The presence of free palmitic acid modulates the effects of lutein on structural and dynamic properties of lipid membranes.

Free fatty acids, like palmitic acid (PA), and xanthophyll pigments, like lutein (LUT) are the natural membrane compounds in plants. To study the effect of PA on LUT and their organization, a model membrane of 1,2-dimyristoyl-sn-glycerol-3-phosphocholine (DMPC) enriched with 2 mol% PA and 1 mol% LUT was formed. Molecular mechanisms underlying the interaction between these two compounds were examined with application of molecular spectroscopy techniques, e.g., visible spectroscopy, electron paramagnetic resonance and Fourier transform infrared. We determined the monomeric/dimeric organization of LUT in the membrane. We proved that the presence of PA in the lipid phase facilitated and stabilized the formation of LUT structures in the membrane. Lutein with PA did not form strong molecular aggregates like H- and J-structures. We presented the simplified model membrane that could be a suitable representation of the physiological process of de-esterification of PA from LUT appearing in natural biomembranes in humans.

The Role of Singlet Oxygen in Photoreactivity and Phototoxicity of Curcumin.

Curcumin is a plant-derived yellow-orange compound widely used as a spice, dye and food additive. It is also believed to have therapeutic effects against different disorders. On the other hand, there are data showing its phototoxicity against bacteria, fungi and various mammalian cells. Since the mechanism of its phototoxic action is not fully understood, we investigated here the phototoxic potential of curcumin in liposomal model membranes and in HaCaT cells. First, detection of singlet oxygen (1 O2 ) luminescence proved that curcumin generates 1 O2 upon blue light irradiation in organic solvent and in liposomes. Then, HPLC-EC(Hg) measurements revealed that liposomal and cellular cholesterol is oxidized by 1 O2 photogenerated by curcumin. Enrichment of liposome membranes with curcumin significantly increased the oxygen photo-consumption rate compared to the control liposomes as determined by EPR oximetry. Cytotoxicity measurements, mitochondrial membrane potential analyses and protein hydroperoxides detection confirmed strong phototoxic effects of curcumin in irradiated HaCaT cells. These data show that since curcumin is advertised as a valuable dietary supplement, or a component of cosmetics for topical use, caution should be recommended especially when skin is exposed to light.

Triplet-driven chemical reactivity of ß-carotene and its biological implications.

The endoperoxides of ß-carotene (ßCar-EPOs) are regarded as main products of the chemical deactivation of 1O2 by ß-carotene, one of the most important antioxidants, following a concerted singlet-singlet reaction. Here we challenge this view by showing that ßCar-EPOs are formed in the absence of 1O2 in a non-concerted triplet-triplet reaction: 3O2 + 3ß-carotene → ßCar-EPOs, in which 3ß-carotene manifests a strong biradical character. Thus, the reactivity of ß-carotene towards oxygen is governed by its excited triplet state. ßCar-EPOs, while being stable in the dark, are photochemically labile, and are a rare example of nonaromatic endoperoxides that release 1O2, again not in a concerted reaction. Their light-induced breakdown triggers an avalanche of free radicals, which accounts for the pro-oxidant activity of ß-carotene and the puzzling swap from its anti- to pro-oxidant features. Furthermore, we show that ßCar-EPOs, and carotenoids in general, weakly sensitize 1O2. These findings underlie the key role of the triplet state in determining the chemical and photophysical features of ß-carotene. They shake up the prevailing models of carotenoid photophysics, the anti-oxidant functioning of ß-carotene, and the role of 1O2 in chemical signaling in biological photosynthetic systems. ßCar-EPOs and their degradation products are not markers of 1O2 and oxidative stress but of the overproduction of extremely hazardous chlorophyll triplets in photosystems. Hence, the chemical signaling of overexcitation of the photosynthetic apparatus is based on a 3chlorophyll-3ß-carotene relay, rather than on extremely short-lived 1O2.

Tauroursodeoxycholic Acid (TUDCA)-Lipid Interactions and Antioxidant Properties of TUDCA Studied in Model of Photoreceptor Membranes.

Tauroursodeoxycholic acid (TUDCA), a hydrophilic bile acid containing taurine conjugated with the ursodeoxycholic acid (UDCA), has been known and used from ancient times as a therapeutic compound in traditional Chinese medicine. TUDCA has recently been gaining significant interest as a neuroprotective agent, also exploited in the visual disorders. Among several mechanisms of TUDCA's protective action, its antioxidant activity and stabilizing effect on mitochondrial and plasma membranes are considered. In this work we investigated antioxidant activity of TUDCA and its impact on structural properties of model membranes of different composition using electron paramagnetic resonance spectroscopy and the spin labeling technique. Localization of TUDCA molecules in a pure POPC bilayer has been studied using a molecular dynamics simulation (MD). The obtained results indicate that TUDCA is not an efficient singlet oxygen (1O2 (1Δg)) quencher, and the determined rate constant of its interaction with 1O2 (1Δg) is only 1.9 × 105 M-1s-1. However, in lipid oxidation process induced by a Fenton reaction, TUDCA reveals substantial antioxidant activity significantly decreasing the rate of oxygen consumption in the system studied. In addition, TUDCA induces slight, but noticeable changes in the polarity and fluidity of the investigated model membranes. The results of performed MD simulation correspond very well with the experimental results.

Effects of Curcumin on Lipid Membranes: an EPR Spin-label Study.

Curcumin is a yellow-orange dye widely used as a spice, food coloring and food preservative. It also exhibits a broad range of therapeutic effects against different disorders such as cancer, diabetes, or neurodegenerative diseases. As a compound insoluble in water curcumin accumulates in cell membranes and due to this location it may indirectly lead to the observed effects by structurally altering the membrane environment. To exert strong structural effects on membrane curcumin needs to adopt a transbilayer orientation. However, there is no agreement in literature as to curcumin's orientation and its structural effects on membranes. Here, we investigated the effects of curcumin on lipid order, lipid phase transition, and local polarity in a model liposome membranes made of DMPC or DSPC using electron paramagnetic resonance (EPR) spin labeling technique. Curcumin affected lipid order at different depths within the membrane: it slightly increased the phospholipid polar headgroup mobility as monitored by spectral parameters of T-PC, while along the acyl chain the ordering effect was observed in terms of order parameter S. Also, rotational correlation times τ2B and τ2C of 16-PC in the membrane center were increased by curcumin. Polarity measurements performed in frozen suspensions of liposomes revealed enhancement of water penetration by curcumin in the membrane center (16-PC) and in the polar headgroup region (T-PC) while the intermediate positions along the acyl chain (5-PC and 10-PC) were not significantly affected. Curcumin at a lower concentration (5 mol%) shifted the temperature of the DMPC main phase transition to lower values and increased the transition width, and at a higher concentration (10 mol%) abolished the transition completely. The observed effects suggest that curcumin adopts a transbilayer orientation within the membrane and most probably form oligomers of two molecules, each of them spanning the opposite bilayer leaflets. The effects are also discussed in terms of curcumin's protective activity and compared with those imposed on membranes by other natural dyes known for their protective role, namely polar carotenoids, lutein and zeaxanthin.

Melanopsin: From a small molecule to brain functions.

Melanopsin, a G family coupled receptor, found within the ganglion cell layer in the retina, plays an important role in non-image-forming visual functions, including hormone secretion, entrainment of circadian rhythms, cognitive and affective processes. Diffuse projections of melanopsin-containing cells to many brain areas suggest that different responses may involve different neural projections, thus different melanopsin cells. Considering the complexity of the melanopsin system, its contribution to so many different biological functions is not surprising. In this review, we summarize the current knowledge about melanopsin in terms of its photophysics, photochemistry, mechanisms of activation, cell signaling, morphology, and physiology. In the last part, the role of melanopsin in image and non-image forming processes and cognitive and affective functioning of animals and humans, are discussed. Although in recent years considerable insight has been gained into the melanopsin system, it still remains an open question of how one protein expressed by several thousand cells in the retina, could be responsible for so many diverse functions and what activation mechanism(s) it uses.

In vitro phototoxicity of rhodopsin photobleaching products in the retinal pigment epithelium (RPE).

Although the primary biological function of retinal photoreceptors is to absorb light and provide visual information, extensive exposure to intense light could increase the risk of phototoxic reactions mediated by products of rhodopsin bleaching that might accumulate in photoreceptor outer segments (POS). The phototoxicity of POS, isolated from bovine retinas, was examined in cultured retinal pigment epithelium cells (ARPE-19) containing phagocytised POS and in selected model systems by determining POS ability to photogenerate singlet oxygen, and photoinduce oxidation of cholesterol and serum albumin. Bleaching of rhodopsin-rich POS with green light resulted in the formation of retinoid products exhibiting distinct absorption spectra in the near-UV. Irradiation of POS-fed ARPE-19 cells with blue light reduced their survival in a dose-dependent manner with the effect being stronger for cells containing prebleached POS. The specific and non-specific phagocytic activity of ARPE-19 cells was inhibited by sub-lethal photic stress mediated by phagocytised POS. The oxidising ability of POS photobleaching products was demonstrated both in a model system consisting of serum albumin and in ARPE-19 cells. Distinct photooxidation of proteins, mediated by POS, was observed using coumarin boronic acid as a sensitive probe of protein hydroperoxides. Irradiation of POS with blue light also induced oxidation of liposomal cholesterol as determined by HPLC-EC(Hg). Time-resolved singlet oxygen phosphorescence demonstrated the efficiency of retinoids, extracted from POS by chloroform-methanol treatment, to photogenerate singlet oxygen. The results indicate that photic stress mediated by POS photobleaching products could inhibit phagocytic efficiency of RPE cells and, ultimately, compromise their important biological functions.

EPR Studies on the Properties of Model Photoreceptor Membranes Made of Natural and Synthetic Lipids.

The membranes of retina photoreceptors have unique lipid composition. They contain a high concentration of polyunsaturated docosahexaenoic acid, with six double bonds, and are enriched in phosphatidylethanolamines. Based on their phospholipid composition and cholesterol content, membranes of photoreceptors can be divided into three types: plasma membrane, young disks membranes, and old disks membranes. High amount of docosahexaenoic acid, abundant illumination, and high respiratory demands make these membranes sensitive to oxidative stress and lipid peroxidation. Human retinas are not easily available for research, therefore most research is done on bovine retinas. However, to follow, in a controlled manner, the changes in membrane properties caused by different factors it seems advisable to apply carefully prepared models of photoreceptor membranes. Using synthetic lipids we prepared liposome models of three types of photoreceptor membranes, and by means of electron paramagnetic resonance spectroscopy and spin labeling technique we compared polarity and fluidity of those model membranes with the properties of membranes consisting of natural lipids extracted from photoreceptor outer segments of bovine retinas. Additionally, we studied the effect of oxidation on the membrane properties in the presence and in the absence of zeaxanthin, which is an antioxidant naturally present in the human retina. The results show that there are significant differences in polarity and fluidity between all investigated membranes, which reflect differences in their lipid composition. The properties of the membranes made of natural photoreceptor outer segment lipids are most similar to the ones of the models of old disks membranes. Oxidation did not change the membrane properties significantly; however, a slight ordering effect was observed in liposomes made of natural photoreceptor outer segment lipids and in the model of old disks membranes. Zeaxanthin affected polarity and fluidity mostly in the model of old disks membranes. The results show that by careful selection and appropriate proportions of lipid mixtures, it is possible to obtain synthetic membranes of the properties similar to the natural ones.

One Piece Immediately Loaded Implants Versus 1 Piece or 2 Pieces Delayed: 3 Years Outcome.

PURPOSE: To compare time-dependent outcome of immediately loaded 1-piece (1P) implants with delayed loaded 1P and 2-piece (2P) implants. MATERIALS AND METHODS: A cohort of 33 patients divided into 3 groups: group A, 13 patients, 49 immediately placed and loaded 1P implants; group B, 11 patients, immediately placed and delayed loaded 1P implants; and group C, 10 patients, 39 2P implants delayed placed and loaded in a two-stage procedure. Marginal bone loss (MBL) was analyzed using x-ray radiography every 6 months, 1 year, and 3 years. RESULTS: A statistically significant mean MBL was observed between baseline, 6 months, 1 year, and 3 years in all groups. There was no statistical significant difference in MBL between immediate and delayed loaded 1P implants. MBL around mandibular implants was lower compared with maxillary implants. CONCLUSIONS: 2P implants showed less MBL compared with 1P implants in both maxilla and mandible. There was no statistical difference in MBL between immediate and delayed loaded 1P implants. Immediate loaded implants show more MBL in maxilla.

One- and Two-Piece Implants Placed in the Same Patients: Clinical Outcomes After 5 Years of Function.

A retrospective evaluation of 24 subjects treated with one-piece (1P) (n = 34) and two-piece (2P) (n = 38) implants placed in contralateral mandibular premolar locations was conducted. Cumulative implant survival was 100% following a 5-year (range = 59.9-72.3 months; mean = 63.1 months) postrestoration monitoring period. Bone loss (P = .1952) and prosthetic complications (P = .3667) did not significantly differ between the two groups. Clinical efficacy was equivalent for both implant designs.

Structural aspects of the antioxidant activity of lutein in a model of photoreceptor membranes.

It was shown that in membranes containing raft domains, the macular xanthophylls lutein and zeaxanthin are not distributed uniformly, but are excluded from saturated raft domains and about ten times more concentrated in unsaturated bulk lipids. The selective accumulation of lutein and zeaxanthin in direct proximity to unsaturated lipids, which are especially susceptible to lipid peroxidation, could be very important as far as their antioxidant activity is concerned. Therefore, the protective role of lutein against lipid peroxidation was investigated in membranes made of raft-forming mixtures and in models of photoreceptor outer segment membranes and compared with their antioxidant activity in homogeneous membranes composed of unsaturated lipids. Lipid peroxidation was induced by photosensitized reactions using rose Bengal and monitored by an MDA-TBA test, an iodometric assay, and oxygen consumption (using EPR spectroscopy and the mHCTPO spin label as an oxygen probe). The results show that lutein protects unsaturated lipids more effectively in membranes made of raft-forming mixtures than in homogeneous membranes. This suggests that the selective accumulation of macular xanthophylls in the most vulnerable regions of photoreceptor membranes may play an important role in enhancing their antioxidant properties and ability to prevent age-related macular diseases (such as age-related macular degeneration (AMD)).

Stress and strain patterns of 1-piece and 2-piece implant systems in bone: a 3-dimensional finite element analysis.

STATEMENT OF PROBLEM: The transition from implant to abutment is solid in 1-piece (1P) and broken in 2-piece (2P) implant designs. This difference may affect occlusal load distribution and marginal bone response. PURPOSE: To determine whether 1P and 2P implants with equivalent geometries exhibited stresses and strains differently under applied loading conditions. MATERIALS AND METHODS: Design software simulated 1P and 2P implants restored with metal copings and embedded in 3 cylindrical bone block models that varied in dimensions, density, and percentage of bone-to-implant contact. Three-dimensional, finite element analysis simulated occlusal loading. Experiments evaluated stresses and strains relative to implant design and (1) periimplant bone thickness, (2) cortical bone thickness, (3) magnitude and direction of occlusal loading, and (4) % bone-to-implant contact. RESULTS: Implants with equivalent dimensions exhibited comparable stresses and strains in all experimental conditions. Implant diameter and periimplant bone thickness influenced stress levels. Only small-diameter (3.0 mm) 1P implants in low-density bone exhibited stress levels that might adversely affect marginal bone stability. CONCLUSIONS: Implant diameter and periimplant bone thickness influenced load distribution in bone, but the type of implant-abutment transition had no significant effect. Small-diameter 1P implants should be limited to dense bone to minimize stress concentrations.

A novel method of preparation of surgical template for guided implant placement: a case report.

Currently, the preparation of surgical templates is a multistage process that engages the restorative dentist, dental laboratory, and external computed tomography center. This new method for production of templates and guides, as well as data transfer protocol, significantly simplifies commonly used methods and removes many of the complications inherent in them. A case report of an implantation procedure carried out with the new protocol is presented. Further studies and comparisons with other guided implant surgery methods are needed to establish the advantage of this novel method.

Morphology of root canal cross-sections of resected roots of first and second lower molars.

Precise preparation of the root canal ostium with a view to hermetical closing with retrograde root filling is an important prognostic factor determining the success of the procedure of tooth resection. Root canals interconnected with a narrow isthmus may cause problems both in endodontic treatment and in retrograde filling. The aim of this work is the research of the transverse cross-section of root canals of first and second lower molars on the resection model. The research encompasses 100 randomly selected molar teeth: 50 first and 50 second lower molars. The tooth root apexes were cut 3 mm below the apex and examined under an electron microscope, special attention being paid to the shape of root canal cross-sections. In the group of first molar teeth, in 20% the presence of an isthmus between canals in the proximal roots was observed; in the group of second molar teeth an isthmus between the canals of proximal roots occurred in 18% of the cases. It seems that the relatively high percentage (20%-18%) of the occurrence of an isthmus, 3 mm below the root apex, between two elongated transverse cross-section proximal root canals of first and second molars should encourage particular caution in the retrograde filling of the above-mentioned canals during the procedure of resection.

The issue of bioresorption of the Bio-Oss xenogeneic bone substitute in bone defects.

Bone grafts and bone substitute biomaterial implemented in guided tissue regeneration should undergo the process of biological decomposition in the recipient's system. The aim of this work is the presentation of current views concerning the issue of Bio-Oss bovine bone bioresorption and their juxtaposition with the results of the author's own research. The work presents histopathological and immunohistochemical tests of the xenogeneic Bio-Oss preparation from biopsies carried out 30 months after implantation. It was observed that the preparations contained correct bone neighbouring remnant particles of Bio-Oss, intratrabecular fibromatosis around the implant, abundant vascularisation, absence of osteoid and of active inflammatory process. A small number of T and B lymphocytes was detected. The results obtained in the above-described case testify to the descending character of the inflammatory infiltration 30 months after the implementation of Bio-Oss and efficient restoration of the bone. The prevalent view in literature is that Bio-Oss is resorbable biomaterial. However, there are also reports questioning this view as remnants of Bio-Oss have been detected even 44 months after implantation into the bone defect. In the author's own cases, Bio-Oss remnants could be observed 30 months after implanting. It seems that although the creation of new bone structure is indisputable, the process of biological decomposition of Bio-Oss should be described as slow bioresorption.

Implementation of the two-stage operation technique for the autotransplantation of teeth with closed apexes--a preliminary report.

Transplantation of teeth with incompletely formed apexes demonstrates a statistically better retention than the transplantation of fully-formed teeth because of the possibilities for reinnervation and revascularisation. However, with regard to the autotransplantation of teeth with completely developed roots, routine endodontic treatment is proposed, as the possibility to retain the vitality of dental pulp is doubtful. In this work the author wishes to present the implementation of the two-stage technique for the transplantation of fully-formed teeth. Observations confirm the possibility for reinnervation and revascularisation occurring after autotransplantation of teeth with closed apexes, which should encourage a cautious approach to the question of routine endodontic treatment following autotransplantation.

Forcing of the root into the maxillary sinus during tooth extraction--and what next?

In the everyday practice of a dental surgeon there may sometimes occur complications during tooth extraction. The anatomical vicinity of the alveolar recess of the maxillary sinus predisposes this area to the occurrence of complications in the form of joining of the oral cavity to the maxillary sinus. However, leaving an accidentally forced root in the maxillary sinus lumen may cause a chronic inflammatory state of the sinus or other complications; hence the vital importance of attempts to remove the root directly after its forcing. This work presents possibilities of root removal, directly after its forcing, through the postextraction dental alveolus or through access in the sinus side wall, similar to the procedure of sinus lifting.