Spectral sensitivities of jumping spider eyes

Summary Spectral sensitivities of the anterior lateral, posterior lateral and anterior median eyes of the jumping spider,Menemerus confusus Boes. et Str. have been studied by recording electroretinograms (ERGs) and receptor potentials. The anterior and posterior lateral eyes have a single type of visual cell with a maximum spectral sensitivity at about 535–540 nm. The anterior median eye has four types of visual cells with maximum sensitivities at about 360, 480–500, 520–540 and 580 nm, respectively. The ERGs recorded from the optic nerve side (posterior part of the retina) were affected greatly by long wave chromatic light and those on the corneal side (anterior part of the retina) by short wave chromatic light, suggesting that each receptor layer contains a different photopigment.     

Precise cytochemical measurement of neotetrazolium formazan by scanning and integrating microdensitometry

Summary The extinction values of the formazan of neotetrazolium, deposited in tissue sections, have been measured using a Vickers M 85 scanning and integrating microdensitometer with a scanning spot of 0.25 diameter. These values have been correlated with the amount of formazan in the same sections as determined by elution and spectrophotometry. The extinction at 520 nm did not increase linearly with increasing amounts of formazan in sections incubated for different times. This has been shown to be due to the presence of two formazan components which have different absorption characteristics; a red diffuse colour with a molar extinction coefficient of 20580 and a more granular purple product with a coefficient of 7370. However, at 585 nm, the isosbestic wavelength, the extinction was directly proportional to the amount of formazan in the section, irrespective of the nature of the end product; at this wavelength the molar extinction coefficient was 7200.These results have shown that scanning and integrating microdensitometry can be used for the precise measurement of the amount of neotetrazolium formazan, in picograms, in a single cell of group or cells in a tissue section.     

Fluorescent pigments in the newly isolated methylotrophs: Pseudomonas J16 and Methylomonas Pl1.

The pigments showing fluorescence maxima at 390, 366, 450-460 and 520 nm at excitation wavelength 254, 366 and 450 nm respectively, were detected in the cells and culture media of the obligate methylotroph Methylomonas Pl1 and facultative methylotroph Pseudomonas J26. The maximum at 520 nm is associated with the occurrence of a flavin pigment enabling growth of Lactobacillus casei E ATCC-7469 on the vitamin B2 deficient medium. The remaining fluorescence maxima are related to the prosthetic group of methanol dehydrogenase.     

Discrete bathochromic shifts exhibited by fluorescein ligand bound to rabbit polyclonal anti-fluorescein Fab fragments

Eleven individual hyperimmune rabbit polyclonal anti-fluorescein Fab fragment preparations were resolved into heterogeneous subfractions based on differential dissociation times from a specific adsorbent. Four Fab subfractions (i.e., 0.1-, 1.0-, 10-, and 100-day elutions) that differed in affinity were characterized and classified according to the extent of the bathochromic shift in the absorption properties of antibody-bound fluorescein ligand. Absorption maxima of bound fluorescein were shifted in all cases to two distinct narrow ranges, namely, 505 to 507 nm or 518 to 520 nm relative to 491 nm for free fluorescein. There was no direct correlation between the two spectral shift populations and antibody affinity, fluorescence polarization, fluorescence quenching, or fluorescence lifetimes of bound ligand. Fluorescence emission maxima varied with the bathochromic shift range. Bound fluorescein ligand, with absorption maxima of 505 to 507 nm and 518 to 520 nm showed fluorescence emission maxima of 519 to 520 nm and 535 nm, respectively. The two spectral shift ranges differed by 14 to 15 nm and/or energies of 1.5 kcal mol^–1 relative to each other and to the absorption maximum for free fluorescein. Spectral effects on the antibody-bound ligand were discussed relative to solvent-water studies and the atomic structure of a high-affinity liganded anti-fluorescein active site.     

Halistaurin, phialidin and modified forms of aequorin as Ca2+ indicator in biological systems.

Two kinds of aequorin-type photoproteins, i.e., halistaurin and phialidin, and four kinds of modified forms of aequorin, i.e., products of acetylation, ethoxycarbonylation, fluorescamine-modification and fluorescein labelling, were prepared. The modified forms of aequorin were more sensitive to Ca2+ than was aequorin in their Ca2+-triggered luminescence reactions, whereas halistaurin and phialidin were less sensitive. The emission maxima of luminescence were all within a wavelength range 450-464 nm, except for fluorescein-labelled aequorin, which emitted yellowish light (lambda max. 520 nm). A new technique of measuring Ca2+ concentration is suggested.     

OSL and thermally assisted OSL response in dental enamel for its possible application in retrospective dosimetry

Dental enamel was studied for its thermoluminescence (TL) and optically stimulated luminescence (OSL) defects. The TL studies showed a wide glow curve with multiple peaks. The thermally assisted OSL (TA-OSL) studies showed that the integrated TA-OSL and thus OSL signal increases with readout temperature between 100 and 250 °C, due to the temperature dependence of OSL. The thermally assisted energy E _A associated with this increase is found to be 0.21 ± 0.015 eV. On the other hand, the signal intensity decreases with temperature between 260 and 450 °C. This decrease could be due to depletion of OSL active traps or possible thermal quenching. The increase of the OSL signal at increased temperature can be used to enhance the sensitivity of dental enamel for ex vivo measurements in retrospective dosimetry. The emission and excitation spectra of its luminescence centers were studied by photoluminescence and were found to be at 412 and 324 nm, respectively. It was found to possess multiple OSL active traps having closely lying photoionization cross sections characterized by continuous wave OSL and nonlinear OSL methods. The investigated dental enamel samples showed a linear OSL dose response up to 500 Gy. The dose threshold was found to be 100 mGy using a highly sensitive compact OSL reader with blue LED (470 nm) stimulation.     

Obelin from the bioluminescent marine hydroid Obelia geniculata: cloning, expression, and comparison of some properties with those of other Ca2+-regulated photoproteins

A cDNA encoding the Ca2+-regulated photoprotein of the bioluminescent marine hydroid Obelia geniculata was cloned and sequenced. The cDNA is a 774 bp fragment containing two overlapping open reading frames, one of which contained 585 bp encoding a 195 amino acid polypeptide which obviously has the primary structure of the apoprotein of a calcium-regulated photoprotein. Many of the residues are identical to those in other Ca2+-regulated photoproteins: 86% compared with that from Obelia longissima, 76% with that from Clytia (Phialidium), 64% with that from Aequorea, and 64% with that from Mitrocoma(Halistaura). The obelin from O. geniculata was overexpressed in Escherichia coli, refolded from inclusion bodies, and purified. The yield of highly purified recombinant protein was 55-80 mg/L of LB medium. O. geniculata obelin has absorption maxima at 280 and 460 nm and a shoulder at approximately 310 nm. The calcium-discharged protein loses visible absorption but exhibits a new absorption maximum at 343 nm. The bioluminescence of the obelin from O. geniculata is blue (lambda(max) = 495 nm). In contrast, the fluorescence of the calcium-discharged protein is yellow-green (lambda(max) = 520 nm; excitation at 340 nm). This is in sharp contrast to aequorin in which the bioluminescence and fluorescence emission spectra of the calcium-discharged protein are almost identical (lambda(max) = 465 nm). The Ca2+ concentration-effect curve for O. geniculata obelin is similar to those of many other photoproteins: at [Ca2+] below approximately 10(-8) M, calcium-independent luminescence is observed, and at [Ca2+] approximately 10(-3) M, the luminescence reaches a maximum. Between these extremes, the curve spans a vertical range of almost 8 log units with a maximum slope on a log-log plot of about 2.5. In the absence of Mg2+ the rate constant for the rise of bioluminescence determined by the stopped-flow technique is about 450 s(-1). The effects of Mg2+ on the kinetics of bioluminescence are complicated, but at all concentrations studied they are relatively small compared to the corresponding effects on aequorin luminescence. At least with respect to speed and sensitivity to Mg2+, the obelins from both O. longissima and O. geniculata would appear to be more suitable than aequorin for use as intracellular Ca2+ indicators.     

Perithecial Formation in Gelasinospora reticulispora: IV. Action Spectra for the Photoinduction

Action spectra for photoinduction of perithecia after different lengths of dark period were determined with apically growing mycelium of a sordariaceous fungus Gelasinospora reticulispora. When hyphae were exposed to monochromatic light in near ultraviolet and visible regions, reciprocity between intensity and exposure time was observed within the range of incident energy used. The resulting action spectrum determined after a dark period of 48 hours showed a peak at 460 nm with shoulders at 420 and 480 nm and another peak at 370 nm, indicating minima at 410, 430, and 470 nm. After 72 hours darkness the spectrum was very similar to the above, except that the major peak shifted to 450 nm and the near ultraviolet region was somewhat less effective. In both cases, wavelengths longer than 520 nm showed no effect.     

Investigation of luminescent banding in solid coral: the contribution of phosphorescence

This study investigates the nature and components of annual luminescent banding in massive Porites coral skeletons, with a view to refining the technique for using this banding to reconstruct past environmental conditions. Three-dimensional excitation-emission-matrix spectroscopy and optical fibre beam delivery have been used to investigate the luminescence properties of the bright and dull bands of solid coral. Six characteristic excitation/emission peaks have been identified: 280/450–600, 340/450, 370/470, 390/485, 420/505 and 450/530 nm. The first peak corresponds to protein-type fluorescence. The others are characteristic of humic acid luminescence. The difference in luminescence intensity between bright and dull bands has been quantified and characterised spectroscopically. The luminescence of the bright bands is up to 25% more intense than their neighbouring dull bands with the greatest increase in relative intensity in the long wavelength emission region, between 500 and 600 nm. The contribution of long-lived phosphorescence to the total luminescence intensity has been determined by time-resolved measurements on the 100 ms timescale. Both bright and dull bands show long-lived phosphorescence with decay times up to 1.5 s. This phosphorescence accounts for about 10% of the total luminescence intensity of bright bands. The difference in phosphorescence intensity between bright and dull bands is substantially greater than the difference in total luminescence intensity: the phosphorescence of bright bands is up to twice as intense as that of dull bands. This suggests that phosphorescence plays an important role in defining luminescent banding in coral. Furthermore, the large observable difference in phosphorescence between bright and dull bands indicates that measurement of phosphorescence profiles across growth bands in corals may prove to be a more sensitive indicator of past environmental conditions than measurements of total luminescence. Received: 18 March 1999 / Accepted: 20 December 1999     

Blue excitable green emitting Ce3+ doped CaS phosphor for w‐LEDs

CaS:Ce³⁺ is an efficient green‐emitting (535 nm) phosphor, excitable with blue light (450–470 nm) and was synthesized via a solid‐state reaction method by heating under a reducing atmosphere. The luminescent properties, photoluminescent (PL) excitation and emission of the phosphor were analyzed by spectrofluorophotometry. The excitation and emission peaks of the CaS:Ce³⁺ phosphor lay in the visible region, which made them relevant for light‐emitting diode (LED) application for the generation of white light. Judd‐Oflet parameters were calculated and revealed that green light emitted upon blue illumination. The prepared phosphor had strong blue absorption at 470 nm and a broad green emission band range from 490–590 nm with the peak at 537 nm. The characteristics of the CaS:Ce³⁺ phosphor make it suitable for use as a wavelength tunable green emitting phosphor for three band white LEDs pumped by a blue LED (470 nm). The Commission International de l'Eclairage co‐ordinates were calculated by a spectrophotometric method using the spectral energy distribution (0.304, 0.526) and confirm the green emission. The potential application of this phosphor is as a phosphor‐converted white light‐emitting diode. Copyright © 2015 John Wiley & Sons, Ltd.     

Separation of two constitutive forms of cytochrome P-450 active in aminopyrine N-demethylation from rabbit intestinal mucosa microsomes

Two constitutive forms of cytochrome P-450, designated P-450ib and P-450ic, were purified from intestinal mucosa microsomes of untreated rabbits. P-450ib and P-450ic have minimal molecular weights of 56 000 and 49 000, respectively, as determined by calibrated sodium dodecyl sulphate polyacrylamide gel electrophoresis. The CO-reduced difference spectral maximum of cytochrome P-450ib is at 450 nm and P-450ic is at 451 nm. Both the cytochromes preferentially demethylate aminopyrine, benzphetamine and N,N-dimethylaniline in the presence of NADPH-cytochrome P-450 reductase. Cytochrome P-450ib has absorption maxima at 417, 535 and 573 nm in the oxidized form, indicating that this cytochrome is in a low-spin state. Ouchterlony double-diffusion studies show that cytochrome P-450ib does not cross-react with antisera against liver cytochrome P-450LM2 purified from phenobarbital-treated rabbits, but P-450ic cross-reacts with spur formation. Unlike cytochrome P-450ib, P-450ic is very similar, if not identical, to liver cytochrome P-450LM2 on the basis of its molecular weight, spectral properties, catalytic activities and immunochemical properties.     

OCTAHEDRAL CRYSTALS IN PHYCOMYCES. II

"Phycomyces blakesleeanus" sporangiophores contain octahedral crystals throughout their cytoplasm and vacuole. More octahedral crystals were found in the wild-type strain G5 (+) than in the β-carotene-deficient mutant C5 (-), and much more than in the mutant C141 (-), which is sensitive to only high light intensity. In the wild type, the number of crystals per sporangiophore increased until the sporangiophore reached stage IV, and then decreased. Stage I contained the most crystals per unit volume. Cultures grown in darkness had the maximum number of crystals. Under high light intensity, there was an overall reduction of crystals. The crystals are regular octahedrons. The crystals were isolated from the sporangiophores by a method of sucrose density-gradient centrifugation. They contain nearly 95% protein, are stable in organic solvents, but can be solubilized in buffer solution above pH 9.5 and below 2.5. The crystals weakly fluoresce with an emission peak at 540 nm, which is affected by irradiation with white light. Absorption spectra of freshly prepared crystals show absorption maxima around 265–285 nm, 350–380 nm, and 450–470 nm. These absorption peaks for the crystals are close to those of the phototropic and light-growth action spectra. These data suggest that the crystals may contain a flavoprotein which may be the photoreceptor pigment of "Phycomyces".     

Enhancement of Cerenkov Luminescence Imaging by Dual Excitation of Er3+, Yb3+-Doped Rare-Earth Microparticles

Cerenkov luminescence imaging (CLI) has been successfully utilized in various fields of preclinical studies; however, CLI is challenging due to its weak luminescent intensity and insufficient penetration capability. Here, we report the design and synthesis of a type of rare-earth microparticles (REMPs), which can be dually excited by Cerenkov luminescence (CL) resulting from the decay of radionuclides to enhance CLI in terms of intensity and penetration. Methods: Yb³⁺- and Er³⁺- codoped hexagonal NaYF₄ hollow microtubes were synthesized via a hydrothermal route. The phase, morphology, and emission spectrum were confirmed for these REMPs by power X-ray diffraction (XRD), scanning electron microscopy (SEM), and spectrophotometry, respectively. A commercial CCD camera equipped with a series of optical filters was employed to quantify the intensity and spectrum of CLI from radionuclides. The enhancement of penetration was investigated by imaging studies of nylon phantoms and nude mouse pseudotumor models. Results: the REMPs could be dually excited by CL at the wavelengths of 520 and 980 nm, and the emission peaks overlaid at 660 nm. This strategy approximately doubled the overall detectable intensity of CLI and extended its maximum penetration in nylon phantoms from 5 to 15 mm. The penetration study in living animals yielded similar results. Conclusions: this study demonstrated that CL can dually excite REMPs and that the overlaid emissions in the range of 660 nm could significantly enhance the penetration and intensity of CL. The proposed enhanced CLI strategy may have promising applications in the future.     

Solution luminescence from chloro(2,2′:6′,2″-terpyridine)platinum(II) chloride in micelles

Previously characterized as being non-luminescent in room-temperature fluid solution, the coordination compound chloro(2,2′:6′,2″-terpyridine)platinum(II) chloride can display two types of luminescence in certain microenvironments. In aqueous solutions of anionic and neutral surfactants having concentrations near or above their critical micelle concentration, [Pt(terpy)Cl]Cl (10-50 μM) displays broad emission centered at ∼610 nm that is characterized as metal-to-ligand charge transfer phosphorescence (³MLCT). In high concentration (10-100 mM) solutions having no surfactant, [Pt(terpy)Cl]Cl aggregates form. Excitation in the 470-540 nm region results in a long-wavelength emission centered at ∼720 nm that is characterized as metal-metal-to-ligand charge transfer phosphorescence (³MMLCT). This emission can also be detected in lower concentration solutions (10-50 μM) with surfactant concentration below its critical micelle concentration. Enhancement of ³MLCT luminescence is also found for the related phenylacetylide complex cation [Pt(terpy)(CCPh)]⁺ in micelles of the anionic surfactant sodium dodecyl sulfate.     

Eye function of Mysidacea (Crustacea) in the northern Baltic Sea

Eye spectral sensitivity, [S(lambda)], was measured in seven northern Baltic mysid species using an electroretinogram technique. Their S(lambda) curves were compared with the spectral distribution of underwater light at their normal habitats. In the littoral species Neomysis integer, Praunus flexuosus and Praunus inermis, the S(lambda) maxima, [S(lambda)(max)], were in the wavelength-bands of 525-535, 505-515 and 520-530 nm respectively. The neoimmigrant species Hemimysis anomala had a S(lambda)(max) around 500 nm and high sensitivity at 393 nm, possibly indicating UV-sensitivity. S(lambda) of the pelagic species Mysis mixta and Mysis relicta sp. II was at about 505-520 nm. M. relicta sp. I from Pojoviken Bay and fresh water humic Lake P??j?rvi had S(lambda)(max) at approximately 550 nm and 570 nm respectively. This is in accordance with a similar long-wavelength shift in light transmittance of the respective waters. The eyes of the latter population were also damaged by strong light. The pontocaspian neoimmigrant H. anomala is clearly adapted to waters transmitting more blue light.     

蓝光对浮游状态和生物膜内铜绿假单胞菌的抗菌作用

目的：探讨450 nm-470 nm可见光（蓝光）是否具有杀灭浮游状态和生物膜内铜绿假单胞菌的作用。方法：分别采用不同能量密度的蓝光照射浮游状态铜绿假单胞菌,与红光对照组、空白对照组相比,将照射后细菌采用平板涂板法评价蓝光杀菌效果;制作铜绿假单胞菌生物膜模型,16 J/cm2能量密度蓝光照射后通过激光共聚焦显微镜和扫描电子显微镜观察生物膜内细菌存活情况以及生物膜结构变化。结果：与空白对照组相比,2 J/cm2及以上能量密度组蓝光照射后,细菌数目明显减少,杀菌率明显增加（P〈0.05）,并呈剂量效应关系;16 J/cm2能量密度光照后生物膜内细菌死亡数较空白对照组明显增加且生物膜结构变稀疏。结论：450 nm-470 nm可见光（蓝光）具有高效杀灭浮游状态和生物膜内铜绿假单胞菌的作用。     

Light absorption patterns of intact Rosa flowers in relation to the flower colour

Absorption curves of fresh, intact petals from 18 rose cultivars and 2 species were measured and compared with visual evaluations of their colours and there was a reasonable correlation. The in vivo maxima of anthocyanin absorption were in the range of 520–560 nm. Five patterns of absorption spectrum in the visible region were recognized: (a) maximum range ca. 520–535 rim (red roses); (b) as (a) but low absorbance (pink roses); (c) absorption pattern varying with age of flowers; (d) absorption at long wavelengths in blue roses due to co-pigmentation of cyanin, flavonols; (e) absorption of carotenoids and anthocyanins together in yellow, orange or orange red flowers.     

On the properties of fluorescing compounds in guard and epidermal cells of Allium cepa L.

Onion guard cells, in contrast to those of Vicia and Pisum, do not require an alkaline treatment in order to fluoresce. Fluorescing compounds of Allium cepa L. were characterized using in-vivo microspectrophotometry; furthermore, invitro chemical analysis for epidermal tissue, intact guard and epidermal cells, and isolated guard-cell protoplasts was performed. The emission intensity (_max 520 nm) decreased when intact onion guard cells were excited with 436 nm light, but increased (_max 470 nm) when excited at 365 nm. This photodecomposition at 436 nm is typical of flavins or flavoproteins whereas an increase in fluorescence intensity with excitation at 365 nm may be explained by the presence of other substances. The presence of flavins could not be unambiguously confirmed from these results. Indeed, the absorption spectra of the vacuolar area of guard cells did not show the peak at 445 nm which is characteristic for flavins. Furthermore, there was no decrease of absorption at the excitation wavelengths of 440 and 330 nm. Since spectral data indicate the presence at high amounts of flavonoids in guard and epidermal cells, this may reduce the sensitivity for the detection of flavins in guard cells. Using thin-layer chromatography and high-performance liquid chromatography together with hydrolytic procedures, flavonol glycosides with kaempferol and quercetin as aglycones substituted with sulphate and glucuronate were identified. Further studies on guard-cell metabolism should consider the presence of flavonoids in stomata of onion and other plants.Abbreviations GCP guard-cell protoplast - HPLC high-performance liquid chromatography - TLC thin-layer chromatography     

Nachweis dichroitischer Absorption des Sehfarbstoffes in den Rhabdomeren des Insektenauges

Zusammenfassung Durch mikrospektrometrische Messung einzelner Rhabdomere im überlebenden Facettenauge von Calliphora erythrocephala Meig. mit linear polarisiertem Licht wird dichroitische Absorption im Bereich von mindestens 450–570 nm nachgewiesen. Das Maximum des Dichroismus bei 500–520 nm fällt mit dem Absorptionsmaximum des Sehfarbstoffs zusammen. Im Gegensatz zur Formdoppelbrechung der Rhabdomere verschwindet der Dichroismus mit der Bleichung des Sehfarbstoffs. Die Befunde werden als Beweis dafür betrachtet, daß das Rhodopsin durch seine orientierte Einlagerung in den Rhabdomeren als Receptorpigment und als Analysator zugleich (Sehfarbstoff-Analysator nach Stockhammer 1956, 1959) funktioniert.

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Summary Microspectrophotometric measurements with linear polarized light in single rhabdomeres of surviving compound eyes of the blow-fly Calliphora erythrocephala Meig. present evidence for dichroic absorption, at least between 450 and 570 nm. The maximum of dichroism at 500 to 520 nm corresponds to the absorption maximum of the visual pigment. In contrast to the structure birefringence of the rhabdomeres, the dichroism disappears during bleaching of the visual pigment. The results are considered to prove the hypothesis that rhodopsin is functioning both as receptor pigment and as analyser (Sehfarbstoff-Analysator after Stockhammer 1956, 1959).

     

Ceruloplasmin-anion interaction. A circular dichroism spectroscopic study.

The effect of anion binding to ceruloplasmin has been studied using absorption and cirbular dichroism spectral data. At anion to ceruloplasmin molar ratios approaching infinite, OCN-, N3- and SCN- bind to ceruloplasmin giving rise to similar alterations in circular dichroism and absorption spectra. The positive bands at 610 and 520 nm in circular dichroism spectra disappear, a negative one apperars at 600 nm and the peak at 450 nm is only slightly modified. There is a new negative band at 410 nm well-defined in OCN- ceruloplasmin spectra. The decrease in absorption at 610 nm is ascribed to the disruption of one type I Cu-S(cysteine) bond owing presumably to the changes induced by anions in the protein secondary structure. The new band at 410 nm is assigned to a charge transfer transition from the ligand replacing cysteine at its binding site. Both absorption and circular dichroism spectra show isobestic points indicating that anion binding to the enzyme, disruption of one of the two type I Cu-S bonds and coordination of this Cu to another protein residue take place simultaneously.