Experimental constraints on theories of high-transition temperature superconductors.

Recent experiments have revealed several key features of the unique nature of the new, high-transition temperature cuprate superconductors. These results provide an easily understandable, physical picture of the structure and behavior of the charge carriers in these materials, and point to the mechanism responsible for their existence. These experiments are now placing strong constraints on possible theoretical models of the phenomenon.

A new immunoassay based on fluorescence excitation by internal reflection spectroscopy.

A new immunoassay technique is described which uses totally internally reflected light to excite the fluorescence of fluorescein labeled antibody which has become bound to a hapten--protein conjugate absorbed on a quartz-plate in the antibody solution. The presence of any free hapten in solution reduces the amount of antibody free to bind to the surface and thus reduces the fluorescence signal. Measurement of the decrease of the fluorescent signal then gives a measure of the concentration of free hapten present. The technique is simple, fast and has high intrinsic sensitivity and specificity. It has been demonstrated for phenyl arsonic acid and morphine. Free morphine at a concentration of 2 X 10(-7) M is readily detected.

Development and in vitro evaluation of an intra-oral controlled-release delivery system for chlorhexidine.

Copolymers of hydroxyethyl methacrylate (HEMA) and methyl methacrylate (MMA) were prepared and used to fabricate a membrane-controlled reservoir-type controlled-release delivery system for chlorhexidine that should be suitable for intra-oral use. The reservoir of the system was prepared by softening an 80:20 mixture of chlorhexidine diacetate and 50:50 HEMA:MMA copolymer with methyl ethyl ketone (MEK), and pressing standard amounts of the resulting dough-like mixture into silicone rubber molds. A membrane was applied to the reservoirs by rotating them through a solution of 30:70 HEMA:MMA copolymer in MEK. The finished oval-shaped controlled-release pellets were approximately 4.7 mm wide, 3.3 mm high, and 7.4 mm long, and contained 45.0 +/- 3.7 mg of chlorhexidine diacetate. The mean in vitro release rate of chlorhexidine diacetate from the pellets into 37 degrees C water was 608 +/- 55 micrograms/24 h for days 2 through 11, and 389 +/- 50 micrograms/24 h for days 15 to 30 of the test period. The chlorhexidine released on day 30 was biologically active, as determined by a serial dilution assay against Streptococcus mutans. The extended release of biologically active chlorhexidine at a controlled rate from this system suggests that it is worthy of further evaluation for the intra-oral therapy of chlorhexidine-treatable oral infections in non-compliant and physically or mentally compromised individuals.

Prevalence of Streptococcus mutans serotypes, Actinomyces, and other bacteria in the plaque of children.

Selected microbial components in dental plaque were determined for children in Biddeford, Maine and Colombia, South America. Using cultural methods, Streptococcus mutans was detected in 51.4% of the Colombian children and 63.3% of the Maine children. Serotype c was predominant in both populations. The greatest difference between the two groups occurred with serotypes d and g which were present in 25% of the Colombian children with S. mutans and were not detected in the Maine children. In the specimens examined with specific FA conjugates. Actinomyces was the predominant genus, present in all individuals and comprising an average of 52% of all cells.

Polymorphisms in the human DNA ligase I gene (LIG1) including a complex GT repeat.

Sequencing of a human DNA ligase I cDNA clone derived from HeLa cells revealed two unreported differences with the published sequence: a single base change and a three-base deletion. Both differences are in exon 6, and were analyzed by amplifying a segment containing exon 5, intron 6, and exon 6. The first finding was that intron 6 is approximately 2.6 kb in size, not the 1 kb reported in the literature. By sequence analysis of amplified segments, the single-base difference in exon 6 was shown to be polymorphic, with HeLa cells heterozygous for the A/C difference. Analysis of 60 unrelated individuals found a frequency of 0.5 for each allele. Primer extension reactions across the exon 5/exon 6 boundary were performed on cDNA obtained from HeLa cells and human thymus. The results show that the three-base deletion is due to a variation in splicing. For both HeLa and thymus, two-thirds of the transcripts are like the published cDNA sequence and one-third have the three-base deletion. Finally, sequencing of part of intron 6 revealed the presence of a complex GT repeat consisting of a 48-50 nucleotide polypurine tract followed by a variable number of GT residues. This entire unit of polypurine tract plus GTs is repeated three times. Detection of the repeated sequences required the development of specialized cloning and PCR conditions. Analysis of a pedigree showed that this complex repeat is polymorphic.

Effects of octenidine on dental plaque and gingivitis in monkeys.

This investigation monitored the effects of daily oral rinses with octenidine on plaque and gingivitis in five monkeys. Formulations containing 0.5% or 1.0% octenidine or the rinse vehicle placebo were provided daily for 2 weeks. Each week the dentition of each monkey was examined, photographed, and sampled for plaque. All responses exhibited a numerical decrease in mean scores following treatments with each concentration of octenidine, whereas the placebo treatment exerted negligible effects. Decreases in plaque mass were observed after 2 weeks of treatment with 1% octenidine (58%) or 0.5% octenidine (55%) compared with the corresponding baseline values. Similar trends were noted in the extent and thickness of supragingival plaque and its ability to decrease the pH of a sucrose solution. Octenidine treatments reduced the proportions of motile forms in samples of subgingival plaque and also restricted its ability to produce H2S. Slight numerical decreases were seen in the Gingival Index and flow rate of the crevicular fluid. These consistent protective trends suggest that octenidine decreases the pathogenic potential of established plaque.

Current aspects of sterilization. The selection and application of various surgical methods of sterilization.

A partial analysis of 4,270 female sterilizations performed during a 7 year time period was described as related to changing patterns of application according to a sterilization program. Special attention was given to such factors as type of procedure, reasons for selection of procedures, ancillary conditions, mortality rate, and follow-up. A plea is made for accuracy when a sterilization is truly used as the primary indication for surgery and for flexibility and individualization in the application of sterilization techniques.

Gap-modulation infrared spectroscopy of high transition temperature superconductors.

Conventional methods of determining the coupling factor alpha(2)(omega)F(omega) for the newly discovered high transition temperature (T(c)) cuprate superconductors by using tunneling and infrared measurements have thus far failed to show the cause of the very high T(c) of these compounds. This is due in part to difficulties in sample preparation for tunneling studies and to difficulties in obtaining good data at relatively high tunneling voltages. Also, in IR (infrared) measurements, small differences in absorptivity between the normal and superconducting state can be masked by changes in the phonon occupation at high and low temperatures. Here we propose a technique for determing the coupling constant, which should be less dependent on the surface quality of the sample than with tunneling and should allow measurements at higher energies with greater precision than do tunneling or simple IR observations. This should make possible a definitive determination of any possible exciton contribution to this coupling term, which would appear at energies well above the range where conventional IR or tunneling measurements are effective.

Antibiotic susceptibility of Streptococcus mutans: comparison of serotype profiles.

A total of 82 strains of Streptococcus mutans representing serotypes a through g were tested for susceptibility to erythromycin, penicillin, methicillin, lincomycin, tetracycline, vancomycin, gentamicin, streptomycin, neomycin, kanamycin, bacitracin, and polymyxin B. Strains included stock cultures and isolates from human and animal dental plaque. Minimal inhibitory concentrations were determined by a broth-microdilution procedure. The major differences in antibiotic susceptibility observed among the serotypes resulted with antibiotics which act on the cell surface. Bacitracin was most active against serotype a strains and polymyxin B against serotype b strains. Serotypes a, d, and g were less susceptible than the other serotypes to methicillin.

Effect of antibody preparations on glucose uptake by a cariogenic Streptococcus.

The effect of antisera to whole cells or cell wall components on glucose uptake by S. mutans 6715 was examined. Early stationary phase 6715 cells were treated with test serum and incubated at 37 degrees C in the presence of 14C-glucose. Samples were removed at timed intervals and measured in a liquid scintillation counter for radioactive uptake. Antisera to both whole cells and components known to be present on the surface of the cells reduced glucose uptake relative to normal serum. It is suggested that inhibition of glucose uptake may be one mechanism by which a caries vaccine may operate.

Comparative recovery of Streptococcus mutans on ten isolation media.

The ability of Streptococcus mutans (Bratthall serotypes a through e) to grow on 10 isolation media was examined. The number and morphology of the colonies were observed to vary on different media. The use of blood-sucrose media consistently produced the highest recoveries. Mitis salivarius agar (MS) and higher recovery values than modified medium 10 (MM10SB), Trypticase-yeast extract-cystine medium (TYC), or MS with 1% tellurite (MST). MST with 40% sucrose (MS40S), MST with 20% sucrose and 0.2 U of bacitracin per ml (MSB), and Carlsson medium with 1% sulfasoxazole (MC), media formulated for the selection of S. mutans, were the most inhibitory for all serotypes. The morphology of several S. mutans strains was atypical on MC and MS40S, making positive identification difficult. Absence of growth of serotype a strains on MSB and serotype d strains on MC were the two major differences observed among the serotypes. Results are discussed in terms of the difficulties in making quantitative determinations from cultural data.

Expression of asparagine independence in variants of ICR 2A haploid frog cells.

Properties of the change from asparagine dependence (asn-) to independence (asn+) were investigated in the androgenetic haploid frog cell line ICR 2A. Two types of asn+ variants arose spontaneously during culture. Glutamine-dependent asparagine synthetase (AS) activity, found to be deficient in asn- cells, was repressed by asparagine in one type of variant and expressed constitutively in the other. No quantitative differences in AS-specific DNA sequences or changes in ploidy were evident between asn+ and asn- cells. The asn+ frequency in ICR 2A populations, not dramatically influenced by chemical mutagens, was increased 130-fold by exposure to 5-azacytidine. The methylation of CCGG sequences at the 5' end of the AS structural gene was found to be reduced equally in both types of asn+ variant. These results indicate that decreased DNA methylation is essential but not necessarily sufficient for the expression of AS activity in this frog cell system.

KvLQT1, a voltage-gated potassium channel responsible for human cardiac arrhythmias.

The clinical features of long QT syndrome result from episodic life-threatening cardiac arrhythmias, specifically the polymorphic ventricular tachycardia torsades de pointes. KVLQT1 has been established as the human chromosome 11-linked gene responsible for more than 50% of inherited long QT syndrome. Here we describe the cloning of a full-length KVLQT1 cDNA and its functional expression. KVLQT1 encodes a 676-amino acid polypeptide with structural characteristics similar to voltage-gated potassium channels. Expression of KvLQT1 in Xenopus oocytes and in human embryonic kidney cells elicits a rapidly activating, K+-selective outward current. The I(Kr)-specific blockers, E-4031 and dofetilide, do not inhibit KvLQT1, whereas clofilium, a class III antiarrhythmic agent with the propensity to induce torsades de pointes, substantially inhibits the current. Elevation of cAMP levels in oocytes nearly doubles the amplitude of KvLQT1 currents. Coexpression of minK with KvLQT1 results in a conductance with pharmacological and biophysical properties more similar to I(Ks) than other known delayed rectifier K+ currents in the heart.