Survival to akinetic mutism state in Japanese cases of MM1-type sporadic Creutzfeldt-Jakob disease is similar to Caucasians.

BACKGROUND: It is not known whether the clinical course of Japanese sporadic Creutzfeldt-Jakob disease (sCJD) cases differs from that of Caucasian sCJD cases. PATIENTS AND METHODS: To investigate the clinical course of Japanese sCJD, clinical findings from 29 patients with Japanese MM1-type sCJD were retrospectively evaluated and compared to Caucasian sCJD findings. RESULTS: Survival of Japanese MM1-type sCJD up to the time of akinetic mutism state is similar to that of Caucasian subjects. However, the total disease duration of Japanese patients was approximately three times longer. CONCLUSIONS: The present observations indicate that Japanese sCJD cases generally show a longer disease duration because of the longer survival period after reaching the akinetic mutism state.

Excitation energy transfer between pigment system II units in blue-green algae.

Efficiency in excitation energy transfer from closed to open reaction center II in blue-green and red algae was estimated by the method developed by Joliot and Joliot (C.R. Acad. Sci. (1964) 258, 4622--4625) after slight modification; the number of open reaction centers II was counted from the mean O2 yield of repetitive short flashes. The efficiency in energy transfer in Chlorella pyrenoidosa was the same in our measurement as that reported by Joliot and Joliot (0.55 +/- 0.02). However, the values obtained with four blue-green algae and one red alga were very small, in a range of 0.00--0.07. The low efficiency was always obtained independently of the size of the apparent photosynthetic unit which was varied by growth conditions. Results indicated that pigment system II forms a unit in which only one reaction center II is operative.

Chlorophyll forms and excitation energy transfer pathways in light-harvesting chlorophyll a/b-protein complexes isolated from the siphonous green alga, Bryopsis maxima.

In this study, examination was made of chlorophyll (Chl) forms and energy transfer pathways in light-harvesting Chl a/b-protein complex (LHC II) isolated from the siphonous green alga, Bryopsis maxima. Three major Chl a forms (Ca664, Ca672 and Ca679) and one minor form (Ca688) were resolved at 15 degrees C. Two Chl b forms were resolved at 648 and 653 nm. Based on the number of Chl bound to an apoprotein, two Chls a were assigned to each of the three major Chl a forms, and three and five Chls b, to Cb648 and cb653, respectively. At 15 degrees C, fluorescence spectra were identical, irrespective of the excitation conditions of Chl a, Chl b and siphonaxanthin. Fluorescence from Chl b was detected in addition to that from all Chl a forms. Very efficient energy transfer from siphonaxanthin or Chl b to Chl a and even uphill transfer from Chl a to Chl b, were noted by measurement of the excitation spectra. At 15 degrees C, the equilibrium of energy distribution was established among pigments. However, Chl b was found not to mediate energy transfer from siphonaxanthin to Chl a. The partial amino acid sequence of Bryopsis LHC II was similar to those of green algae and higher plants. The energy transfer pathway between pigments and molecular organization of Bryopsis LHC II were compared with LHC II isolated from spinach.

Estimation of chlorophyll a distribution in the photosynthetic pigment systems I and II of the blue-green alga Anabaena variabilis.

Chlorophyll a distribution in pigment systems I and II was estimated with the blue-green alga Anabaena variabilis by two methods: first, with intact cells using delayed light emission as an index reaction; second, by measuring the 2,6-dichlorophenolindophenol-Hill reaction and the cytochrome c photooxidation in membrane fragments. The first estimation indicated that 0.053+/-0.014 of total chlorophyll a functions as a component of pigment system II, and the second method 0.086+/-0.012. Though the values were somewhat different in the two methods, both estimations indicated that pigment system II chlorophyll a occupies a very small fraction of total chlorophyll a.

Uphill energy transfer in a chlorophyll d-dominating oxygenic photosynthetic prokaryote, Acaryochloris marina.

The steady-state fluorescence properties and uphill energy transfer were analyzed on intact cells of a chlorophyll (Chl) d-dominating photosynthetic prokaryote, Acaryochloris marina. Observed spectra revealed clear differences, depending on the cell pigments that had been sensitized; using these properties, it was possible to assign fluorescence components to specific Chl pigments. At 22 degrees C, the main emission at 724 nm came from photosystem (PS) II Chl d, which was also the source of one additional band at 704 nm. Chl a emissions were observed at 681 nm and 671 nm. This emission pattern essentially matched that observed at -196 degrees C, as the main emission of Chl d was located at 735 nm, and three minor bands were observed at 704 nm, 683 nm, and 667 nm, originating from Chl d, Chl a, and Chl a, respectively. These three minor bands, however, had not been sensitized by carotenoids, suggesting specific localization in PS II. At 22 degrees C, excitation of the red edge of the absorption band (which, at 736 nm, was 20 nm longer than the absorption maximum), resulted in fluorescence bands of Chl d at 724 nm and of Chl a at 682 nm, directly demonstrating an uphill energy transfer in this alga. This transfer is a critical factor for in vivo activity, due to an inversion of energy levels between antenna Chl d and the primary electron donor of Chl a in PS II.

Fluorescence properties of chlorophyll d-dominating prokaryotic alga, acaryochloris marina: studies using time-resolved fluorescence spectroscopy on intact cells

Antenna components and the primary electron donor of the photosystem (PS) II in the Chlorophyll (Chl) d-dominating prokaryote, Acaryochloris marina, were studied using time-resolved fluorescence spectroscopy in the ps time range. By selective excitation of Chl a or Chl d, differences in fluorescence properties were clearly resolved. At physiological temperature, energy transfer was confirmed by a red shift of emission maximum among PS II antenna components, and the equilibrium of energy distribution among Chl a and Chl d was established within 30 ps. A fluorescence component that can be assigned to delayed fluorescence (DF) was observed at 10 ns after the excitation; however, it was not necessarily resolved by the decay kinetics. At -196 degrees C, a red shift of emission maximum was reproduced but the equilibrium of energy distribution was not detected. DF was resolved in the wavelength region corresponding to Chl a by spectra and by decay kinetics. The lifetime of the DF was estimated to be approx. 15 ns, and the peaks were located at 681 and 695 nm, significantly shorter wavelengths than those of Chl d. These findings strongly suggest that an origin of DF is Chl a, and Chl a is most probably the primary electron donor in the PS II reaction center (RC). These results indicate that the constitution of PS II RC in this alga is essentially identical to that of other oxygenic photosynthetic organisms.

Energy transfer processes in Rhodopseudomonas palustris grown under low-light conditions. Heterogeneous composition of LH 2 complexes and parallel energy flow pathways.

Excitation energy flow in the purple photosynthetic bacterium Rhodopseudomonas palustris grown under a low-light intensity was studied by time-resolved fluorescence spectroscopy in the ps time range. This bacterium synthesized the B824 component under this light condition. Time-resolved spectra at 20 degrees C indicated the sequential energy flow in the order of B803, B856, B882 and B900, long wavelength antenna. An emission from B803 was not observed. A remarkable feature was the emission from B824 throughout the measuring time. After the excitation pulse of 100 ps, the spectra did not change any further, indicating the establishment of an equilibrium among components. Based on the energy distribution after equilibrium, parallel energy transfer pathways to LH 1 were suggested; one including B824 integrated in the B803-824-856 complex, and the other, from the B803-856 complex to B882. The latter was the dominant energy flow pathway in this bacterium.

Presence and significance of minor antenna components in the energy transfer sequence of the green photosynthetic bacterium Chloroflexus aurantiacus.

Antenna components in the energy transfer processes of a green photosynthetic bacterium Chloroflexus aurantiacus were spectrally investigated by time-resolved fluorescence spectroscopy at -196 degrees C on intact cells. Besides major antenna components so far reported, three minor components were resolved; those were Bchl c located at 785 nm, the baseplate Bchl a at 819 nm and Bchl a in the B808-866 complex at 910 nm. The last component was assigned to a longer wavelength antenna closely associated with a reaction center. An additional Bchl c fluorescence component was kinetically suggested to be present, which can be an energy donor to a major Bchl c. Presence of these minor components was signified in terms of (1) increase in the spectral overlap integral and (2) adjustment of the direction of dipole moments in the energy transfer sequence of intact cells.

A novel peridinin-chlorophyll a protein (PCP) from the marine dinoflagellate Alexandrium cohorticula: a high pigment content and plural spectral forms of peridinin and chlorophyll a.

A new type of peridinin-chlorophyll a protein (PCP) was isolated from the marine dinoflagellate Alexandrium cohorticula. Unlike previous studies. PCP was obtained as a single component in the presence of a protease inhibitor. The monomer had a molecular mass of 37 kDa with 12 peridinin molecules associated with 2 chl a molecules. This pigment content was much higher than that reported previously. We observed a partial amino acid sequence of the N-terminus that is novel among photosynthetic pigment-protein complexes. Magnetic circular dichroism clearly indicated that chl a in PCP had monomeric features. Multiple spectral components were suggested for both chl a and peridinin. Based on the high pigment content, the optical properties were compared with those for a reported PCP containing 4 chl a and 1 peridinin.

Energy migration in allophycocyanin-B trimer with a linker polypeptide: analysis by the principal multi-component spectral estimation (PMSE) method.

Energy migration processes in allophycocyanin-B trimer with a linker polypeptide were analyzed using the principal multi-component spectral estimation (PMSE) method, which does not require assumption of component number, decay function, or the spectral band shape. We determined the number of spectral components showing independent kinetic behavior by the eigen-value of an auto-correlation matrix, and further the spectra of the components and their rise and decay curves. Two decay components were resolved at 20 degrees C: one corresponded to the decay of one type of beta-84 chromophore, and the other to the decay from the thermally equilibrated state between another type of beta-84 chromophore and the alpha-allophycocyanin B chromophore. An additional slow decay process was resolved at -196 degrees C. We also compared the component spectra obtained using the PMSE method with the decay-associated spectra obtained using the global analysis.

Absorption spectrum of allophycocyanin isolated from Anabaena cylindrica: variation of the absorption spectrum induced by changes of the physico-chemical environment.

The absorption spectrum of allophycocyanin of Anabaena cylindrica was studied. The extinctions of the main absorption bands (650 and 620 nm) varied depending on the protein concentration, ionic strength, and pH. At higher protein concentrations or higher ionic strength, the 650 nm band became stronger and the 620 nm band became weaker. At pH values lower than 6.0, reverse changes occurred in association with protein dissociation into monomer. Similar spectral variation was also induced by sugars and polyols. Glucose, sucrose, or glycerol (1-5 M) induced an increase in the 650 nm band and a decrease in the 620 nm band without causing any changes in protein conformation. Propylene glycol and ethylene glycol showed a reverse effect and caused protein dissociation into monomer. The difference spectra of all spectral changes were identical, consisting of a sharp and strong peak at 650 nm and a broad and weak one in the reverse direction at a wavelength below 620 nm. The spectral variation probably results from shifts of the electronic state of phycocyanobilin. We postulated that a protein field favorable to the state producing the 650 nm band is established around phycocyanobilin when the protein takes a "tight state" through protein association or by the action of sugar in aqueous environment; in a "relaxed state" in the monomer, the state of phycocyanobilin similar to that in phycocyanin becomes dominant.

Crystallization and preliminary X-ray diffraction studies of C-phycocyanin from a red alga, Porphyra tenera.

C-Phycocyanin from a red alga, Porphyra tenera, has been crystallized by the vapor-diffusion procedure. Both orthorhombic and hexagonal forms were obtained from ammonium sulfate solutions, whereas only the orthohombic form was selectively grown from sodium citrate solutions. The orthorhombic crystals are more suitable for further crystallographic work; their space group is P2(1)2(1)2(1), with unit-cell dimensions of a = 105, b = 121, and c = 184 A. The asymmetric unit comprises two (alpha beta)3 trimer molecules of C-phycocyanin. These crystals diffract X-rays up to about 3 A resolution.

[Screening test for hematopoietic stem cells in umbilical cord blood with the SE-9000 automated hematology analyzer].

The immature information IMI channel on an SE-9000 automated hematology analyzer was used for detection of stem cells in cord blood and the results were compared with other standard methods, including flow cytometry analysis and progenitor assays. After the removal of red blood cells, the IMI count in cord blood samples significantly correlated with the number of CD34+ cells (r = 0.810), CFU-GM (r = 0.606) and BFU-E (r = 0.961). The cell count in hematopoietic progenitor cell (HPC) area also showed a correlation with CD34+ cell number (r = 0.722), but to a lesser extent than the IMI count; no significant correlations were observed with the numbers of progenitor cells. Cord blood contained higher fractions of CD34+/CD38- and CD34+/CD117+ cells representative of immature subclasses of progenitor cells. This might explain the difference in the HPC module results for the cord blood and peripheral blood stem cell samples. A higher coefficient of correlation was obtained with leukocyte suspensions than with whole blood samples. These results demonstrated the usefulness of IMI-positive cell measurement as a stem cell screening test for cord blood banking.

A high molecular weight terminal pigment ("anchor polypeptide") and a minor blue polypeptide from phycobilisomes of the cyanobacterium Nostoc sp. (MAC): Isolation and characterization.

A 94 kD pigment-polypeptide, which is presumed to be involved in anchoring the phycobilisomes to the thylakoids, was isolated from Nostoc phycobilisomes by gel filtration in 63 mM formic acid. The isolation condition did not require detergents or denaturating reagents, as in previous procedures, and enzymatic degradation was not observed at the low pH of 2.5. The "anchor polypeptide" thus obtained had absorption (Abs) and fluorescence maxima (Em) at 658 and 673 nm, respectively, in 63 mM formic acid at room temperature. The maxima shifted to longer wavelengths in 100 mM potassium phosphate (pH 6.8), Abs 665 and Em 683 nm at room temperature, and Abs 665 and Em 684 nm at liquid nitrogen temperature. The fluorescence maxima at both temperatures correspond to the longest wavelength component resolved in phycobilisomes from second derivative spectra. A minor blue polypeptide was also found by this isolation method. The molecular weight of this polypeptide was ca. 18,000 and is probably similar to a polypeptide which has been found in the phycobilisome core of other cyanobacteria.

Fluorescence properties of the allenic carotenoid fucoxanthin: Implication for energy transfer in photosynthetic pigment systems.

The fluorescence spectrum of an allenic carotenoid, all-trans-fucoxanthin isolated from a brown alga, has been reported for the first time. This carotenoid is known to function efficiently as a primary photosynthetic antenna pigment in marine algae. The emission bands were located around 630, 685 and 750 nm in CS2 at 20°C, absorption bands being located at 448, 476 and 505 nm. The energy difference between the 0-0 bands of absorption and emission spectra was about 3900 cm(-1) and location of the emission maximum was less sensitive to the polarizability of solvents than that of the absorption maximum. These clearly indicate that the emission originates from the optically forbidden singlet state (2Ag). This is in contrast to other carotenoids whose emission is assigned to 1Bu state, probably due to the symmetric structure of the conjugated double bond responsible for the absorption in the visible region. A rapid internal conversion from 1Bu to 2Ag state might be facilitated by distorted structure of the conjugated double bond of fucoxanthin. The energy level responsible for the emission is almost identical to the Qy level of the acceptor molecule (Chl a), thus we propose an energy transfer pathway from the optically forbidden 2Ag state of the carotenoid to the Qy transition of Chl a in algal pigment systems.

Deconvolution of C-phycocyanin beta-84 and beta-155 chromophore absorption and fluorescence spectra of cyanobacterium Mastigocladus laminosus.

Absorption and fluorescence spectra of the C-phycocyanin beta-subunit were quantitatively deconvoluted into component spectra of the beta-84 and beta-155 chromophores. The deconvolution procedure was based on a theoretical treatment of polarization properties. Four kinds of spectra (absorption, emission, emission polarization, and excitation polarization) measured on C-phycocyanin isolated from the cyanobacterium Mastigocladus laminosus were used as the experimental data set. Without any assumption of spectral shape, the absorption and fluorescence spectra of both chromophores were unambiguously resolved and their fluorescence quantum yields were evaluated. By combining the spectra of the alpha-subunit, independently measured, with the resolved spectra of the beta-subunit, the fluorescence and fluorescence polarization spectra and the fluorescence quantum yield of the monomer were estimated; they agree with experimental values to within an acceptable error. Further, the matrix of energy transfer rates in the monomer was estimated; it gave a significantly different result (by up to 40%) from previously estimated ones.

Allophycocyanin complexes of the phycobilisome from Mastigocladus laminosus. Influence of the linker polypeptide L8.9C on the spectral properties of the phycobiliprotein subunits.

The following phycobiliproteins and complexes of the allophycocyanin core were isolated from phycobilisomes of the thermophilic cyanobacterium Mastigocladus laminosus: alpha AP, beta AP, (alpha AP beta AP), (alpha AP beta AP)3, (alpha AP beta AP)3L8.9C, (alpha APB alpha AP2 beta AP3)L8.9C. The six proteins and complexes were characterised spectroscopically with respect to absorption, oscillator strength, extinction coefficient, fluorescence emission, relative quantum yield, fluorescence emission polarisation and fluorescence excitation polarisation. The interpretation of the spectral data was based on the three-dimensional structure model of (alpha PC beta PC)3 (Schirmer et al. (1985) J. Mol. Biol. 184, 257-277), which is related to the allophycocyanin trimer. The absorption and CD spectra of the complexes (alpha AP beta AP)3, (alpha AP beta AP)3L8.9C and (alpha APB alpha AP2 beta AP3)L8.9C could be deconvoluted into the spectra of the phycobiliprotein subunits. The assumptions made for the deconvolution could be checked by the synthesis of the spectra of (alpha APB beta AP)3. The synthesised spectra are in good agreement with the corresponding measured spectra published by other authors. Considering the deconvoluted spectra the following influences on the chromophores could be ascribed to L8.9C: L8.9C neither influences the alpha AP nor the alpha APB chromophores. L8.9C shifts the absorption maximum of the beta AP chromophore to longer wavelength than the absorption maximum of the alpha AP chromophore in trimeric complexes. L8.9C increases the oszillator strength of the beta AP chromophores to about the value of the alpha AP chromophores in trimeric complexes. L8.9C turns the beta AP chromophores from sensitizing into weak fluorescing chromophores. By means of the hydropathy plot and the predicted secondary structure, a postulated three-fold symmetry in the tertiary structure of L8.9C could be confirmed.

Chlorophyll b expressed in Cyanobacteria functions as a light-harvesting antenna in photosystem I through flexibility of the proteins.

Photosynthetic pigments bind to their specific proteins to form pigment-protein complexes. To investigate the pigment-binding activities of the proteins, chlorophyll b was for introduced the first time to a cyanobacterium that did not synthesize that pigment, and expression of its function in the native pigment-protein complex of cyanobacterium was confirmed by energy transfer. Arabidopsis CAO (chlorophyll a oxygenase) cDNA was introduced into the genome of Synechocystis sp. PCC6803. The transformant cells accumulated chlorophyll b, with the chlorophyll b content being in the range of 1.4 to 10.6% of the total chlorophyll depending on the growth phase. Polyacrylamide gel electrophoresis analysis of the chlorophyll-protein complexes of transformant cells showed that chlorophyll b was incorporated preferentially into the P700-chlorophyll a-protein complex (CP1). Furthermore, chlorophyll b in CP1 transferred light energy to chlorophyll a, indicating a functional transformation. We also found that CP1 of Chlamydomonas reinhardtii, believed to be a chlorophyll a protein, bound chlorophyll b with a chlorophyll b content of approximately 4.4%. On the basis of these results, the evolution of pigment systems in an early stage of cyanobacterial development is discussed in this paper.