Structure-function relationship between a natural cosmetic active ingredient and the olfactory receptor OR2AT4.

OBJECTIVE: Although the olfactory receptor OR2AT4 was described as involved in epidermal renewal, there is no data about a cosmetic active ingredient activating this receptor. The aim of this research work was thus to identify a natural molecule binding to this receptor in order to stimulate keratinocyte migration. METHODS: For this purpose, natural molecules were extracted from Cocos nucifera flour. Then, efficacy of this natural extract was evaluated on keratinocyte migration in vitro. Molecules of the Cocos nucifera flour extract were then identified by UPLC-MS/MS. Molecular docking was finally conducted to investigate the potential interaction between identified molecules and the olfactory receptor OR2AT4. RESULTS: The Cocos nucifera flour extract significantly increased keratinocyte migration and results demonstrated that this effect was mediated by the olfactory receptor OR2AT4. Metabolomic analysis revealed two molecules, nonioside D and butyl 4-O-alpha-D-glucopyranosyl-beta-D-glucopyranoside, as significantly present in the Cocos nucifera flour extract compared to both Cocos nucifera oil and water. Finally, molecular docking revealed that butyl 4-O-alpha-D-glucopyranosyl-beta-D-glucopyranoside could interact with the extracellular domain 2 of the OR2AT4. CONCLUSION: This study highlighted for the first time a natural molecule, extracted from Cocos nucifera flour, able to interact with the olfactory receptor OR2AT4 and promote the keratinocyte migration and thus the epithelialization.

OBJECTIF: Bien que le récepteur olfactif OR2AT4 a été décrit comme étant impliqué dans le renouvellement de l'épiderme, il n'y a à ce jour aucune donnée concernant un ingrédient actif cosmétique activant ce récepteur. Le but de cette étude est donc d'identifier une molécule naturelle capable de se fixer au récepteur OR2AT4 afin de stimuler la migration des kératinocytes. METHODE: L'efficacité de cet extrait naturel sur la migration des kératinocytes in vitro a ensuite été évaluée. Les molécules présentes dans l'extrait de farine de Cocos nucifera ont ensuite été identifées par UPLC-MS/MS. Enfin, des expérimentations de docking moléculaire ont été réalisées afin d'identifier une potentielle interaction entre les molécules précédemment identifiées et le récepteur olfactif OR2AT4. RESULTATS: L'extrait de farine de Cocos nucifera augmente significativement la migration des kératinocytes via l'activation du récepteur olfactif OR2AT4. Les analyses métabolomiques ont révélées deux molécules, le nonioside D et le butyl 4-O-alpha-D-glucopyranosyl-beta-D-glucopyranoside, comme étant significativement présentes dans l'extrait de farine de Cocos nucifera en comparaison avec l'huile et l'eau de coco. Enfin, le docking moléculaire révèle que le butyl 4-O-alpha-D-glucopyranosyl-beta-D-glucopyranoside, présent dans l'extrait de farine de Cocos nucifera peut interagir avec le domaine extracellulaire 2 du récepteur olfactif OR2AT4. CONCLUSION: Cette étude a identifié pour la première fois une molécule naturelle, extraite de la farine de Cocos nucifera, capable d'interagir avec le récepteur olfactif OR2AT4, afin de stimuler la migration kératinocytaire et ainsi favoriser le processus de ré-épithélialisation.

A critical investigation of NADPH oxidase activity in human spermatozoa.

It has been suggested that human spermatozoa contain an NADPH oxidase that could generate reactive oxygen species involved in signalling pathways to promote fertility. The proposal depends on observations that the addition of NADPH to purified human spermatozoa stimulates chemiluminescence by the superoxide (O2-) probe, lucigenin. We confirmed these observations, but demonstrated that lucigenin increases NADPH consumption by spermatozoa and stimulates artefactual O2- production via a diphenyleneiodonium (DPI) sensitive flavoprotein. In the absence of cytochrome c, DPI-inhibitable NADPH oxidation by permeabilized spermatozoa was 8 times too small to account for the rate of NADPH-stimulated cytochrome c reduction. Thus NADPH can directly reduce cytochrome c by a flavoprotein dependent mechanism making this O2- assay also unreliable in sperm suspensions. We were unable to observe O2- production by 40 x 10(6) spermatozoa/ml using electron paramagnetic resonance spectroscopy but could identify O(2)(-) generation from 2000 4beta-phorbol-12-myristate-13-actetate (PMA)-stimulated leukocytes. Using spectrophotometry, we did not detect the reduced cytochrome b(558) component of the neutrophil NADPH oxidase in human spermatozoa. No hydrogen peroxide generation was observed using a sensitive Amplex Red assay. We conclude that human spermatozoa do not possess significant NADPH oxidase activity and that the mechanism by which NADPH promotes capacitation must be re-evaluated.

A clinical review of non-age-related cataracts.

BACKGROUND: A cataract is any opacity or partial loss of transparency of the lens, whether the absence of transparency is small or complete. Pupil dilation affords a view of a legion of internal variations and abnormalities of the human crystalline lens which often tell us about its events and insults, as well as when in the patient's life these might have occurred. METHODS: In this article, we review the major non-age-related association of cataractogenesis with respect to metabolic, environmental, ocular-specific, infectious, dermatologic, retinal, and toxic etiologies. The data are presented from the clinical perspective of incidence for a given condition and cataract type. RESULTS: Two simplified summary reference sheets are provided: (1) frequency of occurrence vs. etiology and (2) cataract type vs. etiology (color-coded). The busy clinician can refer to both tools chair-side. CONCLUSIONS: The human body has numerous methods of signaling insults and abnormalities. As the crystalline lens is an important gauge of overall health, an argument can be made for routine dilation of all patients. This information is also essential for future neutraceutical and pharmaceutical therapeutic intervention.

Nutritional influences on eye health.

BACKGROUND: As with cardiovascular disease, nutrition and environmental intervention present new clinical practice and research opportunities in eye care. This includes maximizing health potential with food choice, the use of supplementation in high-risk and noncompliant patients, avoiding toxins such as cigarettes, and encouraging exercise. METHOD: The author provides his perspective on viewing biochemical, physiological, and epidemiological research in the context of disease prevention in both common and uncommon applications. In most cases, this approach requires food frequency intake evaluation and application of physiological optics to measure efficacy when there is an alteration in diet and/or addition of supplementation. These are details of ocular integrative medical practice that are ignored or underutilized. RESULTS: There are multiple opportunities for intervention in the age-related diseases of cataract, macular degeneration, and glaucoma. A summary of the application of nutrition to create a model for treatment of age-related macular degeneration is presented. CONCLUSIONS: Economic incentives, now lacking, are required to explore new paradigms in nutritional preventative eye care. Schools and colleges of Optometry could play a vital public health role.

Innervation of dromyosuppressin (DMS) immunoreactive processes and effect of DMS and benzethonium chloride on the Phormia regina (Meigen) crop.

Antibody to the dipteran myosuppressin peptide, dromyosuppressin, TDVDHVFLRFamide, stained cells and fibers in the brain, optic lobes, subesophageal ganglion, and thoracico-abdominal ganglion of the blow fly, Phormia regina (Meigen). Dromyosuppressin-like immunoreactive fibers were detected in the cardiac recurrent nerve, hypocerebral ganglion/corpora cardiaca complex, crop duct, and crop. In order to explore the mechanisms involved in regulating crop movement, we established an in vitro bioassay. The basal rate of crop movement was 50.8 +/- 1.5 contractions per minute. Application of 1 microl of saline to the crop did not significantly affect the rate of movement compared to the basal rate (46.1 +/- 1.1 contractions per minute, P < 0.05). Application of 1 microl 10(-6) M dromyosuppressin or 1 microl 10(-3) M benzethonium chloride to the crop slowed the rate to 2.2 +/- 0.2 and 6.1 +/- 0.7 contractions per minute, respectively. Although other data have previously been interpreted to suggest that dipteran crop contractions do not include a neural component, the neuropeptide dromyosuppressin affected P. regina crop motility. Innervation of the crop and crop duct by dromyosuppressin immunoreactive processes that originated in the central nervous system and the effect of dromyosuppressin on crop muscle contractions suggest that dromyosuppressin is released locally to modulate crop contractions and that crop motility is under neural regulation. Myosuppressins isolated from numerous insects have a high degree of structure identity and reduce spontaneous muscle contractions of the hindgut, oviduct, and heart. Benzethonium chloride, previously identified as a myosuppressin agonist on the cockroach hindgut and locust oviduct, mimicked the effect of dromyosuppressin on the crop. This suggests that structural requirements for myosuppressin receptor binding in the cockroach hindgut, locust oviduct, and fruit fly crop are similar.

A protocol for the evaluation and treatment of atrophic age-related macular degeneration.

BACKGROUND: Age-related macular degeneration (ARMD), the leading cause of vision loss in aging Western societies, is (in part) a nutrition-responsive disease that can be stabilized through environmental intervention. The atrophic form of the disease constitutes nearly 90% of cases, yet a standardized ophthalmic and systemic evaluation protocol does not exist. METHODS: A basic science and epidemiologic rationale for evaluating atrophic ARMD using a standardized low-cost, "low-technology," clinical approach is presented. The protocol is based on optometric physiological optics and ARMD retinal physiology/pathophysiology. RESULTS: The ophthalmic component involves baseline and serial measurements of contrast sensitivity (CSF), low-luminance, low-contrast visual acuity (SKILL), and glare recovery (GR) in each eye. The systemic component includes self-administered evaluation of nutritional status (food and supplement intake), exercise and activities of daily living associated with ARMD (night driving/glare adaptation disturbance and dark green-leafy vegetable/plant food consumption). CONCLUSIONS: The literature supports careful baseline and serial evaluation of high-risk patients using germane parameters of ocular function and systemic health specific to ARMD. This "ARMD workup" is analogous to a glaucoma workup, but involves less equipment and chair time. Any optometrist can obtain these measurements using inexpensive testing protocols. (Part II of this paper presents serial environmental intervention case report data.)

ARMD--pilot (case series) environmental intervention data.

BACKGROUND: A description and justification for using a standardized clinical low-cost, "low-technology," ocular and systemic "ARMD work-up" protocol was presented in the methods paper (Part I). METHODS: Fourteen male patients (70 +/- 9 years), receiving 0.73 +/- 0.45 portions of dark-green, leafy vegetables/day base intake, were placed on an additional portion of 5 ounces sauted spinach 4 to 7 times per week or lutein-based antioxidant (three patients). Detailed "ARMD work-up" data for six of these patients and summary pilot (case series) environmental intervention data are presented. RESULTS: One, Patients demonstrated short-term positive effects in visual function in one or both eyes with this mild therapeutic approach: Amsler grid (87%); Snellen Acuity (71%); Contrast sensitivity (92%); SKILL (65%); Glare recovery (69%); and Activities of Daily Vision Subscale (60%); Two, There was no obvious correlation between ophthalmoscopic appearance of the retina and visual outcome; and Three, Patient symptoms did not necessarily correlate with observed changes in visual function. CONCLUSIONS: The approach to atrophic ARMD presented here warrants informal practitioner replication and formal randomized prospective clinical case-control evaluation.

Water soluble antioxidants in mammalian aqueous humor: interaction with UV B and hydrogen peroxide.

HPLC/electrochemical detection was used to identify five major low MW water soluble electrochemically active molecules from the aqueous humor of three species of mammals: New Zealand White rabbits and humans (diurnal) and Sprague-Dawley rats (nocturnal). These molecules are L-cysteine (CYS), L-ascorbic acid (AA), glutathione (GSH), uric acid (UA) and L-tyrosine (TYR); all of these molecules have known antioxidant properties. Nocturnal rat aqueous humor is concentrated in two thiols: GSH (125 microM; n = 24 pooled eyes) and CYS (63 microM), in contradistinction to diurnal species which have high concentrations of AA. No deterioration of any of these antioxidants occurs in a synthetic aqueous humor mixture irradiated with a physiologically relevant spectral UV B dose of 30 mJ/cm2/h (5.5 UV equivalent sunlight hours). The same result occurred with addition of the endogenous aqueous humor UV B photosensitizer L-tryptophan. In a second set of experiments, human synthetic aqueous humor was subjected to hydrogen peroxide induced oxidant stress. The decay of antioxidants was CYS > GSH > AA > UA > TYR. The second highest concentrated antioxidant in human aqueous humor is TYR. Yet TYR failed to protect AA against H2O2-induced free radical damage in a synthetic aqueous humor model system (P = 0.10; ANOVA). The existence of multiple electrochemically active constituents and their thermodynamic interactions must be recognized when choosing animal models to evaluate human aqueous humor antioxidant defense.

Ocular oxidants and antioxidant protection.

Oxidative damage and antioxidant protection in ocular tissues has not been reviewed recently. Metabolism in the eye is of increasing interest because the organ is highly susceptible to damage by sunlight, oxygen, various chemicals, and pollutants. Interest is expected to increase because of an aging Western world population and a continued depletion of stratospheric ozone. Hydrogen peroxide is discussed because it is both a byproduct and a source of free radical reactions and is normally present in the aqueous humor. The metabolism of reactive oxygen species by enzymes, nutrients, pigments, and low molecular weight scavengers is evaluated. Ascorbic acid, because of its high concentration in the eye, is thought to be a primary substrate in ocular protection; progress in determining the mechanisms by which it is recycled and maintained in the useful, reduced state is discussed. Recent information is included about antioxidants not previously known to be present in the eye, and some importance is placed on the properties of the vitreous humor and tear fluid because of the previous lack of emphasis on these.

Tear fluid content of electrochemically active components including water soluble antioxidants.

PURPOSE: To describe the content of water-soluble antioxidants in tear fluid. METHODS: We collected tear fluid from healthy subjects, either into borosilicate glass tubing or by absorption onto Schirmer strips. High pressure liquid chromatography with electrochemical detection provided data on several components in a single assay. The system was sufficiently sensitive to provide reliable values for components present in tear fluid collected at normal (basal) or stimulated rates of secretion. RESULTS: Tear fluid at basal secretion was found to contain four compounds often considered to function in biology as antioxidants. Ascorbic acid (AA) is found at 665 microM, tyrosine at 45 microM and glutathione (GSH) at 107 microM. Cysteine (48 microM) and uric acid (328 microM) are reported for the first time; the latter is somewhat controversial as a physiologically active antioxidant. One peak on the chromatogram was consistently present but has not been identified. During stimulation of flow by brief inhalation of ammonium hydroxide fumes, the concentration of each compound was lower. CONCLUSION: This work demonstrates the presence of electro-chemically active components that might function as antioxidants at the anterior surface of the cornea against potential damage from radiation, oxygen toxicity, abrasion and environmental chemicals.

Properties of electrochemically active components in mammalian vitreous humor.

Antioxidants are considered to have important roles in protecting the eye against radiation, oxygen toxicity and other threats. Several water-soluble electrochemically-active compounds that might function as antioxidants in vitreous humor of rabbit, rat and bovine eyes were identified and quantitated. In that the rat is a nocturnal animal species and the other two are diurnal, the results allow a tentative evaluation on the comparative physiology of light-dark behavior patterns that influence this ocular fluid. All analyses were performed by HPLC with electrochemical detection. The utility of this approach is that components of vitreous humor can be identified by their retention time and by their characteristic response when the voltages of the two sensing electrodes are progressively brought together. Four compounds regarded as functional antioxidants in the animal kingdom were identified and quantitated. Other antioxidants known to be present in several biological fluids are not present at detectable or significant levels in vitreous humor; metabolites or precursors of these might be present in trace quantities.

A practical guide for differentiating between iron deficiency anemia and anemia of chronic disease in children and adults.

Anemia is a common disorder that should be diagnosed and managed in a primary care setting. Iron deficiency and chronic disease anemia are two subtypes of insidious anemias that may be difficult to differentiate. There are often excessive laboratory studies performed, patient delays, and increased costs while completing the workup. With the rising cost of health care, a greater emphasis is being placed on managed care. This type of system utilizes its own resources in a cost-effective manner and encourages the investigation to be in the domain of the primary practitioner. As the nation focuses on this type of delivery system, outside referrals will be limited. Primary care providers as well as physicians in these types of settings will need to become well versed in the interpretation of these uncomplicated anemias. With appropriate training and experience in the interpretation of blood studies and peripheral smears, clinicians can effectively differentiate between these two classifications while easing the burden on the health care system.

An apparatus for ultraviolet B irradiation of small volumes of biological fluid under controlled oxygen tension.

BACKGROUND: The design, construction, and calibration of a unique apparatus for ultraviolet (UV) B irradiation of small volumes of biological fluid under temperature-controlled open atmospheric or closed cell conditions of defined Po2 is described. METHODS: The apparatus consists of a custom designed UV transmissible quartz silica 3 port spectrophotometric cuvette coupled to a deuterium UV B source by UV transmissible fiber optics. Fluid movement, temperature regulation, dosimetry, and miniaturized biosensors for oxygen, pH, and temperature are described along with a preliminary experiment. RESULTS: The apparatus produces a known photochemical effect using an endogenous aqueous humor UV B photosensitizer-L tryptophan. UV B radiation is contained, thus protecting laboratory personnel. The system provides a continuous spectral UV B dose of 30 mJ/cm2/hr, which is equivalent to approximately 5.5 UV sunlight hours reaching an aliquot of mammalian aqueous humor. CONCLUSIONS: This biophysical apparatus can be used to investigate antioxidant protective systems in ocular fluid (aqueous/vitreous humors and tears), and is also applicable to evaluating the interaction of radiation with ocular and nonocular tissue cell homogenate.

Multicenter ophthalmic and nutritional age-related macular degeneration study--part 2: antioxidant intervention and conclusions.

BACKGROUND: The experimental design, subjects, procedures and baseline data for the prospective double blind dry ARMD-antioxidant intervention study have been described in Part 1. METHODS: At eight DVA medical centers, 32 patients (group one) were assigned a placebo and 39 patients (group two) a "broad spectrum" antioxidant capsule. Data was collected in five areas: demographic; ophthalmic; dietary analysis of daily food intake; serum analysis; and adverse gastrointestinal symptoms. Data was serially acquired at baseline, 6 months, 12 months and 18 months, and was analyzed by univariate repeated factors ANOVA, p = 0.05. RESULTS: Group two (antioxidant po BID) maintained their distance LogMAR visual acuity (p = 0.03), while there was a trend toward both stabilized near M print (p = 0.07) and 6 cycle/degree contrast sensitivity (p approximately 0.10), in left eyes. However, group two (antioxidant) also had increased cortical opacification of the right lens (p = 0.04), compared to group one (placebo). Self perceived stabilization of vision was reported by subjects in group two and supported the objective data (Pearson chi square; p = 0.05). CONCLUSIONS: A specific 14 component antioxidant capsule taken twice daily stabilized but did not improve dry ARMD over the study period of 1.5 years. The ARMD stabilized eyes had less advanced disease functionally but not by fundus appearance. Decreased intake of cardioprotective nutrients (vitamin E, zinc, magnesium, B6 and folate) in ARMD patients remained constant over the course of the trial.

Managing expectations between patient and nurse.

With increasing consumer interest in the cost and quality of healthcare, it is imperative that patients' expectations of nursing care be recognized and managed. This concept is critical to the continued viability of any healthcare institution in the current competitive market. The authors describe the process that one hospital undertook in looking at patient satisfaction with nursing care, the modification of a patient satisfaction tool, and the development of a quality model of care.

Is there a prevention and treatment strategy for macular degeneration?

BACKGROUND: Reduced antioxidant defense of the retina secondary to poor nutrition and smoking combined with increased life expectancy and exposure to solar radiation can provide a triple assault on the aging retina. If left untreated, without prophylactic measures, 7.5 million American adults will suffer vision loss from ARM by the year 2020. METHODS: Better known pathobiologic interrelationships between ARM, nutritional/antioxidant status and UV/blue visible solar exposure risk based on animal research and recent epidemiologic studies is presented, along with an overview of ocular free radical biochemistry. RESULTS: Because ARM begins parafoveally, and confounding ocular lens changes often occur, there have been practical clinical/research difficulties in early diagnosis, and assessing improvement/loss from nutritional intervention. Threshold Amsler grid testing, contrast sensitivity and more recently red foveal flicker fusion frequency testing are presented as promising tests for discerning subtle pathologic and functional changes in vision. CONCLUSIONS: A conservative prevention/treatment strategy is presented involving identification of at-risk patients, solar radiation protection, nutritional counseling based on the new USDA Food Pyramid concept and vitamin/mineral supplementation if necessary for non-compliant high risk patients. Excessive vitamin intake of particular micronutrients has some drawbacks, and the optometrist should be aware of the implications of self-prescribed OTC ocular formulations, the difference between UV risk vs. "blue light hazard" and ongoing clinical trials.

Cutaneous and ocular changes associated with the use of chlorpromazine.

A 57-year-old schizoaffective disorder patient was placed on chlorpromazine because of its sedative properties and low profile for extrapyramidal side effects. After two years of treatment, the patient developed photosensitivity and blue-gray pigmentation of the skin of the face, neck and dorsum of hands. Significant pigment deposits were also observed in eye lens and cornea. An oval translucent lesion lateral to the left angle of the mouth was removed and the biopsy showed a basal cell carcinoma. The cutaneous and eye pigmentation were only partially reversible after the discontinuation of chlorpromazine. There has been no recurrence of basal cell carcinoma.

Case report: prismatic redirection of vision for progressive supranuclear palsy.

Limitation of extraocular movements in progressive supranuclear palsy is a significant and uncomfortable problem. Specially designed mirror-prism spectacles make it possible for these patients to read and feed themselves.