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Response univariance in bull-frog rods with two visual pigments.

Rods in the bull-frog retina contain varying proportions of rhodopsin (lambda max = 502 nm) and porphyropsin (lambda max = 527 nm) in a dorso-ventral gradient from the porphyropsin-rich dorsal rim to the virtually pure rhodopsin fields of the central and ventral retina. We investigated if quantal excitations in the same rod are different depending on whether they are initiated by isomerization of a rhodopsin or a porphyropsin molecule. Current photoresponses were recorded from dark-adapted rods by sucking the outer segment into a recording pipette. The relation between pigment composition and spectral sensitivity was established by comparison with microspectrophotometrically measured absorbance spectra of rods from the same neighbourhood. Rods with suitable porphyropsin: rhodopsin mixtures (ideally between 1:4 and 1:2) were stimulated with flashes of red (608 nm) and blue (465 nm) light, whereby the red light will isomerize porphyropsin much more often than rhodopsin, and the reverse will be true of the blue light. The amplitude and shape of the single-photon response were found to be identical for the "red" and "blue" flash series to within measurement error (ca 10%). This indicates that the quantal responses initiated by the two pigments are identical.

Animals↗

An analysis of two spectral properties of vertebrate visual pigments.

Spectral data are scrutinized on two properties: (1) the chromophore-induced wavelength shift, i.e. the variation in absorbance maximum (lambda max) upon exchanging the vitamin A1-based chromophore with one based on vitamin A2 in the same opsin; (2) the half-band width (HBW) variation as a function of the reciprocal of the peak absorbance (lambda max)-1. It is shown that in an extended spectral range that includes the UV and when data are plotted in wavenumbers, the chromophore-induced shifts can be approximated with a parabola, whose minimum occurs near 23,000 cm-1 (approximately 430 nm). Similarly, HBW variations can also be fitted with parabolas, however, these show maxima near 430 nm. The theoretical implications of the two phenomena concerning lambda max tuning in vertebrate visual pigments are discussed.

Amphibians↗

Ultraviolet visual pigments in marine fishes of the family pomacentridae.

Near-UV visual pigments have been reported in single cones of several freshwater and euryhaline fishes. The presence of UV visual pigments in stenohaline marine fishes have, as yet, not been identified. In the pomacentridae near-UV visual pigments are present in single cones from the three species we examined--the tropical coral fishes Dascyllus trimaculatus and Pomacentrus coelestis, and the temperate Chromis punctipinnis. Maximum absorption of the UV pigments is centered around 360 nm. In juvenile Chromis, however, the UV visual pigment is not present. Instead there is a single cone containing a violet-sensitive pigment absorbing maximally around 420 nm. All three species are obligate diurnal planktivores. The UV sensitivity may function to enhance their ability to forage on zooplankton.

Animals↗

Ultraviolet vision in a passeriform bird: from receptor spectral sensitivity to overall spectral sensitivity in Leiothrix lutea.

A survey of recent results out of spectrophotometric, microspectrophotometric, and behavioral tests concerning the UV vision of the passeriform bird Leiothrix lutea is presented. In the spectrophotometric study it was shown that the ocular media of Leiothrix' eyes are highly transparent to the near UV with lambda T50 at 320 nm. The comparison of the microspectrophotometric and the behavioral data showed a good fit between the peaks of the four single cones' effective sensitivity spectra and the four peaks in the behavioral spectral sensitivity function. The relation further suggests that the behavioral function might be described as the "over-envelope" of the single cone sensitivities. Leiothrix lutea possesses a genuine UV cone type and reveals it's highest sensitivity in the behavioral test to UV light.

Animals↗

A third, ultraviolet-sensitive, visual pigment in the Tokay gecko (Gekko gekko).

Numerous extraction and microspectrophotometric studies have shown that the nocturnal Tokay gecko (Gekko gekko), has two visual pigments: a "green" with lambda max at 521 nm and a "blue" at 467 nm. In addition, similar studies on other nocturnal gecko species have found only the same two classes of visual pigment. With the finding that some diurnal species of gecko have a third visual pigment class with lambda max peaking in the UV, doubts were raised concerning the presence of only two visual pigment classes in nocturnal forms. Therefore, a microspectrophotometric re-examination of the Tokay gecko was undertaken to look specifically for a UV visual pigment. A UV-absorbing pigment (364 nm lambda max) was found in approx. 20% of the thin outer segments of type C double rods, thought previously to contain only the 467 nm pigment. That this UV-absorbing pigment was truly a visual pigment was confirmed by its dichroism, behaviour following exposure to UV radiation and "nomogram" fit. It is suggested that this visual pigment had been seen in previous microspectrophotometric studies, but its similarity to known photoproducts peaking in the same spectral region resulted in a case of mistaken identity.

Animals↗

Ultraviolet photoreception in mantis shrimp.

An UV-sensitive class of photoreceptors exists in all regions of the retinas of mantis shrimps. UV photosensitivity apparently resides in rhabdomeres of the eighth retinular cell (R8) that lies atop each rhabdom; and in ommatidia where the R8 rhabdomere consists of microvilli parallel in a single direction, sensitivity is maximal when the e-vector of plane-polarized light is parallel to the microvilli. Spectral sensitivity of the UV photoreceptor peaks at 345 nm and is best explained by the presence of a photopigment with lambda max near 325 nm overlain by material that absorbs UV light at wavelengths below approximately 350 nm. Rhabdomeres of R8 cells in several different retinal regions of a variety of species examined contain a photopigment absorbing maximally below 340 nm. Under appropriate conditions, a metapigment with lambda max near 460 nm can be formed. UV vision may be useful for enhancing the visual contrast of midwater predators or prey.

Animals↗

Specialization of retinal function in the compound eyes of mantis shrimps.

Visual function and its specialization at the level of the retina were studied in 13 species of stomatopod crustaceans, representing three superfamilies: Gonodactyloidea, Lysiosquilloidea, and Squilloidea. We measured attenuation and irradiance spectra in the environment of each species, at the actual depths and times of activity where we observed individuals. We also characterized the intrahabdomal filters of all study species and determined the absolute spectral sensitivity functions and approximate photon capture rates of all photoreceptor classes below the level of the 8th retinular cell in seven of these species. Shallow-water gonodactyloid species have four distinct classes of intrarhabdomal filters, producing photoreceptors that are relatively insensitive but which have the broadest spectral coverage of all. Deep-water gonodactyloids and all lysiosquilloids have filters that are spectrally less diverse. These species often discard the proximal filter classes of one or more receptor types. As a result, their retinas are more sensitive but have reduced spectral range or diversity. The single squilloid species has the most sensitive photoreceptors of any we observed, due to the lack both of intrarhabdomal filters and tiered photoreceptors. Photon absorption rates, at the times of animal activity, were similar in most photoreceptor classes of all species, whether the receptors were tiered or untiered, or filtered or unfiltered. Thus, the retinas of stomatopods are specialized to operate at similar levels of stimulation at the times and depths of actual use, while evidently maintaining the greatest possible potential for spectral coverage and discrimination.

Adaptation, Physiological↗

Characterization of Ca2+ signals generated by extracellular nucleotides in supporting cells of the organ of Corti.

ATP has been demonstrated to act as a co-transmitter or neuromodulator in various physiological processes. There is recent evidence that ATP receptors, characterized as P2 purinergic receptors, are expressed in the sensory hair cells of the auditory organ. The aim of the present study was to know whether other cell types of the organ of Corti, the supporting cells, were also sensitive to external ATP. In both types of supporting cells considered in this study, Deiters' cells (DCs) and Hensen's cells (HEs), extracellular ATP at sub-micromolar concentrations evoked a transient increase in [Ca2+]i as monitored with fluorescence microscopy using the calcium probe Indo-1. An apparent Kd of 0.5 and 0.9 microM was determined for DCs and HEs, respectively. Furthermore, we demonstrated that ATP stimulated Ca2+ release from internal stores in DCs, but not in HEs. Dynamic calcium imaging by confocal laser scanning microscopy of ATP induced Ca2+ mobilization demonstrated a calcium wave propagation in the cell body of DCs which originated in the phalangeal processes, suggesting a functional organization of Ca2+ sequestering stores in DCs.

Adenosine Triphosphate↗

Localization and quantification of the nonspecific esterase in injured skin for timing of wounds.

Histochemical quantification of the nonspecific esterase (NSE) in injured skin was performed using histochemical demonstration of the enzyme and a microspectrophotometric scanning technique on specimens taken from 32 Hartley guinea-pigs and 8 cases of human skin wounds. In all antemortem incisions and lacerations, including those made at the agonal stage, NSE activity could be observed in the dermal tissue of the wound edge. The enzyme activity increases with the antemortem duration of the injuries. Both total content and mean concentration of NSE in the wound edge between antemortem and postmortem wound groups differ significantly (less than 0.01). Multiple range test shows that significant differences (P less than 0.05 or P less than 0.01) of total content of NSE in the wound edge also exist in 0-5 min, 15-30 min, 1-h, 2-h and 4-h antemortem incised wound groups and in 0-min, 5-min, 15-30 min, 1-h and 4-h antemortem laceration groups. The positive NSE reactions in 8 cases of human skin wounds were similar. The study indicates that histochemical quantification of NSE in injured skin is very useful in timing wounds and is exactly applicable in medicolegal practice. According to the different influences of inhibitors on enzymes, it was inferred that the enzyme activity in wound edges was due to B-esterase.

Animals↗

Correlation between CT and DNA contents in peripheral lung cancers.

The DNA contents of 30 lung cancers were correlated with preoperative chest CT findings. Tumors with diameters less than 4 cm had lower DNA contents than larger ones, and tumors with spiculated margins had lower DNA than those with lobulated or smooth margins. As DNA contents are correlated with prognosis, the border configuration of peripheral lung cancers may be enough to predict prognosis.

Aged↗

Photofrin-induced fluorescence in progressive and regressive murine colonic cancer cells: correlation with cell photosensitivity.

Microspectrofluorometry and fluorescence imaging were used to investigate the intracellular fluorescence of two murine colonic cancer cell lines--a progressive cell line (PROb) and a regressive cell line (REGb)--incubated with Photofrin. These two cell lines, which were initially cloned from the same chemically induced colonic murine cancer, differ in their metastatic properties and have been considered as models to mimic the tumoral cell heterogeneity. The fluorescence from cytoplasmic area of cells incubated with Photofrin appeared as a complex emission, with two maxima at 632 and 695 nm assigned to monomer species, and a poorly resolved band around 665 nm assigned to aggregates. The spectral distribution was shown to depend on the incubation time, with an aggregate contribution increasing for extended periods. The amount of Photofrin uptake, as determined from the total fluorescence intensity, was found for PROb to be twice that for REGb. However, the phototoxicities were quite similar for both cell lines, suggesting that drug concentration may not be the only determining factor in photobiological efficiency.

Animals↗

Structural conformation and leaching from in vitro aged and retrieved Invisalign appliances.

The objectives of this study were to investigate the structure of Invisalign appliances (Align Technology, Santa Clara, Calif) after intraoral exposure, and to qualitatively and quantitatively characterize the substances leached from the aligners after accelerated in vitro aging. Samples of Invisalign appliances were randomly selected from 10 patients before intraoral placement and after retrieval, and the prepared specimens were subjected to (1) bright-field optical reflection microscopy to study the surface morphology; (2) Fourier transform infrared microspectroscopy to characterize the in vivo changes in molecular composition induced on appliance surfaces, (3) scanning electron microscopy and energy dispersive X-ray microanalysis to identify the elemental composition of integuments formed on the surface, and (4) Vickers hardness (HV 200) testing. Another set of reference and retrieved appliances was subjected to artificial aging for 2 weeks, and the extracts were subjected to gas chromatography-mass spectroscopy. The retrieved appliances demonstrated substantial morphological variation relative to the as-received specimens involving abrasion at the cusp tips, adsorption of integuments, and localized calcification of the precipitated biofilm at stagnation sites. Buccal segments of retrieved appliances showed an increase in hardness, which might be attributed to mastication-induced cold work; however, the clinical implication of this effect on mechanotherapy is unknown. In vitro aged and retrieved appliances were found to leach no traceable amount of substances in an ethanol aging solution.

Adsorption↗

Spatial resolution in infrared microspectroscopic imaging of tissues.

Spatial resolution is one of the most critical measurement parameters in infrared microspectroscopy. Due to the distinct levels of morphologic heterogeneity in cells and tissues the spatial resolution in a given IR imaging setup strongly affects the character of the infrared spectral patterns obtained from the biomedical samples. This is particularly important when spectral data bases of reference microspectra from defined tissue structures are collected. In this paper we have also pointed out that the concept of spatial resolution in IR imaging is inseparable from the contrast. Based on infrared microspectroscopic transmittance data acquired from an USAF 1951 resolution target we have demonstrated how the spatial resolution can be determined experimentally and some numbers for the spatial resolution of popular IR imaging systems are provided. Finally, we have presented a new computational procedure which is suitable to improve the spatial resolution in IR imaging. A theoretical model of 3D-Fourier self-deconvolution (FSD) is given and advantages or pitfalls of this method are discussed. Based on synchrotron IR microspectroscopic data we have furthermore demonstrated that the technique of 3D-FSD can be successfully applied to increase the spatial resolution in a real IR imaging setup.

Animals↗

Micro-XRD study of beta-titanium wires and infrared soldered joints.

OBJECTIVE: The purpose of this study was to investigate the metallurgical phases in beta-titanium soldered joints prepared by infrared soldering, using the Micro X-ray diffraction technique (Micro-XRD), and to characterize the Vickers hardness in the soldered beta-titanium wires. METHODS: Beta-titanium wires with cross-section dimensions of 0.032in.x0.032in. (TMA, Ormco), and both titanium-based solder (Ti-30Ni-20Cu, Selec) and silver-based solder (Ag-22Cu-17Zn-5Sn, Tomy) were selected. Soldering was performed using infrared radiation (RS-1, Morita) under argon atmosphere. Micro-XRD analyses were performed at room temperature. Micro-XRD spectra were obtained for the boundary region of the soldered beta-titanium wires using 50microm and 10microm diameter analysis regions. Hardness was measured at 30microm intervals from boundary of the diffusion layer and beta-titanium wire. The Kruskal-Wallis test with the Bonferroni and Wilcoxson Mann-Whitney tests for nonparametric means were employed as statistical methods (P<0.05). RESULTS: For both types of soldered beta-titanium samples, the Micro-XRD spectra contained four major peaks for body-centered cubic (bcc) beta-titanium. Additional peaks at about 41 and 45 degrees are attributed to Cu-Ti intermetallic phase(s), which may be metastable under soldering conditions. The diffusion layer had greater hardness than bulk beta-titanium for both types of soldered specimens (P<0.05). SIGNIFICANCE: Soldering of beta-titanium orthodontic wire by infrared radiation may be acceptable for clinical use, since Micro-XRD spectra revealed that both types of soldered specimens largely retained the bcc beta-titanium structure. Further research is needed to investigate the mechanical properties and corrosion behavior of infrared-soldered beta-titanium wire.

Alloys↗

Thermal behavior and thermal decarboxylation of 10-hydroxycamptothecin in the solid state.

In order to investigate the thermal-related properties and thermal stability of 10-hydroxycamptothecin (10-HCPT) in the solid state, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) and Fourier transform infrared (FT-IR) microspectroscopy were used. A novel combination of FT-IR microspectroscopy with thermal analyzer was applied simultaneously to monitor the dehydration and rehydration processes of the 10-HCPT sample. The thermal-induced decomposition of the 10-HCPT sample was also determined by using electrospray-ion trap mass spectrometry (ES-ITMS). The results indicated that the 10-HCPT sample used in this study was a monohydrate in structure, this form that can dehydrate to an anhydrate form if the temperature goes beyond 90 degrees C. The 10-HCPT anhydrate was first suggested to have two polymorphs, in which the form I might transform to form II when the 110 degrees C-preheated sample was cooled to 30 degrees C. The polymorphic transformation temperature was shown within 90-120 degrees C with 10.46 kcal/mol of enthalpy. The peak at 1723 cm(-1) found in the IR spectrum of 10-HCPT monohydrate might correspond to the hydrogen-bonded CO stretching vibration of lactone, which shifted to 1750 cm(-1) assigned to a free CO group of lactone after the destruction of hydrogen bonding via dehydration. This suggests that monohydrate seems to interact intramolecularly with 10-HCPT by hydrogen bonding. However, the rehydration process of the 10-HCPT anhydrate might cause it to return to being a monohydrate, depending on the storage condition. In addition, the thermal-induced decarboxylation of the solid-state 10-HCPT when the temperature is beyond 226 degrees C was proven by the appearance of a new IR peak at 1701 cm(-1) and one major mass spectral peak at m/z 321. This unique IR spectral peak at 1701 cm(-1) was due to the conjugated carbonyl group in the degraded product of 10-HCPT. The m/z 321 assigned to the decarboxylation of 10-HCPT was equal to the molecular weight loss of 44 from mass spectra; which was consistent with the weight loss of 11.9% (molecular weight of 43.3) from TGA curve of 10-HCPT anhydrate.

Antineoplastic Agents, Phytogenic↗

Analysis of pigments and coverings by X-ray diffraction (XRD) and micro Raman spectroscopy (MRS) in the cemetery of Tutugi (Galera, Granada, Spain) and the settlement convento 2 (Montemayor, Córdoba, Spain).

This paper focuses on the identification of the composition of the coverings and pigments of two archaeological sites. The sites researched here lie in Andalusia and show two contexts, which have a highly symbolic and ritual meaning. The first, Convento 2 (Montemayor, Córdoba), dates back to the period of formation of the Iberian Culture (VIIth century b.c.). The second is the cemetery of Tutugi (Galera, Granada), of the mid-Iberian period (IVth century b.c.). The analytical procedure consisted in combined and complementary use of XRD and MRS. This allowed to identify the materials used, namely hematite, goethite, coal, gypsum and calcite. Identification of these materials proves essential for the restoration and musealization of both archaeological sites.

Archaeology↗

Chemistry at level of individual aerosol particle using multivariate curve resolution of confocal Raman image.

Airborne particles with aerodynamic diameter in the 10-1 microm range have been collected in an industrial/urban zone by impaction and have been investigated by automated confocal Raman microspectrometry. The computer-microcontrolled XY scanning and Z focusing of Raman images provided many pixel Raman spectra which are characteristics of complex mixture at level of individual particle. The large heterogeneity was not resolved by the spatial resolution of the instrument which is limited by the optical diffraction. The severe spectral overlaps generated by heterogeneity were resolved by multivariate curve resolution (MCR) methods. The purity based method (SIMPLISMAX) was used to resolve both luminescence spectra and pure Raman spectra without prior information. The MCR-alternating least square (ALS) was used as a refined method of both spectra and spectral concentrations. The reconstructing Raman images of the respective spectral contribution supply a versatile potential to characterize the chemistry of atmospheric aerosols at the level of the individual particles.

Air Pollutants↗