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Disrupted in schizophrenia 1 (DISC1) is a multi-functional scaffolding protein that has been associated with neuropsychiatric disease. The role of DISC1 is to assemble protein complexes that promote neural development and signaling, hence tight control of the concentration of cellular DISC1 in neurons is vital to brain function. Using structural and biochemical techniques, we show for we believe the first time that not only is DISC1 turnover elicited by the ubiquitin proteasome system (UPS) but that it is orchestrated by the F-Box protein, FBXW7. We present the structure of FBXW7 bound to the DISC1 phosphodegron motif and exploit this information to prove that disruption of the FBXW7-DISC1 complex results in a stabilization of DISC1. This action can counteract DISC1 deficiencies observed in neural progenitor cells derived from induced pluripotent stem cells from schizophrenia patients with a DISC1 frameshift mutation. Thus manipulation of DISC1 levels via the UPS may provide a novel method to explore DISC1 function.

The usefulness of micronutrient antioxidant therapy for recurrent (non-gallstone) pancreatitis has recently been endorsed by a 20-week double-blind double-dummy cross-over trial in 20 patients. Treatment was delivered as two types of tablets, providing daily doses of 600 micrograms organic selenium, 9000 i.u. beta-carotene, 0.54 g vitamin C, 270 i.u. vitamin E and 2 g methionine. We report antioxidant profiles in blood samples collected before entry, at the cross-over stage and upon completion of trial. Baseline serum concentrations of selenium, beta-carotene and vitamin E in the patients were significantly lower than in healthy controls, were unaltered by placebo and normalized by active treatment, but reverted to basal values in the subgroup that received placebo subsequently. The baseline serum concentration of a free radical marker--the 9-cis, 11-trans isomer of linoleic acid--was significantly higher in the patients than in controls, fell inexplicably in the placebo phase and fell further upon active treatment. Discriminant analysis eliminated the overlap in free radical marker and selenium concentrations between control sera on the one hand and baseline or post-placebo samples from the patients on the other: antioxidant treatment normalized the relationship between these biochemical parameters. Subnormal baseline serum levels of S-adenosylmethionine drifted downwards upon active treatment whereas a sharp rise was noted when a relapse of pancreatitis occurred during the placebo phase. The results confirm that adequate exposure to antioxidants in the active treatment phase was associated with amelioration of oxidative stress, and that there was no residual effect 10 weeks after switching over to placebo treatment. Furthermore, the paradoxical behaviour of S-adenosylmethionine may imply that the beneficial effect of micronutrient antioxidants in recurrent pancreatitis is linked with preservation of the methionine trans-sulfuration pathway in pancreatic acinar cells.

A total of 2691 single-lesion isolates of Phytophthora infestans was established from samples of late-blight disease from 354 commercial and garden/allotment sites in Scotland, England and Wales over four growing seasons, 1995–98. The A2 mating type was rare (3·0% of isolates) and was detected at only 34 sites. In vitro tests of sensitivity to the phenylamide fungicide metalaxyl showed that 316 sites yielded isolates with some insensitivity (resistant and/or intermediate); these were more often commercial sites than garden/allotment sites. Over the four seasons, the frequency of isolates with intermediate fungicide sensitivity increased, while the frequency of resistant phenotypes decreased. Resistant isolates were always of A1 mating type. A subset of 1459 isolates from 326 sites was analysed for molecular diversity. Mitochondrial DNA (mtDNA) haplotype Ia predominated (91·0% of isolates); haplotype IIa was present at 54 sites and both haplotypes at 33 sites. The multilocus RFLP probe RG57 detected 30 fingerprints. Four fingerprints were particularly common (RF002, RF006, RF039 and RF040) and 10 were unique to a single site in a single year. The three commonest fingerprints (RF039 > RF002 > RF006) were of A1 mating type and the fourth (RF040) was A2. RF002 isolates were resistant to the phenylamide metalaxyl and were more common in Scotland than in England and Wales. Small sample sizes limited the usefulness of estimates of diversity. Although approximately half of all sites appeared to be colonized by RF039 genotypes, some sites (both commercial and garden/allotment) showed a higher diversity, having both common and unique genotypes. The genotypic diversity within isolates collected from commercial sites and those from garden/allotment sites were similar. The contributions of sexual reproduction and alternatives to sex to the generation of variation are discussed.

In a survey of Scottish potato late blight (Phytophthora infestans) populations from 1995 to 1997, nearly 500 isolates were collected from over 80 disease outbreaks in commercial potato crops and gardens/allotments. The isolates were characterized by mating type, resistance to the fungicide metalaxyl and almost 300 were examined by DNA-based AFLP fingerprinting. These data were examined alongside cropping details to determine the population structure in the context of existing disease management strategies. A1 and A2 mating type isolates were present in both commercial potato crops and gardens or allotments although they coexisted more frequently in the latter sites. One-fifth of the isolates collected were of the A2 mating type and the frequency was similar over the 3 years and amongst sites. In 1995 the proportions of isolates that were sensitive and resistant to metalaxyl were equal (∼40%) but, over the following 2 years, the frequency of resistant isolates decreased and that of intermediate isolates increased. The mating type response to metalaxyl differed markedly, with 52% of A1 and only 5% of A2 isolates being resistant. Considerable molecular diversity was observed, with over half of the isolates having unique AFLP patterns. Analysis of the molecular and phenotypic data revealed a broad clustering of the population into three groups. Many factors point to an A2 population restricted by its sensitivity to phenylamides. The majority of the A2 isolates were found in a single AFLP group, but the presence of mixed mating type samples, an increasing frequency of isolates of intermediate metalaxyl resistance and the extent of the AFLP diversity suggest occasional sexual recombination, and thus gene flow, between groups.

Anthropogenic contributions of lead to the urban environment havebeen dominated by combustion of leaded gasoline. A number of studies have used lead concentrations in road deposited sediments(RDSs) to infer automobile contributions. However, few studies have combined concentration data, enrichment ratios, and lead isotope ratio data into a comprehensive picture of lead contamination of road sediments. An urban, non-industrialized basin, in Oahu, Hawaii, was selected for investigation. Twenty RDSsamples were collected throughout the 11 km2 system, with anundisturbed soil profile sampled to a depth of 30 cm to documentbackground lead levels.Median lead concentrations from a weak (cold) HCl digestion and a hot nitric acid digestion were 3 and 7 mg kg-1 for thebackground soil, and 256 and 303 mg kg-1 for RDSs. The median Pb enrichment ratio (HCl digestion) for RDSs was 129, witha range from 24 to 883. All the data collected point to a highlycontaminated environment.Lead isotope ratios from potential sources were examined relativeto those observed for RDSs in the system. Host geological rocks,paint, and long-range aerosol transport were ruled-out as significant sources based on an examination of isotope ratios andpotential loadings to road sediments. Leaded gasoline wasidentified as the major contributor to present-day road sedimentsbased on their radiogenic nature, with mean 206Pb/204Pb = 18.787 ± 0.096 (95% confidence interval), 207Pb/204Pb = 15.847± 0.074, 208Pb/204Pb= 38.836 ± 0.221, and 206Pb/207Pb = 1.184 ± 0.009. The contribution of gasoline additives to RDS for two periods, pre-1968 and post-1968, were estimated using 206Pb/207Pb ratios. The average contribution of post-1968 lead to RDSs was 59%, with a range from 32 to 81%. To explain the mixed age of lead in the RDSs, we suggest that erosional processes have mobilized sediment from roadside reservoirs in the basin that have accumulated automobile emissions primarily since the 1930s. The significant shift in useof radiogenic (J-type) ores, mostly from Missouri, USA, have allowed us to fingerprint and apportion lead in RDSs of thissystem.

Observant dans le domaine des medias une revendication excessive de la liberte d'expression, l'A. revise son approche de la definition de ce principe chez Mill, en montrant que le droit inalienable a une liberte d'expression absolue est contraire au principe meme de la liberte individuelle qui consiste a ne pas porter prejudice a autrui. Examinant la definition des pensees comme incitation a l'action, l'A. etudie la justification des libertes chez Mill et met en evidence l'incompatibilite de la liberte d'expression (orale et ecrite) avec le principe d'utilite.

Studies of the biological chemistry of aluminium can gain significantly from the use of the long-lived isotope 26Al as a tracer, although the cost of the isotope often precludes its determination by radiochemical counting techniques. Accelerator mass spectrometry (AMS) provides an ultra-sensitive method of determination, free from isobaric interference from atomic (26Mg) or molecular species. The source materials for AMS can be aluminium oxide or phosphate, both of which can be readily prepared at a sufficient level of purity from biological substrates. Natural aluminium (27Al, 100%) is added to the preparations as a chemical yield monitor and to provide the reference for the isotope ratio measurement. 26Al/27Al ratios can be determined over the range 10(-14)-10(-7), implying a limit of detection for 26Al of around 10(-18) g. The precision of measurement and long-term reproducibility are5% and7% (RSD), respectively. Chemical methodologies for routine measurements on blood and urine samples have been developed.

Etude des mecanismes immoraux de la corruption et de la subornation, fondes sur le principe de la tentation qui consiste a mettre une personne a l'epreuve d'une promesse. L'A. montre que celle-ci n'est pas fautive de l'action realisee sous la contrainte, mais qu'elle est responsable de son defaut de caractere, pour avoir accepte les pots-de-vin

The prevention and treatment of aluminum (Al) accumulation and toxicity are reviewed. Recommendations to further our understanding of desferrioxamine (deferoxamine, DFO) treatment and to develop more effective chelation approaches are provided. Reduction of Al accumulation and toxicity may benefit end-stage renal disease (ESRD) patients and perhaps those suffering from specific neurodegenerative disorders as well as workers with Al-induced neurocognitive disorders. The clearance of Al may be increased by extracorporeal chelation, renal transplantation, perhaps complexation with simple ligands such as silicon (Si), and systemic chelation therapy. The abilities of extracorporeal chelation and Si to reduce Al accumulation require further evaluation. Although it may not be possible to design Al-specific chelators, chelators with greater Al selectivity are desired. Aluminum-selective chelation might be achieved by targeted chelator distribution or by the use of adjuvants with the chelator. The ability of carboxylic acids to facilitate Al elimination, under specific conditions, warrants further study. Desferrioxamine does not produce significant biliary Al excretion. A chelator with this property may be useful in ESRD patients. The necessity for an Al chelator to distribute extravascularly to be effective is unknown and should be determined to guide the selection of alternatives to DFO. The lack of oral efficacy and occasional side effects of DFO encourage identification of orally effective, safer Al chelators. The bidentate 3-hydroxypyridin-4-ones are currently the most encouraging alternatives to DFO. They have been shown to increase urinary Al excretion in rats and rabbits, but to have toxicity comparable to, or greater than, DFO. Their toxicity may relate to incomplete metal complexation. The ability of orally effective chelators to increase absorption of chelated metal from the gastrointestinal (Gl) tract needs to be evaluated. Orally effective, safe Al chelators would be of benefit to peritoneal dialysis patients and those with neurodegenerative disorders, if Al chelation therapy is indicated. The reduction of Alzheimer's disease (AD) progression and the reversal of Al-induced behavioral deficits and neurofibrillary tangles by DFO encourage further study of Al chelation therapy for selected neurodegenerative disorders.

As a "hard" trivalent metal ion, Al 3+ binds strongly to oxygen-donor ligands such as citrate and phosphate. The aqueous coordination chemistry of Al is complicated by the tendency of many Al complexes to hydrolyze and form polynuclear species, many of which are sparingly soluble. Thus there is considerable variation among the Al stability constants reported for several important ligands. The complexity in the aqueous chemistry of Al has also affected Al toxicity studies, which have often utilized poorly characterized Al stock solutions. Serum fractionation studies show that most Al is protein bound, primarily to the serum iron transport protein transferrin. Albumin appears to play little, if any, role in serum transport. There is little agreement as to the speciation of the remaining low-molecular-mass fraction of serum Al. The lability of the Al 3+ ion precludes the simple separation and identification of individual Al complexes. Computational methods are available for detailed computer calculations of ...

Mill devotes more than one-third of On Liberty' to 'the liberty of thought and discussion.' (I, 36-71). He asserts that there ought to be 'absolute freedom of opinion and sentiment on all subjects', and a corresponding absolute 'liberty of expressing and publishing opinions', since such expression and publication of thoughts is 'practically inseparable' from thinking. (I, 33) R is the author of a review of Mill's essay which was published in the British Quarterly Review, Vol. 31, 1860, pp. 173-195.2 We have to know this author as 'R', because his or her review is anonymous, and because it is the only substantial response in Pyle's anthology the author of which it has been impossible to identify with the aid of the Wellesley Index to Victorian Periodicials. (II, v, xx) This is a pity, since one would like to know the authorship of this powerful and perspicacious essay. R is concerned with Mill's view on 'social tyranny' rather than on 'legal tyranny'. (II, 191) His exposition of this thesis is as follows: 'Mr Mill ... seems to think that the alleged tyranny of society against obnoxious opinions ... can be put down ... by inculcating the duty of listening ... to anything that everybody has to say for his opinions.' (II, 194-195, italics added) Mill inculcates the duty of listening in his account of 'the causes which make diversity of opinion advantageous' (I, 63-9). An extreme expression of this thesis is that '... it is [social] tyranny, if a man is allowed to publish a book, unless men will also read it ...' (II, 196) R's objections to this thesis are the following three: (a) What is sauce for the goose (the communicator) is sauce for the gander (the communicatee). As there is a moral obligation not to restrain a person from thinking or expressing his thoughts-an

The relative pathogenicity of isolates of Macrophomina phaseolina was comparable both on sunflower tissue cultures and on mature plants and seedlings. The relative susceptibility of the different sunflower lines in vitro showed a similar pattern for cotyledon callus cultures, but not for immature embryo cultures. Although these protocols appear to be unsuitable for the selection of novel disease resistance, they do offer the potential for a rapid, non-destructive screen for resistant material.

Fuel particles released during the accident at the Chernobyl reactor (1986) and during the operation at the Windscale (UK) pile (1952–57) were investigated with respect to structure using scanning electron microscopy and composition using X-ray microanalysis and γ-, β- and α-spectrometry. Particles from Chernobyl released at a relatively high temperature have in general a compact crystalline-like structure with a relatively uniform surface layer of uranium. Aggregates consisting of small-sized (1–2 µm) spherical uranium granulates were also identified. These granulates were similar to particles identified in Norway. About 25% of the particles investigated were covered by lead, which is attributed to a release during the last stage of the accident. Particles from Windscale released under low-temperature conditions have a flake-like layered structure, varying in size (10–250 µm) and activity levels. Thus the travelling distance for these particles may be substantially longer than previously anticipated for spheres. The surfaces of these particles were also composed of uranium. The mobility of radionuclides associated with fuel particles of different structure is believed at least initially to be low when compared with those associated with condensed particles or with ionic tracers. However, mobilization of especially 90Sr due to weathering of hot particles should be expected in the years to come.

The iron-chelating agent desferrioxamine now finds extensive use in the treatment and diagnosis of aluminum-related diseases in renal patients. We review the chemistry and pharmacokinetics of desferrioxamine in chelation therapy for patients on hemodialysis.

A simple method for the determination of cesium, rubidium and scandium in soil, vegetation and animal tissues by neutron activation analysis has been developed and evaluated. The methodology was designed to enable the comparison of fallout radiocesium from Chernobyl with stable elements in radioecological studies. Sample materials are dried and homogenized, and aliquots are irradiated, in sealed polyethylene vials (ca. 0.5 cm3), for 7 hours at a reactor flux of ca. 3·1012 n·cm−2·s−1. The activation products of interest have long half-lives, and are analyzed by high resolution γ-spectrometry after a cooling time of at least 12 days. The practical lower limits for determination of Cs, Rb and Sc are 3, 7 and 0.13 μg·kg−1 for soils and 10, 20, 0.4 μ·kg−1 for biological materials, respectively. Measurement of scandium levels in vegetation allows a straightforward correction to be made for soil contamination. Various applications to soil-grass-animal ecosystems are described.

This paper is a consideration of how an agent ought to proceed who faces a moral dilemma. It completes a previous discussion of this question.1 Philosophers have discussed moral dilemmas at least since Kant's day, and never more intensively than in recent times. But the questions that they treat of differ from my questions. They ask: Can there be moral dilemmas? Does the existence of feelings of guilt and regret prove the existence of moral dilemmas? Are there irresolvable moral dilem

We have examined the possibility that selenium deficiency may underlie one or more of the following peculiarities of chronic pancreatitis in tropical as compared to temperate zones: much higher prevalence, propensity for pancreatic calculi, and high frequency of diabetes. Selenium was measured by graphite furnace atomic absorption spectrometry in sera from 20 healthy volunteers, 36 patients with chronic pancreatitis (calcific 35, diabetic 32), and 23 patients with primary forms of diabetes, from Madras, South India; results were compared with data from 41 controls and 37 patients with chronic pancreatitis (calcific 13, diabetes 8) from Manchester, North West, England. We conclude that (a) bioavailability of selenium is equally high in each geographic area; (b) decrement in serum selenium (p less than 0.001) is of a similar order in Manchester and Madras patients, which denies a connection with calculi formation or pancreatic exocrine failure (since the incidence of these two problems was substantially higher in the Madras series); and (c) selenium levels do not account for accelerated course to diabetes in tropical chronic pancreatitis.

Moral dilemmas, or moral conflicts, present a leading problem in Ethics. Ross calls them the problem of conflicting prima facie moral obligations. Lemmon calls them ‘moral dilemmas’, and Sinnott-Armstrong in his recent book discusses them thoroughly and provides extensive references to relevant literature.

Selenium status and its relationship to lowered platelet glutathione peroxidase activity was investigated in 18 subjects with aspirin (ASA)-induced asthma and in asthmatic and non-asthmatic ASA-tolerant subjects. Mean serum selenium concentration in ASA-tolerant asthmatics (1.25 microns/l; 98.5 micrograms/l) was significantly higher than that in ASA-induced asthma subjects (1.14 microns/l; 89.7 micrograms/l) and than that in healthy controls (1.15 microns/l; 91 micrograms/l). Although there was a correlation between serum selenium concentration and platelet glutathione peroxidase activity, enzyme activity was significantly lower in the ASA-induced asthma group compared to other groups even after correcting for selenium status. These results indicate that lowered platelet glutathione peroxidase activity in ASA-induced asthma is a product of both selenium availability and an unidentified syndrome-specific (possibly genetic) factor.

A major source of environmental lead, particularly in urban areas, has been from the combustion of leaded petrol. Street dust has previously been used to assess urban lead contamination, and the dust itself can also be a potential source of lead ingestion, particularly to children. The progressive reduction of lead in petrol, in recent years, would be expected to have been reflected in a reduction of lead in urban dust. We have tested this hypothesis by repeating an earlier survey of Manchester street dust and carrying out a comparable survey in Paris. Samples were collected from streets and parks, lead was extracted by digestion with concentrated nitric acid and determined by electrothermal atomic absorption spectrometry. Lead isotope ratios were measured by inductively coupled plasma mass spectrometry. Results for Manchester show that lead concentrations have fallen by about 40% (street dust averages, 941 micrograms g-1 (ppm) in 1975 down to 569 ppm in 1997). In Paris, the lead levels in street dust are much higher and significant differences were observed between types of street (not seen in Manchester). Additionally, lead levels in parks were much lower than in Manchester. Samples collected under the Eiffel Tower had very high concentrations and lead isotope ratios showed that this was unlikely to be fallout from motor vehicles but could be due to the paint used on the tower. Isotope ratios measurements also revealed that lead additives used in France and the UK come from different sources.

Exponentially growing cells cultured in medium containing bromodeoxyuridine, then exposed to UVA light in the presence of the dye Hoechst 33258, show significant levels of DNA strand breaks and base damage. This dye-bromodeoxyuridine-UVA photolysis treatment is markedly cytotoxic. We now demonstrate that exposure of cells to the agents used in photolysis leads directly to the formation of chromosome aberrations. Furthermore, we demonstrate that this photochemical treatment induces delayed chromosomal instability in clonal populations derived from single progenitor cells surviving photolysis. These results suggest that photolysis-induced DNA damage leads to chromosome rearrangements that could account for the observed cytotoxicity. Furthermore, in those cells surviving photolysis, the delayed effects of this treatment can be observed several generations after exposure and are manifested as compromised genomic integrity.

Aluminium uptake from blood into tissues of control and homozygous hypotransferrinaemic (hpx/hpx) mice, following continuous intravenous infusion of 26Al and 67Ga, has been compared with that of gallium, a proposed tracer for aluminium. 26Al uptake into tissues of control (hpx/+ and +/+) mice occurred in the order (expressed as a space): bone 464.7 ml 100 g-1; renal cortex 102.9 ml 100 g-1; liver 13.0 ml 100 g-1; spleen 8.4 ml 100 g-1 and brain 0.8 ml 100 g-1. 67Ga uptakes were similar in liver, spleen and brain, but smaller in the renal cortex and bone, at one-third and one-fifth of the values for 26Al, respectively. In the hypotransferrinaemic mice, uptake of 67Ga into all tissues was increased, especially in renal cortex (ninefold) and bone (twentyfold) as compared with the controls. Increases in 67Ga uptakes into cerebral hemisphere, cerebellum and brain stem of the hypotransferrinaemic mice were 3.8, 4.2 and 2.8 fold, respectively. 26Al uptake into tissues of the hypotransferrinaemic mice was similar to control values except in bone where it was three times greater. Pre-treatment of control animals with the anti-transferrin receptor antibody, RI7 208, enhanced 67Ga uptake in all tissues, the effect being greatest in renal cortex (tenfold) and bone (ninefold). 67Ga uptakes into cerebral hemisphere, cerebellum and brain stem in the mice pre-treated with RI7 208 were 6.4, 6 and 10 times greater than in untreated mice, respectively. No influence of antibody on 26AI uptake into mouse tissues was observed except in spleen where it was three times greater than in untreated mice. Hence, transport of aluminium and gallium into mouse tissues is not similar under all conditions. Non-transferrin mediated transport of each metal can occur into all tissues, especially in renal cortex and bone, where gallium may be a suitable marker for aluminium.

In this study of the toxicokinetics of aluminum we have examined some of the fundamental issues that currently define our understanding of the toxicology of aluminum in humans. There is a vast literature on this subject, and it was not our aim to review this literature but to use it to develop our understanding of the toxicokinetics of aluminum and to identify critical and unresolved issues related to its toxicity. In undertaking this task we have chosen to define the term toxicokinetics to encompass those factors that influence both the lability of aluminum in a body and the sites at which aluminum is known to accumulate, with or without consequent biological effect. We have approached our objective from the classical pharmacological approach of ADME: the absorption, distribution, metabolism, and excretion of aluminum. This approach was successful in identifying several key deficits in our understanding of aluminum toxicokinetics. For example, we need to determine the mechanisms by which aluminum crosses epithelia, such as those of the gastrointestinal tract and the central nervous system, and how these mechanisms influence both the subsequent transport and fate of the absorbed aluminum and the concomitant nature and severity of the biological response to the accumulation of aluminum. Our hope in highlighting these unresolved issues (summarized in Table 1) is that they will be addressed in future research.

The reported geographical association between Alzheimer's disease and levels of aluminium (Al) in water supplies may reflect the inverse relation between Al and silicon (Si) concentrations in water, and the potential for Si to reduce the bioavailability of the metal. We tested this hypothesis using isotopic 26Al tracer administered orally to five healthy volunteers in the presence and absence of Si. Dissolved Si, at a concentration found in some water supplies (100 mumol/L), reduced the peak plasma 26Al concentration to 15% of the value obtained in the absence of Si. The results indicate that dissolved Si is an important factor in limiting the absorption of dietary Al.

Lead toxicity is known to be subject to individual susceptibility. This study compares two lead-exposed subjects, one (A; blood Pb 1800 micrograms/L) who remained totally asymptomatic, the other (B; blood Pb 1610 micrograms/L) who showed symptoms of toxicity. We have assessed the speciation of lead in the intra-erythrocyte proteins in these patients and have examined its significance in relation to clinical toxicity. Chromatographic separations of erythrocyte haemolysates from these patients showed a metallothionein-like lead containing protein. It was demonstrated that in patient A, most (approximately 70%) of the erythrocyte lead was associated with this protein, whilst in patient B the protein only contained about 20% of the total lead, with significant amounts bound to high molecular weight proteins, including Hb. Further purification of this protein from each patient showed it to contain a number of constituents, one in particular being the major lead-binding species. This component was more abundant in patient A and, relative to patient B, contained a higher proportion of lead. These results suggest that this protein may act to sequester lead into a non-bioavailable form, hence protecting the body from lead toxicity as with patient A.

The x-ray structure of the EcoRI endonuclease-DNA complex (3) suggests that hydrogen bonds between amino acids, glutamic acid 144, arginine 145, and arginine 200, and major groove base moieties are the molecular determinants of specificity. We have investigated residue 144 using aspartate and glutamine substitutions introduced by site-directed mutagenesis. Substitution with glutamine results in a null phenotype (at least a 2000-fold reduction in activity). On the other hand, the aspartic acid mutant (ED144) retained in vivo activity. Substrate binding and catalytic studies were done with purified ED144 enzyme. The affinity of the ED144 enzyme for the canonical sequence 5'-GAATTC-3' is about 340-fold less than the wild-type (WT) enzyme, while its affinity for nonspecific DNA is about 50 times greater. The ED144 enzyme cleaves one strand in the EcoRI site in plasmid pBR322 with a kcat/Km similar to WT. In contrast to the WT enzyme, the ED144 enzyme dissociates after the first strand cleavage. Partitioning between cleavage and dissociation at the first and second cleavage steps for the ED144 enzyme is extremely salt-sensitive. The altered partitioning results largely from a destabilization of the enzyme-DNA complex, particularly the enzyme-nicked DNA complex, with only small changes in the respective cleavage rates. The hydrogen bonds of Glu-144 are critical, they appear to act cooperatively with other specificity contacts to stabilize the enzyme-DNA complex.

Esophageal disease has been reported in 70% to 90% of patients with scleroderma, of whom nearly 50% will have reflux esophagitis. The combined motility disorder of low LES pressure and aperistalsis of the esophageal body makes scleroderma patients especially susceptible to severe gastroesophageal reflux disease (GERD). Symptomatic GERD is a common problem in pregnancy, affecting 30% to 50% of women. Hormonal effects of estrogen and progesterone likely promote GERD by compromising LES function. Fortunately, the problem is usually relieved with delivery of the baby. Although difficult to quantitate, the reflux of both acid and especially alkaline material may be a common sequela of many types of gastric surgery. Medical therapy binding bile salts usually does not bring relief. The Rouxen-Y biliary diversion operation is the best solution for this problem. GERD complicates the treatment of achalasia after 10% of Heller myotomies and 2% of pneumatic dilatations. Nearly 50% of patients with the Zollinger-Ellison syndrome have esophagitis, which may be more difficult to treat than their ulcer disease.