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PubMed1990

[Dental sac and its role in periodontal development].

The present study examined the role of the innermost layer of dental sac tissue in periodontal development. Empty bony crypts were filled with Hydroxylapatite ceramic granules of porous consistency in 5-day-old rats. After 4.5 and 24 weeks the specimens were processed for light and transmission electron microscopy. Only 4 weeks later cement regeneration was seen on ceramic surfaces with bundles of oriented collagen fibers inserting in the cementoid matrix. Ultrastructurally the newly formed cementum-like tissue was in intimate contact to the synthetic hydroxylapatite crystals. A consistent finding was the presence of an interface between cementum and the ceramic surface. The present findings question the unique role of the "investing layer" in periodontal development and the existence of cement-inducting properties of dentin.

Animals↗

PubMedDate not supplied

Liquid chromatographic-atomic absorption spectrophotometric method for determination of methyl mercury in seafood: collaborative study.

A previously developed method that uses a simplified sample preparation procedure and atomic absorption detection of liquid chromatographic eluates for the determination of methyl mercury in seafood has been collaboratively studied. The unique feature of the method involves the use of a specially designed interface for the generation of mercury vapor. Methyl mercury is isolated from the blended sample by chloroform elution from a diatomaceous earth-hydrochloric acid column. The organomercury compound is then extracted into a small volume of 0.01M sodium thiosulfate solution. An aliquot of this solution is injected onto a Zorbax ODS column and eluted with methanol-ammonium acetate solution (3 + 2), pH 5.7, containing 0.01% mercaptoethanol. Mercury is detected by flameless atomic absorption spectrophotometry using the interface. The samples analyzed in the study were unspiked swordfish, unspiked and spiked lobster, and unspiked and spiked tuna. The spiked samples contained methyl mercury both above and below the U.S. Food and Drug Administration guideline level of 1 microgram Hg/g. Reproducibility relative standard deviations ranged from 10.5% at 1 microgram Hg/g to 18.2% at about 0.1 microgram Hg/g. Accuracy, measured by comparison to reference values obtained by the Associate Referee, ranged from 94.4 to 99.6%. The method has been adopted official first action.

Animals↗

PubMed2005

A single Mms2 "key" residue insertion into a Ubc13 pocket determines the interface specificity of a human Lys63 ubiquitin conjugation complex.

Human Ubc13 and Mms2 (or its homolog, Uev1) form a unique ubiquitin-conjugating enzyme (Ubc) complex that generates atypical Lys(63)-linked ubiquitin conjugates. Such conjugates are attached to specific targets that modulate the activity of various cellular processes including DNA repair, mitotic progression, and nuclear factor-kappaB signaling. Whereas Ubc13 is a typical Ubc, Mms2 is a non-catalytic Ubc variant. Substantial biochemical evidence has revealed a mechanism whereby Mms2 properly orients ubiquitin to allow for Lys(63) conjugation by Ubc13; however, how this specific Ubc13-Mms2 complex is formed and why Mms2 does not form a complex with other Ubcs have not been reported. In order to address these questions, we used a structure-based approach to design mutations and characterize the human Ubc13-Mms2 interface. We used the yeast two-hybrid assay, glutathione S-transferase pull-downs, and surface plasmon resonance to test in vivo and in vitro binding. These experiments were paired with functional complementation and ubiquitin conjugation studies to provide in vivo and in vitro functional data. The results in this study allowed us to identify important residues of the Ubc13-Mms2 interface, determine a correlation between heterodimer formation and function, and conclude why Mms2 forms a specific complex with Ubc13 but not other Ubc proteins.

Amino Acid Sequence↗

PubMed2006

Two-dimensional self-assembly and phase behavior of an alkoxylated sandwich-type bisphthalocyanine and its phthalocyanine analogues at the liquid-solid interface.

Lanthanide bisphthalocyanine complexes are interesting objects for the construction of two-dimensional (2D) ordered arrays with prospective applications in molecular electronics due to their unique semiconductor and electrochromic properties as well as their double-decker architecture. The 2D self-assembly of dodecyloxy-substituted (bisphthalocyaninato)erbium(III) has been studied at the solid (highly oriented pyrolytic graphite)-liquid interface by scanning tunneling microscopy. The results show that the bisphthalocyanine molecules form an ordered quadratic 2D lattice (a = b = 3.0 +/- 0.1 nm, gamma = 90 +/- 3 degrees), which is nearly identical to that formed by its (phthalocyaninato)cobalt(II) analogue bearing alkoxy chains of the same length. This clearly shows that sandwich molecules adsorb to the graphite surface by one of the phthalocyanine disks and its eight alkoxy groups. Despite the very similar mode of interaction with the surface, mixtures of alkoxylated (bisphthalocyaninato)erbium(III) with its (phthalocyaninato)cobalt(II) analogue show partial phase separation on the nanoscale: areas are enriched in one of the compounds. A much clearer phase separation between (bisphthalocyaninato)erbium(III) and (phthalocyaninato)cobalt(II) molecules was achieved by mixing molecules containing alkoxy groups of different length. The results provide insight for the development of well-ordered nanostructures of bisphthalocyanines in the presence of phthalocyanines, which could be of importance for future nanometer-scale functional materials.

Journal Article↗

PubMed2004

New approaches to liquid interfaces through changes in the refractive index and nonlinear susceptibility utilizing ultrashort laser pulses.

Molecules in inhomogeneous liquid environments, such as air/liquid, liquid/liquid, solid/liquid interfaces interact with each other specifically, and sometimes form characteristic structures and emerge unique properties. Here, we introduce two newly developed spectroscopic techniques, the total-internal-reflection ultrafast transient lens method (TIR-UTL) and second harmonic generation-coherent vibrational spectroscopy (SHG-CVS), to investigate the characteristic behaviors of molecules in such inhomogeneous environments. TIR-UTL probes the refractive-index change with sub-picosecond resolution and provides information on ultrafast changes in the population, density, and thermal properties, such as temperature increase and energy transfer from the solute molecules to the surrounding solvent molecules. On the other hand, SHG-CVS probes nonlinear susceptibility changes at the interfacial areas, and is expected to provide spectroscopic information on the low-frequency vibrational modes that reflect the corrective motion of the molecules in such an inhomogeneous environment. These new approaches are based on pump-probe techniques utilizing (ultra) short laser pulses. They are expected to provide further information on inhomogeneous environments from the viewpoints of solute-solvent interactions, changes in the molecular orientation, and the corrective motion of molecules at liquid interfaces.

Journal Article↗

PubMed2001

Regulated gene expression dictates enamel structure and tooth function.

Enamel is a complex bioceramic tissue. In its final form, enamel is a reflection of the unique molecular and cellular activities occurring during organogenesis. From the ectodermal origins of ameloblasts, their gene activity and protein expression profiles exist for the sole purpose of producing a mineralized shell, almost entirely devoid of protein, deposited over the 'bone-like' dentine. The interface between enamel and dentine is referred to as the dentine enamel junction and it is also unique in its biology. This review article is narrow in its scope. We restrict our review to selected advances in our understanding of the genetic, molecular and structural aspects of enamel biology. We present a model of enamel formation that relates gene expression to the assembly of an extracellular protein matrix that in turn controls the structural hierarchy and mechanical aspects of enamel and the tooth organ.

Amelogenin↗

PubMed2001

Superficial lymphangitis with interface dermatitis occurring shortly after a minor injury: possible involvement of a bacterial infection and contact allergens.

BACKGROUND: Linear supralymphatic eruptions with epidermal involvement have rarely been reported. OBJECTIVE: A search was made for apparent anatomical reasons and for external factors to explain the unique distribution pattern and clinical course in three cases in which the linear lesions occurred shortly after a minor injury. METHODS: Efforts to search for its etiology include careful outlining of the localization, bacterial culture from the site of traumatic injury, patch tests, and skin biopsies. RESULTS: Linear lesions developed along superficial lymphatic vessels and the presence of eczematous conditions around the injured sites and isolation of Staphylococcus aureus from the site were observed concomitantly. The histopathological findings showed interface dermatitis. CONCLUSION: Our cases provide a unique example of the combined effects of a bacterial infection and contact allergens in the development of the linear supralymphatic eruptions.

Adult↗

PubMed1994

New surface-hardened, low-modulus, corrosion-resistant Ti-13Nb-13Zr alloy for total hip arthroplasty.

To optimize the performance of total hip replacement, scientists and clinicians are seeking new materials and noncemented, press-fit designs that can improve load transfer to the bone and reduce the incidence of loosening and thigh pain. Currently used Co-Cr-Mo alloy has a relatively high elastic modulus (E = 227 GPa), which limits its ability to transfer load to the surrounding bone in the proximal calcar region. Thus to improve load transfer, designs are considered with less cross-sectional area to increase flexibility, but at the expense of fit and fill, and thus stability of the implant within the bone. Should stem loosening occur, the stem stresses may exceed the relatively low fatigue strength of the Co-Cr-Mo alloy and lead to stem breakage. To improve these conditions, lower modulus Ti-6Al-4V alloy (E = 115 GPa) is being used. More recently, a new lower-modulus (E = 79 GPa) Ti-13Nb-13Zr alloy has been developed which does not contain any elemental constituents associated with adverse cell response (i.e., Co, Cr, Mo, Ni, Fe, Al, V), and which possesses comparable or superior strength and toughness to existing Ti-6Al-4V alloy. The carefully selected Nb and Zr constituents improve bone biocompatibility and corrosion resistance compared to that of currently used implant metals. Additionally, a unique diffusion hardening (DH) treatment can be conducted during the age-hardening process of this near-beta alloy to produce a hardened surface with abrasion resistance superior to that of Co-Cr-Mo alloy. This also provides an improvement in the micro-fretting tendencies that may occur within femoral head-neck taper regions and modular interfaces of other implant designs. The present study describes the metallurgy and mechanical properties of this unique low modulus Ti-13Nb-13Zr alloy, and the heat treatments used to obtain the high strength, corrosion resistance, and surface hardening that renders this biocompatible alloy well-suited for press fit hip replacement applications. Because of the relatively lower beta transus (735 degrees C), this alloy is also much easier to net shape forge into more complex stem designs.

Alloys↗

PubMed2006

Adsorption of mixtures of nonionic sugar-based surfactants with other surfactants at solid/liquid interfaces I. Adsorption of n-dodecyl-beta-D-maltoside with anionic sodium dodecyl sulfate on alumina.

Sugar-based surfactants can be synthesized from renewable materials and are environmentally benign. They have some unique solution and interfacial properties and have potential applications in a wide variety of processes, and there is a need for corresponding information on their behavior at various interfaces. In this study, co-adsorption of nonionic sugar-based n-dodecyl-beta-D-maltoside (DM) and anionic sodium dodecyl sulfate (SDS) on alumina was studied as a function of mixing ratios and solution pHs. It is found that at solid-liquid interface, depending on the solid type and the solution conditions, there are various interactions that dictate synergy or antagonism. At pH 6 where alumina is positively charged, marked synergistic effects between DM and SDS were observed, while at pH 11 where alumina is negatively charged, SDS shows antagonistic adsorption effects with DM. The ratios of surfactant components on solids change as a function of surfactant structure and concentrations as well, indicating various interactions at solid/liquid interface under different conditions that can be utilized for many industrial processes.

Adsorption↗

PubMed1995

Materno-fetal immunobiology in normal pregnancy and its possible failure in recurrent spontaneous abortion?

Many features contributing to the success of pregnancy in humans have been identified over the last 20 years. Trophoblast cells (which form the interface of fetal tissue with the mother) have specialized immunological features which may confer unique transplantation protection for the fetus throughout pregnancy. Both syncytiotrophoblast and cytotrophoblast cells do not express classical class I [human leucocyte antigen (HLA)-A or -B] or II (HLA-DP, -DQ or -DR) major histocompatibility complex (MHC) alloantigens, and the regulation of these cell surface glycoproteins appears to be at the transcriptional level. In contrast, extravillous cytotrophoblast cells express the non-classical class I MHC molecule HLA-G. One form of HLA-G (HLA-G1) is potentially capable of presenting a variety of peptide antigens to T cells. Alternatively, HLA-G may act as a cell surface class I MHC molecule, protecting cytotrophoblast from maternal MHC non-restricted natural killer (NK) cell attack; the expression of HLA-G by otherwise HLA-null cell transfectants has been shown to decrease their sensitivity to NK cell-mediated cytolysis. All fetal trophoblast populations throughout gestation express high levels of cell surface complement regulatory proteins, providing protection from complement-mediated damage at the materno-fetal interface. Analyses of trials with allogeneic leucocytes for the treatment of recurrent spontaneous abortion show little evidence of an improved success rate in immunized patients.

Abortion, Habitual↗

PubMed1997

Care of the terminally ill cachectic cancer patient: interface between nursing and psychological anthropology.

The wasted appearance of the terminally ill cachectic cancer patient is a major source of concern for both patients and families. To date, this experience has mainly been understood from a biomedical perspective. However, considering this experience occurs within unique social, historical and cultural contexts, a more holistic approach to capturing this experience is warranted. This paper will argue that the interface between psychological anthropology and nursing provides an alternative approach from which the experience of 'wasting away' might be examined. Special attention to the use of explanatory models of illness and illness narratives in capturing this interface will be explored.

Anthropology, Cultural↗

PubMed1993

Cross-sectional AEM preparation technique for ceramic-coated WC-Co cutting tools.

The preparation of cross-sectional specimens for AEM studies of materials such as ceramic coated tungsten carbide presents some unique problems. Pieces joined by the use of epoxides often separate at the interface between the WC and ceramic coating during the initial mechanical grinding and subsequent thinning process as a result of the vibration and physical strain placed on the sample. These problems have been overcome through the use of a preparation process which essentially encapsulates the sample within the confines of an epoxy filled quartz tube. This preparation process has allowed for facile AEM cross-sectional analysis of TiN/TiCN coatings on WC-Co substrates, and has revealed two distinct grain morphologies within the TiCN coating.

Ceramics↗

PubMed2001

The 1.85 A resolution crystal structures of tissue factor in complex with humanized Fab D3h44 and of free humanized Fab D3h44: revisiting the solvation of antigen combining sites.

The outstanding importance of the antigen-antibody recognition process for the survival and defence strategy of higher organisms is in sharp contrast to the limited high resolution structural data available on antibody-antigen pairs with antigenic proteins. The limitation is the most severe for structural data not restricted to the antigen-antibody complex but extending to the uncomplexed antigen and antibody. We report the crystal structure of the complex between tissue factor (TF) and the humanized Fab fragment D3h44 at a resolution of 1.85 A together with the structure of uncomplexed D3h44 at the same resolution. In conjunction with the previously reported 1.7 A crystal structure of uncomplexed TF, a unique opportunity is generated to explore details of the recognition process. The TF.D3h44 interface is characterised by a high number of polar interactions, including as may as 46 solvent molecules. Conformational changes upon complex formation are very small and almost exclusively limited to the reorientation of side-chains. The binding epitope is in complete agreement with earlier mutagenesis experiments. A revaluation of two other antibody-antigen pairs reported at similar resolutions, shows that all these complexes are very similar with respect to the solvation of the interface, the number of solvent positions conserved in the uncomplexed and complexed proteins and the number of water molecules expelled from the surface and replaced by hydrophilic atoms from the binding partner upon complex formation. A strategy is proposed on how to exploit this high resolution structural data to guide the affinity maturation of humanised antibodies.

Amino Acid Sequence↗

PubMed2001

Dynamics-function correlation in Cu, Zn superoxide dismutase: a spectroscopic and molecular dynamics simulation study.

A single mutation (Val29-->Gly) at the subunit interface of a Cu, Zn superoxide dismutase dimer leads to a twofold increase in the second order catalytic rate, when compared to the native enzyme, without causing any modification of the structure or the electric field distribution. To check the role of dynamic processes in this catalytic enhancement, the flexibility of the dimeric protein at the subunit interface region has been probed by the phosphorescence and fluorescence properties of the unique tryptophan residue. Multiple spectroscopic data indicate that Trp83 experiences a very similar, and relatively hydrophobic, environment in both wild-type and mutant protein, whereas its mobility is distinctly more restrained in the latter. Molecular dynamics simulation confirms this result, and provides, at the molecular level, details of the dynamic change felt by tryptophan. Moreover, the simulation shows that the loops surrounding the active site are more flexible in the mutant than in the native enzyme, making the copper more accessible to the incoming substrate, and being thus responsible for the catalytic rate enhancement. Evidence for increased, dynamic copper accessibility also comes from faster copper removal in the mutant by a metal chelator. These results indicate that differences in dynamic, rather than structural, features of the two enzymes are responsible for the observed functional change.

Amino Acid Sequence↗

PubMed2003

Brain barrier systems: a new frontier in metal neurotoxicological research.

The concept of brain barriers or a brain barrier system embraces the blood-brain interface, referred to as the blood-brain barrier, and the blood-cerebrospinal fluid (CSF) interface, referred to as the blood-CSF barrier. These brain barriers protect the CNS against chemical insults, by different complementary mechanisms. Toxic metal molecules can either bypass these mechanisms or be sequestered in and therefore potentially deleterious to brain barriers. Supportive evidence suggests that damage to blood-brain interfaces can lead to chemical-induced neurotoxicities. This review article examines the unique structure, specialization, and function of the brain barrier system, with particular emphasis on its toxicological implications. Typical examples of metal transport and toxicity at the barriers, such as lead (Pb), mercury (Hg), iron (Fe), and manganese (Mn), are discussed in detail with a special focus on the relevance to their toxic neurological consequences. Based on these discussions, the emerging research needs, such as construction of the new concept of blood-brain regional barriers, understanding of chemical effect on aged or immature barriers, and elucidation of the susceptibility of tight junctions to toxicants, are identified and addressed in this newly evolving field of neurotoxicology. They represent both clear challenges and fruitful research domains not only in neurotoxicology, but also in neurophysiology and pharmacology.

Animals↗

PubMed1997

Use of mutator cells as a means for increasing production levels of a recombinant antibody directed against Hepatitis B.

A mutation strategy which utilises phage display technology and the Escherichia coli mutator strains, mutD5-FIT and XL1-RED, was applied to a Hepatitis B (HepB) specific single-chain Fv (scFv) to incorporate random mutations throughout the gene. Messenger RNA from a hybridoma producing antibodies against HepB was isolated, reverse transcribed and used as template for the production of scFv. Following production of the scFv protein using an E. coli expression vector (pGC), the scFv gene was recloned into a phage display vector (pHFA). This gene construct was introduced into E. coli mutator cells and the transformed cells were used as an inoculum for liquid cultures. After five cycles of growth at 37 degrees C, each followed by dilution and re-inoculation of fresh media, recombinant phage were recovered. Nucleotide sequence analysis of the scFv gene in phage selected on HBsAg-coated magnetic beads identified amino acid substitutions which produced an increase of greater than 10-fold in apparent production levels. Competitive ELISA studies showed that the selected scFv mutants appeared to have similar affinity to HBsAg as the parent scFv. The apparent increase in production was not the result of improved surface characteristics of regions uniquely exposed in scFvs, as the sites did not correlate with the variable/constant interface of the scFv variable region normally masked in Fabs or IgGs.

Amino Acid Sequence↗

PubMed2006

One functional switch mediates reversible and irreversible inactivation of a herpesvirus protease.

Distinct mechanisms have evolved to regulate the function of proteolytic enzymes. Viral proteases in particular have developed novel regulatory mechanisms, presumably due to their comparatively rapid life cycles and responses to constant evolutionary pressure. Herpesviruses are a family of human pathogens that require a viral protease with a concentration-dependent zymogen activation involving folding of two alpha-helices and activation of the catalytic machinery, which results in formation of infectious virions. Kaposi's sarcoma-associated herpesvirus protease (KSHV Pr) is unique among the herpesvirus proteases in possessing an autolysis site in the dimer interface, which removes the carboxyl-terminal 27 amino acids comprising an alpha-helix adjacent to the active site. Truncation results in the irreversible loss of dimerization and concomitant inactivation. We characterized the conformational and functional differences between the active dimer, inactive monomer, and inactive truncated protease to determine the different protease regulatory mechanisms that control the KSHV lytic cycle. Circular dichroism revealed a loss of 31% alpha-helicity upon dimer dissociation. Comparison of the full-length and truncated monomers by NMR showed differences in 21% of the protein structure, mainly located adjacent to the dimer interface, with little perturbation of the overall protein upon truncation. Fluorescence polarization and active site labeling, with a transition state mimetic, characterized the functional effects of these conformational changes. Substrate turnover is abolished in both the full-length and truncated monomers; however, substrate binding remained intact. Disruption of the helix 6 interaction with the active site oxyanion loop is therefore used in two independent regulatory mechanisms of proteolytic activity.

Binding Sites↗

PubMed1996

Raman spectroscopy of the Ff gene V protein and complexes with poly(dA): nonspecific DNA recognition and binding.

Raman spectra of crystals and solutions of the single-stranded DNA binding protein of bacteriophage Ff (gene V protein, gVp) and of solution complexes of gVp with single-stranded poly-(deoxyadenylic acid) [poly(dA)] reveal the following: (i) The gVp secondary and tertiary structures are similar in solution and in the crystal and are dominated by beta-sheet domains, in agreement with NMR and X-ray findings. (ii) Subunit conformation and side chain environments of gVp are virtually unchanged over a wide range of salt concentration (0 < [NaCl] < 100 mM); however, the solution conformation of poly(dA) exhibits sensitivity to added salt. The perturbed Raman markers indicate subtle changes in helix backbone geometry with accompanying small differences in base stacking as the concentration of NaCl is changed. (iii) In complexes with poly(dA), neither the conformation of gVp nor its side chain environments are altered significantly in comparison to the free protein. This is the case at both high salt (nucleotide-to-subunit binding stoichiometry n = 4) and low salt (n = 3). (iv) The Raman signature of poly(dA) undergoes small perturbations upon gVp binding, indicative of small changes in base stacking and phosphodiester backbone conformation. The present results show that the different stoichiometric binding modes of gVp to poly(dA) are accomplished without significant changes in gVp subunit structure and with only modest changes in the single-stranded poly(dA) ligand. This contrasts sharply with sequence-specific double-stranded DNA binding proteins, such as the phage lambda and D108 repressors, which undergo substantial structural changes upon DNA binding, and which also alter more dramatically the Raman fingerprints of their DNA target sites. Thus, nonspecific and specific nucleic acid recognition modes are distinguishable by Raman spectroscopy. The Raman signature of gVp also allows examination of hydrogen bonding interactions of unique side chains within the hydrophobic core (cysteine 33) and at the binding interface (tyrosine 41). These are discussed in relation to the recently published gVp crystal structure.

Amino Acid Sequence↗