The effects of antipsychotics on presynaptic dopaminergic functions, such as dopamine synthesis capacity, might also be related to their therapeutic efficacy. Positron emission tomography (PET) was used to examine the effects of the partial agonist antipsychotic drug aripiprazole on presynaptic dopamine synthesis in relation to dopamine D2 receptor occupancy and the resulting changes in dopamine synthesis capacity in healthy men. On separate days, PET studies with [11C]raclopride and L-[β-11C]DOPA were performed under resting condition and with single doses of aripiprazole given orally. Occupancy of dopamine D2 receptors corresponded to the doses of aripiprazole, but the changes in dopamine synthesis capacity were not significant, nor was the relation between dopamine D2 receptor occupancy and these changes. A significant negative correlation was observed between baseline dopamine synthesis capacity and changes in dopamine synthesis capacity by aripiprazole, indicating that this antipsychotic appears to stabilize dopamine synthesis capacity. The therapeutic effects of aripiprazole in schizophrenia might be related to such stabilizing effects on dopaminergic neurotransmission responsivity.Dopamine D Recently, a new atypical antipsychotic drug acting as a partial agonist of dopamine D2 receptors, aripiprazole, has been widely used for the treatment of schizophrenia 2 receptors can modulate the dopaminergic neurotransmission as functional agonists or functional antagonists Effects of antipsychotic drugs with antagonistic property mediated by blockade of postsynaptic dopamine D2 receptors using [3H]DOPA, L-[β-11C]DOPA, and 6-[18F]fluoro-L-DOPA, respectively Effects of antipsychotics on presynaptic dopaminergic functions, e.g., dopamine synthesis capacity, might also be related to their therapeutic effects. The regional activity of aromatic L-amino acid decarboxylase (AADC) in brain, indicating dopamine synthesis capacity, can be estimated using radiolabeled L-DOPA 18F]fluoro-L-DOPA revealed a significant increase in dopamine synthesis capacity in healthy human subjects 18F]fluoro-L-DOPA Effects of antipsychotics with antagonistic property on dopamine synthesis capacity have been studied in brains of human subjects. The acute administration of the antipsychotic drug haloperidol and the use of PET with 6-[2 receptor bindings and dopamine synthesis capacities at resting condition and after oral administration of a single dose of aripiprazole were measured in the same human subjects by PET with [11C]raclopride and L-[β-11C]DOPA, respectively, to determine changes in dopamine synthesis capacity by this antipsychotic in relation to the occupancy of dopamine D2 receptors. Similar experimental protocol as previous our work with risperidone was used, and results were compared In the present study, dopamine D2 receptors for each dose of aripiprazole as measured by PET with [11C]raclopride ranged from 53% to 79% in the caudate and from 51% to 77% in the putamen  and 10.4±1.5 ng/mL for 3 mg of oral administration dose of aripiprazole, 29.0±2.1 ng/mL and 25.6±2.1 ng/mL for 6 mg, and 39.6–40.4 ng/mL and 38.2–39.7 ng/mL for 9 mg, respectively. The plasma concentrations of dehydroaripiprazole during [11C]raclopride and L-[β-11C]DOPA PET studies were 0.4±0.2 ng/mL (mean ± SD) and 0.5±0.2 ng/mL for 3 mg of oral administration dose of aripiprazole, 0.9±0.3 ng/mL and 1.1±0.4 ng/mL for 6 mg, and 1.1–1.6 ng/mL and 1.4–2.4 ng/mL for 9 mg, respectively.The plasma concentrations of aripiprazole during [i of L-[β-11C]DOPA in the caudate and putamen, indicating the dopamine synthesis capacity for baseline and drug challenge studies, are shown in i were observed between the two studies (paired t-test). Typical images of L-[β-11C]DOPA for baseline study are shown in The uptake rate constants k2 receptor occupancy and percentage changes in ki by the drug challenge. There were no significant correlations. No dose dependency was observed in percentage changes in ki by the drug challenge. The relations between ki in the baseline study and percentage change in ki by the drug challenge for each administration dose of aripiprazole are shown in P = 0.005, putamen: P = 0.027).2 receptors 2 receptors corresponding to the dose of aripiprazole was observed 2 receptors and changes in dopamine synthesis capacity by aripiprazole. These findings are similar to our previous observation in healthy human subjects using risperidone 2 receptors were observed in animal studies 2 receptors might reduce presynaptic activity through feedback regulation The present study was performed using similar experimental protocol as previous our work with the antipsychotic risperidone, an antagonist for dopamine D2 receptors ranged from about 50% to 80% in the present study, there might be some kind of threshold of occupancy by aripiprazole for the stabilizing effect of dopamine synthesis capacity to emerge. Further investigations about such threshold should be considered.In the present study, significant negative correlations were observed between baseline dopamine synthesis capacity and the percentage changes in dopamine synthesis capacity by aripiprazole. This indicates that aripiprazole administration causes either increase or decrease in dopamine synthesis capacity in subjects with low or high baseline dopamine synthesis capacity, respectively, and the degrees of increase and decrease in dopamine synthesis capacity depend on the baseline dopamine synthesis capacities. These findings are similar to our previous observation in healthy human subjects using the antagonist antipsychotic risperidone 2 receptors in this study might be relatively lower than in previous reports regarding drug challenge studies being performed after daily administration of aripiprazole for more than ten days The occupancy of dopamine D2A receptors and a partial agonistic action on 5-HT1A receptors with relatively high affinity 2A receptor antagonists have been reported to modulate endogenous dopamine release 2A receptors, it may modulate endogenous dopamine release. These reports suggest that changes in dopamine synthesis capacity by the administration of aripiprazole might be due not only to pharmacological effects on dopaminergic autoreceptors, but also on serotonin 5-HT2A receptors similar to our previous report on risperidone 2 receptors, such as sulpiride, should be performed Aripiprazole also has an antagonistic action on serotonin 5-HT2 receptor bindings and dopamine synthesis capacities at resting condition and after oral administration of a single dose of the partial agonist antipsychotic aripiprazole were measured in the same human subjects. While dose-corresponding occupancy of dopamine D2 receptors was observed, no significant changes in dopamine synthesis capacity by aripiprazole administration were observed. In addition, no significant correlation between occupancy of dopamine D2 receptors and changes in dopamine synthesis capacity by aripiprazole was observed. On the other hand, a significant negative correlation was observed between baseline and aripiprazole-induced changes in dopamine synthesis capacities, indicating that the partial agonist antipsychotic aripiprazole can be considered as having a stabilizing effect on dopamine synthesis capacity, the same as antagonist antipsychotic drugs. This suggests that the therapeutic effects of aripiprazole in schizophrenia are possibly related to the stabilizing effects on dopaminergic neurotransmission responsivity.In conclusion, dopamine DThe study was approved by the Ethics and Radiation Safety Committees of the National Institute of Radiological Sciences, Chiba, Japan. Twelve healthy men  were recruited and written informed consent was obtained. The subjects were free of somatic, neurological and psychiatric disorders according to their medical history and magnetic resonance (MR) imaging of the brain. No histories of current or previous drug abuse were revealed by interviews.68Ge-68Ga line source was performed for attenuation correction. A head fixation device with thermoplastic attachments for individual fit was used to minimize head movement during the PET measurements.All PET studies were performed with a Siemens ECAT Exact HR+ system, providing 63 sections with an axial field of view of 15.5 cm 11C]raclopride and L-[β-11C]DOPA were performed sequentially. Dynamic PET scanning was performed for 60 minutes following an intravenous rapid bolus injection of [11C]raclopride. Then, one hour later, dynamic PET scanning was performed for 89 minutes after intravenous rapid bolus injection of L-[β-11C]DOPA. The frame sequence consisted of twelve 20-sec frames, sixteen 1-min frames, and ten 4-min frames for [11C]raclopride, and seven 1-min frames, five 2-min frames, four 3-min frames, and twelve 5-min frames for L-[β-11C]DOPA. The radioactivity injected was 218–237 MBq and 364–392 MBq in the baseline studies, and 199–233 MBq and 364–415 MBq in the drug challenge studies for [11C]raclopride and L-[β-11C]DOPA, respectively. Specific radioactivity was 162–239 GBq/µmol and 24–124 GBq/µmol in the baseline studies, and 125–253 GBq/µmol and 17–273 GBq/µmol in the drug challenge studies for [11C]raclopride and L-[β-11C]DOPA, respectively. A venous blood sample was taken at the beginning of L-[β-11C]DOPA PET scanning to measure natural neutral amino acid (NAA) concentration in plasma. The NAA concentration was measured by HPLC . The amino acids are phenylalanine, tryptophan, leucine, methionine, isoleucine, tyrosine, histidine, valine and threonine, which are transported via the same carrier at the blood-brain barrier as L-DOPA PET studies were performed under resting condition (baseline study) and oral administration of aripiprazole  on separate days. The interval between the 2 studies was 7 days in 7 subjects, and 14 days in 5 subjects. In each study, both PET scans with [11C]raclopride. The aripiprazole dose was 3 mg in 5 subjects, 6 mg in 5 subjects, and 9 mg in 2 subjects. To estimate the plasma concentration of aripiprazole and its active metabolite, dehydroaripiprazole, venous blood sampling was performed at the start and end of each PET scan In the drug challenge studies, aripiprazole at 3–9 mg was orally administered 3.5 hours before the start of PET scanning with [All MR imaging studies were performed with a 1.5-Tesla MR scanner . Three-dimensional volumetric acquisition of a T1-weighted gradient echo sequence produced a gapless series of thin transverse sections .All MR images were coregistered to the PET images with the statistical parametric mapping (SPM2) system 11C]raclopride, the binding potential (BPND) was calculated by the reference tissue model method i is the radioactivity concentration in a brain region; Cr is the radioactivity concentration in the reference region; RI is the ratio of K1/K1′ ; k2 is the efflux rate constant for the brain region; ND, RI, and k2) were estimated by non-linear least-squares curve fitting. The cerebellum was used as reference region. Dopamine D2 receptor occupancy by aripiprazole was calculated as follows:ND(baseline) is the BPND value in the baseline study, and BPND(drug) is the BPND value in the drug challenge study.For PET studies with [11C]DOPA, indicating the dopamine synthesis capacity, was estimated by graphical analysis i using time-activity data in a reference brain region with no irreversible binding. The ki values can be estimated by simple linear least-squares fitting as follows:i and Ci′ are the total radioactivity concentrations in a brain region with and without irreversible binding, respectively, and t* is the equilibrium time of the compartment for unchanged radiotracer in brain tissue. Plotting t*, yields a straight line with the slope ki and intercept F. In the present study, the occipital cortex was used as reference region with no irreversible binding, because this region is known to have the lowest dopamine concentration t* was set to be 29 min, and data plots of 29 to 89 min were used for linear least-squares fitting i by oral administration of aripiprazole was calculated as follows:i(baseline) is the ki value in the baseline study, and ki(drug) is the ki value in the drug challenge study.The uptake rate constant for L-[β-


Molecular symmetry is intimately connected with the classical concept of three-dimensional molecular structure. In a non-classical theory of wave-like interaction in four-dimensional space-time, both of these concepts and traditional quantum mechanics lose their operational meaning, unless suitably modified. A required reformulation should emphasize the importance of four-dimensional effects like spin and the symmetry effects of space-time curvature that could lead to a fundamentally different understanding of molecular symmetry and structure in terms of elementary number theory. Isolated single molecules have no characteristic shape and macro-biomolecules only develop robust three-dimensional structure in hydrophobic response to aqueous cellular media. Most quantum-theoretical concepts of chemistry have their origin in spectroscopy. The atomic spectroscopy of Kirchhoff, Bunsen, Fraunhofer and others resulted in the formulation of Balmer’s mysterious formula, the direct stimulus for the development of quantum mechanics. From molecular spectroscopy, which also dates from the 19th century, developed the concept of molecular symmetry, based on point-group theory. The concept of molecular structure was conjectured to bridge the gap between the two branches of spectroscopy. It is noted in passing that the discipline of quantum molecular theory is based entirely on this postulated, but unproven, concept of molecular structure.The analysis of molecular crystals by X-ray diffraction is routinely accepted as final justification of the molecular-structure hypothesis, on assuming that individual molecules maintain their integrity and structure in the solid state. This assumption is demonstrably unjustified. The molecular unit projected from crystallographic analysis is best described as a rigid three-dimensional framework that connects point atoms and is stabilized by crystal-packing interactions. In the absence of such constraints the crystallographic point group no longer represents the shape of the more flexible molecule. Related techniques such as gas-phase electron diffraction define no more than a radial distribution function. Spectroscopists are well aware of the complications associated with defining the symmetry group of non-rigid molecules, but the chemist naïvely accepts that their primitive concept of molecular structure is consistent with the most fundamental theories of physics. It is not. A recently published history of quantum chemistry  never meThe classic example of a non-rigid molecule is ammonia, which appears as a bipyramidal arrangement of interconverting polyhedra that reflects by tunneling through the plane of the central N atom . It is pSimilar situations can be constructed for all molecules.As far as the quantum-mechanical formalism is concerned, ammonia is by far no exception. Neither chemical structures nor chemical bonds do exist any more.They do not fit into the molecular-structure theme of traditional chemistry and are considered as “strange” states from a chemical point of view. On the other extreme, molecules such as the adamantane-derived bispidine ligands have been demonstrated in synthetic studies to operate as rigid units with apparently well-defined classical structure . We contMolecular symmetry is a classical concept based on the hypothetical model of point atoms held together by rigid chemical bonds. There is no experimental evidence for the existence of either point atoms nor chemical bonds. Both of these concepts are postulates to support the assumption of molecules with three-dimensional geometrical structure and molecular symmetry is the mathematical description of this structure. Alternatively molecular symmetry could be defined in line with more recent views on atomic and molecular structure.A more appropriate theory, starting from the nature of electrons and atomic nuclei as wave-like distortions of four-dimensional space-time, characterizes a molecule as a 4D standing-wave structure in which individual particles do not have independent existence. The only meaningful symmetry of such a construct must be four- and not three-dimensional. It will be argued that classical molecular structure is the three-dimensional projection of non-classical four-dimensional molecular geometry.mn is a remnant of the classically assumed ultimate philosophical limit to the subdivision of matter. It has been used in this form by Newton, Dalton, Mendeleév and others, until the end of the 19th century. Following Nagaoka, Rutherford and Bohr the concept of a structureless atom was abandoned and replaced by the planetary model of point-like electrons, with mass me in orbit around a point-like heavy nucleus. With the advent of quantum mechanics the Bohr–Sommerfeld orbits gave way to chemical shells of probabilistic charge density—now defining the electron as a point particle! To account for the behaviour of electrons in diffraction experiments the kabbalistic notion of wave-particle duality was concocted, notwithstanding the wave-mechanical alternative proposed by Schrödinger.The point atom with mass As the particle model found its way into textbooks the electronic wave structure was completely forgotten, although the quantum-mechanical model never made any logical sense. In order to rationalize the probability function as representing a single electron it is necessary to integrate over all space. The electron associated with a given hydrogen atom, although likely to occupy the ground-state, has a finite probability to be on the moon. The chemists of the world have been content to live with this assumption for a hundred years. They have been brainwashed so thoroughly as not to tolerate any alternative suggestion that makes logical sense.The most bothersome aspect of electronic quantum mechanics is the failure to account for the most prominent attribute of an electron, known as its spin. Efforts to artificially add a spin function to the three-dimensional state function of an electron have only been partially successful. A common remedy is often proposed through the assertion that spin is a relativistic phenomenon, but a conclusive proof has never been formulated. What has been demonstrated is that the spin function, known as a spinor, occurs naturally as an inherent feature of four-dimensional motion. In order to define an electron with spin it is therefore necessary to describe it as an entity in four-dimensional space-time. This would automatically rule out its definition as a point particle, noting that a point in four-dimensional space-time has, not only zero extension but also zero duration. It cannot exist.With this realization the interminable debate about particle or wave becomes redundant and the remaining option is to consider the electron as a four-dimensional wave structure. In fact, Schrödinger’s equation is a modification of the general wave Equation. It is fashionable to state  thatplausible; they cannot be used to derive or prove this equation.… analogies between geometrical optics and classical mechanics on the one hand, and wave optics and quantum mechanics on the other hand … can only make the Schrödinger equation seem Statements like this are standard textbook fare that derives from the Copenhagen campaign to discredit Schrödinger’s wave interpretation ):x0 = ict, etc., this equation rearranges intoBy defining i.e.,The solution of i, j, k are generalizations of i2 + j2 + k2 = ijk = −1.known as a quaternion or the spin function. The coefficients J = L + S, a solution of In order to obtain Schrödinger’s solution it is necessary to separate the space and time variables on solving In the quaternion representation an electron can no longer be considered as an entity that moves through space, but rather as a distortion of space-time, or the vacuum. A vivid visualization was proposed by Weingard … conceiving of the motion of particles on analogy with the motion of the letters on the illuminated news sign in Times Square. The letters move, not by anything moving, but by the sequential blinking of the lights of which the sign is composed. Similarly, we can conceive of the motion of a particle as the motion of a geometrical property. Regions of space do not move, but the geometry of adjacent regions change in such a way that a pattern of geometrical properties—the pattern we identify with a certain particle—moves.In quaternion terminology such a geometrical pattern may be described as a standing four-dimensional spin wave. Such a wave packet is flexible, by definition, and adapts to the geometry of its local environment. The size of a wave packet in space-time is restricted by the limited time component that prevents indefinite extension. The hydrogen electron can no longer be on the moon. The topology of the wave packet manifests, in addition to spin, also as characteristic charge and mass.On interaction with a proton, a spin wave of opposite charge and high mass, the electron envelops the proton in a hyperspherical shroud. Assuming that waves of similar topology readily coalesce into larger composite waves, the formation of heavier atoms appears as a natural process. All interactions and rearrangements consist of finding an equilibrium among merging patterns. The only feature to explain is the origin of the elementary patterns such as electron, proton and neutrino.The answer is provided by the theory of general relativity. We note in passing that the Lorentz transformation that defines special relativity amounts to a complex rotation in four-dimensional space-time, with the same structure as the quaternion spin function . Wave meGμν balances the energy-stress tensor Tμν, which may be interpreted as distortions (matter) in space-time. This means that Euclidean (flat) space-time has Tμν = 0 and hence contains no energy or matter. As space-time curves, elementary wrinkles develop and coalesce into atomic matter and eventually into macroscopic massive objects that cause increased local curvature of space-time, recognized as a gravitational field.In words, the curvature tensor The empirical Periodic Table of the Elements is widely accepted as the major contribution of chemistry to science, and correctly so. Whereas the Bohr–Sommerfeld atomic model could be demonstrated to be consistent with the periodic model by invoking the empirical exclusion principle of Pauli, the Schrödinger wave-mechanical model can only partially account for elemental periodicity. Two reasons for the discrepancy are the neglect of spin and the assumption of zero interaction with the environment as in Euclidean space-time.A few years ago it was demonstrated  that, onOf more immediate relevance is that appearance of the golden mean indicates a self-similar relationship between atomic structure, botanical growth, patterns in the solar system and spiral galaxies. In response it could be shown ,11 that The classical straight-line representation of a chemical bond poorly describes the mode of interaction between spherical wave structures. A more likely model is pictured in The consecutive frames may be interpreted as schematic representations of homonuclear covalent interactions of increasing order. To distinguish between different modes of overlap, frames 1 and 3 could represent integer and frame 2 half-integer order.The relative nuclear positions implied by the two-dimensional drawing of π to restore the initial configuration and represents an element of four-dimensional symmetry, which does not occur in three dimensions.There are two possible modes of rotation, shown schematically in In this mode the cyclic disposition of atomic cores fluctuates with the spin function, consistent with the interaction between wave structures being subject to spin pairing. The hyperspherical molecular symmetry may be visualized as the interpenetration of two wave packets in spherical rotation such that the equilibrium separation of their centres of mass defines a vibrating interatomic distance. Generalization of this pattern suggests that atomic cores in multi-atomic molecules are confined, with restricted freedom, within a sea of valence electrons.In the case of heteronuclear interaction the interference patterns must obviously be of lower symmetry, but in four dimensions it has the same hyperspherical symmetry as before. As more atoms participate in the formation of larger molecules a unique holistic interference pattern characterizes each equilibrium arrangement. The removal or displacement of any atom in the molecule from/by another affects the entire wave pattern, and hence the molecular symmetry, in a non-trivial way.Atoms do not interact spontaneously to form molecules under all circumstances. Chemical theory assumes that interaction in a reacting system only commences under specific thermodynamic conditions that define a characteristic valence state for the system. This happens when a single activated electron becomes decoupled from its atomic core and free to interact with a similarly activated electron from another atom, as shown in r0, known as the characteristic ionization radius (not to be confused with ionic radius) [r0. The valence electron in the activated stationary state with spherical symmetry has neither kinetic nor classical potential energy and its energy ofThe decoupled valence electron behaves like an elementary charge, which is smeared out uniformly over a sphere of radius  radius)  of the a radius)  is distrwhich represents quantum-potential energy  ofEg in eV.is simply related to the classical electronegativity , χ=Eg, r0. Steric distortion of the equilibrium molecular arrangement is resisted by the interference pattern that requires either integer or half-integer bond orders. Such resistance should correlate with spectroscopically measured harmonic stretching force constants. In principle all characteristics of covalent interaction could therefore be calculated directly from atomic ionization radii as the only parameter.All covalence interaction parameters such as interatomic distance, bond order and dissociation energy should therefore be simple functions of d, of homonuclear diatomic interactions of the same order are linear functions of ionization radius,Although the definition of bond order as the extent of overlap between interfering waves is completely different from the classical concept, interatomic distances predicted by the wave model are remarkably similar to experimental estimates of empirically assumed bond orders. In terms of the wave model all interatomic distances, kb is characteristic for order b. In the same way that atomic electron distribution, as a standing wave, is predicted by logarithmic-spiral optimization, bond-order coefficients, kb, are specified in the same way, as shown in kb varies from unity to τ for b = 0 to 4.where τn, of the golden ratio.For heteronuclear interaction the relationship between interatomic distance and bond order is more complicated, but in all cases the golden ratio is involved. By the use of predicted bond orders and interatomic distance, dissociation energy and stretching force constants are next calculated directly from ionization radii and integral powers, bondons to represent the electron pairs of covalent interaction. Final synthesis of these ideas into a working model of covalence seems to converge towards an understanding based on the role of the quantum potential and the golden ratio [Other alternative approaches to the problem of covalence have appeared in the recent literature –22. The en ratio .divine proportion.To cut a long story short we repeat that the characteristics of electrons, atoms and molecules are intimately related to the golden parameter and in order to understand molecular structure and symmetry we find it necessary to establish why this should be the case. At the most fundamental level we suspect the golden ratio to function as a critical characteristic of the general curvature of space-time. In order to explore this possibility it is necessary to consider a few relevant aspects of cosmology. We start from classical cosmology with a known link to the golden ratio, which Johannes Kepler, on cosmological grounds, referred to as the The cosmology of Pythagoras ,25  which interact through a third irrational diagonal component that contains the sum of the first male and female numbers (3 + 2), divides the four-element  world in the divine proportion of The cosmic Translated into modern terms :τ, which puts the universe into harmonious perspective of projective geometry.The cosmic whole consists of a world and an anti-world, separated by an interface that creates all matter as it curves through four-dimensional space-time, with an involution, described by the golden ratio For the benefit of prospective readers not familiar with projective geometry, a short introductory primer summarizes some details and Kepler’s contribution in the πr/2) to the length of the diagonal chord . It proWe emphasize the similitude between Pythagorean and projective cosmologies to highlight the importance of self-similarity for the understanding of molecular symmetry. Noting that … space itself has a structure that influences the shape of every existing thing,molecules, like space-time itself, would be shaped in four dimensions. This is referred to as non-classical molecular shape as distinct from the traditionally assumed three-dimensional classical shape. The closest accord between classical and non-classical molecular structures occurs in close confinement as in a crystal lattice. The biggest difference is in free interstellar space where any stable molecule is of highly symmetrical shape. As such a molecule finds itself in more crowded environments, as in a molecular cloud, each interaction reduces its symmetry according to Goldstone’s theorem . LikewisIt becomes problematic to decide which of these is the true molecular symmetry. The 0 K classical structure which is used invariably in spectroscopic analysis is probably the worst possible choice. The reason why it is a good practical model is because spectroscopic analysis relies mainly on internal parameters such as interatomic distance, described correctly by the classical structure. Molecular shape and symmetry are not predicted by molecular spectra.Without this knowledge the vibrational symmetry of molecules becomes as meaningless as the feckless caricature of statically rigid three-dimensional molecules.Nowhere in nature is there exact symmetry, only equilibrium. The reason is that the symmetry of any object depends on the symmetry of its environment, which is nowhere rigorously isotropic. It is safe to conclude that there are no symmetrical molecules except at 0 K, which is unreachable. As a mathematical concept however, symmetry is precisely defined by group theory and we hear that … the number and kinds of energy levels which an atom or molecule may have are rigorously and precisely determined by the symmetry of the molecule or of the environment of the atom.This statement can only refer to a hypothetical molecule considered as a static array of point atoms. Knowing the number and kinds of energy levels of such a molecule is of no chemical interest. In fact, there is only the single ground-state energy level. In systems of chemical significance group theory has no function unless molecular symmetry is already known empirically. One may ask, as in the classic hamburger commercial: “ where is the beef?”via the techniques of classical molecular mechanics. In principle it is possible to progress from here to situations of chemical importance by modelling the effects of thermodynamics on classical bond parameters. Considerable progress has been achieved by simulations of this type, known as molecular dynamics. However, commentators [Without any knowledge of molecular shape or symmetry all that remains is the connectivity pattern which is directly commensurate with the 0 K classical structure and symmetry entators  who procChemistry has a long way to go towards an understanding of the four-dimensional shape and symmetry of molecules. A useful first step would be to admit the inadequacy of three-dimensional quantum mechanics, which cannot deal with four-dimensional wave phenomena such as the electromagnetic field and the solutions of … it is not unusual when it comes to quantum philosophy, to find the very best physicists and mathematicians making sharp emphatic claims, almost of a mathematical character, that are trivially false and profoundly ignorant.sp hybrids the orthogonality of s and p functions, which means zero overlap, is simply ignored.The performance of chemists is even worse—they accept the claims of the physicist at face value, then insert their classical model as a quantum concept, by the simple device of describing complex functions as real hybrid orbitals . The impEqually remarkable is the way in which molecular spectroscopists keep up the pretence of a quantum theory, despite the well-known fact that the assignment of group frequencies is independent of the assumed molecular structure and symmetry. Without classical structure there is precious little left of molecular quantum theory and symmetry. To fill this hiatus would require a complete reformulation of the problem.It is, first of all, necessary to appreciate the four-dimensional nature of molecules. The symmetry of such a molecule is to be understood, by analogy with the three-dimensional VSEPR model, as resulting from the equilibrium arrangement of various wave structures that minimizes the spin function. Molecules with non-zero residual angular momentum are optically active. All others project into three dimensions with spherical symmetry. It should be clear that this 3D projection cannot be static as the time coordinate of space-time, as perceived in projection, would be non-zero. This means that the shape of a free achiral molecule represents the average of a spherically pulsating configuration.The spherical symmetry could break down if the molecule, when set into axial rotation, as in a microwave field, exhibits a dipole moment. In an applied electric field Stark modulation permits spectroscopic measurement of the dipole moment. Infrared and Raman spectra arise from molecular vibrations induced by further breakdown of symmetry. Molecular spectra simply do not reveal the structure or symmetry that molecules do not possess. Any symmetry argument that enters into the interpretation of molecular spectra arises from structure due to interaction with electromagnetic fields in the environment, becoming more pronounced in condensed phases, including solutions.Three-dimensional structures inferred from NMR spectra and X-ray crystallography are purely classical and confirms no more than chemical connectivity patterns. The frequent occurrence of molecular rearrangements shows that these patterns are not necessarily invariant. It is of interest to note that the complete molecular eigenstate, even of the Born–Oppenheimer Hamiltonian of complex molecules, signifies spherical symmetry . StructuBefore the ideas outlined here can be incorporated into a meaningful theory of chemistry it is necessary to contemplate the nature of a non-classical four-dimensional molecule. For most chemists this would imply a major paradigm shift; away from their traditional view of rigid three-dimensional molecular quantum objects. The enormity of the proposed paradigm shift is underlined by statements from leading theoreticians such as :There is no such thing as spacetime in the real world of physics … It is an approximation idea, an extremely good approximation under most circumstances, but always only an approximation.Rydberg molecule, in which the distance scale between interacting units is highly inflated. This implies an increase in atomic (and molecular) size, ionization radius and interatomic distance, with concomitantly decreased dissociation energy and molecular stability. Not surprisingly only the most stable diatomic molecules H2, CO, N2, etc., have been observed [The first obstacle to overcome is the counter-intuitive notion of space-time entanglement that implies a temporal component to space variables. Projection into three-dimensional space separates space and time coordinates to create the illusion of an object in isotropic three-dimensional motion. This means that a hyperspherical object appears as a pulsating spheroid in three-dimensional space, if all environmental effects are ignored. This model describes a molecule in intergalactic space. By analogy with Rydberg atoms  it shoulobserved . MacromoTo develop these ideas into useful computational models is not a straightforward exercise; for the time being it is difficult to see how it could be developed beyond the classical model of pairwise interaction. The only topic of immediate interest appears to be the spontaneous folding of macromolecules with biological function into characteristic single conformations. As revealed by crystallographic analyses the overall structure of proteins is remarkably compact, at about the same level as the crystals of small organic molecules , which mProbably the most remarkable feature of protein folding is the observation  that the… simple estimates of the number of conformations possible with even a small polypeptide chain of, say, only 50 amino acid residues imply that an astronomical length of time, many orders of magnitude longer than the age of the universe, would be required for all of them to be sampled randomly. Consequently, the folding process must be directed in some way …α–helix, β–sheet, etc.) may be predicted from the primary sequence and embedded structural elements that cause reverse turns in the surface of the protein ensure the eventual globular structure. However, since all biomolecules operate in aqueous environments, the influence of space-time curvature on molecular shape is effectively masked by hydrophobic interaction, which means that all non-polar amino-acid side chains are buried in the interior of the globule, with polar groups exposed in the hydrophilic surface; in exact analogy with the formation of micelles in soap solutions. The crucial consideration towards elucidation of protein folding therefore is the distribution of amino acids with the alternation of polar and non-polar regions required for the formation of globular micelles. The key to this is encoded in the primary amino-acid sequence, as generated by the evolution of life. Similar conclusions, from the opposite point of view [In our biased opinion the curvature of space-time is one possible factor that could direct such specific action. It is noted that in many proteins the primary polypeptide chain pursues a moderately straight course across the entire breadth of the structure and then turns to the other side, without getting knotted or entangled. Domains of uniform secondary structure ( of view , are sumWater is an integral part of biomolecular structural organization, and is central to the assembly and three-dimensional shape of proteins and nucleic acids.We have to conclude that the shape and symmetry of free molecules cannot be related to space-time topology. Molecules, small enough to persist in free space, are too small to develop a characteristic shape and macromolecules, large enough to exhibit three-dimensional structure, adopt their characteristic shape in response to interaction in condensed phases. It is only in those cases where such molecules are incorporated as building blocks in biological growth structures that the familiar Fibonacci patterns, characteristic of space-time topology, become noticeable ,29. The


Electrospinning technique is commonly used to produce micro- and/or nanofibers, which utilizes electrical forces to produce polymeric fibers with diameters ranging from several micrometers down to few nanometers. Desirably, electrospun materials provide highly porous structure and appropriate pore size for initial cell attachment and proliferation and thereby enable the exchange of nutrients. Composite nanofibers consisting of silk and hydroxyapatite nanoparticles (HAp) (NPs) had been considered as an excellent choice due to their efficient biocompatibility and bone-mimicking properties. To prepare these nanofiber composites, it requires the use of acidic solutions which have serious consequences on the nature of both silk and HAp NPs. It is ideal to create these nanofibers using aqueous solutions in which the physicochemical nature of both materials can be retained. However, to create those nanofibers is often difficult to obtain because of the fact that aqueous solutions of silk and HAp NPs can precipitate before they can be ejected into fibers during the electrospinning process. In this work, we had successfully used a three-way stopcock connector to mix the two different solutions, and very shortly, this solution is ejected out to form nanofibers due to electric fields. Different blend ratios consisting HAp NPs had been electrospun into nanofibers. The physicochemical aspects of fabricated nanofiber had been characterized by different state of techniques like that of FE-SEM, EDS, TEM, TEM-EDS, TGA, FT-IR, and XRD. These characterization techniques revealed that HAp NPs can be easily introduced in silk nanofibers using a stopcock connector, and this method favorably preserves the intact nature of silk fibroin and HAp NPs. Moreover, nanofibers obtained by this strategy were tested for cell toxicity and cell attachment studies using NIH 3 T3 fibroblasts which indicated non-toxic behavior and good attachment of cells upon incubation in the presence of nanofibers. Recent advances in tissue-engineering techniques had enabled scientists in fabricating the novel scaffolds with multi-functional properties to overcome the problems faced in the existing one. The 2D and/or 3D scaffolds used for tissue-engineering applications had greatly influenced the present scenario for scaffold construction. Although there are lots of advances made in tissue-engineering, the scientific community is still facing a major challenge to select the perfect strategy and choice of materials while considering the fabrication of scaffolds. The most uniquely used biopolymer made from silk fibroin proteins are obtained from silkworms and had a long history of applications in the human body as sutures. Silk fibroin contains peptides composed of RGD sequences that can promote cell adhesion, migration, and proliferation ,2. TheseOn the other hand, hydroxyapatite (HAp) is a major solid component of the human bone which can be used as a vital implant due to its excellent biocompatibility, bioactivity, non-immunogenicity, non-inflammatory behavior, and osteoconductive nature . HoweverAmong available methods, to create a suitable scaffold in which these biologically important materials can be incorporated is the electrospinning technique, which had emerged as a versatile technique to convert biologically significant polymers into nanofibers, so as to use them as potential candidate for tissue-engineering -12. The The use of electrospinning to fabricate the silk-based nanofibers and HAp nanoparticles (NPs) had been exploited to create 2D scaffolds. For instance, efforts to modify silk fibroin nanofibers to attribute properties of HAp was done by soaking in stimulated body fluid (SBF) by Kim et al., and this similar mineralization approach had been also frequently used by other researchers ,19. HoweIn this work, for the first time, we presented the use of aqueous regenerated silk fibroin solution blended with HAp NPs using a three-way stopcock connector. In our system, the aqueous silk solution and HAp NPs colloidal suspension combine together at the center of the three-way connector for a short time without giving enough time to precipitate, and this blend solution is immediately ejected out to form nanofibers. Different weight ratios of 10%, 30%, and 50% of HAp NPs were used as blend solution to electrospun nanofibers. The obtained nanofibers were characterized for various psychochemical characterizations, and interaction of these nanofibers with fibroblasts was done to study the cell toxicity and cell attachment of nanofibers incorporated with HAp NPs.Silkworm cocoons were obtained from the Rural Development Administration . Poly(ethylene oxide) (PEO) with an average molecular weight of 200,000  was used as sacrificial polymer to electrospun silk solution and to make HAp/PEO colloid solutions. HAp rod-shaped NPs measuring 30 to 60 nm were obtained from Dae Jung, Siheung, Gyeonggi, Korea. NIH 3 T3 fibroblasts were purchased from ATCC . Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fatal bovine serum, cocktail of 1% penicillin-streptomycin, Trypsin, were obtained from Welgene, Fresh Media™ . Trypan Blue Stain 0.4% was obtained from Gibco® . 3--2,5-diphenyltetrazolium bromide (MTT) reagent used to check the cell viability was purchased from Duchefabiochemie, Haarlem, The Netherlands. Dimethyl sulfoxide (DMSO) with high purity grade of 99.9% was acquired from Sigma-Aldrich. Tissue culture flasks and microplates for cell seeding and growth were purchased from BD Falcon™, Winston-Salem, NC, USA and SPL Life Sciences, Pocheon-si, Gyeonggi-do, Korea.λ = 1.540 A) radiation over Bragg angle ranging from 10° to 60°. To identify the vibrations caused due to functional groups in nanofibers, Fourier transform infrared spectroscopy (FT-IR) analysis was done using BIO-RAD . The samples were directly loaded on ATR window, and spectra were collected using Excaliber Series by averaging 32 scans with the resolution of 4 cm−1. The thermal analysis of the synthesized nanofibers was carried out with a thermal analysis system,  by ramping the samples at 10°C/min, and heating was started from 30°C to 700°C. Heating was followed under a continuous nitrogen purge of 100 mL/min, and spectra were collected using Q600 Software (TA Instruments). For checking the cell attachment on nanofibers by FE-SEM, the images were captured with an accelerating voltage of 3 KV with magnifications of 1 K.Variable pressure field emission scanning electron microscope (FE-SEM) EVO® LS10 equipped with energy-dispersive X-ray spectroscopy (EDS) obtained from Carl Zeiss SMT., Ltd., Oberkochen, Germany, was used to investigate the morphology and elemental detection of nanofibers. Before viewing, the samples were pasted on a carbon tape and sputter-coated using a thin layer of gold palladium for 120 s for two consecutive cycles at 45 mA with the Ion Sputter 1010, Hitachi, Chiyoda-ku, Japan. After sample coating, the micrographs from each samples were taken at an accelerating voltage of 2 KV and with magnifications of 15 K. The EDS images were captured at an accelerating voltage of 10 KV and with magnifications of 15 K. The average nanofiber diameters were calculated using the software Innerview 2.0, Dong, Bundang Daeduk Plaza, Korea, after measuring 100 diameters per sample from FE-SEM images. Transmission electron microscopy (TEM) was done by JEOL JEM-2200FS operating at 200 KV, JEOL Ltd., Akishima-shi, Japan. The samples for TEM were prepared by dispersing 10 mg of nanofibers in 200 μl of ethanol and subsequently dispersed by bath sonicator using locally supplied ultrasonic cleaner  for 120 s. After dispersing the nanofibers, 20 μl of dispersion was pipetted out by micropipette and carefully poured on 200 mesh copper grid. The extra solution was removed using Kimwipes supplied by Kimberly-Clark Professional, GA, USA, and the grid was allowed to dry overnight at room temperature. Information about the phases and crystallinity was obtained using PANalytical diffractometer  with Cu, Cr (Bombyx mori cocoons into suitable pieces and were boiled in 0.02 M Na2CO3 for an hour and subsequently washed with de-ionized water (2 to 3 times) to remove the unbound sericin. Later on, the samples were dried at room temperature for 1 day. After drying, 60 g of degummed silk was dissolved in ternary solvent composed of CaCl2/Ethanol/H2O  at 98°C for 40 min in round-bottomed flasks. Following this, protein mixture was filtered through miracloth  to remove small aggregates. Furthermore, this solution was dialyzed against deionized water using a dialysis tubing with molecular weight cutoff 12,000 to 14,000 Da  for 3 days, and water was exchanged once a day. The yielding aqueous silk fibroin solution was calculated to be 8 wt.% (which was determined by weighing the remaining solid weight after drying). Finally, the aqueous silk fibroin solutions were stored in a refrigerator and used within 15 days of time to avoid denaturation and/or precipitation.The aqueous silk solutions to be used for electrospinning were prepared by the following procedure. Firstly, degumming was achieved by cutting Before using the HAp NPs for modifying the nanofibers, the NPs were characterized for shape and size. In this regard, the morphology of obtained HAp NPs was checked by TEM. Figure For preparing solution to electrospun pristine silk nanofibers, 20 ml of 8 wt.% of aqueous silk solution was removed from the refrigerator. To give appropriate viscosity to this solution, so as to have proper bending instability for fiber formation, 4 ml of previously prepared 30 wt.% PEO solution was added as a ‘sacrificial polymer.’ The resultant blend solutions were loaded in syringes and used for electrospinning. For preparing solutions to fabricate silk fibroin nanofibers containing HAp NPs, a stepwise methodology was adopted. On one hand, silk solution was prepared in the same way as mentioned for the preparation of pristine silk nanofibers and subsequently loaded in syringes. On the other hand, PEO/HAp colloidal solution was prepared by adding 2 g of PEO in 20 ml of 0.001 molar solution of phosphate buffer saline (PBS), and this solution was mixed well to solubilize. To this solution, HAp NPs were added to give the final concentration of 10%, 30%, and 50% HAp with respect to 8% of aqueous silk fibroin solution. After adding HAp NPs in PEO solution, the HAp NPs were agitated using an ultrahigh sonication device. This was achieved using Sonics Vibra-cell model VCX 750, Newtown, CT, USA, operating at 20 kHz with an amplitude of 20%. The ultrasonic agitation was allowed to continue for a period of 1 min. After complete sonication, the samples were viewed as homogeneously dispersed and well stabled without being precipitated at the bottom. Further on, these dispersed HAp/PEO solutions were filled into the syringes to be used for electrospinning.The electrospinning of nanofibers was carried out using an electrospinning instrument purchased from eS-robot®, ESR-200R2D, NanoNC, Geumcheon-gu, Seoul, Korea. For fabricating the pristine nanofibers by electrospinning, the silk/PEO solutions were injected using 10 ml disposable plastic syringe fitted with a 22needle gauge (0.7 mm OD × 0.4 mm ID). The syringes were mounted on an adjustable stand, and flow rate of 0.8 mL/min was adjusted using a multi-syringe pump to keep the solution at the tip of the needle without dripping. The high power supply capable of generating +30 kV and −30 kV for positive and negative voltages was used to eject out the nanofibers from the needle tip. A metallic wire originating from the positive electrode (anode) with an applied voltage of +20 kV was connected to the needle tip through alligator clips, and a negative electrode (cathode) with an applied voltage of −1 kV was attached to the flat bed metallic collector ,25. The 2 environment at 37°C for 1, 2, and 3 days. The cell viability was evaluated by MTT reduction assay. After desired days of incubation, the media from 96-well were suctioned out and treated with 200 μl of the MTT solution, by mixing the contents by side-tapping, and further on, these plates were incubated at 37°C for 2 h. After incubation, MTT solution was suctioned out and added with 200 μl of DMSO, which was subsequently rocked to form purplish blue-colored formazan solution. The solubilized formazan appearing from each well were transferred to fresh wells of 96-well plate for spectrophotometric analysis at 540 nm in an ELISA microplate reader . The cell viability was obtained by comparing the absorbance of cells cultured on the nanofiber scaffolds to that of the control well containing DMSO. For cell checking attachment on nanofibers, the cells were allowed to grow for 3 and 12 days’ time, and media was changed after every 3 days. To check the cell morphology, cell fixation and dehydration was done by rinsing the samples twice with PBS followed by fixation with a 2.5 vol.% glutaraldehyde solution for 4 h. After cell fixation, the samples were rinsed with PBS and then dehydrated with graded concentrations of ethanol  for 10 min each. Finally, the samples were kept overnight in a vacuum oven and observed in FE-SEM to determine cell attachment. The samples for FE-SEM were coated by keeping the same conditions as described previously in the ‘Characterization’ section. However, the micrographs of each sample were taken at an accelerating voltage of 2 KV and with magnifications of 15 K.The frozen ampules of NIH 3 T3 fibroblasts removed from liquid nitrogen tank were incubated at 37°C for 1 to 2 min to form a semisolid suspension. The cells from these ampules were taken out and added with fresh media, centrifuged to get cell debris, and enriched with fresh media allowed to incubate at 37°C for 3 days for the completion of the first subculture. In this study, cells were used after two subcultures to check the cell viability, and cell attachment with renewal of culture media was done after 3 days. The nanofiber samples used for checking cell viability and cell attachment studies were pierced into disk shapes using biopsy punchers  forming 6-mm round disks, giving it an appropriate diameter to fit in a 96 well plate. Each nanofiber disk was sterilized by dipping it in 70% ethanol in 6-well plate for 30 min. The excess of ethanol on nanofibers after sterilization was rinsed by dipping the samples in 10 mL of DMEM. Further on, the nanofiber samples were transferred on 96-well plates in triplicates. A 100 μl of cell suspension containing 25,000 cells/mL was counted using cell counting method, and the cells were carefully seeded over the top of sterilized nanofiber disks in the 96-well plate. The seeded scaffolds were incubated at 37°C for 30 min to allow cell adhesion. Following this, 100 μl of fresh medium was added in each well, and the plates were incubated in a humidified incubator with 5% COThe three-way stopcock connector was used as the solution blending tool before ejecting the solution into nanofibers. In this regard, Figure To ensure that the added HAp particles are really present in/on nanofibers, FE-SEM equipped with EDS analysis was utilized for a comparative study of pristine and one of the modified nanofibers containing HAp NPs; the results are presented in Figure Due to the poor resolution of scanning electron microscopy, it can only reveal the surface architect of materials, while internal contents often remain untracked. For this reason, we could not find the exact location of HAp NPs on nanofiber by FE-SEM. Therefore, we used TEM to investigate the location of HAp NPs inside the nanofibers. In this context, Figure To investigate the location and chemical nature of HAp NPs over the nanofibers more precisely, TEM combined with the energy dispersive X-ray (EDS) analysis was done. Figure θ values of 31.77°, 32.90°, 34.08°, 40.45°, and 46.71° that correspond to the crystal planes (211), (300), (202), (310), and (222), respectively, which are in proper agreement with the JCPDS database [θ values from 10° to 28°. The broad diffraction peak in this inset shows the scatter peak with 2θ values of 21.9° which is indicating typical amorphous scattering pattern of amorphous silk [XRD can be utilized as a highly stable technique to investigate the crystalline nature of any material. Figure database ,28. Unfodatabase ,28. Furt planes 2, (300), ous silk . Interes−1, 1,516 cm−1, and 1,232 cm−1 which confirm the nature of the silk fibroin in the nanofibers. Moreover, nanofibers modified with HAp also showed the presence of these amide bands; however, there was a downshift of 1 to 2 units for amide Ι and amide II bands. The reason is to show that this shift can be attributed to conformational changes occurred in the silk fibroin from random coil structure to β-sheet confirmation due to the incorporation of HAp NPs [−1, which indicates the existence of a Ca-O phase. The peaks appearing at 1,059 cm−1 and 1,097 cm−1 can be attributed due to the asymmetric stretching mode vibration in PO4−3, and a medium intensity band at about 962 cm−1 results from P-O asymmetric stretching of the stretching vibrations in PO4−3[−1 is assigned to the O-H bending deformation mode due to the presence of HAp NPs in the nanofibers. The intensity of these peaks increases as the amount of original HAp used to make colloidal solution for electrospinning increases.FT-IR can be used as an efficient tool to investigate the structural confirmations because of the knowledge of the vibration origins of the amide bonds, the sensitivity of some of these band positions to conformation, and the possibility of predicting band positions for a given helical or extended conformation . The cha HAp NPs ,32. The  in PO4−3. Also, aFigure Figure To find out the cell attachment on nanofibers, the results after culturing the fibroblast for 3 and 12 days is presented in Figures In conclusion, a highly trustable technique which employs the use of stopcock connector can be used to electrospun a blend solution of fibroin and HAp together in aqueous solutions, which is impossible if simple mixing procedure is followed. Without the use of any toxic chemical, this technique can yield nanofibers with desirable properties. The FE-SEM and TEM techniques can be used to figure out the location of HAp in nanofibers and simultaneously support the use of stopcock connector to electrospun silk fibroin and HAp NPs. Fourier transform infrared spectroscopy analysis indicated the chemical interaction occurring between HAp NPs and silk fibroin, which resulted in the transformation of random coil to β-sheet confirmation of silk fibroin. It can also be concluded that HAp NPs enhanced the β-sheet conformation of fibroin and resulted in the improvement of the properties of nanofibers. Generally, it is believed that the presence of highly favorable components likeHAp could improve the cell attachment; however, in our case, we find that the attachment of fibroblast on nanofibers is independent to the presence of HAp. Moreover, the cell viability of nanofibers can be improved by this technique.The authors declare that they have no competing interests.FAS designed the study, carried out the experiments, and prepared the manuscript. HWJ, BMM, and HJP maintained the cell lines and provided vital information about the cell culture studies. OJL and JHK maintained the paperwork for obtaining the chemicals and arranging the facility to perform the characterization of materials. CHP supervised the whole work and attributed important part in the discussions of this manuscript. All authors read and approved the final manuscript.


Environmental Health Perspectives (EHP) was born. The journal published monographs on topics related to environmental human health and research papers based on conference proceedings for more than 20 years before adopting its current format in 1993 to include news, correspondence, editorials, reviews, commentaries, and research articles. In 2004 EHP became one of the first open-access scientific journals, with all news and research content available for free on the journal’s website , determined there was a need for a journal dedicated to research on human health and the environment, and  website . In keepEHP, which is how the vast majority of our readers access the journal now. In 2011 the EHP website had more than 1.2 million unique visitors and 2.7 million page views, and there were another 1.4 million visits to EHP content via PubMed Central. In contrast, there were approximately 600 paid subscribers to the print version of the journal and a total print circulation of 2,500 copies. Clearly, EHP already reaches a much larger audience online than in print, and our time, effort, and resources should be directed accordingly.There are several reasons for going to an online-only format at this time. First and foremost, going paperless will allow us to focus on optimizing electronic delivery of EHP website and expanding options for electronic delivery. EHP is currently available via a free iPhone application. An Android application is under development, and we are planning tablet and mobile site applications as well. A fully digital format will increase opportunities to include video and audio material in both news and research articles. In addition, we are planning a more dynamic format for the EHP News section, which will help attract additional readers and increase the breadth of our audience.Going paperless also will increase the flexibility of the journal and allow us to concentrate on enhancing the EHP, because of storage space and costs, and our number of individual print subscribers is decreasing as readers become increasingly comfortable with the free online version of the journal. Consequently, our revenue from print subscriptions is decreasing, while costs associated with printing and distributing the journal continue to increase. Of course, there will still be significant costs associated with producing a paperless version of the journal. This is especially true since EHP is committed to maintaining its high-quality layout and graphics, continuing to publish superior news content, and upgrading its website capabilities. Therefore, we will continue to receive support from the NIEHS and fees from authors to help pay for the cost of processing, reviewing, editing, and publishing EHP online.There are also economic advantages to going paperless. Libraries and research institutions are cancelling print subscriptions to journals, including EHP’s use of paper and ink, eliminate the need to transport and distribute print copies of the journal, and reduce the journal’s contribution to the waste stream. Going paperless is also consistent with the goals of Executive Order 13589  involves some degree of environmental impact, moving to an online-only format will substantially reduce er 13589 , which wEHP to continue to broaden its dissemination and increase the impact of its environmental health information without compromising its high standards of peer review and rigorous editorial oversight. We believe scientific publishing is headed toward a future in which paperless journals are the industry standard. For EHP, the future is now.There are many advantages to moving to a paperless format, but above all, it will allow


EHP relies on the diligence and integrity of experts to help determine the quality and impact of papers submitted for possible publication. In 2010 EHP received over 1,450 papers, and about 550 of those papers were sent by our Associate Editors to at least two anonymous peer reviewers for evaluation. EHP published 268 papers in 12 issues during 2010, and the journal is very grateful for the time and effort of the more than 1,000 reviewers who assisted us last year. A list of those reviewers is available on the journal’s website (http://ehponline.org/article/info:doi/10.1289/ehp.119-a59). In this issue, EHP recognizes its top 12 Reviewers of the Year. These are individuals who reviewed at least five papers during the year and received excellent ratings for the timeliness and quality of their reviews by the Associate Editor who handled the peer-review process. They are Carol Angle, Adrian Barnett, Joe Braun, Jane Clougherty, Adrienne Ettinger, Matthew Longnecker, John Meeker, Sumi Mehta, David Savitz, Leonardo Trasande, Roberta White, and Judith Zelikoff. We congratulate these reviewers and thank the hundreds of others who contributed to the success of EHP in 2010.Like all peer-reviewed journals,


In this study, the clinical and morphological features of vesiculobullous lesions observed in Kaposi sarcoma are analyzed, and the features of bullous Kaposi sarcoma cases are emphasized.A total of 178 biopsy materials of 75 cases diagnosed as classic-type cutaneous Kaposi sarcoma were reviewed. Twenty-five cases showing vesiculobullous features were included in the study. Tumor, epidermis, dermis, and clinical data regarding these cases was evaluated.Vesicular changes were observed in 21 (12%) out of 178 lesions of the 75 cases, while bullous changes were present in only 4 (2%). In all cases where vesicular and bullous changes were detected, tumor, epidermis, and dermis changes were similar. All cases were nodular stage KS lesions, whereas hyperkeratosis and serum exudation in the epidermis, marked edema in the dermis, and enlarged lymphatic vessels and chronic inflammatory response were observed.Our findings suggest that changes in vascular resistance occurring during tumor progression are the most important factors comprising vesiculobullous morphology.http://www.diagnosticpathology.diagnomx.eu/vs/1646397188748474The virtual slide(s) for this article can be found here:  Kaposi sarcoma (KS) is composed of four different epidemiological forms—classic, endemic, iatrogenic, and AIDS-associated (Acquired Immune Deficiency Syndrome) etiology comprising HHV-8 (Human Herpes Virus-8) . The mosBullous KS [BKS] defines KS lesions in which a bullous appearance is clinically dominant. Bullous changes are rare in KS, but they are important in that they may be confused with other vesiculobullous skin disorders. The evolutionary mechanisms of bullous lesions in KS are still controversial, including whether or not they should be assessed as a different morphological subgroup -21.In this study, the clinical and morphological features of vesiculobullous lesions observed in classic KS are analyzed, and the features of bullous KS lesions are emphasized in association with the relevant literature.The study sample included 178 classic cutaneous KS cases in 75 patients who were diagnosed in the pathology departments of the University of Karaelmas, and Dr. Lütfi Kırdar Training and Research Hospital between 2001 and 2010. The clinical data was obtained from patient files. There was no history of immunosuppressive drug use, transplantation, or human immunodeficiency virus-1 [HIV-1] infection in any of the patients. All patients with KS were HIV-1 seronegative. All of the cases were classified as classic KS.The study was performed in accordance with the Helsinki Declaration, and the privacy of the patients was protected through the decoding of data, according to the privacy regulations of the Zonguldak Karaelmas University Hospital , and Dr. Lütfi Kırdar Training and Research Hospital .The histopathological diagnoses of all KS cases were confirmed through the examination of archived hematoxylin and eosin (H&E) slices, and immunohistochemical survey results. Histological sections of biopsy materials were assessed in terms of the properties of the tumors , epidermis , and dermis . Features were graded semi-quantitatively between 0–3 in a grading of histopathological findings, i.e., 0 – negative, 1 – mild, 2 – moderate, 3 – marked.Blisters are fluid-filled cavities located within or beneath the epidermis. Blisters equal to or greater than 5 mm are called bullae, whereas blisters smaller than 5 mm are called vesicles. We also used these criteria to differentiate between vesicles and bullae .The histological progression of KS lesions is composed of three phases, which can also be detected clinically: early (patch), intermediate (plaque), and late stages (nodule). The following criteria have been used to determine the histological stages of lesions:Early phase: Lesions which are composed of inflammatory cells and newly-formed vascular structures, and contain no spindle cells.Intermediate phase: Lesions with scattered spindle cells and cleft-like structures filled with erythrocytes, in addition to neoplastic vascular structures.Late phase: Lesions in which spindle cells form nodular structures and blood-filled pseudovascular structures ,2.Immunohistochemical analysis with endothelial indicators  and HHV-8 (latent nuclear antigen-1 [LNA-1]) monoclonal antibodies were performed for confirmation of the histopathlogical diagnosis in each case. Immunohistochemical studies were performed using archival materials. A set of 4-μm-thick tissue sections was prepared from formalin-fixed and paraffin-embedded blocks, deparaffinized in xylene, rehydrated, and microwaved for 10 min at 30% power in a citrate buffer (pH 6.0) for antigen retrieval. Endogenous peroxidase activity was blocked using 0.3% hydrogen peroxide. The slides were then incubated at room temperature for 1 hour with D2-40 , CD31 , CD34 , and HHV-8 (LNA- 1) , using the biotin-streptavidin complex-diaminobenzidine (BSA-DAB) technique for detection. Negative controls were processed in parallel without primary antibodies. The D2-40 immunoreactivity of lymphatic endothelial cells, and the CD31 and CD34 immunoreactivity of capillaries in the upper dermis were used as internal positive controls.Vesiculobullous histomorphology was observed in 25 14%) out of 178 lesions of 75 cases. Vesicular formation was detected in 21 (12%) and bullous formation was detected in 4 (2%) cases in microscopic evaluation. Clinical vesicular changes were not detected in any of lesions in which vesicular formation microscopically whereas bullae were detected clinically in all lesions in which bullous formation was detected microscopically  and 6 female 29%) patients, and the mean age was found to be 72.50 ± 11.09 min. 35–max. 84). Of all the lesions, 11 were seen in the lower extremities (52%), while 6 were in the upper extremities (29%) and 4 on the trunk (19%). All lesions were nodular stage KS lesions and, when epidermal changes were analyzed, hyperkeratosis was detected in 100% of the cases, serum exudation into the keratin layer in 64%, intraepidermal splitting in 84%, subepidermal splitting in16%, ulceration in 10%, papillomatous changes in 10%, and a rise in pigmentation on the basal membrane in 5%. Marked dermal edema and enlarged lymphatic vessels were observed in all cases, while chronic-type inflammatory response was seen in 86%. In cases in which ulceration was observed within the epidermis, neutrophil infiltration was detected in varying amounts, in addition to chronic-type inflammatory response , and these cases were evaluated as BKS. In cases in which bulla morphology, hence named BKS, was observed clinically, the mean age was found to be 66.75 ± 10.14 (min. 57-max. 72), and the female:male ratio was 1:3. The lesions were observed on the lower extremities (n = 2), the upper extremities (n = 1), and the head and neck region (n = 1). All cases were found to be nodular stage lesions, and serum exudation into the keratin layer was detected in 4 cases, intraepidermal (IE) and subepidermal (SE) bulla were detected in 3 and 1 cases, respectively, and ulceration in 2 cases. Papillomatosis was observed in 1 case and pigmentation increase in the basal layer was observed in 2 cases. Atrophy in the epidermal layer was not displayed in any of the cases. Marked edema, enlarged lymphatic vessels, and chronic-type inflammatory response were detected in all cases when the dermal properties were analyzed , and HHV-8 (LNA-1) (nuclear) immunoreactivities. The non-neoplastic lymphatic endothelium, lymphangioma-like channels, proliferative capillaries, and spindle cells were reactive with D2-40, whereas the non-neoplastic blood vessels and thick-walled, mature vascular structures were non-reactive. The reaction was positive in non-neoplastic blood vessels with CD31 and CD34, while the non-neoplastic lymphatic vessels were non-reactive. There was a reaction with CD31 in all cellular components forming KS. There was a homogenous reaction with CD34 in neoplastic capillary structures, and spindle cells. A diffuse, strong nuclear reaction was observed with HHV-8 (LNA-1) in neoplastic vascular structures and spindle cells forming KS.KS is classically characterized by blue-purple colored vascular lesions -5. It isBullous lesions in KS were first defined by Ronchese and Kern in 1957 . To dateGrayson et al. reported that bullous changes were seen more frequently among classic KS cases . When thBlister formation in skin lesions develops by various mechanisms. The best-known mechanisms are spongiosis, achantholysis, ballooning degeneration, cytolysis, and basal membrane destruction. Among the mechanisms leading to blister formation are extracellular and intracellular edema, physical and chemical agents, infections, and immunological mechanisms. Spongiosis is the disintegration of keratinocytes because of accumulation of extracellular fluid in the epidermis. As a result, fluid-filled vesicles and bullae are formed in some cases. Spongiotic changes are accompanied by dermal edema, lymphatic dilatation, and perivascular inflammation .Various theories regarding the etiopathogenesis of vesiculobullous lesions in KS have been suggested. Some researchers advocate that these lesions develop over chronic lymphedema ground, and that it is a specific entity, similar to lymphangioma circumscriptum ,9,16. HoIn our series, all of the lesions showing bullous pattern were nodular stage lesions, and marked edema plus enlarged lymphatic vessels were observed within the papillary dermis. Intraepidermal and subepidermal bullae were detected in 3 and 1 cases, respectively. In all cases, serum exudation within the keratin layer was observed. Lymphedema was not detected clinically in any of the cases. Our findings suggest that lymphangiectasia and peritumoral edema secondary to tumor play a vital role in the etiology of bullous KS.Hyperkeratosis is frequently observed, and in some cases, necrosis plus ulceration can be seen on the bulla ceiling in bullous KS cases. Dermal edema and enlarged lymphatic vessels are prominent in bullous KS cases, and fibrosis can be observed in those patients who receive radiotherapy -13. In oCossu et al. reported that bullous changes are more frequent in KS cases showing lymphangioma-like features . HoweverVesicular changes  were more common than bullous changes  in KS lesions in our study. No vesicular formation was observed clinically in lesions in which microscopic vesicle formation was observed. On the other hand, a clinical bullous formation was observed in cases with microscopic bullous formation. Vesicle and bulla differentiation is associated with blister size. Growth in vesicle size by fluid accumulation results in the formation of bullae . Thus, iIn conclusion, the results of our study found that the demographic data of KS cases, which demonstrated a bullous pattern, was similar to that of other classic KS cases. Histologically, these lesions were closely associated with high stage, marked dermal edema and enlarged lymphatic vessels. Our findings support the idea that the vesiculobullous patterns observed during KS develop secondary to tumorogenesis. Therefore, the term “bullous KS” should be used as a clinical definition, and it should be remembered that it may be present in different histological subtypes. Clinical and histopathological recognition of vesiculobullous changes in KS is important so that accurate data can be assimilated for further studies.KS: Kaposi sarcoma; HHV-8: Human herpesvirus 8; AIDS: Acquired Immune Deficiency Syndrome; PGLKS: Pyogenic granuloma-like KS; LLKS: Lymphangiom-like KS; UKS: Usual KS; BKS: Bullous KS; HIV-1: Human Immunodeficiency Virus-1; H&E: Hematoxylin and eosin; F: Female; M: Male; IE: Intraepidermal; SE: Subepidermal; LNA-1: Latent nuclear antigen-1; BSA-DAB: Biotin-Streptavidin complex- Diaminobenzidine.The authors declare that they have no competing interests.NOK conducted the design of the study, performed microscopic evaluation, and drafted the manuscript. FB and NST participated in the design of the study and performed the selection of appropriate cases and data collection and helped to draft the manuscript. BDG and SHK participated in the design of the study and immunohistochemical evaluation. ŞOÖ conceived of the study, and participated in its design and coordination and helped to draft the manuscript. SHK participated in the design of the study and performed statistical analysis. All authors read and approved the final manuscript.


Molecular chaperone or heat shock proteins (HSP) are vital proteins that increase cell survival by allowing it to combat stress caused by injurious stimuli through certain cyto-protective mechanisms . These cStabilization of the structure of important agents in malignant transformation, such as kinases (Src and Met tyrosine kinases) and transcription factors  allows molecular chaperones to stimulate angiogenesis by promoting endothelial cell proliferation and permitting growth of cancer beyond the oxygen capacity of tissue diffusion . MoleculThe new therapeutic agents or Heat Shock Protein inhibitors function by blocking the intrinsic ATPase activity of molecular chaperones allowing oncogenic proteins  to be taDespite effective results in phase 1 of clinical trials , HSP inhAlthough still in phase 2 of clinical trial, the development of HSP inhibitors provides an exciting alternative for molecular-based therapy in cancer . HSP inh


Life-review interventions (LRI) are psychotherapeutic techniques originally derived from gerontology, which can be distinguished from other biographical and reminiscing techniques. They have been systematically implemented and investigated not only in elderly clients with depression, cognitive decline, in oncology units and in hospices but also in adolescents with various mental problems. LRI are mainly based on the elaboration of the autobiographical memory as well as on personal identity consolidation. This bears the potential for the systematic introduction, use, and evaluation of LRI within the field of psychotraumatology.This article gives a general overview and outlines a structured LRI by means of a case example of a World War II-traumatised patient. Other applications and implementations of LRI in psychotraumatology and other related areas are presented.So far, only uncontrolled or controlled LRI case studies have been investigated with traumatized samples.The importance of further randomized controlled studies is emphasized. Psychotraumatology has given rise to new inventions in the treatment of traumatic stress syndromes such as prolonged exposure, eye movement desensitization and reprocessing (EMDR) or narrative exposure. Recently, treatment methods for specific client or patient groups have been proposed, including phase-based treatment for complex PTSD  interventions. In psychodynamics or psychoanalysis, these interventions merge more or less seamlessly into standard methods of particular therapeutic orientations, when used with elderly patients and when therapists have patients talk about their lives, starting with childhood  , was introduced by Knaevelsrud and colleagues  . Positive memories  should dominate over negative ones .Find meaning: Negative experiences, including trauma, can be given a meaning. Even if trauma or death of a spouse remains a negative fact, the subjective experience of having been altered by the event in a positive sense, enables and supports a new extended view on one's own life.Elaboration of memory, that is, greater detail of what is remembered actively. In depressed patients not only negative aspects are remembered, but also positive ones. In the case of trauma, memories should be elaborated and processed into a narrative.On a pragmatic level, three active principles of LRI can be described . Moreover, LRI with PTSD-patients exceeds common trauma focusing by aiming for a broader and more adequate conception of the traumatic experience. The structured character of chronological recall and evaluation of life stages is also assumed to enhance the specificity of non-traumatic memories  are in focus.In the following, a typical approach will be outlined. For PTSD patients, LRI is divided intoinitial phase, goals and actual processing are discussed with the patient. Depending on the level of education or comprehension of the patient, the reasons/rationales for the following process vary. In patients with good comprehension, all of the above suggested three active principles  are discussed.In the Remembering childhood, often gives great pleasure and brings people in a good mood, which is useful for those who sometimes have problems. This can sometimes help with problems. I think that many people's lives are very interesting, and I would be glad if for the next hours you told me a few detailed stories you remember. In patients with lower comprehension, more simple sentences are selected, for example, The patient is informed that between 10 and 15 sessions are needed to discuss the important stages of their life. Furthermore, they are asked to bring appropriate personal memorabilia to each session .Please imagine as vividly as possible that you are doing gardening or cleaning a room….). An initial resistance of telling memories  may be due to depressive symptoms. With the help of general psychotherapeutic relationship work or by providing positive feedback for the first memories expressed, therapists can frequently overcome this resistance.For some patients, it is extremely difficult to engage in a life review based on concrete experiences. In this case, guided imagination may be helpful so that first an exercise can be carried out Adulthood I: working life, partnerships, until first child birthAdulthood II: children from birth until moving out of familyAdulthood III: from age 50 to retirementRetirement age to presentIn the subsequent case study, two sessions for early childhood were set, because it is vital that enough LRI sessions  have taken place before reviewing the trauma in adolescence (see details below). In this specific case, only one session was set for the time of “reminiscence bump” (see above) of 15–25 years (youth to career start) —more sessions can be implemented at this point for other cases.Today we will start talking about your life. Let us start more or less at the beginning. It is best to proceed chronologically, recounting the earliest memories first. At the moment, it is not so important how far we get with remembering. What are some of your earliest memories? It is not only important that the patient describes the memories but that reflection over these memories is stimulated (What did this mean to you then?).The discussion of life cycle begins with childhood, such as How were you then? What did you value? What was important to you? What were your strengths? Did you enjoy your work? How important was this activity for you?Questions about the life stage of adulthood, are for example, How much time is left? What are the things to be done?). The therapist can approach the gaps with questions .It is not possible to conduct an extensive review within 5–8 sessions, neither is it necessary in order to meet the objectives of LRI. However, if the therapist notices large time gaps or missed issues , a more in-depth exploration of these periods is indicated . Other elements of trauma narration prompt questions to go into more detail, personal thoughts and feelings, as it is important not to rush through these memories. However, in contrast to prolonged or narrative exposure procedures, the patient is not prompted to focus on sensory details if he or she does not bring them up spontaneously.If the patient has experienced a trauma or—as it is the case of complicated grief—experienced the loss of an important person, a separate session is reserved to address this issue. The discussion takes place before the session on the phase of life in which the trauma happened, which is of special importance. The therapist expresses his/her empathy and explains that he knows how difficult it can be for the patient to deal with the terrible experience in full detail. At first, the narrative of the patient is not interrupted by questions for positive aspects . This can be done afterwards or in the subsequent session with a focus on integration of life balance. The existential severity of the trauma is appreciated by the therapist (Have you noticed that you have learned something positive from this experience of life?). In case of denial the theme of drawing a positive conclusion will be addressed at this point, Did you find a conclusion for yourself? What does it look like or could look like? It is essential to continue discussing the subsequent stages of life in the following sessions, since this implicitly supports a key purpose of life review (The trauma is only one part of life).At the end of the session, the therapist asks for positive changes related to overcoming the trauma  or to positively reframe the patients’ experiences . These sessions can extend therapeutic discussions of post-traumatic growth issues  were diagnosed.The life-review therapy began with a single session on pre-traumatic childhood, which she described as happy. Already in the second session, the trauma of the bombing was discussed. First, she described the events in a very summary form and she went into more detail only on request. The courage she had proved in protecting her younger siblings was appreciated in detail and confirmed. .In the third session, the typhoid fever and the patient's fear that she would die at age 22 were discussed. She was enabled to express her grief that years of her life had been taken from her because of the war. When asked about positive aspects in this phase of life, she replied that she had then begun to plan her life and to live consciously.In the therapeutic conversations, the patient stated repeatedly that she had been thinking about writing a book on her traumatic experiences in war for a long time. Although the traumatic experiences have been very relevant in every phase of life, she had been delaying writing the book again and again for 30 years already. Instead, she took part in various charitable activities that she initiated.In the course of therapy, rapid improvement was achieved. In particular, the involuntary stressful memories (intrusions) decreased. The psychological examination of her PTSD symptoms before, during and after treatment showed a sustained reduction of the values . Her difficulties of falling asleep and sleeping through the night did not improve, however. The patient stated that a reason for the continuing insomnia could be the heavy burden of her voluntary work.For this patient, it was important that her reports of overcoming the traumatic experiences, her motivation to write a book about them and her present and former honorary posts were confirmed by the therapist.In an effort to provide new relevant client groups with LRI, various New Media applications such as the internet, e-mail or computer software have been developed . Their treatment services are aimed at people over 65 who are currently suffering from psychological long-term consequences of traumatic experiences during, and shortly after, World War II.As mentioned above, Knaevelsrud et al.  have useClients and their constantly assigned e-therapists communicate exclusively via the internet, and in exceptional cases via post or fax. The e-therapists follow a specified treatment manual deriving both from the LRI approach and online psychotherapy of PTSD  on LRI has been conducted with traumatized samples. However, a large number of RCT have proven that the highly structured LRI is a very effective treatment against depressive symptoms. A previous meta-analysis found nine RCTs with LRI in which 520 patients were treated. The mean effect size of LRI was 0.92, which can be considered a very good therapeutic effect  conducted by Pinquart and Forstmeier , these rg between 0.40 and 0.64) to small (g between 0.24 and 0.39): Ego-integrity (g=0.64), purpose in life (g=0.48), death preparation (g=0.40), mastery (g=0.40), mental health (g=0.33), positive well-being (g=0.33), social integration (g=0.31) and cognitive performance (g=0.23) were significantly improved when LRI or reminiscing techniques and story-telling were applied.While most meta-analysis mainly investigated depressive symptoms, Pinquart and Forstmeier  includedPinquart and Forstmeier  provide To date, only single case studies reporting high effects are available for LRI-effects on PTSD (see above). There is a need for quantitative studies examining the effect of LRI on PTSD-symptoms as well as other aspects of mental health and well-being. Many of the existing studies evaluating the connection between LRI and depression do not include follow-up measurements and consequently little is known about the long-term effects of LRI.Future research could examine whether certain groups of people, for example, individuals with unresolved biographical conflicts, profit to a higher degree from LRI compared to individuals with little or no unresolved biographical conflicts (Pinquart & Forstmeier, d=0.58 underscores the potential of LRI as a preventive technique for elderly people with an increased risk for depression.The applicability of LRI as a preventive method for depressive disorders in older adults was demonstrated by Pot et al.  who evalLRI have a positive effect on various aspects of human experiencing and behavior: when structured LRI are applied in a psychotherapeutic setting with depressive patients, considerable therapeutic effects can be achieved. Not only do the depressive symptoms improve but also a range of other variables such as well-being and ego integrity. As outlined, we regard the potential for a broader introduction of LRI into psychotraumatology a promising setting for further advancement and differentiation of PTSD treatments.