The influence of the occlusal vertical dimension on masticatory muscle activities and hyoid bone position in complete denture wearers.

PURPOSE: Long lasting usage of complete dentures causes lower occlusal vertical dimension, producing potential detrimental consequences. The aim of this study was to investigate how changes in vertical dimension during denture exchange affect muscular activity and hyoid bone position. MATERIAL AND METHODS: Twenty-five edentulous, otherwise healthy patients (14 females, 11 males) aged 70.5 +/- 9 years, wearing their dentures over 5 years but no longer than 15 years (mean 9.8 +/- 5.2), were studied. New dentures were fabricated and the occlusal vertical dimension was recorded on cephalometric radiographs. Cephalograms were analyzed according to Ricketts. The relation of the hyoid bone position to the mandible was estimated. An evaluation of muscular activity was performed using the Biopak Electromyography Recording System synchronized with the T-Scan II Occlusal Analysis System. RESULTS: The occlusal vertical dimension was higher with the new dentures compared with the old dentures. The transition to new dentures was accompanied by a change of the vertical position of the hyoid bone. Digastric muscle activity was lower with the new dentures in comparison with the old dentures. CONCLUSIONS: Increase of the occlusal vertical dimension in complete denture wearers affects the hyoid bone position and masticatory muscle activity.

Ca2+-dependent and Ca2+-independent excretion modes of salicylic acid in tobacco cell suspension culture.

14C-salicylic acid (SA) was used to monitor SA metabolism and its regulation in tobacco cell suspension culture. Two SA concentrations (20 microM and 200 microM) were used for comparison. SA was quickly taken up in both treatments, and the 200 microM-treated cells absorbed approximately 15 times that of 20 microM-treated cells within 5 min. More than 85% and 50% of the absorbed SA were excreted in free form to the culture medium within 5 h from cells treated with 200 microM and 20 microM SA, respectively. SA excretion was significantly inhibited by EGTA and the inhibition could be reversed by the addition of exogenous Ca2+ to the culture medium in the 200 microM SA treatment. However, EGTA had little or no effect on SA excretion in the 20 microM SA treatment. The data suggest that tobacco suspension-cultured cells may contain both Ca2+-dependent and Ca2+-independent pathways for SA excretion. Reduced glutathione (an active oxygen species scavenger), staurosporine (a protein kinase inhibitor), and cycloheximide (an inhibitor of de novo protein synthesis) also blocked intracellular SA excretion to the culture medium in the 200 microM but not in the 20 microM SA treatment. These data support the existence of alternative SA excretion pathways in tobacco suspension-cultured cells. Tobacco cells may use both Ca2+-dependent and Ca2+-independent excretion pathways to cope with different intracellular SA status, and the pathway influenced by EGTA, reduced glutathione, staurosporine, and cycloheximide is activated by SA at 200 microM, but not at 20 microM.

Mutation analysis of the DCX gene and genotype/phenotype correlation in subcortical band heterotopia.

Subcortical band heterotopia (SBH) comprises part of a spectrum of phenotypes associated with classical lissencephaly (LIS). LIS and SBH are caused by alterations in at least two genes: LIS1 (PAFAH1B1) at 17p13.3 and DCX (doublecortin) at Xq22.3-q23. DCX mutations predominantly cause LIS in hemizygous males and SBH in heterozygous females, and we have evaluated several families with LIS male and SBH female siblings. In this study, we performed detailed DCX mutation analysis and genotype-phenotype correlation in a large cohort with typical SBH. We screened 26 sporadic SBH females and 11 LIS/SBH families for DCX mutations by direct sequencing. We found 29 mutations in 22 sporadic patients and 11 pedigrees, including five deletions, four nonsense mutations, 19 missense mutations and one splice donor site mutation. The DCX mutation prevalence was 84.6% (22 of 26) in sporadic SBH patients and 100% (11 of 11) in SBH pedigrees. Maternal germline mosaicism was found in one family. Significant differences in genotype were found in relation to band thickness and familial vs sporadic status.

The location and type of mutation predict malformation severity in isolated lissencephaly caused by abnormalities within the LIS1 gene.

Lissencephaly is a cortical malformation secondary to impaired neuronal migration resulting in mental retardation, epilepsy and motor impairment. It shows a severity spectrum from agyria with a severely thickened cortex to posterior band heterotopia only. The LIS1 gene on 17p13.3 encodes a 45 kDa protein named PAFAH1B1 containing seven WD40 repeats. This protein is required for optimal neuronal migration by two proposed mechanisms: as a microtubule-associated protein and as one subunit of the enzyme platelet-activating factor acetylhydrolase. Approximately 65% of patients with isolated lissencephaly sequence (ILS) show intragenic mutations or deletions of the LIS1 gene. We analyzed 29 non-deletion ILS patients carrying a mutation of LIS1 and we report 15 novel mutations. Patients with missense mutations had a milder lissencephaly grade compared with those with mutations leading to a shortened or truncated protein (P = 0.022). Early truncation/deletion mutations in the putative microtubule-binding domain resulted in a more severe lissencephaly than later truncation/deletion mutations (P < 0.001). Our results suggest that the lissencephaly severity in ILS caused by LIS1 mutations may be predicted by the type and location of the mutation. Using a spectrum of ILS patients, we confirm the importance of specific WD40 repeats and a putative microtubule-binding domain for PAFAH1B1 function. We suggest that the small number of missense mutations identified may be due to underdiagnosis of milder phenotypes and hypothesize that the greater lissencephaly severity seen in Miller-Dieker syndrome may be secondary to the loss of another cortical development gene in the deletion of 17p13.3.

An optimized set of human telomere clones for studying telomere integrity and architecture.

Telomere-specific clones are a valuable resource for the characterization of chromosomal rearrangements. We previously reported a first-generation set of human telomere probes consisting of 34 genomic clones, which were a known distance from the end of the chromosome ( approximately 300 kb), and 7 clones corresponding to the most distal markers on the integrated genetic/physical map (1p, 5p, 6p, 9p, 12p, 15q, and 20q). Subsequently, this resource has been optimized and completed: the size of the genomic clones has been expanded to a target size of 100-200 kb, which is optimal for use in genome-scanning methodologies, and additional probes for the remaining seven telomeres have been identified. For each clone we give an associated mapped sequence-tagged site and provide distances from the telomere estimated using a combination of fiberFISH, interphase FISH, sequence analysis, and radiation-hybrid mapping. This updated set of telomeric clones is an invaluable resource for clinical diagnosis and represents an important contribution to genetic and physical mapping efforts aimed at telomeric regions.

Subcortical band heterotopia in rare affected males can be caused by missense mutations in DCX (XLIS) or LIS1.

Subcortical band heterotopia (SBH) are bilateral and symmetric ribbons of gray matter found in the central white matter between the cortex and the ventricular surface, which comprises the less severe end of the lissencephaly (agyria-pachygyria-band) spectrum of malformations. Mutations in DCX (also known as XLIS ) have previously been described in females with SBH. We have now identified mutations in either the DCX or LIS1 gene in three of 11 boys studied, demonstrating for the first time that mutations of either DCX or LIS1 can cause SBH or mixed pachygyria-SBH (PCH-SBH) in males. All three changes detected are missense mutations, predicted to be of germline origin. They include a missense mutation in exon 4 of DCX in a boy with PCH-SBH (R78H), a different missense mutation in exon 4 of DCX in a boy with mild SBH and in his mildly affected mother (R89G) and a missense mutation in exon 6 of LIS1 in a boy with SBH (S169P). The missense mutations probably account for the less severe brain malformations, although other patients with missense mutations in the same exons have had diffuse lissencephaly. Therefore, it appears likely that the effect of the specific amino acid change on the protein determines the severity of the phenotype, with some mutations enabling residual protein function and allowing normal migration in a larger proportion of neurons. However, we expect that somatic mosaic mutations of both LIS1 and DCX will also prove to be an important mechanism in causing SBH in males.

Purification and characterization of an acidic beta-1,3-glucanase from cucumber and its relationship to systemic disease resistance induced by Colletotrichum lagenarium and tobacco necrosis virus.

An acidic beta-1,3-glucanase was detected in cucumber leaves inoculated with either Colletotrichum lagenarium or tobacco necrosis virus (TNV) as well as in the leaves above those inoculated with the pathogens. The enzyme is extracellular and migrates in native polyacrylamide gel electrophoresis (PAGE) together with a Class III chitinase, a bifunctional chitinase/lysozyme. The beta-1,3-glucanase was separated by ultra-narrow pH range IEF-PAGE or by SDS_PAGE and was purified to apparent homogeneity. Only one isoform of the enzyme was detected. Its apparent molecular mass in 38 kDa as estimated by SDS-PAGE, its isoelectric point is 3.6 and the specific activity is approximately 26 micromol glucose equivalents liberated from laminarin min(-1)mg(-1) protein. Partial amino acid (five peptide fragments with a total of 65 amino acids) sequencing of the beta-1,3-glucanase revealed similarities of 49% to 72% to sequences of published beta-1,3-glucanases from tobacco, tomato, soybean, barley, and rice plants. A time course study indicated that the increase of the beta-1,3-glucanase activity was associated with induced resistance against C. lagenarium. The implications of these results to coordinate defense responses in plant-microbe interactions are discussed.

Management of acute myocardial infarction in the postanesthesia care unit.

Surgery and anesthesia can compromise the delicate homeostasis between myocardial oxygen supply and demand in the patient with preexisting cardiac disease. This article identifies those patients at risk for developing a myocardial infarction. It also focuses on patient care assessment of the patient with chest pain in the PACU, and the implementation measures to manage chest pain and limit infarction size.

Compounds from plants that regulate or participate in disease resistance.

Disease resistance is multifactorial. The response phase includes: synthesis of phytoalexins, i.e. low molecular weight antimicrobial compounds which accumulate at sites of infection; systemically produced enzymes which degrade pathogens, e.g. chitinases, beta-1,3-glucanases and proteases; systemically produced enzymes which generate antimicrobial compounds and protective biopolymers, e.g. peroxidases and phenoloxidases; biopolymers which restrict the spread of pathogens, e.g. hydroxyproline-rich glycoproteins, lignin, callose; and compounds which regulate the induction and/or activity of the defence compounds, e.g. elicitors of plant and microbial origin, immunity signals from immunized plants and compounds which release immunity signals. Disease resistance in plants is not determined by the presence or absence of genes for resistance mechanisms, it is determined by the speed and degree of gene expression and the activity of the gene products. It is likely, therefore, that all plants have the genetic potential for resistance. This potential can be expressed systemically (immunization) after restricted inoculation with pathogens, attenuated pathogens or selected non-pathogens, or treatment with chemical substances that are produced by immunized plants or chemicals which release such signals. Immunization is effective against diseases caused by fungi, bacteria and viruses, and it has been successfully tested in the laboratory and field. Advances in science have provided information and technology to enhance resistance to plant pests. Pesticides are part of this technology, but they also contribute to a complex world problem which threatens our environment and hence our survival. The future will see the restriction of pesticide use and a greater reliance on resistant plants generated using immunization and other biological control technologies, genetic engineering and classical plant breeding. However, as with past and current technology, we will have created unique problems. The survival of our planet depends upon anticipating these problems and meeting the challenge of their solution.

Direct detection of beta-1,3-glucanase isozymes on polyacrylamide electrophoresis and isoelectrofocusing gels.

A procedure to assay isozymes of beta-1,3-glucanase directly on polyacrylamide gel electrophoresis (PAGE) and isoelectrofocusing (IEF) gels by using 2,3,5-triphenyltetrazolium chloride is described. The reagent reacts with reducing sugars released by beta-1,3-glucanases from the substrate laminarin. Acidic and neutral isozymes of beta-1,3-glucanase were detected and quantified on 17.5% native PAGE gels run with an anodic buffer system. A significant linear relationship (alpha = less than 0.01, R = 0.991) was observed between amounts of beta-1,3-glucanase loaded and intensity of bands stained with the reagent on native PAGE gels. A full isozyme pattern was obtained on 7.5% IEF gels with a pH range of 3.5-9.5. The IEF gels were heated in a microwave oven during the staining process to minimize diffusion.

Inhibition by Salicylhydroxamic Acid, BW755C, Eicosatetraynoic Acid, and Disulfiram of Hypersensitive Resistance Elicited by Arachidonic Acid or Poly-l-Lysine in Potato Tuber.

The hypothesis that arachidonic acid (AA) induction of sesquiterpene accumulation and browning in potato (Solanum tuberosum) is mediated by a lipoxygenase metabolite of AA was tested using lipoxygenase inhibitors. Salicylhydroxamic acid (SHAM) and 3-amino-1-(3-trifluoromethylphenyl)-2-pyrazoline hydrochloride (BW755C) delayed the response to AA. Inhibition by eicosatetraynoic acid (ETYA) was more persistent. These results are consistent with previous reports that SHAM and BW755C are reversible inhibitors of lipoxygenase and easily oxidized by potato while ETYA acts as an irreversible inhibitor. Disulfiram (tetraethylthiuram disulfide) also inhibited AA elicitor activity. SHAM was most effective if applied at the time of AA treatment, having no effect if applied 6 hours afterward. SHAM was effective in the presence of MES or MOPS buffers but not in acetate-buffered or unbuffered solutions; neither BW755C nor ETYA exhibited this restriction. However, SHAM, BW755C, and ETYA also were inhibitors of browning and sesquiterpene accumulation elicited in potato by poly-l-lysine, which, unlike AA, is not a lipoxygenase substrate. SHAM effectiveness also was restricted to 6 hours after treatment with poly-l-lysine. While the results with AA support a role for lipoxygenase, those with poly-l-lysine may be evidence that these compounds are having other effects in potato tissue.

A role for ca in the elicitation of rishitin and lubimin accumulation in potato tuber tissue.

Calcium and strontium ions enhanced rishitin but not lubimin accumulation in tuber tissue of potato (Solanum tuberosum cv Kennebec) treated with arachidonic acid (AA). The same cations in the presence of poly-l-lysine (PL) enhanced the accumulation of lubimin more than rishitin. In contrast, Mg(2+) did not affect AA-elicited rishitin and lubimin accumulation and inhibited the accumulation of these compounds following application of PL. AA-elicited potato tuber tissue remained sensitive to the stimulatory effects of Ca(2+) and Sr(2+) up to 24 h after application of AA, but PL-elicited tuber tissue was sensitive to Ca(2+) and Sr(2+) for only 6 hours after PL application. Ethyleneglycol-bis (beta-aminoethyl ether)-N,N'-tetraacetic acid and La(3+) both inhibited rishitin and lubimin accumulation elicited by AA. The inhibition by either agent was overcome by the addition of Ca(2+). Calcium was more effective in overcoming lanthanum inhibition when applied simultaneously than when applied 12 hours later. Lanthanum was only effective in inhibiting rishitin and lubimin accumulation when applied within 3 hours of the application of AA. Inhibition of phytoalexin accumulation was greater when La(3+) was applied simultaneously with AA compared to La(3+) application after AA application to discs. These observations suggest that the mobilization of calcium may play a central regulatory role in the expression of phytoalexin accumulation following elicitation in potato tissue.

Isolation and Characterization of an Elicitor of Necrosis Isolated from Intercellular Fluids of Compatible Interactions of Cladosporium fulvum (Syn. Fulvia fulva) and Tomato.

Intercellular fluids of compatible race-cultivar interactions of Cladosporium fulvum and tomato contain specific elicitors of necrosis. These elicitors which are of fungal origin induce chlorosis and necrosis in resistant but not in susceptible plants. With the tomato cultivar Sonatine (carrying resistance gene Cf9, resistant to the fungal races 0, 4, 5, 2, 2.4, and 2.4.5 but susceptible to race 2.4.5.9) as the test plant for assaying necrosis-inducing activity, we isolated and partially characterized an elicitor of necrosis on this cultivar. The elicitor bound to CM-Sephadex but not to DEAE-Sephadex; it was stable to heat (10 minutes at 100 degrees C), HCl (0.01 normal), NaOH (0.01 normal), and NaIO(4) (0.02 molar), sensitive to pronase and protease (from Bacillus polymyxa) but not to other proteases such as alpha-chymotrypsin and trypsin. After electrophoresis of partially purified elicitor preparations under low pH conditions, the necrosis-inducing activity was association with a peptide with an apparent molecular weight of 5500. Races 0, 4, 5, 2.4, and 2.4.5 but not race 2.4.5.9 produced this elicitor in high yields. The elicitor is probably a product of avirulence gene A9 which is present in all races except in race 2.4.5.9 and induces necrosis in cultivars carrying resistance gene Cf9.

Phytoalexins.

Plants respond to infection by accumulating low-molecular-weight antimicrobial stress metabolites called phytoalexins. The phytoalexins are generally lipophilic substances that are products of a plant's secondary metabolism, and they often accumulate at infection sites to concentrations which are inhibitory to the development of fungi and bacteria. Resistance and susceptibility in plants are not determined by the presence or absence of genetic information for resistance mechanisms, including biosynthetic pathways for phytoalexin synthesis, but, rather, by the speed with which the information is expressed, the activity of the gene products, and the magnitude of the resistance response. Unlike the antibody-antigen component of the immune system in animals, low specificity is the general rule for the induction of phytoalexin accumulation and their activity against microorganisms. Annual plants can be systemically immunized against diseases caused by fungi, bacteria, and viruses by restricted infection with the pathogens, avirulent forms of pathogens, or compounds formed in immunized plants. Immunization induces plants to respond rapidly to infection with a multicomponent resistant response. The biosynthesis and accumulation of phytoalexins is one component of this resistant response. Resistance may be elicited by components in the walls and cell surfaces of fungi and bacteria and by compounds liberated from cells, their walls, or surfaces. Resistance can be enhanced or suppressed by products produced by the pathogen, the host, or by their interaction. The successful pathogen avoids recognition by the plant as nonself, suppresses the resistance response, or detoxifies its products. The actors in this play for survival on the metabolic level include the shikimate, acetate-malonate, and acetate-mevalonate pathways; glucans; oligogalacturonates; glycoproteins; lipopolysaccharides; and poly-unsaturated fatty acids. The play is directed by the genetic information of host and pathogen, and this direction is at the level of recognition and not by the presence or absence of mechanisms to contain the development of infectious agents.

Arachidonic acid-related elicitors of the hypersensitive response in potato and enhancement of their activities by glucans from Phytophthora infestans (Mont.) deBary.

The dose response for elicitation of the hypersensitive reaction in potato tuber discs by arachidonic acid (AA) suggested saturation at higher concentrations. Glucans from Phytophthora infestans, inactive themselves as elicitors of the hypersensitive reaction, enhanced sesquiterpene accumulation and hypersensitive browning elicited by AA. Significant activity (seven times control values) was observed with 33 pmol AA/3.0-cm potato disc in the presence of glucans. Glucans did not affect accumulation of steroid glycoalkaloids, influence the timing or relative amounts of sesquiterpenes which accumulate, or affect recovery of AA added to potato discs. Glucans enhanced activity whether added to potato discs 18 h prior to AA, at the same time as AA, or 18 h after AA. Elicitor activity in the presence of glucans was evident with 20-carbon unsaturated fatty acids that had little or no elicitor activity in the absence of glucans. The position of double bonds had considerable influence on the specific activity of unsaturated fatty acids. The most active had a minimum of three double bonds in a methylene-interrupted series beginning with delta 5, e.g., delta 5,8,11. A delta 5 double bond conferred significant activity even if it was not part of a methylene-interrupted series. The 20-carbon chain length appeared optimal for elicitor activity. The 22-carbon chain acids had low activity, and 16- and 18-carbon acids were inactive. A free carboxyl group or easily transesterified group appeared necessary for activity. Arachidonyl alcohol had very low activity and arachidonyl cyanide was inactive. AA-containing phosphatidylcholine, lysophosphatidylcholine and monoacylglycerol were at least as active as free AA, AA-containing diacylglycerols were slightly less active than free AA, and triarachidonyl glycerol was inactive.

Phytoalexins and disease resistance mechanisms from a perspective of evolution and adaptation.

Plants respond to cellular injury and infection by accumulating low molecular weight antimicrobial stress metabolites called phytoalexins. The accumulation of phytoalexins, together with lignification, suberization, callose formation and the production of agglutinins and inhibitors of extracellular microbial hydrolases, appears to be part of a multi-component response mechanism associated with disease resistance and wound repair. Compared to the antibody-antigen response in animals, the phytoalexin response in plants has low specificity for induction and activity of the phytoalexins. Plants also contain preformed antimicrobial chemical and physical barriers to infection in their external tissues. The successful pathogen has evolved to cope with preformed inhibitors and barriers and either avoids eliciting the response mechanism, or suppresses the mechanism, or detoxifies its antimicrobial components. Annual plants can be systemically immunized against diseases caused by viruses, bacteria and fungi by limited infection with any one of the respective organisms. As with animals, disease resistance in plants depends on the rate and magnitude of response rather than on the ability to respond. The genetic information for disease resistance is found in all organisms, and disease resistance is the rule in nature. The interactions of plants with microorganisms in their environment are nature's example of diplomacy--compromise, adjustment to change and avoidance of deadly conflict.

Factors affecting the elicitation of sesquiterpenoid phytoalexin accumulation by eicosapentaenoic and arachidonic acids in potato.

Eicosapentaenoic and arachidonic acids in extracts of Phytophthora infestans mycelium were identified as the most active elicitors of sesquiterpenoid phytoalexin accumulation in potato tuber slices. These fatty acids were found free or esterified in all fractions with elicitor activity including cell wall preparations. Yeast lipase released a major portion of eicosapentaenoic and arachidonic acids from lyophilized mycelium. Concentration response curves comparing the elicitor activity of the polyunsaturated fatty acids to a cell-free sonicate of P. infestans mycelium indicated that the elicitor activity of the sonicated mycelium exceeded that which would be obtained by the amount of eicosapentaenoic and arachidonic acids (free and esterified) present in the mycelium. Upon acid hydrolysis of lyophilized mycelium, elicitor activity was obtained only from the fatty acid fraction. However, the fatty acids accounted for only 21% of the activity of the unhydrolyzed mycelium and the residue did not enhance their activity. Centrifugation of the hydrolysate, obtained from lyophilized mycelium treated with 2n NaOH, 1 molarity NaBH(4) at 100 degrees C, yielded a supernatant fraction with little or no elicitor activity. Addition of this material to the fatty acids restored the activity to that which was present in the unhydrolyzed mycelium. The results indicate that the elicitor activity of the unsaturated fatty acids is enhanced by heat and base-stable factors in the mycelium.

Eicosapentaenoic and Arachidonic Acids from Phytophthora infestans Elicit Fungitoxic Sesquiterpenes in the Potato.

Mycelial extracts from Phytophthora infestans caused necrosis and elicited the accumulation of antimicrobial stress metabolites in potato tubers. A portion of the material with elicitor activity could be extracted from the mycelium by a mixture of chloroform and methanol. The most active elicitors of stress metabolites in these extracts were eicosapentaenoic and arachidonic acids. These fatty acids were found in either free or esterified form in all active fractions of the mycelial extracts.