Associations among Emdogain®, human oral carcinoma cells and oral proteolytic enzymes

The Enamel matrix derivative Emdogain® (EMD) is a commercially available tissue extract preparation of porcine enamel origin. Studies have shown EMD to be clinically useful in promoting periodontal regeneration. EMD has been widely used in periodontal therapy for over ten years, but the mechanism of its action and the exact composition are not completely clear. EMD is predominantly amelogenin (>90%). However, unlike amelogenin, EMD has a number of growth factor-like effects and it has been shown to enhance the proliferation, migration and other cellular functions of periodontal ligament fibroblasts and osteoblasts. In contrast, the effects of EMD on epithelial cell lines and in particular on oral malignant cells have not been adequately studied. In addition, EMD has effects on the production of cytokines by several oral cell lines and the product is in constant interaction with different oral enzymes. Regardless of the various unknown properties of EMD, it is said to be clinically safe in regenerative procedures, also in medically compromised patients. The aim of the study was to examine whether gingival crevicular fluid (GCF), which contains several different proteolysis enzymes, could degrade EMD and alter its biological functions. In addition, the objective was to study the effects of EMD on carcinogenesis-related factors, in particular MMPs, using in vitro and in vivo models. This study also aimed to contribute to the understanding of the composition of EMD. GCF was capable of degrading EMD, depending on the periodontal status, with markedly more degradation in all states of periodontal disease compared to healthy controls. EMD was observed to stimulate the migration of periodontal ligament fibroblasts (PLF), whereas EMD together with GCF could not stimulate this proliferation. In addition, recombinant amelogenin, the main component of EMD, decreased the migration of PLFs. A comparison of changes induced by EMD and TGF-β1 in the gene profiles of carcinoma cells showed TGF-β1 to regulate a greater number of genes than EMD. However, both of the study reagents enhanced the expression of MMP-10 and MMP-9. Furthermore, EMD was found to induce several factors closely related to carcinogenesis on gene, protein, cell and in vivo levels. EMD enhanced the production of MMP-2, MMP-9 and MMP-10 proteins by cultured carcinoma cells. In addition, EMD stimulated the migration and in vitro wound closure of carcinoma cells. EMD was also capable of promoting metastasis formation in mice. In conclusion, the diseased GCF, containing various proteases, causes degradation of EMD and decreased proliferation of PLFs. Thus, this in vitro study suggests that the regenerative effect of EMD may decrease due to proteases present in periodontal tissues during the inflammation and healing of the tissues in vivo. Furthermore, EMD was observed to enhance several carcinoma-related factors and in particular the production of MMPs by benign and malignant cell lines. These findings suggest that the clinical safety of EMD with regard to dysplastic mucosal lesions should be further investigated.

Embedding Of Non-Simple Lie-Groups, Coupling-Constant Relations And Non-Uniqueness Of Models Of Unification

A general derivation of the coupling constant relations which result on embedding a non-simple group like SU L (2) @ U(1) in a larger simple group (or graded Lie group) is given. It is shown that such relations depend only on the requirement (i) that the multiplet of vector fields form an irreducible representation of the unifying algebra and (ii) the transformation properties of the fermions under SU L (2). This point is illustrated in two ways, one by constructing two different unification groups containing the same fermions and therefore have same Weinberg angle; the other by putting different SU L (2) structures on the same fermions and consequently have different Weinberg angles. In particular the value sin~0=3/8 is characteristic of the sequential doublet models or models which invoke a large number of additional leptons like E 6, while addition of extra charged fermion singlets can reduce the value of sin ~ 0 to 1/4. We point out that at the present time the models of grand unification are far from unique.

Unsteady nonsimilar natural convection over a vertical flat plate in a thermally stratified fluid

Abstract is not available.

Dynamics of dense sheared granular flows. Part 1. Structure and diffusion

Shear flows of inelastic spheres in three dimensions in the Volume fraction range 0.4-0.64 are analysed using event-driven simulations.Particle interactions are considered to be due to instantaneous binary collisions, and the collision model has a normal coefficient of restitution e(n) (negative of the ratio of the post- and pre-collisional relative velocities of the particles along the line joining the centres) and a tangential coefficient of restitution e(t) (negative of the ratio of post- and pre-collisional velocities perpendicular to the line Joining the centres). Here, we have considered both e(t) = +1 and e(t) = e(n) (rough particles) and e(t) =-1 (smooth particles), and the normal coefficient of restitution e(n) was varied in the range 0.6-0.98. Care was taken to avoid inelastic collapse and ensure there are no particle overlaps during the simulation. First, we studied the ordering in the system by examining the icosahedral order parameter Q(6) in three dimensions and the planar order parameter q(6) in the plane perpendicular to the gradient direction. It was found that for shear flows of sufficiently large size, the system Continues to be in the random state, with Q(6) and q(6) close to 0, even for volume fractions between phi = 0.5 and phi = 0.6; in contrast, for a system of elastic particles in the absence of shear, the system orders (crystallizes) at phi = 0.49. This indicates that the shear flow prevents ordering in a system of sufficiently large size. In a shear flow of inelastic particles, the strain rate and the temperature are related through the energy balance equation, and all time scales can be non-dimensionalized by the inverse of the strain rate. Therefore, the dynamics of the system are determined only by the volume fraction and the coefficients of restitution. The variation of the collision frequency with volume fraction and coefficient of estitution was examined. It was found, by plotting the inverse of the collision frequency as a function of volume fraction, that the collision frequency at constant strain rate diverges at a volume fraction phi(ad) (volume fraction for arrested dynamics) which is lower than the random close-packing Volume fraction 0.64 in the absence of shear. The volume fraction phi(ad) decreases as the coefficient of restitution is decreased from e(n) = 1; phi(ad) has a minimum of about 0.585 for coefficient of restitution e(n) in the range 0.6-0.8 for rough particles and is slightly larger for smooth particles. It is found that the dissipation rate and all components of the stress diverge proportional to the collision frequency in the close-packing limit. The qualitative behaviour of the increase in the stress and dissipation rate are well Captured by results derived from kinetic theory, but the quantitative agreement is lacking even if the collision frequency obtained from simulations is used to calculate the pair correlation function used In the theory.

Low-frequency dielectric constant measurements in the critical polar + non-polar binary liquid system: Methanol + n-heptane

The low-frequency (5–100 kHz) dielectric constant epsilon (Porson) has been measured in the temperature range 7 × 10−5 < t = (T − Tc)/Tc < 8 × 10−2. Near Tc an exponent ≈0.11 characterizes the power law behaviour of Image consistent with the theoretically predicted t−α singularity. However, over the full range of t an exponent ≈0.35 is obtained.

Intrinsic equations of steady rotating, incompressible viscous fluid flow

The equations governing the flow of a steady rotating incompressible viscous fluid are expressed in intrinsic form along the vortex lines and their normals. Using these equations the effects of rotation on the geometric properties of viscous fluid flows are studied. A particular flow in which the vortex lines are right circular helices is discussed.

Photocytotoxic 3d-Metal Scorpionates with a 1,8-Naphthalimide Chromophore Showing Photoinduced DNA and Protein Cleavage Activity

Ternary 3d-metal complexes of formulation [M(Tp(Ph))(py-nap)](ClO4)(1-3), where M is Co(II) (1), Cu(II) (2), and Zn(II) (3); Tp(Ph) is anionic tris (3-phenylpyrazolyl)borate; and py-nap is a pyridyl ligand with a conjugated 1,8-naphthalimide moiety, have been prepared from the reaction of metal perchlorate with KTp(Ph) and py-nap in CH2Cl2. The complexes have been characterized from analytical and physicochemical data. The complexes are stable in solution as evidenced from the electrospray ionization mass spectrometry data. The complexes show good binding propensity with calf thymus (CT) DNA, giving binding constant (K-b) values of similar to 10(5) M-1 and a molecular ``light-switch'' effect that results in an enhancement of the emission intensity of the naphthalimide chromophore on binding to CT DNA. The complexes do not show any hydrolytic cleavage of DNA. They show poor chemical nuclease activity in the presence of 3-mercaptopropionic acid or hydrogen peroxide (H2O2). The Co(II) and Cu(II) complexes exhibit oxidative pUC19 DNA cleavage activity in UV-A light of 365 rim. The Zn(II) complex shows moderate DNA photocleavage activity at 365 nm. The Cu(II)complex 2 displays photoinduced DNA cleavage activity in red light of 647.1 nm and 676 rim and near-IR light of >750 rim. A mechanistic studyin UV-A and visible light suggests the involvement of the hydroxyl radical as the reactive species in the DNA photocleavage reactions. The complexes also show good bovine serum albumin (BSA) protein binding propensity, giving K-BSA values of similar to 10(5) M-1. Complexes 1 and 2 display significant photoinduced BSA cleavage activity in UV-A light. The Co(II) complex 1 shows a significant photocytotoxic effect in HeLa cervical cancer cells on exposure to UV-A light of 365 nm, giving an IC50 value of 32 mu M. The IC50 value for the py-nap ligand alone is 41.42 mu m in UV-A light. The IC50 value is >200 mu M in the dark, indicating poor dark toxicity of 1. The Cu(II) complex 2 exhibits moderate photocytotoxicity and significant dark toxicity, giving IC50 values of 18.6 mu m and 29.7 mu m in UV-A light and in the dark, respectively.

Steady motion of a rotating stratified fluid bounded by porous disks

Abstract is not available.

Unsteady MHD mixed convection flow over an impulsively stretched permeable vertical surface in a quiescent fluid

The unsteady mixed convection flow of an incompressible laminar electrically conducting fluid over an impulsively stretched permeable vertical surface in an unbounded quiescent fluid in the presence of a transverse magnetic field has been investigated. At the same time, the surface temperature is suddenly increased from the surrounding fluid temperature or a constant heat flux is suddenly imposed on the surface. The problem is formulated in such a way that for small time it is governed by Rayleigh type of equation and for large time by Crane type of equation. The non-linear coupled parabolic partial differential equations governing the unsteady mixed convection flow under boundary layer approximations have been solved analytically by using the homotopy analysis method as well as numerically by an implicit finite difference scheme. The local skin friction coefficient and the local Nusselt number are found to decrease rapidly with time in a small time interval and they tend to steady-state values for t* >= 5. They also increase with the buoyancy force and suction, but decrease with injection rate. The local skin friction coefficient increases with the magnetic field, but the local Nusselt number decreases. There is a smooth transition from the unsteady state to the steady state. (C) 2010 Elsevier Ltd. All rights reserved.

Spectral and kinetic characterization of 7,8-diaminopelargonic acid synthase from Mycobacterium tuberculosis

The indispensability of biotin for crucial processes like lipid biosynthesis coupled to the absence of the biotin biosynthesis pathway in humans make the enzymes of this pathway, attractive targets for development of novel drugs against numerous pathogens including M. tuberculosis. We report the spectral and kinetic characterization of the Mycobacterium tuberculosis 7,8-Diamino-pelargonic acid (DAPA) synthase, the second enzyme of the biotin biosynthesis pathway. In contrast to the E. coli enzyme, no quinonoid intermediate was detected during the steady state reaction between the enzyme and S-adenosyl-L-methionine (SAM). The second order rate constant for this half of the reaction was determined to be 1.75 +/- 0.11 M-1 s(-1). The K-m values for 7-keto-8-aminopelargonic acid (KAPA) and SAM are 2.83 mu M and 308.28 mu M, respectively whereas the V-max and k(cat) values for the enzyme are 0.02074 mu moles/min/ml and 0.003 s(-1), respectively. Our initial studies pave the way for further detailed mechanistic and kinetic characterization of the enzyme.

Interrelations between Dry Eye Syndrome and Tear Fluid Phospholipid Transfer Protein

The simplified model of human tear fluid (TF) is a three-layered structure composed of a homogenous gel-like layer of hydrated mucins, an aqueous phase, and a lipid-rich outermost layer found in the tear-air interface. It is assumed that amphiphilic phospholipids are found adjacent to the aqueous-mucin layer and externally to this a layer composed of non-polar lipids face the tear-air interface. The lipid layer prevents evaporation of the TF and protects the eye, but excess accumulation of lipids may lead to drying of the corneal epithelium. Thus the lipid layer must be controlled and maintained by some molecular mechanisms. In the circulation, phospholipid transfer protein (PLTP) and cholesteryl ester transfer protein (CETP) mediate lipid transfers. The aim of this thesis was to investigate the presence and molecular mechanisms of lipid transfer proteins in human TF. The purpose was also to study the role of these proteins in the development of dry eye syndrome (DES). The presence of TF PLTP and CETP was studied by western blotting and mass spectrometry. The concentration of these proteins was determined by ELISA. The activities of the enzymes were determined by specific lipid transfer assays. To study the molecular mechanisms involved in PLTP mediated lipid transfer Langmuir monolayers and asymmetrical flow field-flow fractionation (AsFlFFF) was used. Ocular tissue samples were stained with monoclonal antibodies against PLTP to study the secretion route of PLTP. Heparin-Sepharose affinity chromatography was used for PLTP pull-down experiments and co-eluted proteins were identified with MALDI-TOF mass spectrometry or Western blot analysis. To study whether PLTP plays any functional role in TF PLTP-deficient mice were examined. The activity of PLTP was also studied in dry eye patients. PLTP is a component of normal human TF, whereas CETP is not. TF PLTP concentration was about 2-fold higher than that in human plasma. Inactivation of PLTP by heat treatment or immunoinhibition abolished the phospholipid transfer activity in tear fluid. PLTP was found to be secreted from lacrimal glands. PLTP seems to be surface active and is capable of accepting lipid molecules without the presence of lipid-protein complexes. The active movement of radioactively labeled lipids and high activity form of PLTP to acceptor particles suggested a shuttle model of PLTP-mediated lipid transfer. In this model, PLTP physically transports lipids between the donor and acceptor. Protein-protein interaction assays revealed ocular mucins as PLTP interaction partners in TF. In mice with a full deficiency of functional PLTP enhanced corneal epithelial damage, increased corneal permeability to carboxyfluorescein, and decreased corneal epithelial occludin expression was demonstrated. Increased tear fluid PLTP activity was observed among human DES patients. These results together suggest a scavenger property of TF PLTP: if the corneal epithelium is contaminated by hydrophobic material, PLTP could remove them and transport them to the superficial layer of the TF or, alternatively, transport them through the naso-lacrimal duct. Thus, PLTP might play an integral role in tear lipid trafficking and in the protection of the corneal epithelium. The increased PLTP activity in human DES patients suggests an ocular surface protective role for this lipid transfer protein.

Cooling of a composite slab in a two-fluid medium

A mixed boundary value problem associated with the diffusion equation that involves the physical problem of cooling of an infinite parallel-sided composite slab in a two-fluid medium, is solved completely by using the Wiener-Hopf technique. An analytical solution is derived for the temperature distribution at the quench fronts being created by two different layers of cold fluids having different cooling abilities moving on the upper surface of the slab at constant speedv. Simple expressions are derived for the values of the sputtering temperatures of the slab at the points of contact with the respective layers, assuming the front layer of the fluid to be of finite width and the back layer of infinite extent. The main problem is solved through a three-part Wiener-Hopf problem of a special type and the numerical results under certain special circumstances are obtained and presented in the form of a table.

Study Of Fluid-Flow And Residence-Time Distribution In A Continuous Slab Casting Mold

The fluid-flow pattern and residence-time distribution (r.t.d.) of the fluid in a continuous casting mould have been studied using a water model. The two recirculating zones below the discharge ports have been found to be asymmetric. The effect of casting speed, discharge port diameter, shroud well depth and the immersion depth on r.t.d. have been investigated. The r.t.d. curve has been well represented by a model of two backmix cells of equal volume in series. The exist of the fluid has been found to be non-uniform across the cross-section of the mould. The fluid-flow pattern has been observed to change with time in a random fashion. Dead volume of upto 31.8% has been found with smaller discharge ports.

Channel Volatiles Of South Indian Cordierites As Indicators Of Metamorphic Fluid Composition

The channel volatiles in cordierites of the Precambrian high-grade metapelites from southern and eastern Karnataka northern Tamil Nadu and southern Kerala were analyzed in an attempt to use them as metamorphic fluid fugacity indicators. Infrared powder absorption spectra, used to characterize the channel volatiles, showed that all the 21 analyzed cordierites have H2O and CO2 as the channel volatiles, indicating the predominantly H2O-CO2 composition of the metamorphic fluids. The H2O fraction in the metamorphic fluid was computed using a published thermodynamic method in conjunction with gravimetrically determined cordierite channel H2O content, available P - T estimates and an appropriate equation of state for the H2O - CO2 fluids. The IR data and these calculated X(H2O) values indicate an overall correlation between the variation in the relative proportion of H2O and CO2 in the fluids and the metamorphic grade. The average computed X(H2O) values are: 0.78 for the amphibolite facies eastern Karnataka pelites, 0.36 for the amphibolite facies southern Karnataka pelites, 0.19 for the southern Karnataka transitional zone rocks and 0.13 for the northern Tamil Nadu granulites. Consistently low X(H2O) values, at about 0.2, were obtained for the orthopyroxene-bearing assemblages.

Cooling of an infinite slab in a two-fluid medium

A mixed boundary-valued problem associated with the diffusion equation, that involves the physical problem of cooling of an infinite slab in a two-fluid medium, is solved completely by using the Wiener-Hopf technique. An analytical solution is derived for the temperature distribution at the quench fronts being created by two different layers of cold fluids having different cooling abilities moving on the upper surface of the slab at constant speed. Simple expressions are derived for the values of the sputtering temperatures of the slab at the points of contact with the respective layers, assuming one layer of the fluid to be of finite extent and the other of infinite extent. The main problem is solved through a three-part Wiener - Hopf problem of a special type, and the numerical results under certain special circumstances are obtained and presented in the form of a table.