Indicators of dental anxiety in children just prior to treatment.

OBJECTIVES: We evaluated the relationship between child dental anxiety and selected child and parental characteristics. STUDY DESIGN: Children and their parents were interviewed at the New York University, College of Dentistry, Pediatric Dentistry Clinic. The Children's Fear Survey Schedule - Dental Subscale (CFSS-DS) evaluated child self-reported anxiety; the Modified Dental Anxiety Scale (MDAS) measured self-reported parental anxiety when the parent received dental treatment. RESULTS: Ninety-three children and their parents completed the questionnaires. Mean CFSS-DS scores were higher for girls than boys (32.5 vs. 26.3, p=0.003) and for children whose accompanying parents had MDAS scores of 11+ vs. ≥ 11 (32.8 vs. 26.6, p=0.001). There was little difference in mean CFSS-DS scores among those aged 6-10 yrs. vs. 11-14 yrs. (30.1 vs. 29.3). Significant correlations were found between CFSS-DS and both gender (Spearman's rho, rs=0.31) and MDAS scores (rs=0.33), but not between CFSS-DS and child age (rs=-0.05). Controlling simultaneously for gender, MDAS score and child age, a high CFSS-DS score (38+ vs. ≥ 38) was positively associated with girls (ORadj=3.76, 95% CI: 1.13-12.54) and an MDAS score of ≤ 15 vs. ≥ 11 (ORadj=2.50, 0.73-8.54), but weakly and inversely associated with age (ORadj=0.80, 0.25-2.52). CONCLUSION: Child gender and parental anxiety are indicators of child dental anxiety.

In situ observation of fluoride-ion-induced hydroxyapatite-collagen detachment on bone fracture surfaces by atomic force microscopy.

The topography of freshly fractured bovine and human bone surfaces was determined by the use of atomic force microscopy (AFM). Fracture surfaces from both kinds of samples exhibited complex landscapes formed by hydroxyapatite mineral platelets with lateral dimensions ranging from ∼90 nm × 60 nm to ∼20 nm × 20 nm. Novel AFM techniques were used to study these fracture surfaces during various chemical treatments. Significant topographical changes were observed following exposure to aqueous solutions of ethylenediaminetetraacetic acid (EDTA) or highly concentrated sodium fluoride (NaF). Both treatments resulted in the apparent loss of the hydroxyapatite mineral platelets on a timescale of a few seconds. Collagen fibrils situated beneath the overlying mineral platelets were clearly exposed and could be resolved with high spatial resolution in the acquired AFM images. Time-dependent mass loss experiments revealed that the applied agents (NaF or EDTA) had very different resulting effects. Despite the fact that the two treatments exhibited nearly identical results following examination by AFM, bulk bone samples treated with EDTA exhibited a ∼70% mass loss after 72 h, whereas for the NaF-treated samples, the mass loss was only of the order of ∼10%. These results support those obtained from previous mechanical testing experiments, suggesting that enhanced formation of superficial fluoroapatite dramatically weakens the protein-hydroxyapatite interfaces. Additionally, we discovered that treatment with aqueous solutions of NaF resulted in the effective extraction of noncollagenous proteins from bone powder.

Atomic force microscopy study of living diatoms in ambient conditions.

We present the first in vivo study of diatoms using atomic force microscopy (AFM). Three chain-forming, benthic freshwater species -Eunotia sudetica, Navicula seminulum and a yet unidentified species - are directly imaged while growing on glass slides. Using the AFM, we imaged the topography of the diatom frustules at the nanometre range scale and we determined the thickness of the organic case enveloping the siliceous skeleton of the cell (10 nm). Imaging proved to be stable for several hours, thereby offering the possibility to study long-term dynamic changes, such as biomineralization or cell movement, as they occur. We also focused on the natural adhesives produced by these unicellular organisms to adhere to other cells or the substratum. Most man-made adhesives fail in wet conditions, owing to chemical modification of the adhesive or its substrate. Diatoms produce adhesives that are extremely strong and robust both in fresh- and in seawater environments. Our phase-imaging and force-pulling experiments reveal the characteristics of these natural adhesives that might be of use in designing man-made analogues that function in wet environments. Engineering stable underwater adhesives currently poses a major technical challenge.

Bone indentation recovery time correlates with bond reforming time.

Despite centuries of work, dating back to Galileo, the molecular basis of bone's toughness and strength remains largely a mystery. A great deal is known about bone microsctructure and the microcracks that are precursors to its fracture, but little is known about the basic mechanism for dissipating the energy of an impact to keep the bone from fracturing. Bone is a nanocomposite of hydroxyapatite crystals and an organic matrix. Because rigid crystals such as the hydroxyapatite crystals cannot dissipate much energy, the organic matrix, which is mainly collagen, must be involved. A reduction in the number of collagen cross links has been associated with reduced bone strength and collagen is molecularly elongated ('pulled') when bovine tendon is strained. Using an atomic force microscope, a molecular mechanistic origin for the remarkable toughness of another biocomposite material, abalone nacre, has been found. Here we report that bone, like abalone nacre, contains polymers with 'sacrificial bonds' that both protect the polymer backbone and dissipate energy. The time needed for these sacrificial bonds to reform after pulling correlates with the time needed for bone to recover its toughness as measured by atomic force microscope indentation testing. We suggest that the sacrificial bonds found within or between collagen molecules may be partially responsible for the toughness of bone.

A novel fluorescent silica tracer for biological silicification studies.

BACKGROUND: Biological silica production has drawn intense attention and several molecules involved in biosilicification have been identified. Cellular mechanisms, however, remain unknown mainly due to the lack of probes required for obtaining information on live specimens. RESULTS: The fluorescence spectra of the compound 2-(4-pyridyl)-5-((4-(2-dimethylaminoethylaminocarbamoyl)methoxy)phenyl)oxazole (PDMPO) are affected by the presence of >3.2 mM silicic acid. Increase in intensity and shift in the fluorescence coincide with the polymerization of Si. The unique PDMPO-silica fluorescence is explored here to visualize Si deposition in living diatoms. The fluorophore is selectively incorporated and co-deposited with Si into the newly synthesized frustules (the outer silica shells) showing an intense green fluorescence. CONCLUSIONS: We suggest that a fluorescence shift is due to an interaction between PDMPO and polymeric silicic acid. PDMPO is an excellent probe for imaging newly deposited silica in living cells and has also a potential for a wide range of applications in various Si-related disciplines, including biology of living organisms as diatoms, sponges, and higher plants, clinical research (e.g. lung fibrosis and cancer, bone development, artificial bone implantation), and chemistry and physics of materials research.

Annual per capita apparent consumption of tobacco products in the United States: 1900-1990.

BACKGROUND: Analyses of secular trends in tobacco consumption can provide evidence of potential tobacco-disease relationships and have utility in terms of public health projections and policy. The purpose of this project was to provide a unique set of continuous apparent tobacco product consumption estimates for the United States over the period spanning 1900 through 1990. METHODS: Two U.S. Department of Agriculture data sources provide information on long-term apparent tobacco consumption in the United States; however, differences exist between these data sets. The consumption estimates in these reports were adjusted to a common population base. A 9-year overlap of the data sets was then used to calibrate one data series to the other using inverse regression. Predicted tobacco consumption estimates for the years 1900 through 1944 were then combined with the adjusted 1945-1990 data. RESULTS: Inverse regression showed a strong linear relationship between the two U.S. Department of Agriculture summaries for each tobacco product during the 9-year overlap period. A continuous set of annual per capita tobacco consumption estimates is reported by product for the United States. CONCLUSIONS: The two U.S. Department of Agriculture reports can be combined to provide a history of tobacco product consumption in the United States over the period 1900-1990.

Direct observation of the transition from calcite to aragonite growth as induced by abalone shell proteins.

The mixture of EDTA-soluble proteins found in abalone nacre are known to cause the nucleation and growth of aragonite on calcite seed crystals in supersaturated solutions of calcium carbonate. Past atomic force microscope studies of the interaction of these proteins with calcite crystals did not observe this transition because no information about the crystal polymorph on the surface was obtained. Here we have used the atomic force microscope to directly observe changes in the atomic lattice on a calcite seed crystal after the introduction of abalone shell proteins. The observed changes are consistent with a transition to (001) aragonite growth on a (1014) calcite surface.

Food group intake and the risk of oral epithelial dysplasia in a United States population.

OBJECTIVES: Oral epithelial dysplasia (OED) is a histopathologic diagnosis associated with an increased risk of oral cancer. The paper explores the relationship between OED risk and food group intake. METHODS: In this case-control study, incident cases of OED were identified through two oral pathology laboratories. Controls, pair-matched 1:1 to cases on age (+/- 5 years), gender, appointment date (+/- 1 year), and surgeon, were identified through the office in which the respective case was biopsied. Exposure data were obtained via a telephone interview and mailed food-frequency questionnaire. Conditional logistic regression was used to obtain odds ratio point estimates. RESULTS: Based upon 87 matched pairs and after controlling for smoking, drinking, and other potential covariates there was an apparent inverse relationship between OED risk and the consumption of fruits and vegetables, with the intake of these foods being associated with a strong attenuating effect among smokers. OED risk decreased with increased poultry consumption, but increased modestly with bread/cereal and dairy food intake. CONCLUSIONS: This investigation provides evidence that some aspects of diet may be associated with the risk of OED. It also suggests that in oral carcinogenesis the role of diet is not simply one of a late effect.

Head and neck in situ carcinoma: incidence, trends, and survival.

This analysis describes the epidemiology of in situ head and neck carcinomas (anatomic sites of lip, oral cavity, pharynx, larynx) about which there is limited knowledge. Data were derived from nine population-based cancer registries participating in the National Cancer Institute's Surveillance, Epidemiology and End Results (SEER) Program. SEER annual age-adjusted incidence rates for in situ head and neck carcinomas increased from 6.33/1,000,000 person-years (PY) in 1976 to 8.04/1,000,000 PY in 1995 [percent change (PC)=35%, P<0. 001]. From 1976 to 1995 age-adjusted changes in incidence by anatomic site ranged from 53% PC (larynx) to -11% PC (lip) (both P<0. 005). Incidence and survival associated with in situ head and neck carcinomas varied by anatomic site, age, sex, and race and did so in a pattern similar to that seen for invasive carcinomas of this region. However, the climbing incidence of in situ carcinoma, which may be related to increased surveillance, contrasts sharply with the declining incidence of invasive carcinoma.

Biomimetic synthesis of ordered silica structures mediated by block copolypeptides.

In biological systems such as diatoms and sponges, the formation of solid silica structures with precisely controlled morphologies is directed by proteins and polysaccharides and occurs in water at neutral pH and ambient temperature. Laboratory methods, in contrast, have to rely on extreme pH conditions and/or surfactants to induce the condensation of silica precursors into specific morphologies or patterned structures. This contrast in processing conditions and the growing demand for benign synthesis methods that minimize adverse environmental effects have spurred much interest in biomimetic approaches in materials science. The recent demonstration that silicatein-a protein found in the silica spicules of the sponge Tethya aurantia--can hydrolyse and condense the precursor molecule tetraethoxysilane to form silica structures with controlled shapes at ambient conditions seems particularly promising in this context. Here we describe synthetic cysteine-lysine block copolypeptides that mimic the properties of silicatein: the copolypeptides self-assemble into structured aggregates that hydrolyse tetraethoxysilane while simultaneously directing the formation of ordered silica morphologies. We find that oxidation of the cysteine sulphydryl groups, which is known to affect the assembly of the block copolypeptide, allows us to produce different structures: hard silica spheres and well-defined columns of amorphous silica are produced using the fully reduced and the oxidized forms of the copolymer, respectively.

Expression of a Scr/Hox5 gene in the larval central nervous system of the gastropod Haliotis, a non-segmented spiralian lophotrochozoan.

Hox genes encode a set of evolutionarily conserved transcription factors that regulate anteroposterior patterning mechanisms in insects and vertebrates and are expressed along this axis in a range of bilaterians. Here we present the developmental expression of a Scr/Hox5 gene in the gastropod mollusc Haliotis. In Haliotis, embryogenesis yields a non-feeding trochophore larva that subsequently develops into the veliger larva, which possesses many of the characteristics of the adult body plan. Quantitative RT-PCR analysis reveals that this gene, which is called Hru-Hox5, is first expressed in the trochophore larva. Hru-Hox5 transcript prevalence increases continually through larval development until metamorphic competence develops in the veliger and then again over the first four days of metamorphosis. In situ hybridization reveals that larval expression of Hru-Hox5 is restricted primarily to the primordial and newly formed branchial ganglia, located between the anterior cerebral-pleuropedal ganglionic complex and the posterior visceral ganglia. The expression of Hru-Hox5 in the central region of the abalone CNS is similar to that observed for its orthologue (Lox20) in the leech, suggesting that Hox5 genes were used, along with other Hox genes, to pattern the CNS of the ancestral spiralian lophotrochozoan.

Trends in the incidence of lip, oral, and pharyngeal cancer: Connecticut, 1935-94.

Using Connecticut Tumor Registry data we explored trends in age-adjusted (AARs) and age-specific (ASRs) incidence rates for lip, oral, and pharyngeal cancer over the 60-year period 1935-94. Particular attention was given to findings from the most recent series of 5-year periods that have not been previously analyzed. There was a long-term decline in lip cancer AARs by period, and ASRs generally fell over time and with successive birth cohorts. This notable decrease in rates continued through 1990-94 and with the more recent cohorts, particularly among males. Oral cancer AARs for males peaked in the early 1960s and the late 1970s while rates for pharyngeal cancer increased into the late 1970s. For each of these sites, rates began to decline in 1980-84 and have continued to fall into the first half of the 1990s. Among females AARs for oral and pharyngeal cancer increased more than threefold between 1935-39 and 1980-84; however, encouragingly, and in sharp contrast to the earlier trend, rates fell during the most recent 10-year period. ASRs for oral and pharyngeal cancer were increasing by the birth cohort of 1900 and tended to increase through the cohort of 1920 for males and the cohorts of 1920-30 for females. ASRs for subsequent cohorts have remained relatively unchanged or decreased modestly. It is doubtful that the observed trends in lip, oral and pharyngeal cancer incidence are primarily artifactual, but more likely represent secular changes in exposure to environmental risk factors.

Silicatein filaments and subunits from a marine sponge direct the polymerization of silica and silicones in vitro.

Nanoscale control of the polymerization of silicon and oxygen determines the structures and properties of a wide range of siloxane-based materials, including glasses, ceramics, mesoporous molecular sieves and catalysts, elastomers, resins, insulators, optical coatings, and photoluminescent polymers. In contrast to anthropogenic and geological syntheses of these materials that require extremes of temperature, pressure, or pH, living systems produce a remarkable diversity of nanostructured silicates at ambient temperatures and pressures and at near-neutral pH. We show here that the protein filaments and their constituent subunits comprising the axial cores of silica spicules in a marine sponge chemically and spatially direct the polymerization of silica and silicone polymer networks from the corresponding alkoxide substrates in vitro, under conditions in which such syntheses otherwise require either an acid or base catalyst. Homology of the principal protein to the well known enzyme cathepsin L points to a possible reaction mechanism that is supported by recent site-directed mutagenesis experiments. The catalytic activity of the "silicatein" (silica protein) molecule suggests new routes to the synthesis of silicon-based materials.

Silicatein alpha: cathepsin L-like protein in sponge biosilica.

Earth's biota produces vast quantities of polymerized silica at ambient temperatures and pressures by mechanisms that are not understood. Silica spicules constitute 75% of the dry weight of the sponge Tethya aurantia, making this organism uniquely tractable for analyses of the proteins intimately associated with the biosilica. Each spicule contains a central protein filament, shown by x-ray diffraction to exhibit a highly regular, repeating structure. The protein filaments can be dissociated to yield three similar subunits, named silicatein alpha, beta, and gamma. The molecular weights and amino acid compositions of the three silicateins are similar, suggesting that they are members of a single protein family. The cDNA sequence of silicatein alpha, the most abundant of these subunits, reveals that this protein is highly similar to members of the cathepsin L and papain family of proteases. The cysteine at the active site in the proteases is replaced by serine in silicatein alpha, although the six cysteines that form disulfide bridges in the proteases are conserved. Silicatein alpha also contains unique tandem arrays of multiple hydroxyls. These structural features may help explain the mechanism of biosilicification and the recently discovered activity of the silicateins in promoting the condensation of silica and organically modified siloxane polymers (silicones) from the corresponding silicon alkoxides. They suggest the possibility of a dynamic role of the silicateins in silicification of the sponge spicule and offer the prospect of a new synthetic route to silica and siloxane polymers at low temperature and pressure and neutral pH.

The relationship between bottle usage/content, age, and number of teeth with mutans streptococci colonization in 6-24-month-old children.

OBJECTIVES: Mutans streptococci (MS) are the primary pathogens involved in the development of early childhood caries. However, factors that may affect their acquisition in the mouths of young children are not well understood, and the period of initial colonization remains controversial. This study investigated the relationship of age, number of teeth, and bottle usage/content with regard to the isolation of MS in 6-24-month-old children. METHODS: A total of 122 children from low-income families attending a nutritional supplement program, and their mothers, participated in this study. Children were examined for dental caries and number of erupted teeth and were sampled for MS. Mothers were administered a questionnaire to obtain details of baby bottle use, including what food items were put in the bottle during the last week. RESULTS: MS was detected in more than one-third of the 6-24-month-olds. Unlike some studies that suggest a later period of infectivity, approximately 20% of children under 14 months of age, including 4 of 22 infants aged 6-9 months, were colonized with MS. When examined separately, age, number of teeth, and bottle usage/content were each found to be related to the presence of MS. Mutans streptococci colonization was more likely with increasing age and number of teeth, and children whose bottles contained sweetened beverages were more likely to be colonized than children whose bottles contained milk. Logistic regression models that controlled for both age and number of teeth indicated that children who consumed sweetened beverages in their baby bottle had a statistically significant, four-fold increase in the odds of colonization by MS relative to children who consumed milk. CONCLUSIONS: The finding that approximately 20% of the children under 14 months of age were infected with MS indicates that colonization in this sample of low-income preschool children may begin earlier than suggested by some investigations. Additionally, the risk of MS colonization appears lower among infants who consume milk rather than sweetened beverages in the bottle.

A Mox homeobox gene in the gastropod mollusc Haliotis rufescens is differentially expressed during larval morphogenesis and metamorphosis.

We have isolated a homeobox-containing cDNA from the gastropod mollusc Haliotis rufescens that is most similar to members of the Mox homeobox gene class. The derived Haliotis homeodomain sequence is 85% identical to mouse and frog Mox-2 homeodomains and 88.9% identical to the partial cnidarian cnox5-Hm homeodomain. Quantitative reverse transcription-polymerase chain reaction analysis of mRNA accumulation reveals that this gene, called HruMox, is expressed in the larva, but not in the early embryo. Transcripts are most prevalent during larval morphogenesis from trochophore to veliger. There are also transient increases in transcript prevalence 1 and 3 days after the intitiation of metamorphosis from veliger to juvenile. The identification of a molluscan Mox homeobox gene that is more closely related to vertebrate genes than other protostome (e.g. Drosophila) genes suggests the Mox class of homeobox genes may consist of several different families that have been conserved through evolution.

Modification of calcite crystal growth by abalone shell proteins: an atomic force microscope study.

A family of soluble proteins from the shell of Haliotis rufescens was introduced over a growing calcite crystal being scanned in situ by an atomic force microscope (AFM). Atomic step edges on the crystal surface were altered in shape and speed of growth by the proteins. Proteins attached nonuniformly to the surface, indicating different interactions with crystallographically different step edges. The observed changes were consistent with the habit modification induced by this family of proteins, as previously observed by optical microscopy. To facilitate further studies in this area, AFM techniques and certain AFM imaging artifacts are discussed in detail.

Effects of ddC and AZT on locomotion and acoustic startle. I: Acute effects in female rats.

Several synthetic nucleoside analogues, including AZT(RETROVIR), ddC (HIVID), ddI (VIDEX), and d4T (ZERIT), are currently being used in the treatment of HIV infection. Unfortunately, in clinical use the appearance of severe and sometimes debilitating peripheral neuropathy and pain has been associated with the long-term use of several of these drugs (i.e., ddC, ddI and d4T), although not with AZT. To date, standard pre-clinical animal toxicity studies have failed to reveal any adverse neurologic effects of these compounds. However, previously reported preliminary findings suggest that ddC may alter several neuro-behavioral parameters (including locomotor activity, acoustic startle responding, and aggression) in rats and mice following presentation in the animals' drinking water for 7 days. The current series of experiments examined effects of acutely administered ddC and AZT on spontaneous locomotor activity and acoustic startle responses (with and without pre-pulse) in female Sprague-Dawley rats. Following intragastric administration, ddC reduced locomotion at all but the highest dose, whereas AZT had no significant effect on locomotor activity. Acutely administered ddC had no effect on ASR, whereas AZT increased ASR at the highest stimulus intensity. These data support the use of behavioral testing in the development of the antiviral nucleoside analogues, as behavioral testing may be more effective in identifying the neurologically active agents than is standard toxicity testing.

Mouthwash use and dentures in relation to oral epithelial dysplasia.

This case-control study investigated the potential association between oral epithelial dysplasia (OED) and both mouthwash and denture use. Incident OED cases aged 20-79 years were identified through two oral pathology laboratories. Controls were pair-matched (1:1) to cases on age (+/- 5 years), gender, appointment date and surgeon. A telephone interview was used to obtain exposure information. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were generated using conditional logistic regression. Based upon 127 case-control pairs and after adjusting for smoking, drinking, education and either denture or mouthwash use, the OR for OED and regular mouthwash use (1+ uses/week for 6+ months) was 0.8 (95% CI, 0.4-1.5) while the OR for OED and wearing a denture was 0.7 (95% CI, 0.4-1.3). There were no clear trends of increased OED risk with increased mouthwash use or years of denture wearing. Our findings suggest that neither mouthwash nor denture use are associated positively with OED risk.

Molecular cloning and characterization of lustrin A, a matrix protein from shell and pearl nacre of Haliotis rufescens.

A specialized extracellular matrix of proteins and polysaccharides controls the morphology and packing of calcium carbonate crystals and becomes occluded within the mineralized composite during formation of the molluscan shell and pearl. We have cloned and characterized the cDNA coding for Lustrin A, a newly described matrix protein from the nacreous layer of the shell and pearl produced by the abalone, Haliotis rufescens, a marine gastropod mollusc. The full-length cDNA is 4,439 base pairs (bp) long and contains an open reading frame coding for 1,428 amino acids. The deduced amino acid sequence reveals a highly modular structure with a high proportion of Ser (16%), Pro (14%), Gly (13%), and Cys (9%). The protein contains ten highly conserved cysteine-rich domains interspersed by eight proline-rich domains; a glycine- and serine-rich domain lies between the two cysteine-rich domains nearest the C terminus, and these are followed by a basic domain and a C-terminal domain that is highly similar to known protease inhibitors. The glycine- and serine-rich domain and at least one of the proline-rich domains show sequence similarity to proteins of two extracellular matrix superfamilies (one of which also is involved in the mineralized matrixes of bone, dentin, and avian eggshell). The arrangement of alternating cysteine-rich domains and proline-rich domains is strikingly similar to that found in frustulins, the proteins that are integral to the silicified cell wall of diatoms. Its modular structure suggests that Lustrin A is a multifunctional protein, whereas the occurrence of related sequences suggest it is a member of a multiprotein family.