Silk oak flowers as a source of β-carotene

The pigment of the yellow flowers of the silk oak (Grevillea robusta, Cunningham) does not appear to have been investigated heretofore. If the dried material is extracted with ether, the solution shows typical absorption maxima at 483 and 453 mµ,corresponding to the spectrum of p-carotene. The rather blurred borders of these bands indicate, however, the presence of other polyenic pigments in small quantities. After saponification a photometric analysis of the total extract gave values which would correspond to 270 mg. of β-carotene in 1 kilo of the dry flowers if no other pigments were present. After a chromatographic separation the true β-carotene content was found to be about 215 mg. per kilo. Two-thirds of this amount was isolated as crystals; lycopene or γ- and α-carotene were not present [1]. The non-carotene fraction is a complicated xanthophyll mixture in which no single compound predominates. From this fraction two very small amounts of crystalline material were isolated, one of which was kryptoxanthin and the other a new carotenoid possessing a remarkably short wave-length spectrum. For the separation and study of carotenoids contained in extracts we suggest the systematic use of the ultraviolet lamp which has been so helpful in the chromatography of colorless substances (2). Plant pigments are frequently accompanied by large amounts of colorless material which prevent the formation of sharp pigment zones in the Tswett column and thus a satisfactory separation of the components. Furthermore, the crystallization of some carotenoids may be hindered. Fortunately many such colorless substances show an intense fluorescence (3). An observation made in ultraviolet light during the chromatographic separation of the pigments may furnish a good indication of the best method and optimum extent of developing the chromatogram. The distribution of the fluorescence may also indicate the lines at which it is best to cut the column. By sacrificing small amounts of pigment large portions of colorless associated material may be eliminated in this simple way

The carotenoid and provitamin A content of the watermelon

10 years ago lycopene, C40H56, and carotene, C40H56, were isolated from the pulp of the European watermelon (Citrullus vulgaris, Schrad. = Cucumis citrullus, L.) by Zechmeister and Tuzson (l), and it was shown that the chief pigment, lycopene, is responsible for the red color. As the chromatographic method was not available at that time, no precise information as to the composition of the pigment was obtained. In some new experiments described below we have carried out a quantitative analysis of the components and have estimated the provitamin A content of the California watermelon. 1 kilo of the pulp examined contained 1.0 mg. of a complicated xanthophyll mixture, 6.1 mg. of lycopene, 0.06 mg. of γ-carotene, 0.16 mg. of unknown carotenoids (located in the column between γ- and β-carotene), 0.46 mg. of β-carotene, 0.01 mg. of α-carotene. The figures include the fractions of lycopene, and γ- and β-carotene which underwent isomerization during the experimental procedure (2). The calorimetric value of the total extract of 1 kilo of pulp corresponded to 7 to 8 mg. of “lycopene”; some samples were, however, considerably richer in pigment. Our material, picked in California in September, contained, according to the above figures, 0.5 mg. of provitamin A in 1 kilo of pulp, or about one-fifth to one-sixth of the daily β-carotene requirement of an adult person. It is interesting to note that a considerable number of yellow and pink unidentified oxygen-containing carotenoids were found in minute quantities; i.e., to the extent of about 0.01 mg. per kilo of pulp. Even with the use of chromatography 1000 or more kilos of melon would be needed for a satisfactory study of these pigments. One of them is spectroscopically identical with torulene, detected by Lederer in red yeast (3)

Soma size distinguishes projection neurons from neurokinin 1 receptor-expressing interneurons in lamina I of the rat lumbar spinal dorsal horn

Lamina I of the spinal dorsal horn contains neurons that project to various brain regions, and ∼80% of these projection cells express the neurokinin 1 receptor (NK1r), the main receptor for substance P. Two populations of NK1r-immunoreactive neurons have been identified in lamina I: small weakly immunoreactive cells and large cells with strong immunolabelling [Cheunsuang O and Morris R (2000) Neuroscience 97:335–345]. The main aim of this study was to test the hypothesis that the large cells are projection neurons and that the small cells are interneurons. Projection neurons were identified by injection of tracers into the caudal ventrolateral medulla and lateral parabrachial area, and this was combined with immunostaining for NK1r. We found a bimodal size distribution for NK1r-immunoreactive neurons. The small cells (with somatic cross-sectional areas <200 μm2) showed weak immunoreactivity, while immunostaining intensity was variable among the large cells. Virtually all (99%) of the immunoreactive cells with soma areas >200 μm2 were retrogradely labelled, while only 10% of retrogradely labelled cells were smaller than this. Soma sizes of retrogradely labelled neurons that lacked NK1r did not differ from those of NK1r-expressing projection neurons. It has been suggested that a population of small pyramidal projection neurons that lack NK1r may correspond to cells activated by innocuous cooling, and we therefore assessed the morphology of retrogradely labelled cells that were not NK1r-immunoreactive. Fifteen percent of these were pyramidal, but these did not differ in size from pyramidal NK1r-immunoreactive projection neurons. These results confirm that large NK1r-immunoreactive lamina I neurons are projection cells, and suggest that the small cells are interneurons. Since almost all of the NK1r-immunoreactive cells with soma size >200 μm2 were retrogradely labelled, cells of this type can be identified as projection cells in anatomical studies

A quantitative study of neurochemically-defined populations of inhibitory interneurons in the superficial dorsal horn of the mouse spinal cord

Around a quarter of neurons in laminae I-II of the dorsal horn are inhibitory interneurons. These play an important role in modulating somatosensory information, including that perceived as pain or itch. Previous studies in rat identified four largely non-overlapping neurochemical populations among these cells, defined by expression of galanin, neuropeptide Y (NPY), neuronal nitric oxide synthase (nNOS) or parvalbumin. The galanin cells were subsequently shown to coexpress dynorphin. Several recent studies have used genetically-modified mice to investigate the function of different interneuron populations, and it is therefore important to determine whether the same pattern applies in mouse, and to estimate the relative sizes of these populations. We show that the neurochemical organisation of inhibitory interneurons in mouse superficial dorsal horn is similar to that in the rat, although a larger proportion of these neurons (33%) express NPY. Between them, these four populations account for ∼75% of inhibitory cells in laminae I-II. Since ∼25% of inhibitory interneurons in this region belong to a novel calretinin-expressing type, our results suggest that virtually all inhibitory interneurons in superficial dorsal horn can be assigned to one of these five neurochemical populations. Although our main focus was inhibitory neurons, we also identified a population of excitatory dynorphin-expressing cells in laminae I-II that are largely restricted to the medial part of the mid-lumbar dorsal horn, corresponding to glabrous skin territory. These findings are important for interpretation of studies using molecular-genetic techniques to manipulate the functions of interneuron populations to investigate their roles in somatosensory processing

Ínfű fajok és a lucerna másodlagos kémiai anyagai és hatástani vizsgálatai rovarokon = Allelochemicals from alfalfa and Ajuga plants and those of biological activity on different insect orders

Különböző Ajuga fajok teljes növényi őrleményét (Plodia interpunctella), teljes és frakcionált metanolos kivonatait (Acyrthosiphon pisum, Dysdercus cingulatus, Aedes aegypti, Daphnia magna), valamint tisztított ekdiszteroidok hatékonyságát vizsgáltuk. A Plodia-teszben az A. reptans 8 %-os dózisa azon túl, hogy kétszeresére növelte a posztembrionális kifejlődéshez szükséges időt és csökkentette a bábok súlyát - számottevő mortalitást (77 %) is okozott. Az A. aegypti esetében jelentős hatékonyságot csak az A. reptans 100%-os metanolos frakciója mutatott (0,1 %-os dózisban ~80 %-os mortalitás). A szívó szájszervű rovaroknál (A. pisum, D. cingulatus) A. chamaepitys 100%-os és az A. bracteosa 60%-os frakciói bizonyultak a leghatékonyabbak (0,1 %-os koncentrációban 100 %-os mortalitás). A tisztított ekdiszteroidok közül a 20E és a Polipodin B voltak hatékonyak a levéltetű tesztben (LC50 = 1,07 és 0,21 ppm). Az A. reptans-ból 10 különböző ekdiszteroidot (20-hidroxiekdizon, 20-hidroxiekdizon 3-acetát, ciaszteron, ciaszteron 3-acetát, 29-nor-ciaszteron, ajugaszteron B', ajugalakton, szengoszteron, 23-hidroxi-kapitaszteron, 24-etil-24,26-dihidroxi-taxiszterol 26-metilát) sikerült izolálnunk. Ezek közül két vegyület (23-hidroxi-kapitaszteron, 24-etil-24,26-dihidroxi-taxiszterol 26-metilát) új természetes ekdiszteroidnak bizonyult. Az A. reptans-ból eddig izoláltak közül a 20-hidroxiekdizon 3-acetátot és a ciaszteron 3-acetátot elsőként nyertük ki ebből az Ajuga fajból. | We studied the efficacy of whole plant grist (Plodia interpunctella), crude and fractionated methanolic extracts (Acyrthosiphon pisum, Dysdercus cingulatus, Aedes aegypti, Daphnia magna) as well as pure ecdysteroids from different Ajuga spp. In the Plodia-test the strongest effects were found at 8% concentration of A. reptans grist, which reduced the pupal weight, doubled the postembryonic developmental time, and caused 77% mortality until emergence of adults. In the Aedes-test the crude as well as the 100% methanolic fraction from A. reptans showed high efficacy (80% mortality at 0.1% concentration). In the case of sucking insect species (A. pisum, D. cingulatus) the 100% methanolic fraction of A. chamaepitys and the 60% methanolic fraction of A. bracteosa were highly effective on A. pisum (100% mortality at 0.1% concentration) Polipodine B and 20-hydroxyecdysone were the most effective pure phytoecdysteroids on aphids (LC50 = 0.21 ppm and 1.07 ppm). We have identified 10 different ecdysteroids from the Ajuga reptans var. reptans (20-hydroxyecdysone, 20-hydroxyecdysone acetate, cyasterone, cyasterone 3 acetate, 29-nor-cyasterone, ajugasterone B', ajugalactone, sengosterone, 23-hydroxi-kapitasterone, 24-ethyl-24,26-dihydroxi-taxisterol 26-metilat). Among these 23-hydroxi-capitasterone and 24-ethyl-24,26-dihydroxi-taxisterol 26-metilat are new phytoecdysteroids while, 20-hydroxyecdysone acetate and cyasterone 3 acetate were firstly isolated from A. reptans

Preprotachykinin A (PPTA) is expressed by a distinct population of excitatory neurons in the mouse superficial spinal dorsal horn including cells that respond to noxious and pruritic stimuli

The superficial dorsal horn, which is the main target for nociceptive and pruritoceptive primary afferents, contains a high density of excitatory interneurons. Our understanding of their roles in somatosensory processing has been restricted by the difficulty of distinguishing functional populations among these cells. We recently defined three non-overlapping populations among the excitatory neurons, based on the expression of neurotensin, neurokinin B (NKB) and gastrin-releasing peptide (GRP). Here we identify and characterise another population: neurons that express the tachykinin peptide substance P. We show with immunocytochemistry that its precursor protein (preprotachykinin A, PPTA) can be detected in ~14% of lamina I-II neurons, and these are concentrated in the outer part of lamina II. Over 80% of the PPTA-positive cells lack the transcription factor Pax2 (which determines an inhibitory phenotype), and these account for ~15% of the excitatory neurons in this region. They are different from the neurotensin, NKB or GRP neurons, although many of them contain somatostatin, which is widely expressed among superficial dorsal horn excitatory interneurons. We show that many of these cells respond to noxious thermal and mechanical stimuli, and to intradermal injection of pruritogens. Finally, we demonstrate that these cells can also be identified in a knock-in Cre mouse line (Tac1Cre), although our findings suggest that there is an additional population of neurons that transiently express PPTA. This population of substance P-expressing excitatory neurons is likely to play an important role in transmission of signals that are perceived as pain and itch

Neuronal circuitry for pain processing in the dorsal horn

Neurons in the spinal dorsal horn process sensory information, which is then transmitted to several brain regions, including those responsible for pain perception. The dorsal horn provides numerous potential targets for the development of novel analgesics and is thought to undergo changes that contribute to the exaggerated pain felt after nerve injury and inflammation. Despite its obvious importance, we still know little about the neuronal circuits that process sensory information, mainly because of the heterogeneity of the various neuronal components that make up these circuits. Recent studies have begun to shed light on the neuronal organization and circuitry of this complex region

Multi-objective genetic algorithm applied to spectroscopic ellipsometry of organic-inorganic hybrid planar waveguides

The applicably of multi-objective optimization to ellipsometric data analysis is presented and a method to handle complex ellipsometric problems such as multi sample or multi angle analysis using multi-objective optimization is described. The performance of a multi-objective genetic algorithm (MOGA) is tested against a single objective common genetic algorithm (CGA). The procedure is applied to the characterization (refractive index and thickness) of planar waveguides intended for the production of optical components prepared sol-gel derived organic-inorganic hybrids, so-called di-ureasils, modified with zirconium tetrapropoxide, Zr(OPr(n))(4) deposited on silica on silicon substrates. The results show that for the same initial conditions, MOGA performs better than the CGA, showing a higher success rate in the task of finding the best final solution. (C) 2010 Optical Society of AmericaFCTFEDERPTDC/CTM/72093/2006SFRH/BD/41943/2007MP070

A putative relay circuit providing low-threshold mechanoreceptive input to lamina I projection neurons via vertical cells in lamina II of the rat dorsal horn

Background: Lamina I projection neurons respond to painful stimuli, and some are also activated by touch or hair movement. Neuropathic pain resulting from peripheral nerve damage is often associated with tactile allodynia (touch-evoked pain), and this may result from increased responsiveness of lamina I projection neurons to non-noxious mechanical stimuli. It is thought that polysynaptic pathways involving excitatory interneurons can transmit tactile inputs to lamina I projection neurons, but that these are normally suppressed by inhibitory interneurons. Vertical cells in lamina II provide a potential route through which tactile stimuli can activate lamina I projection neurons, since their dendrites extend into the region where tactile afferents terminate, while their axons can innervate the projection cells. The aim of this study was to determine whether vertical cell dendrites were contacted by the central terminals of low-threshold mechanoreceptive primary afferents. Results: We initially demonstrated contacts between dendritic spines of vertical cells that had been recorded in spinal cord slices and axonal boutons containing the vesicular glutamate transporter 1 (VGLUT1), which is expressed by myelinated low-threshold mechanoreceptive afferents. To confirm that the VGLUT1 boutons included primary afferents, we then examined vertical cells recorded in rats that had received injections of cholera toxin B subunit (CTb) into the sciatic nerve. We found that over half of the VGLUT1 boutons contacting the vertical cells were CTb-immunoreactive, indicating that they were of primary afferent origin. Conclusions: These results show that vertical cell dendritic spines are frequently contacted by the central terminals of myelinated low-threshold mechanoreceptive afferents. Since dendritic spines are associated with excitatory synapses, it is likely that most of these contacts were synaptic. Vertical cells in lamina II are therefore a potential route through which tactile afferents can activate lamina I projection neurons, and this pathway could play a role in tactile allodynia