevated liver enzymes, low platelets (HELLP) syndrome. In this respect, we studied the prevalence of factor V Leiden in 18 women who were consecutively diagnosed at our Department of Obstetrics and Gynaecology as having HELLP syndrome, between 1995 and 1999. Women were tested either at the time of diagnosis or months or years after delivery for coagulation parameters, protein C (PC), protein S (PS), antithrombin III, lupus-like anticoagulant, anticardiolipin antibodies (ACA), activated protein C (APC) resistance and detection of the G1691A mutation (factor V Leiden). In all women, the parameters studied were normal and in none of the investigated cases was the G1691A mutation found. HELLP being a severe form of preeclampsia, we think that the reported association between factor V Leiden and HELLP may reflect the well-known association with preeclampsia.

Inflammatory bowel diseases are not associated with major hereditary conditions predisposing to thrombosis.

Patients with inflammatory bowel diseases (IBD) are at increased risk for thromboembolic complications. Our aim was to evaluate whether the increased risk for thrombosis in IBD could be due to a genetic association of IBD with hereditary prothrombotic conditions. In all, 102 IBD patients (51 with ulcerative colitis and 51 with Crohn's disease) with no history of thrombosis and 204 matched normal subjects were enrolled. DNA specimens were evaluated by PCR and restriction fragment length polymorphism for factor V Leiden, methylene tetrahydrofolate reductase (MTHFR) and prothrombin gene mutations. In IBD patients and matched controls the observed allele frequencies were similar, being 1.5% and 1.2% for factor V Leiden gene mutation, 1.1% and 0.7% for prothrombin gene mutation, and 45.1% and 47.4% for MTHFR gene mutation, respectively. These rates also were not significantly different when patients were analyzed according to age and sex distribution, diagnosis, and extension and clinical type of disease. In conclusion, our study shows no association between IBD and the most frequent hereditary prothrombotic conditions. Other factors should be evaluated in order to understand the mechanisms underlying the thrombotic risk of IBD.

A novel polymorphism in intron 1a of the human factor VII gene (G73A): study of a healthy Italian population and of 190 young survivors of myocardial infarction.

We have identified a novel polymorphism located in intron 1a of the human factor VII gene, caused by the nucleotide change G to A at position + 73. In a population of 128 healthy individuals from northern Italy, the variant A73 allele had a frequency of 0.21, whereas the frequency of the previously reported 10 bp insertion allele located at -323 in the promoter region was 0.17 and that of the Q353 allele in the catalytic region of the factor VII gene was 0. 20. In 75% of the healthy individuals, the A73 allele was present together with the 10 bp insertion and the Q353 alleles, indicating a strong linkage disequilibrium. The concomitant presence of A73 with both the 10 bp and the Q353 alleles was associated with the lowest factor VII levels, measured as coagulant activity, activated factor VII and factor VII antigen. The G73A polymorphism was also evaluated in 190 survivors of myocardial infarction who had experienced the event before the age of 45 years and in 179 individuals with a negative exercise test matched with patients for sex, age and geographical origin. Patients carrying the A73 allele associated with lower factor VII levels tended to have a lower risk of myocardial infarction (adjusted odds ratio 0.54; 95% confidence intervals 0.29-0.99). In conclusion, we found a novel variant allele in intron 1a of the human factor VII gene that is often associated in healthy individuals with the 10 bp and Q353 alleles in the promoter and catalytic region of the same gene. This intronic mutation, alone or in association with other factor VII gene polymorphisms, might confer protection against myocardial infarction in the young.

High risk of cerebral-vein thrombosis in carriers of a prothrombin-gene mutation and in users of oral contraceptives.

BACKGROUND: Idiopathic cerebral-vein thrombosis can cause serious neurologic disability. We evaluated risk factors for this disorder, including genetic risk factors (mutations in the genes encoding factor V and prothrombin) and nongenetic risk factors (such as the use of oral contraceptive agents). We compared the prevalence of these risk factors in 40 patients with cerebral-vein thrombosis, 80 patients with deep-vein thrombosis of the lower extremities, and 120 healthy controls. The G1691A mutation in the factor V gene and the G20210A prothrombin-gene mutation, which are established genetic risk factors for venous thrombosis, were studied. We also assessed the use of oral contraceptives and other risk factors for thrombosis. RESULTS: The prevalence of the prothrombin-gene mutation was higher in patients with cerebral-vein thrombosis (20 percent) than in healthy controls (3 percent; odds ratio, 10.2; 95 percent confidence interval, 2.3 to 31.0) and was similar to that in patients with deep-vein thrombosis (18 percent). Similar results were obtained for the mutation in the factor V gene. The use of oral contraceptives was more frequent among women with cerebral-vein thrombosis (96 percent) than among controls (32 percent; odds ratio, 22.1; 95 percent confidence interval, 5.9 to 84.2) and among those with deep-vein thrombosis (61 percent; odds ratio, 4.4; 95 percent confidence interval, 1.1 to 17.8). For women who were taking oral contraceptives and who also had the prothrombin-gene mutation (seven patients with cerebral-vein thrombosis but only one control), the odds ratio for cerebral-vein thrombosis rose to 149.3 (95 percent confidence interval, 31.0 to 711.0). CONCLUSIONS: Mutations in the prothrombin gene and the factor V gene are associated with cerebral-vein thrombosis. The use of oral contraceptives is also strongly and independently associated with the disorder. The presence of both the prothrombin-gene mutation and oral-contraceptive use raises the risk of cerebral-vein thrombosis further.

PIP: The role of the prothrombin-gene mutation in idiopathic cerebral-vein thrombosis and its interaction with other risk factors was investigated in a study of 40 patients (9 men and 31 women) 15-64 years of age who presented to a thrombosis center in Milan, Italy, in 1991-97 after a first episode of this thrombosis. Also enrolled were 80 men and women randomly selected from patients screened at the same center during the study period after a first documented episode of proximal deep-vein thrombosis of the lower extremities. 120 healthy controls were matched to cerebral-vein thrombosis patients by sex, age, geographic origin, and education. 20% of patients with cerebral-vein thrombosis (odds ratio (OR), 10.2; 95% confidence interval (CI), 2.3-31.0), 18% of those with deep-vein thrombosis, and 3% of controls were carriers of the prothrombin-gene mutation. Factor V mutation was more prevalent in patients with cerebral-vein thrombosis (15%) than controls (3%) (OR, 7.8; 95% CI, 1.8-34.1), but the thrombotic risks associated with these two mutations were independent of each other. Oral contraceptive (OC) ever-use was more frequent among women with cerebral-vein thrombosis (96%) (OR, 22.1; 95% CI, 5.9-84.2) and deep-vein thrombosis (61%) (OR, 4.4; 95% CI, 1.1-17.8) compared with controls (32%). For the 7 women with cerebral-vein thrombosis who were both OC ever-users and had the prothrombin-gene mutation, the thrombotic risk rose to 149.3 (95% CI, 31.0-711.0). These findings show that there is a hypercoagulable state in 35% of patients with idiopathic cerebral-vein thrombosis. Although screening for the prothrombin-gene mutation in young women before they are prescribed OCs is unlikely to be cost-effective, carriers of the mutation who have had a thrombosis episode should discontinue OC use.

A factor V genetic component differing from factor V R506Q contributes to the activated protein C resistance phenotype.

Factor V gene polymorphisms were investigated to detect components that may contribute to the activated protein C (APC) resistance phenotype in patients with venous thromboembolism. A specific factor V gene haplotype (HR2) was defined by six polymorphisms and its frequency was found to be similar in normal subjects coming from Italy (0.08), India (0.1), and Somalia (0.08), indicating that it was originated by ancestral mutational events. The relationship between the distribution of normalized APC ratios obtained with the functional assay and haplotype frequency was analyzed in patients heterozygous for factor V R506Q (factor V Leiden). The HR2 haplotype was significantly more frequent in patients with ratios below the 15th percentile than in those with higher ratios or in normal controls. Moreover, the study of 10 patients with APC resistance in the absence of the factor V R506Q mutation showed a 50-fold higher frequency of HR2 homozygotes. The HR2 haplotype was associated with significantly lower APC ratios both in patients with venous thromboembolism and in age- and sex-matched controls. However, the two groups showed similar HR2 haplotype frequencies. Plasma mixing experiments showed that an artificially created double heterozygote for the factor V R506Q mutation and the HR2 haplotype had an APC ratio lower than that expected for a simple R506Q heterozygote. Time-course experiments evaluating the decay of factor V in plasma showed the normal stability of the molecule encoded by the factor V gene marked by the HR2 haplotype, which ruled out the presence of a pseudo-homozygous APC resistance mechanism. Our results provide new insights into the presence of factor V genetic components other than the factor V R506Q that are able to contribute to the APC resistance phenotype in patients with venous thromboembolism.

Gene polymorphisms predicting high plasma levels of coagulation and fibrinolysis proteins. A study in centenarians.

Gene polymorphisms associated with the plasma levels of fibrinogen, factor VII, and plasminogen activator inhibitor 1 (PAI-1)-hemostasis proteins that help to predict the risk of atherothrombotic disease-were compared in 124 healthy individuals > or = 100 years old and 130 young, healthy individuals to identify genetic influences on extreme longevity. We investigated the restriction fragment length polymorphism G/A-455 located in the promoter of the beta-fibrinogen gene, the guanine insertion/deletion polymorphism 4G/5G in the promoter of the PAI-1 gene, and the R353Q substitution polymorphism in exon 8 of the factor VII gene. Alleles and genotypes associated with elevated plasma levels of fibrinogen and factor VII were found with similar frequencies in centenarians and in the comparison group. However, in centenarians there was a significantly higher frequency of the 4G allele and of the homozygous 4G4G genotype associated with high PAI-1 levels. Since high PAI-1 is considered a predictor of recurrent myocardial infarction in young men, it is intriguing that the corresponding genetic marker is more frequent in centenarians who have escaped major age-related atherothrombotic disease and reached the extreme limits of human life. Homozygosity for the 4G allele, despite its association with impaired fibrinolysis, is compatible with successful aging.