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A light-emitting diode light standard for photo- and videomicroscopy.

A light calibration system consisting of a compact light-emitting diode (LED) source with feedback control of intensity is described. The source is positioned in the focal plane of the microscope objective and produces flat-field illumination of up to 31 microW. The source can be easily used to determine the performance of microscope optics and camera response. It can also be used as a standard light source for calibration of experimental systems. Selectable light intensities are produced by controlling the LED input power via a feedback circuit consisting of a photodiode that detects output light intensity. Spectral coverage extends between 550 and 670 nm using green, yellow and red LEDs mounted side by side, which are selected individually. The LED chips are encapsulated in plastic diffusers which homogenize the light, and a flat field of illumination is obtained through a thin 1-mm-diameter aperture positioned directly over each chip. Provision is made for insertion of Ronchi rulings over the aperture to enable measurements of contrast modulation in a uniform field. The light may be pulse-modulated to assess camera response times and the device can be synchronized with video frames. Narrow bandpass interference filters can be placed between the objective lens and the LED source to produce monochromatic light without affecting the spacing of controlled light intensities since emission spectra do not shift appreciably over the range of LED powers chosen in this design. Results of tests using controlled light intensity and uniform illumination are presented.

Calibration↗

Light-induced dopamine release from teleost retinas acts as a light-adaptive signal to the retinal pigment epithelium.

In the retinal pigment epithelium (RPE) of lower vertebrates, melanin pigment granules migrate in and out of the cells' long apical projections in response to changes in light condition. When the RPE is in its normal association with the retina, light onset induces pigment granules to disperse into the apical projections; dark onset induces pigment granules to aggregate into the cell bodies. However, when the RPE is separated from the retina, pigment granule movement in the isolated RPE is insensitive to light onset. It thus seems likely that a signal from the retina communicates light onset to the RPE to initiate pigment dispersion. We have examined the nature of this retina-to-RPE signal in green sunfish, Lepomis cyanellus. In isolated retinas with adherent RPE, light-induced pigment dispersion in the RPE is blocked by treatments known to block Ca2+-dependent transmitter release in the retina. In addition, the medium obtained from incubating previously dark-adapted retinas in the light induces light-adaptive pigment dispersion when added to isolated RPE. In contrast, the medium obtained from incubating dark-adapted retinas in constant darkness does not affect pigment distribution when added to isolated RPE. These results are consistent with the idea that RPE pigment dispersion is triggered by a substance that diffuses from the retina at light onset. The capacity of the conditioned medium from light-incubated retinas to induce pigment dispersion in isolated RPE is inhibited by a D2 dopamine antagonist, but not by D1 or alpha-adrenergic antagonists. Light-induced pigment dispersion in whole RPE-retinas is also blocked by a D2 dopamine antagonist.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Light-evoked changes in [K+]o in proximal portion of light-adapted cat retina.

1. The M-wave is a light-adapted response of proximal retina consisting of phasic negative field potentials at light onset and offset that are spatially tuned for small stimuli. We measured light-dependent changes in extracellular K+ concentration ([K+]o) in proximal retina to investigate the hypothesis that the M-wave originates from Müller cell responses to changes in [K+]o. 2. Extracellular field potentials, and changes in [K+]o evoked in response to circular spots of light flashed on steady backgrounds, were recorded with double-barreled K(+)-sensitive electrodes placed in the retina at different depths. 3. Increases in [K+]o during illumination and at light offset were maximal in proximal retina, with the On [K+]o increase located more proximally than the Off increase. The [K+]o increase during illumination consisted of a phasic and sustained response, whereas the Off [K+]o increase was predominantly phasic. The spatial tuning of the [K+]o increases was similar to the tuning of the field potentials. 4. The Off-field potential was larger than the On potential; it tended to be maximal more distally and was more sharply localized in retinal depth. Stimulus-response characteristics of the field potentials were not altered by intravitreal tetrodotoxin (TTX; 3.8 microM) sufficient to block retinal ganglion cell action potentials. 5. There were no rod contributions to the proximal [K+]o increases and field potentials recorded at the background illuminations used in this study (9.5-11.5 log q.deg-2.s-1). 6. An intravitreal injection of L- or DL-2-amino-4-phosphonobutyric acid (APB; 1 mM) was used to block On-system neuronal responses in proximal retina and isolate Off-system responses. After APB the [K+]o response consisted of a sustained decrease in [K+]o during illumination followed by an overshoot at light offset, while the field potential was a sustained positive response at light onset followed by an initially phasic negative response at light onset followed by an initially phasic negative response at light offset. These responses retained spatial tuning. To isolate the On-system components, the APB-isolated responses were subtracted from the controls. The [K+]o response now consisted of a sustained increase during illumination followed by an undershoot at light offset. The field potential was a sustained negative potential with an initial phasic peak that decayed at Off. Results with kynurenate (KYN; 5 mM) and (+/-)cis-2,3-piperidine (PDA; 5 mM) confirmed the sustained nature of the On component.(ABSTRACT TRUNCATED AT 400 WORDS)

Adaptation, Physiological↗

Evaluation of blue-light hazards from various light sources.

Visible light of short wavelength (blue light) may cause a photochemical injury to the retina, called photoretinitis or blue-light hazard. In this study, various light sources were evaluated for blue-light hazard. These sources include the sun, the arc associated with arc welding and plasma cutting, molten steel, iron and glass, the interior of furnaces, the arc or envelope of discharge lamps, the filament or envelope of incandescent lamps, the envelope of fluorescent lamps and light-emitting diodes. The spectral radiance of each light source was measured, and blue-light effective radiance and the corresponding permissible exposure time per day were calculated in accordance with the ACGIH (American Conference of Governmental Industrial Hygienists) standard. The sun, arc welding, plasma cutting and the arc of discharge lamps were found to have extremely high effective radiances with corresponding permissible exposure times of only 0.6-40 s, suggesting that viewing these light sources is very hazardous to the retina. Other light sources were found to have low effective radiances under the study conditions and would pose no hazard, at least for short exposure times.

Dose-Response Relationship, Radiation↗

The role of extraocular photoreceptors in newt magnetic compass orientation: parallels between light-dependent magnetoreception and polarized light detection in vertebrates.

Theoretical models implicating specialized photoreceptors in the detection of the geomagnetic field have been the impetus for studying the effects of light on magnetic compass orientation. Magnetic orientation in flies, amphibians and birds has been found to be influenced by light, and in all these groups a shift of approximately 90 degrees in the direction of magnetic compass orientation has been observed under certain wavelengths and/or intensities of light. In the eastern red-spotted newt Notophthalmus viridescens, wavelength-dependent effects of light on magnetic compass orientation appear to result from an antagonistic interaction between short-wavelength (< or = 450 nm) and long-wavelength (> or = 500 nm) photoreception mechanisms. We have demonstrated that at least the short-wavelength input to the newt's magnetic compass is mediated by extraocular photoreceptors located in or near the pineal organ, and here we present new findings that indicate that the putative long-wavelength mechanism is also associated with pineal photoreceptors. Interestingly, the amphibian pineal organ mediates orientation to both the e-vector of plane-polarized light and the magnetic field. Although the wavelength-dependence of the polarized light orientation in amphibians has not been studied, polarization sensitivity in fishes appears to be mediated by two antagonistic photoreception mechanisms that have similar spectral characteristics to those of the newts' magnetic compass response. These parallels, along with similarities in the types of receptors that are expected to be involved in light-dependent magnetoreception and polarized light detection, suggest that similar photoreception mechanisms may mediate the light-dependent magnetic and polarized light compasses.

Animals↗

A comparison of polymerization by light-emitting diode and halogen-based light-curing units.

BACKGROUND: Light-emitting diode, or LED, technology provides certain advantages over halogen-based light polymerization of resin-based composites. The authors investigated the adequacy of cure of LED light-curing units, or LCUs. METHODS: The authors used two halogen-based light-curing units (Optilux 400 and 501, Demetron Research Corp., Danbury, Conn.) and two commercially available LED LCUs (LumaCure, LumaLite, Spring Valley, Calif., and VersaLux, Centrix, Shelton, Conn.) to polymerize top surfaces of hybrid (Filtek Z-250, 3M, St. Paul, Minn.) and microfilled (Renamel, Cosmedent, Chicago) resin-based composite specimens. Specimens were indented on their top and bottom surfaces with a Knoop hardness tester and measured for hardness. Bottom:top hardness ratios determined the percentage of cure. The authors separated the data into eight groups (two composites cured with four different lights) with 15 observations per group, for a total sample size of 120. RESULTS: The authors compared composites and curing lights by a two-way analysis of variance, and results indicated significant main effects. The main effect of composite was statistically significant (P < .0001) when microfilled composite was compared with hybrid composite, regardless of curing light, for all top and bottom hardness measurements, with the hybrid producing much higher hardness measurements overall. The main effect of light was significant as well (P < .0001), regardless of composite type, with the two halogen-based lights producing harder top and bottom composite surfaces than the two LED LCUs. CONCLUSIONS: The light output of commercially available diodes for resin-based composite polymerization still requires improvement to rival the adequacy of cure of halogen-based LCUs. Additional studies are necessary. CLINICAL IMPLICATIONS: Commercially available LED LCUs were introduced just in the past year. However, they may not adequately polymerize resin-based composites, which can lead to restoration failures and adverse pulpal responses to unpolymerized monomers.

Analysis of Variance↗

Hypericin-mediated photocytotoxic effect on HT-29 adenocarcinoma cells is reduced by light fractionation with longer dark pause between two unequal light doses.

The present study demonstrates the in vitro effect of hypericin-mediated PDT with fractionated light delivery. Cells were photosensitized with unequal light fractions separated by dark intervals (1 or 6 h). We compared the changes in viability, cell number, survival, apoptosis and cell cycle on HT-29 cells irradiated with a single light dose (12 J/cm(2)) to the fractionated light delivery (1 + 11 J/cm(2)) 24 and 48 h after photodynamic treatment. We found that a fractionated light regime with a longer dark period resulted in a decrease of hypericin cytotoxicity. Both cell number and survival were higher after light sensitization with a 6-h dark interval. DNA fragmentation occurred after a single light-dose application, but in contrast no apoptotic DNA formation was detected with a 6-h dark pause. After fractionation the percentage of cells in the G1 phase of the cell cycle was increased, while the proportion of cells in the G2 phase decreased as compared to a single light-dose application, i.e. both percentage of cells in the G1 and G2 phase of the cell cycle were near control levels. We presume that the longer dark interval after the irradiation of cells by first light dose makes them resistant to the effect of the second illumination. These findings confirm that the light application scheme together with other photodynamic protocol components is crucial for the photocytotoxicity of hypericin.

Adenocarcinoma↗

Effectiveness of composite resin polymerization using light-emitting diodes (LEDs) or halogen-based light-curing units.

The clinical performance of composite resins is greatly influenced by the quality of the light-curing unit used. The aim of this study was to compare the efficiency of a commercial light-emitting diode (LED) with that of a halogen-based light-curing unit by means of dye penetration of a micro hybrid composite resin. The composite resin evaluated was Filtek Z250 (3M Dental). The composite was filled into acrylic moulds that were randomly polymerized for 40 seconds by each of the light-emitting systems: light-emitting diode Ultraled (Dabi Atlante) or halogen light Degulux (Degussa Hülls) curing units. Immediately after polymerization, each specimen was individually immersed in 1 ml of 2% methylene blue solution at 37 degrees C +/- 2 degrees C. After 24 hours, the specimens were rinsed under running distilled water for 1 minute and stored at 37 degrees C +/- 2 degrees C at relative humidity for 24 hours. The composite resins were removed from the moulds and individually triturated before being immersed in new test tubes containing 1 ml of absolute alcohol for 24 hours. The solutions were filtered and centrifuged for 3 minutes at 4,000 rpm and the supernatant was used to determine absorbance in a spectrophotometer at 590 nm. To verify the differences between groups polymerized by LED or halogen light t-test was applied. No significant differences were found between composite resins light-cured by LED or halogen light-curing unit (p > 0.05). The commercially LED-based light-curing unit is as effective to polymerize hybrid composite resins as the halogen-based unit.

Composite Resins↗

Light chain cardiomyopathy. Structural analysis of the light chain tissue deposits.

Cardiomyopathy due to monoclonal light chain deposits is a complication of plasma cell disorders. The deposits may be either fibrillar as in light chain amyloid or nonfibrillar as in light chain deposition disease. The reasons for these structural differences are still unknown. We characterized the myocardial deposits by immunohistochemical examination of sections and extraction and biochemical analysis of the tissue deposits in a patient (MCM) who died of myeloma and systemic light chain deposition disease. Amino acid sequence analysis of the extracted nonfibrillar MCM kappa-light chain reveals that it belongs to the L12a germline subset of the kappa(I) protein and contains five distinctive amino acid substitutions (three in the framework region III and two in the complementarity-determining region III) that have not been reported previously in the same positions in other kappa(I) light chains. The theoretically determined isoelectric point (pI 8.21) of the MCM light chain is high compared with the low isoelectric point of other Bence Jones proteins from subjects without light chain deposition disease. The diffuse binding to basement membranes and the high isoelectric point of the MCM kappa-light chain suggest electrostatic interaction as a possible mechanism of tissue deposition. The spatial locations of the five distinctive residues and a sixth rare substitution of the MCM protein modeled on the backbone structure of REI, a kappa(I)-soluble Bence Jones light chain of known three-dimensional structure, may be responsible for protein destabilization, partial unfolding, and aggregation leading to tissue deposition.

Adult↗

Factors affecting light transmission of single-use, plastic light-curing tips.

Recently, manufacturers introduced presterilized, single-use, plastic light-curing tips to be used either routinely or on patients with known or questionable communicable health concerns. The purpose of this study was to examine the effect of these single-use tips on light transmission compared to conventional fiber-optic bundles in a variety of commercial light-curing units. Also, the effects of surface contact with the plastic tips (human tissues, reflective or opaque media, and barrier films) were evaluated. Where applicable, single-use tips from two sources (Caulk/Dentsply and Demetron) were placed in commercial curing units (Optilux 150 and 500, MAX 100, Spectrum Curing Light, and 3M XL-3000), and the intensity was compared to that of the conventional glass curing tip used with that specific curing unit. Intensity readings were also made for 6 continuous minutes using plastic tips in a high-intensity curing unit to simulate veneer bonding. If the sides of the plastic tip came in contact with the operator's fingers or the patient's tongue and/or cheek during a clinical procedure, a lowering of transmitted light intensity resulted. The glow emitted from the sides of the tip when in use may be annoying to the operator. To prevent this glare, the operator may be tempted to treat the sides of the tip by painting, applying a thin polymer barrier, abrasion, or wrapping in an opaque reflective material (aluminum foil). A significant decrease in light intensity can result if plastic curing tips contact oral tissues or bare hands. Application of thin polymer barriers was found to significantly reduce light transmission value. Also, surface modification (coating with paint or surface scratches) was found to greatly reduce light intensity levels, while wrapping the tip in aluminum foil produced a very small increase. Results indicated that transmitted light intensity with use of plastic tips was dependent upon both the brand of plastic tip tested and the different photocuring units. Either a slight increase or a slight decrease in intensity was noted. Plastic tips did not degrade in transmitted intensity when exposed to the heat produced during a simulated veneering scenario. In summary, use of plastic, single-use light-curing tips can provide adequate intensity for photoactivated restorative techniques; however, the clinician must be aware of specific, clinically relevant limitations with their use. Clinicians must also note that these tips are not designed for re-use.

Analysis of Variance↗

Differential Transcription of Pea Chloroplast Genes during Light-Induced Leaf Development (Continuous Far-Red Light Activates Chloroplast Transcription).

Plastid gene expression was analyzed in pea (Pisum sativum L.) plants grown in darkness, continuous far-red light, and white light. Responses induced by continuous far-red light were most likely mediated by PHYA. Plastid transcription activity was low in dark-grown plants. In contrast, plastids of plants grown in white or far-red light showed a 10-fold increase in transcription activity between 4 and 6 d postimbibition (dpi) and a decrease between 6 and 9 dpi. Plastid RNAs accumulated in illuminated plants from 5 to 7 dpi. In far-red-light-illuminated plants, plastid mRNA levels remained elevated until 14 dpi. In white-light-grown plants, most plastid RNAs decreased in abundance after 7 dpi to very low levels by 14 dpi. This indicates that white light induces a general decrease in plastid RNA stability compared to far-red-light-illuminated seedlings. PsbA mRNA accumulated in older, dark-grown, far-red, and white-light-illuminated seedlings, consistent with this RNA having high stability. Transcription of genes encoding the plastid's transcription and translation apparatus increased relative to rbcL and other genes encoding proteins of the photosynthetic apparatus from 4 to 5 dpi and then declined 10-fold from 5 to 9 dpi. These data document dynamic modulation of plastid gene transcription and mRNA stability during light-induced chloroplast development in pea.

Journal Article↗

Far-red light-induced changes in intracellular potentials of spinach mesophyll cells: interaction with red light.

In green plants, the large bioelectric changes that photosynthetically active light stimulates make it difficult to observe electrical potential changes related to phytochrome photoconversion. As a first step towards distinguishing between photosynthetic and phytochrome effects, we showed that red light enhances far-red stimulated intracellular potential changes in spinach (Spinacia oleracea) leaf mesophyll cells.For a dark-adapted leaf, the response to far-red light increased during the first 10 to 30 exposures of 2.5 minutes, after which it was constant. The intracellular potential depolarized by an average of 0.3 millivolts during each 2.5-minute far-red light period, and returned to the resting value during each subsequent dark period. Continuous supplementary red light (at 1-5% of the fluence rate of the far-red light that stimulated the depolarizations) increased the response to far-red 2- to 3-fold. Supplementary red light did not amplify the response to alternating 702 nanometers light and dark periods. The Emerson enhancement effect thus does not seem to explain amplification of the response to 730 nanometers light by supplementary red light. This does not prove that photosynthetic pigments are not involved in some other way.

Journal Article↗

Biogenesis and light regulation of the major light harvesting chlorophyll-protein of diatoms.

The apoprotein of the major light harvesting pigment-protein complex from the diatom Phaeodactylum tricornutum (UTEX 646) is composed of two similar polypeptides of 17.5 and 18.0 kilodaltons (kD). The in vivo synthesis of these polypeptides is inhibited by the 80s protein synthesis inhibitor cycloheximide, but not by the 70s ribosome inhibitor chloramphenicol. When total poly(A)(+) RNA was used in in vitro protein synthesis, a number of polypeptides were synthesized with a dominant product at 22 kD. When the polypeptides were immunoprecipitated with monospecific antibodies to the 17.5 and 18.0 polypeptides, a single protein zone of 22 kD was detected. Immunoprecipitation with preimmune serum failed to precipitate detectable levels of protein at any relative molecular weight (M(r)). These findings indicate that the two apoprotein polypeptides of the diatom light harvesting pigment-protein are translated from polyadenylated message on cytoplasmic ribosomes as either a single or two (or more) similar M(r) precursor proteins. These findings also suggest that this protein is encoded in the nucleus.Photosynthetic light adaptation features of P. tricornutum UTEX 646 indicate that it responds to low light by increasing cell size and numbers of photosystem I and II reaction centers per cell, but does not change photosynthetic rate per cell or photosynthetic unit sizes significantly. When low light cells are exposed to higher photon flux densities, the in vivo incorporation of label into the apoprotein of the light harvesting complex decreases. In contrast, high light grown cells show rapid (<3 hour) increases in apoprotein synthesis when exposed to low light levels. This is the first demonstration of a specific role of photon flux density in regulating the synthesis of a major light harvesting pigment-protein during photosynthetic light adaptation.

Journal Article↗

Monte Carlo and Lambertian light guide models of the light output from scintillation crystals at megavoltage energies.

A new model of the light output from single-crystal scintillators in megavoltage energy x-ray beams has been developed, based on the concept of a Lambertian light guide model (LLG). This was evaluated in comparison with a Monte Carlo (MC) model of optical photon transport, previously developed and reported in the literature, which was used as a gold standard. The LLG model was developed to enable optimization of scintillator detector design. In both models the dose deposition and light propagation were decoupled, the scintillators were cuboids, split into a series of cells as a function of depth, with Lambertian side and entrance faces, and a specular exit face. The signal in a sensor placed 1 and 1000 mm beyond the exit face was calculated. Cesium iodide (CSI) crystals of 1.5 and 3 mm square cross section and 1, 5, and 10 mm depth were modeled. Both models were also used to determine detector signal and optical gain factor as a function of CsI scintillator thickness, from 2 to 10 mm. Results showed a variation in light output with position of dose deposition of a factor of up to approximately 5, for long, thin scintillators (such as 10 X 1.5 x 1.5 mm3). For short, fat scintillators (such as 1 X 3 X 3 mm3) the light output was more uniform with depth. MC and LLG generally agreed to within 5%. Results for a sensor distance of 1 mm showed an increase in light output the closer the light originates to the exit face, while a distance of 1000 mm showed a decrease in light output the closer the light originates to the exit face. For a sensor distance of 1 mm, the ratio of signal for a 10 mm scintillator to that for a 2 mm scintillator was 1.98, whereas for the 1000 mm distance the ratio was 3.00. The ratio of quantum efficiency (QE) between 10 and 2 mm thicknesses was 4.62. We conclude that these models may be used for detector optimization, with the light guide model suitable for parametric study.

Journal Article↗

Reciprocal expression of the TNF family receptor herpes virus entry mediator and its ligand LIGHT on activated T cells: LIGHT down-regulates its own receptor.

The TNF receptor (TNFR) family plays a central role in the development of the immune response. Here we describe the reciprocal regulation of the recently identified TNFR superfamily member herpes virus entry mediator (HVEM) (TR2) and its ligand LIGHT (TL4) on T cells following activation and the mechanism of this process. T cell activation resulted in down-regulation of HVEM and up-regulation of LIGHT, which were both more pronounced in CD8(+) than CD4(+) T lymphocytes. The analysis of HVEM and LIGHT mRNA showed an increase in the steady state level of both mRNAs following stimulation. LIGHT, which was present in cytoplasm of resting T cells, was induced both in cytoplasm and at the cell surface. For HVEM, activation resulted in cellular redistribution, with its disappearance from cell surface. HVEM down-regulation did not rely on de novo protein synthesis, in contrast to the partial dependence of LIGHT induction. Matrix metalloproteinase inhibitors did not modify HVEM expression, but did enhance LIGHT accumulation at the cell surface. However, HVEM down-regulation was partially blocked by a neutralizing mAb to LIGHT or an HVEM-Fc fusion protein during activation. As a model, we propose that following stimulation, membrane or secreted LIGHT binds to HVEM and induces receptor down-regulation. Degradation or release of LIGHT by matrix metalloproteinases then contributes to the return to baseline levels for both LIGHT and HVEM. These results reveal a self-regulating ligand/receptor system that contributes to T cell activation through the interaction of T cells with each other and probably with other cells of the immune system.

CD4-Positive T-Lymphocytes↗

Isolation of a human plasmin-derived, functionally active, light (B) chain capable of forming with streptokinase an equimolar light (B) chain-streptokinase complex with plasminogen activator activity.

A functionally active human plasmin light (B) chain derivative, stabilized by the streptomyces plasmin inhibitor leupeptin, was isolated from a partially reduced and alkylated enzyme preparation by an affinity chromatography method with a L-lysine-substituted Sepharose column. This light (B) chain derivative was found to be relatively homogeneous by electrophoretic analysis in both an acrylamide gel/dodecyl sulfate system and on cellulose acetate. It possessed approximately 3% of the proteolytic activity (casein substrate) of the original enzyme, and it incorporated 0.09 mol of [3H]diisopropyl phosphorofluoridate per mol of protein. It contained 3.1 +/- 0.3 carboxymethylated cysteines per mol of protein and can be designated as a CmCys5-light (B) chain (CmCys)3. When this isolated light (B) chain derivative was mixed in equal molar amounts with streptokinase, the mixture developed both human and bovine plasminogen activator activities; the bovine activator activity was approximately 66% of the bovine activator activity of the equimolar human plasmin-streptokinase complex. Although this complex now incorporated 0.50 mol of [3H]diisopropyl phosphorofluoridate per mol of protein, its proteolytic activity, on a molar basis, was the same as the proteolytic activity of the isolated light (B) chain derivative. It was shown by electrophoretic analysis in both an acrylamide gel/epsilon-aminocaproic acid system and on cellulose acetate that the light (B) chain derivative and streptokinase forms an equimolar light (B) chain-streptokinase complex, indicating that the binding site for streptokinase is located on the light (B) chain of the enzyme. A functionally active equimolar light (B) chain-streptokinase complex was also isolated from a partially reduced and alkylated equimolar human plasmin-streptokinase complex by the affinity chromatography method. The plasminogen activator activities (human and bovine) of this light (B) chain-streptokinase complex were similar to those of the plasmin-streptokinase complex from which it was derived. Although this complex incorporated 0.70 mol of [3H]diisopropyl phosphorofluoridate per mol of protein, its proteolytic activity, on a molar basis, was only 14% of proteolytic activity of the plasmin-streptokinase complex.

Amino Acids↗

Novel human light chain V kappa segment: serologic and structural analyses of the kappa III-like Bence Jones protein and IgG kappa light chain REE.

Immunochemical and sequence analyses of kappa light chain REE (Bence Jones protein REE and the light chain isolated from IgG kappa myeloma protein REE) revealed antigenic and structural features not previously described for human kappa-chains. Although closely related to proteins of the V kappa III subgroup, light chain REE is readily distinguished from light chains classified serologically as members of the kappa IIIa or kappa IIIb sub-subgroups. Light chains REE (Bence Jones protein REE and light chain REE) are identical in sequence and differ from kappa III proteins by at least 10 uncommon amino acid substitutions in the first three framework regions. Further, kappa-chain REE is unique by virtue of a four-residue deletion in the third complementarity-determining region. The deletion encompasses the three carboxyl-terminal residues in the V kappa-encoded segment and the first residue at the site of V-J recombination. Urine specimens from patient REE also contained a light chain fragment that lacked the first (amino-terminal) 85 residues of the native light chain but otherwise was identical in sequence to the light chain REE. The extensive amino acid differences and unique length of the V kappa segment in light chain REE indicate that this kappa-chain is the product of an unusual V kappa III gene or, alternatively, represents a rarely expressed and novel human V kappa gene.

Amino Acid Sequence↗

Light-induced trimer to monomer transition in the main light-harvesting antenna complex of plants: thermo-optic mechanism.

The main chlorophyll a/b light-harvesting complex of photosystem II, LHCIIb, has earlier been shown to be capable of undergoing light-induced reversible structural changes and chlorophyll a fluorescence quenching in a way resembling those observed in granal thylakoids when exposed to excess light [Barzda, V., et al. (1996) Biochemistry 35, 8981-8985]. The nature and mechanism of this unexpected structural flexibility has not been elucidated. In this work, by using density gradient centrifugation and nondenaturing green gel electrophoresis, as well as absorbance and circular dichroic spectroscopy, we show that light induces a significant degree of monomerization, which is in contrast with the preferentially trimeric organization of the isolated complexes in the dark. Monomerization is accompanied by a reversible release of Mg ions, most likely from the outer loop of the complexes. These data, as well as the built-in thermal and light instability of the trimeric organization, are explained in terms of a simple theoretical model of thermo-optic mechanism, effect of fast thermal transients (local T-jumps) due to dissipated photon energies in the vicinity of the cation binding sites, which lead to thermally assisted elementary structural transitions. Disruption of trimers to monomers by excess light is not confined to isolated trimers and lamellar aggregates of LHCII but occurs in photosystem II-enriched grana membranes, intact thylakoid membranes, and whole plants. As indicated by differences in the quenching capability of trimers and monomers, the appearance of monomers could facilitate the nonphotochemical quenching of the singlet excited state of chlorophyll a. The light-induced formation of monomers may also be important in regulated proteolytic degradation of the complexes. Structural changes driven by thermo-optic mechanisms may therefore provide plants with a novel mechanism for regulation of light harvesting in excess light.

Arabidopsis Proteins↗