Germline Mutations in RASA1 Are Not Found in Patients with Klippel-Trenaunay Syndrome or Capillary Malformation with Limb Overgrowth.

The RASA1 gene encodes p120RASGAP, a multidomain cytoplasmic protein that acts as a negative regulator of the RAS signalling pathway. Heterozygous loss-of-function RASA1 mutations were identified in patients with Parkes Weber syndrome and multifocal capillary malformations. This syndrome is characterised by a capillary blush on an extremity, arteriovenous microfistulas, and bony and soft tissue hypertrophy. The aim of this study was to test RASA1 in 2 disorders characterised by asymmetric limb enlargement and vascular malformations, namely Klippel-Trenaunay syndrome and regional capillary malformation with overgrowth. We did not identify any clear pathogenic change in these patients. Thus, besides clinical and radiological criteria, RASA1 testing constitutes an additional tool to differentiate Parkes Weber syndrome of capillary malformation-arteriovenous malformation (CM-AVM) from overlapping disorders.

Gab2 signaling in chronic myeloid leukemia cells confers resistance to multiple Bcr-Abl inhibitors.

Grb2-associated binder 2 (Gab2) serves as a critical amplifier in the signaling network of Bcr-Abl, the driver of chronic myeloid leukemia (CML). Despite the success of tyrosine kinase inhibitors (TKIs) in CML treatment, TKI resistance, caused by mutations in Bcr-Abl or aberrant activity of its network partners, remains a clinical problem. Using inducible expression and knockdown systems, we analyzed the role of Gab2 in Bcr-Abl signaling in human CML cells, especially with respect to TKI sensitivity. We show for the first time that Gab2 signaling protects CML cells from various Bcr-Abl inhibitors (imatinib, nilotinib, dasatinib and GNF-2), whereas Gab2 knockdown or haploinsufficiency leads to increased TKI sensitivity. We dissected the underlying molecular mechanism using various Gab2 mutants and kinase inhibitors and identified the Shp2/Ras/ERK and the PI3K/AKT/mTOR axes as the two critical signaling pathways. Gab2-mediated TKI resistance was associated with persistent phosphorylation of Gab2 Y452, a PI3K recruitment site, and consistent with this finding, the protective effect of Gab2 was completely abolished by the combination of dasatinib with the dual PI3K/mTOR inhibitor NVP-BEZ235. The identification of Gab2 as a novel modulator of TKI sensitivity in CML suggests that Gab2 could be exploited as a biomarker and therapeutic target in TKI-resistant disease.

Treatment of health complaints attributed to amalgam.

The aim of the present study was to compare the reduction of subjective complaints by 3 treatment strategies in 90 "amalgam patients" whose complaints could not be explained by a medical or psychological disorder. The individuals were randomly assigned either to removal of dental amalgam only (removal group), or removal in combination with a "biological detoxification" therapy with high doses of vitamins and trace elements (removal-plus group), or participation in a health promotion program without removal of dental amalgam (no-removal group). Between baseline and month 12, the sum score of main complaints decreased by 3.5 (SD=2.2) points on average in the removal group as well as in the removal-plus group, and by 2.5 (SD=2.4) points in the no-removal group (p=0.152). Both removal groups showed a significant decrease in steady-state levels of inorganic mercury compared with the no-removal group. Thus, all 3 interventions were associated with clinically relevant improvements.

Blood and urine mercury levels in adult amalgam patients of a randomized controlled trial: interaction of Hg species in erythrocytes.

UNLABELLED: Parts of the population are permanently exposed to low levels of Hg degrees and Hg(II) from dental amalgam. It was the aim (1) to investigate the internal exposure to amalgam-related mercury from the kinetics of inorganic Hg in plasma and erythrocytes after amalgam removal, and (2) to estimate the amalgam-related absorbed dose. Dietary coexposure was monitored by determination of blood organic-Hg. Postremoval steady-state Hg concentrations were measured for 18 months. Eighty-two patients had been randomized into three groups: (A) removal of the fillings; (B) removal and non-specific detoxification, and (C) a health promotion program without removal. After amalgam removal, inorganic Hg dropped rapidly in plasma and red cells, stabilizing at 27% of preremoval levels after 60 days. Concentrations of organic Hg in plasma remained unchanged, indicating no change in dietary uptake of organic Hg. The concentration of organic Hg in red cells of group A was in the early postremoval phase lower and in the late postremoval phase higher than the preremoval control (p<0.01 for low-high difference). A protracted increase in organic Hg was also found in red cells of group B after 60 days. Thus, the effect of removal on organic Hg levels in the combined group A+B was compared with the values of group C in a linear mixed effects (LME) model which showed a significant increase with time in group A+B (p=0.028). In all groups, time profiles of urinary concentration and excretion of total-Hg were very similar to those of inorganic-Hg levels in plasma. From extrapolations of blood and urine data it was estimated that the amalgam-related inhalation and ingestion of Hg species were within the limits proposed by WHO, ATSDR and EPA. The integrated daily Hg dose absorbed from amalgam was estimated up to 3 microg for an average number of fillings and at 7.4 for a high amalgam load. CONCLUSIONS: This is the first study on adult amalgam patients which continuously monitored the postremoval decline of inorganic Hg and the coexposure from dietary organic Hg in a randomized-controlled-trial design. The integrated daily dose of 7.4 microg absorbed from a high amalgam load is well below the tolerable dose of 30 microg (WHO, 1990). The unexpected postremoval increase in erythrocyte organic Hg, which is associated with the depletion of cellular inorganic Hg, might result from binding of organic Hg to cellular sites previously occupied by inorganic Hg.

The beta 3-adrenergic receptor gene and obesity in a population sample of African Americans.

OBJECTIVE: To examine the role of the Trp64Arg polymorphism in the beta 3-adrenergic receptor gene and the beta 3-adrenergic receptor gene locus in obesity-related traits in African Americans. SUBJECTS: A total of 687 individuals representing 193 African American families who were residents of metropolitan Chicago. MEASUREMENTS: Genotyping of the Trp64Arg polymorphism in the beta 3-adrenergic receptor gene and three microsatellite markers flanking the beta 3-adrenergic receptor gene (ADRB3) locus and measuring various obesity-related traits, including body mass index (BMI), fat-free mass, fat mass, percentage fat mass, waist circumference and serum lipid levels. RESULTS: The prevalence of obesity (defined as body mass index > or = 30 kg/m(2)) in the population was 27.3% and 51.2% in men and women, respectively. The frequency of the Arg64 allele was 10.0%. Multivariate regression analyses confirmed the existence of a significant contribution of familial variance to each of the five obesity-related traits noted above. Likelihood ratio statistics computed in a multivariate regression analysis failed to demonstrate a significant association between the Arg64 allele and any of the five obesity-related traits. Single and multipoint analyses using extended Haseman--Elston regression analyses failed to demonstrate suggestive evidence of linkage of three microsatellite markers that flank the beta 3-adrenergic receptor gene to BMI, percentage body fat, waist circumference or serum leptin levels. CONCLUSION: Given the contribution of familial variance to obesity-related traits in this population, neither the null finding for the Arg64 allele nor the lack of evidence of linkage of the ADRB3 locus to obesity-related traits could be attributed to lack of transmissibility of the traits suggesting that neither the Arg64 variant of the beta 3-adrenergic receptor gene nor another genetic variant in or near the ADRB3 locus contribute significantly to familial aggregation of obesity-related traits in African Americans. International Journal of Obesity (2001) 25, 54-60

Steady-state transfer and depletion kinetics of mercury from amalgam fillings.

In 29 volunteers with a low amalgam load, the number of amalgam-covered tooth surfaces and the occlusal area of the fillings were determined. Before and at select times after removal of all amalgams, concentrations of total mercury were measured by cold-vapor atomic absorption in plasma and erythrocytes as well as in urine together with the excretion rate. Absorbed daily doses were estimated from intraoral Hg emission by two separate methods. The transfer of Hg from the fillings via the oral cavity and blood to urinary excretion was evaluated according to the most representative combination of parameters. This consisted of occlusal area (1), absorbed dose (2), Hg concentration in plasma (3) and urinary excretion (4). Pairwise correlation coefficients were 0.49 for parameters 1 vs. 2, and 0.75 each for parameters 2 vs. 3 and 3 vs. 4. Within 9 days after removal of the fillings, a transient increase in Hg levels was observed in plasma only; in the group without a rubber dam, concentrations increased significantly above pre-removal values at days 1 and 3, whereas they decreased significantly below pre-removal values at day 30 in the rubber-dam group and at day 100 in both groups. Excretion rates decreased significantly at day 100 in the protected group. Peak plasma-Hg was 0.6 ng/ml on average at day 1 and decreased with halftimes of 3 and 43 days in subjects protected by a rubber dam. The results indicated that concentrations of total mercury in plasma responded rapidly to changes in the amalgam status and reflected the actual absorption most reliably. Notably, plasma-Hg levels were sensitive enough to detect a transient attenuation of the additional exposure after using a rubber dam during the removal of only a few fillings. However, being small in magnitude and lasting 100 days at best, the rubber-dam effect had minor toxicological relevance.

Ethnic differences in the rates of low birth weight attributable to differences in early motherhood: a study from the Third National Health and Nutrition Examination Survey.

OBJECTIVE: To estimate the impact of early motherhood (being a mother at < 20 years of age) on ethnic differences in the risk of low birth weight (LBW) in a representative sample (n = 9141) of American infants and children. METHODS: Risks for LBW and the population-attributable fraction due to early motherhood were estimated adjusting for maternal smoking and education in logistic regression models. The contribution of early motherhood to ethnic differences in the risks of LBW was determined using a relative attributable risk estimate that compared Hispanics and Blacks with Whites. RESULTS: Early motherhood was independently associated with increased risk of LBW in each of the three ethnic groups, adjusting for maternal smoking during pregnancy and education. Hispanic and Black ethnicity were each associated with 15% and 123% increased risk of LBW relative to Whites. The population-attributable fractions of LBW due to early motherhood were 6.2%, 7.4%, and 2.3%, for Whites, Hispanics, and Blacks, respectively. The responses of early motherhood for LBW were different among the three ethnic groups (p < 0.05). Adjusting for maternal smoking and education, 4.8% and 7.4% of the differences in the risk of LBW between Whites and Hispanics and between Whites and Blacks, respectively, were due to differences in early motherhood. CONCLUSION: The result of this study underscores the risk of LBW due to early motherhood. Because early motherhood is preventable and avoidable, appropriate public health strategies to educate young women on the need to delay childbearing in these ethnic groups, particularly among Hispanics and Blacks, are warranted.

Effect of rubber dam on mercury exposure during amalgam removal.

It was the aim of this investigation to treat 20 volunteers with maximally 5 amalgam fillings by the same comprehensive protocol in which all removals with (n = 8) and without (n = 12) rubber dam had been performed within a few months. Nine amalgam-related parameters indicated a close matching of both groups before removal. In the group without rubber dam, mercury (Hg) levels in plasma increased significantly above preremoval values at days 1 and 3 after removal; they decreased significantly below preremoval values at day 30 in the rubber-dam group and at day 100 in both groups. Excretion rates did not increase significantly in either group, but decreased significantly at day 100 in the protected group. Peak plasma-Hg was 0.6 ng/mL on average at day one and decreased with halftimes of 3 and 43 d in subjects protected by rubber dam. The results indicated that concentrations of total mercury in plasma responded rapidly to changes in the amalgam status and reflected the actual absorption most reliably. Notably, plasma-Hg levels were sensitive enough to detect a transient attenuation of the additional exposure by using rubber dam during the removal of only a few fillings. However, being small in magnitude and lasting 100 d at best, the rubber-dam effect had minor toxicological relevance.

Systemic transfer of mercury from amalgam fillings before and after cessation of emission.

In 29 volunteers with a low amalgam load, the number of amalgam-covered tooth surfaces and the occlusal area of the fillings were determined. Concentrations of total mercury were measured in plasma and erythrocytes as well as in urine together with the excretion rate. Absorbed daily doses were estimated from intraoral Hg emission by two separate methods. The transfer of Hg from the fillings via the oral cavity and blood to urinary excretion was evaluated according to the most representative combination of parameters. This consisted of urinary excretion (1), Hg concentration in plasma (2), absorbed dose (3), and occlusal area (4). Pairwise correlation coefficients were 0.75 for parameters 1 vs 2 and 2 vs 3 and 0.49 for parameters 3 vs 4. Within 9 days after removal of the fillings, a transient increase was observed in plasma Hg levels only. This was reduced in those volunteers to whom a rubber dam had been applied during removal. Peak plasma Hg was 0.6 ng/ml on average and decreased with halftimes between 5 and 13 days. A significant decrease in Hg excretion was noted not before 100 days after removal. Being relatively insensitive to dietary mercury, the determination of total mercury in plasma and of its urinary excretion rate appears, under practical aspects, most suitable for the investigation of Hg uptake from amalgam.

Environmental issues in dentistry--mercury. FDI Commission.

One of the consequences of placing amalgam restorations is that mercury is required for the trituration process. In turn, this raises the issue of the possible environmental impact of mercury. This report considers ways in which any impact can be modified and reduced by careful attention to mercury usage and hygiene in the dental practice, the use of filters and separators in waste water pipes and the appropriate disposal of waste contaminated with amalgam. The total amount of mercury discharged into the environment varies considerably in different parts of the world due to both natural and human activities. The extent to which dentistry adds to this total also varies according to local circumstances and requirements. Recommendations are given for further development of ways to reduce mercury discharge and for further research into the environmental impact of the metal.

Compartmental transfer of mercury released from amalgam.

The number of amalgam-covered surfaces and the occlusal area of the fillings, the concentrations of total mercury in plasma, erythrocytes and urine, the urinary excretion rate, and the absorbed daily doses estimated by two separate methods from intra-oral Hg emission were determined in 29 volunteers with a low amalgam load. The transfer of Hg from the fillings via the oral cavity and blood to urinary excretion was evaluated by multiple correlations between these variables. In addition, the combination of variables most representative of the entire compartmental transfer of amalgam Hg was determined. Urinary excretion (1), Hg concentration in plasma (2) and absorbed dose (3) were most closely correlated to each other, followed by correlations with the variables of the fillings (4). Correlation coefficients were 0.75 for variables 1 vs 2 and 2 vs 3, and 0.49 for variables 3 vs 4. It was concluded that variables 1-3 best reflected the transfer of mercury from amalgam fillings throughout the organism and that they were relatively insensitive to dietary mercury. The determination of total mercury in plasma and of its urinary excretion rate appears, under practical aspects, most suitable for the investigation of Hg uptake from amalgam.

Combined estimation of mercury species released from amalgam.

Amalgam fillings constitute, after food, the main source of exposure to mercury for the general population. Banning amalgam would incur huge costs for additional dental treatment. An evaluation of potential health risks must be based on the mercury dose released from fillings. In this study, dose is estimated by a new procedure of mercury speciation which elutes the released elemental and inorganic mercury with solvents of differing polarity. We tested the procedure by incubating spherical amalgam pellets in a mixture of light paraffin oil and saline (0.9% NaCl). Release of mercury into paraffin and saline was linear in relation to both amalgam surface area and exposure time. Measurements with this method were then extended to a group of 21 amalgam-bearing volunteers. The absorbed dose averaged 4.8 micrograms/day compared with 3.7 measured conventionally in intra-oral air from the same persons. With both methods, the dose was significantly correlated to the number of amalgam-covered tooth surfaces. This dose, combined with the nearly equal mercury uptake from food, is far below the acceptable daily intake limit.

Estimation of mercury dose by a novel quantitation of elemental and inorganic species released from amalgam.

Amalgam fillings constitute, after food, the main source of exposure to mercury for the general population. An evaluation of potential health risks has to be based on the dose of mercury released from the fillings. This dose is estimated by a new procedure of mercury speciation which elutes the released elemental and inorganic mercury with solvents of different polarity (paraffin and saline). In vitro tests with spherical amalgam pellets have shown that mercury release into the solvents is linearily correlated to time and amalgam surface area. Doses estimated in volunteers by this method average 4.5 micrograms/day (range 0.3-13.9), as compared to a dose of 3.4 micrograms/day (range 0.1-11.8) measured conventionally in the oral air. The aforementioned dose, combined with the nearly equal mercury uptake from food, is below the acceptable daily intake of 40 micrograms for all forms of mercury.

Amalgam tooth fillings and man's mercury burden.

Next to nutrition, amalgam fillings represent the main source for exposure of the general population to mercury. Toxicological considerations focus on the dose of mercury resulting from such exposure. Various approaches to estimate this dose are reviewed. Introducing the dose into the known toxicokinetic model for mercury, tissue and blood and urine concentrations related to mercury release from the fillings can be predicted. These agree well with autopsy and in vivo observations. An assessment of the health hazard for individuals with amalgam fillings shows that the combined mercury intake from food and amalgam does not exceed the acceptable daily intake. In addition, blood and urine mercury concentrations of amalgam bearers are below one tenth of the critical values associated with the onset of early symptoms or of subclinical effects attributable to mercury.

Mercury compounds: lipophilicity and toxic effects on isolated myocardial tissue.

Lipophilicity is suggested to modulate the diffusion and the cytotoxic effects of mercury compounds. To investigate this, the positive inotropic effect of four Hg compounds (HgCl2, CH3HgCl, chlormerodrin, bromomercurihydroxypropane) was studied in catecholamine-depleted isolated heart muscle preparations. The rate of development of the positive effect was inversely correlated to the concentration in the case of HgCl2 and chlormerodrin, i.e. the product of concentration (c) and time to half-maximal effect (t50) remained constant. This was in accordance with the assumption of a permeation-controlled rate of action, as was shown earlier for p-chloromercuriphenyl-sulfonic acid. In addition, the c X t50 values of the individual mercurials decreased hyperbolically with the increase in lipophilicity as measured by the octanol/water partition. The results support the view that the toxicity of mercurials increases with their lipid solubility. In conjunction with the previously reported negative inotropic effect of Hg compounds, a model is proposed allocating thiol groups responsible for the negative inotropic action to lipid compartments within the cell membrane, while SH groups conveying the increase in contraction force are thought to be situated at the internal surface of the sarcolemma.

The action of organic mercury compounds on the function of isolated mammalian heart muscle.

The effects of four organic mercury compounds (methylmercuric chloride; bromomercurihydroypropane, BMHP; chlormerodrin; p-chloromercuribenzoic acid, PCMB) on mechanical and electrical functions of guinea-pig papillary muscles were investigated. An initial decline in contraction force was followed by a transient positive inotropic response. The first was accompanied by a shortening of the action-potential duration and by a reduction of the depolarization velocity and the duration of the Ca2+-dependent slow response. The latter was characterized by an indirect component (release of noradrenaline) and by a direct component, which was dependent on the stimulation rate and on the extracellular concentration of Na+ and K+. The direct positive effect, therefore, was likely to have resulted from inhibition of the sarcolemmal Na+ + K+-ATPase. This notion was confirmed by experiments with isolated membrane particles. The prevalence of the negative or positive inotropic action of these compounds could be ascribed to their lipophilic or hydrophilic properties, respectively.