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PubMed2006

Performance analysis of LVQ algorithms: a statistical physics approach.

Learning vector quantization (LVQ) constitutes a powerful and intuitive method for adaptive nearest prototype classification. However, original LVQ has been introduced based on heuristics and numerous modifications exist to achieve better convergence and stability. Recently, a mathematical foundation by means of a cost function has been proposed which, as a limiting case, yields a learning rule similar to classical LVQ2.1. It also motivates a modification which shows better stability. However, the exact dynamics as well as the generalization ability of many LVQ algorithms have not been thoroughly investigated so far. Using concepts from statistical physics and the theory of on-line learning, we present a mathematical framework to analyse the performance of different LVQ algorithms in a typical scenario in terms of their dynamics, sensitivity to initial conditions, and generalization ability. Significant differences in the algorithmic stability and generalization ability can be found already for slightly different variants of LVQ. We study five LVQ algorithms in detail: Kohonen's original LVQ1, unsupervised vector quantization (VQ), a mixture of VQ and LVQ, LVQ2.1, and a variant of LVQ which is based on a cost function. Surprisingly, basic LVQ1 shows very good performance in terms of stability, asymptotic generalization ability, and robustness to initializations and model parameters which, in many cases, is superior to recent alternative proposals.

Algorithms↗
PubMed1999

Persistent expression of foreign genes in cultured hepatocytes: expression vectors.

We have optimized a liposome-based transfection method that mediated highly efficient stable expression of foreign genes in hepatocytes. Moreover, we have observed that the metallothionein 1 promoter in the bovine papilloma virus-based expression vector drove the highest expression of foreign genes in hepatocytes as compared with the cytomegalovirus and the human polypeptide chain elongation factor 1alpha (EF-1alpha) promoters in the pcDNA 3-based expression vector. The cytomegalovirus promoter failed to yield significant expression in these cells. Furthermore, expression of foreign genes persisted up to at least 15 passages when expression was under the control of either the EF-1alpha or the metallothionein 1 promoter. Thus, these two promoters led to comparable stability of foreign genes in hepatocytes, with the metallothionein 1 promoter yielding a higher level of expression of foreign genes in these cells.

Animals↗
PubMed1999

Continuous indirect electrochemical regeneration of galactose oxidase.

The development of an indirect anaerobic electrochemical regeneration of galactose oxidase (GOase) allows the prevention of the undesired production of the enzyme inhibitor hydrogen peroxide, which is generated under aerobic regeneration conditions during synthetic applications of GOase. The pH optimum for the electrochemical regeneration of GOase with polyethyleneglycol-modified ferrocene mediators in carbonate buffer is 10.8. Total turnover numbers achieved by either electrochemical or aerobic regeneration of GOase are almost the same. The electrochemical regeneration is half as fast as the aerobic regeneration. It is not necessary to work under anaerobic conditions, because at pH 10.8 the aerobic regeneration of GOase is prevented. The enzyme can be stabilized most effectively by immobilization on an aminopropylated polysiloxane (DELOXAN) via the glutaric dialdehyde procedure with good activity yields up to 37%. Buffers containing amino groups proved to be fatal for long-term GOase stability.

Buffers↗
PubMed2006

Iron(III)-nitro porphyrins: theoretical exploration of a unique class of reactive molecules.

DFT(PW91/TZP) calculations, including full geometry optimizations, have been carried on [FeII(P)(NO2)]-, Fe(III)(P)(NO2), [Fe(II)(P)(NO2)(py)]-, Fe(III)(P)(NO2)(py), [Fe(III)(P)(NO2)2]-, and Fe(III)(P)(NO2)(NO), where P is the unsubstituted porphine dianion, as well as on certain picket fence porphyrin (TPivPP) analogues. The bonding in [Fe(II)(P)(NO2)]- and Fe(III)(P)(NO2), as well as in their pyridine adducts, reveals a sigma-donor interaction of the nitrite HOMO and the Fe dz2 orbital, where the Fe-Nnitro axis is defined as the z direction and the nitrite plane is identified as xz. Both molecules also feature a pi-acceptor interaction of the nitrite LUMO and the Fe dyz orbital, whereas the SOMO of the Fe(III)-nitro complexes may be identified as dxz. The Fe(III)-nitro porphyrins studied all exhibit extremely high adiabatic electron affinities, ranging from about 2.5 eV for Fe(III)(P)(NO2) and Fe(III)(P)(NO2)(py) to about 3.4 eV for their TPivPP analogues. Transition-state optimizations for oxygen-atom transfer from Fe(III)(P)(NO2) and Fe(III)(P)(NO2)(py) to dimethyl sulfide yielded activation energies of 0.45 and 0.77 eV, respectively, which is qualitatively consistent with the observed far greater stability of Fe(III)(TPivPP)(NO2)(py) relative to Fe(III)(TPivPP)(NO2). Addition of NO to yield {FeNO}6 nitro-nitrosyl adducts such as Fe(P)(NO2)(NO) provides another mechanism whereby Fe(III)-nitro porphyrins can relieve their extreme electron affinities. In Fe(P)(NO2)(NO), the bonding involves substantial Fe-NO pi-bonding, but the nitrite acts essentially as a simple sigma-donor, which accounts for the relatively long Fe-Nnitro distance in this molecule.

Journal Article↗
PubMed1992

NK cell activity in treated prostate cancer patients as a probe for circulating tumor cells: hormone regulatory effects in vivo.

Natural killer (NK) cell activity was studied together with tumor marker serotests (PSA, PAP) and blood testosterone, estradiol, cortisol, and prolactin concentrations in treated prostate cancer patients. NK cell activity data were correlated with tumor stage (stage D0 + D1 versus stage D2) and showed statistically insignificant differences. Both tumor progression and stabilization of metastatic disease, triggered by the application of more appropriate therapy in progressive subjects, yielded low NK activity data. By contrast, normal NK activity was found during both partial remission of stage D2 tumor and stabilization of the same disease, after an initial period of tumor remission. Differences between NK activity data from the aforementioned two groups are statistically significant (P less than 0.01). In subjects examined, the application of NK activity assay to those with advanced disease reflected changes in the outcome of the treatment more closely than it did routine tumor marker assessment. The activity of NK cells seems unaffected by changes in basal blood estradiol, cortisol, testosterone, and prolactin concentrations that occur during therapy with pharmacological agents (estradiol, cyproterone acetate, diethylstilbestrol, and flutamide) and during surgical castration. The reported NK activity recordings in treated prostate cancer patients might be indicative of the presence of tumor cells in the circulation. If this holds true, the measurement of NK activity would appear to furnish urological oncology with a new tool for early, rapid recognition of progressive metastatic tumors.

Acid Phosphatase↗
PubMed2004

Fully automated synthesis system of 3'-deoxy-3'-[18F]fluorothymidine.

We developed a new fully automated method for the synthesis of 3'-deoxy-3'-[18F]fluorothymidine ([18F]FLT), by modifying a commercial FDG synthesizer and its disposable fluid pathway. Optimal labeling condition was that 40 mg of precursor in acetonitrile (2 mL) was heated at 150 degrees C for 100 sec, followed by heating at 85 degrees C for 450 sec and hydrolysis with 1 N HCl at 105 degrees C for 300 sec. Using 3.7 GBq of [18F]F- as starting activity, [18F]FLT was obtained with a yield of 50.5 +/- 5.2% (n = 28, decay corrected) within 60.0 +/- 5.4 min including HPLC purification. With 37.0 GBq, we obtained 48.7 +/- 5.6% (n = 10). The [18F]FLT showed the good stability for 6 h. This new automated synthesis procedure combines high and reproducible yields with the benefits of a disposable cassette system.

Automation↗
PubMed1999

Implications of cytosine methylation on (+)-anti-Benzo[a]pyrene 7, 8-dihydrodiol 9,10-epoxide N(2)-dG adduct formation in 5'-d(CGT), 5'-d(CGA), and 5'-d(CGC) sequence contexts of single- and double-stranded oligonucleotides.

Covalent binding of (+)-anti-benzo[a]pyrene 7,8-dihydrodiol 9, 10-epoxide (anti-BPDE) to the N(2)-amino group of deoxyguanine in the oligonucleotides 5'-d(CCTATCGXTATCC) and 5'-d(CCTATm(5)CGXTATCC) (X being T, A, or C) has been studied. The extent of formation of the (+)-trans-anti-BPDE-N(2)-dG adduct in single-stranded 13-mer oligonucleotides with 5'-d(m(5)CGT) and 5'-d(m(5)CGA) sequence contexts was significantly higher (1.5- and 2.4-fold, respectively) relative to that of the nonmethylated sequences. With the 5'-d(CGC) sequence context, m(5)dC had no significant effect on adduct formation. When the reaction was allowed to proceed in the presence of oligonucleotide duplexes (composed of a 13-mer parent strand and a 9-mer complement), a significant increase in the extent of adduct formation was observed with 5'-d(m(5)CGT)/d(CGA) and 5'-d(m(5)CGA)/d(CGT), but not with 5'-d(CGC)/d(GCG), relative to those of the nonmethylated duplexes. Independent of sequence context, no clear effect of m(5)dC on diol epoxide binding to the opposite dG in the complementary strand was observed. The level of diol epoxide binding to the dG target in the 13-mer oligonucleotides is in general higher in single-stranded sequences than in the duplexes. With 5'-d(CGA) and 5'-d(m(5)CGA), for instance, adduct yields were 3- and 4-fold higher, respectively. The thermodynamic stability of the (+)-trans-anti-BPDE-N(2)-dG adduct in the 5'-d(m(5)CGT)-containing duplex (composed of a 13-mer parent strand and a full complement) was substantially higher than in the 5'-d(CGT)/d(GCA) sequence context. The stimulating effect of cytosine methylation on the formation of DNA adducts of anti-BPDE has previously been demonstrated in other experimental systems. The increase in yield could possibly be rationalized in terms of prestacking of the pyrenyl ring system with the nucleobases prior to the nucleophilic addition reaction of the exocyclic amino group. The results from induced circular dichroism studies with the (+)-trans-anti-BPDE-N(2)-dG adduct in the 5'-d(m(5)CGT)-containing duplex are consistent with substantial heterogeneity of adduct conformations, including both external minor groove-localized and intercalated structures.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗
PubMed1996

A comparative study of N.C.A. fluorine-18 labeling of proteins via acylation and photochemical conjugation.

Three methods for 18F-labeling of proteins were evaluated with respect to conjugation yields, suitability for remote-controlled routine synthesis, and in vivo stability of the conjugates-i.e., photochemical conjugation (PCC) using 4-azidophenacyl-[18F]fluoride ([18F]APF) as well as classical conjugation using 4-nitrophenyl 2-[18F]fluoropropionate ([18F]NPFP) and N-succinimidyl 4-[18F]fluorobenzoate ([18F]SFB). For this purpose, [18F]APF was synthesized in one step with a radiochemical yield (RCY) of up to 70% within about 15 min. The 18F-labeling was performed by photogeneration of the corresponding [18F]arylnitrene by irradiating [18F]APF with UV light in presence of the protein in aqueous buffered solution. Using this procedure, human serum albumin (HSA), transferrin, IgG, and avidin were labeled. The [18F]NPFP was synthesized according to a recently published method. Preparation of [18F]SFB was achieved within 35 min with radiochemical yields of 55 +/- 10% by an improved method using O-(N-succinimidyl)-N-N,N',N'-tetramethyluronium tetrafluoroborate (TSTU) as activating reagent. Compared to [18F]APF, protein labeling with [18F]NPFP and [18F]SFB gave rise to considerably higher RCY, of up to 90%. Labeling studies showed that conjugation yields using [18F]NPFP depend on the lysine, tyrosine, and histidine content of the proteins used, whereas conjugation with [18F]APF and [18F]SFB predominantly depends on the Lys content. Owing to competing O-acylation of Tyr residues, [18F]fluoropropionylated HSA was partially unstable under slightly basic conditions. Biodistribution studies with 18F-labeled HSA in NMRI mice revealed the highest in vivo stability for the [18F]SFB conjugate. Based on these results, [18F]SFB seems to be the most suitable 18F-labeling agent for proteins, particularly for the labeling of antibodies.

Acylation↗
PubMed1992

Characterization of acatalasemia detected in two Hungarian sisters.

Acatalasemia was detected in 2 sisters of a Hungarian family. The pedigree of the family showed hypocatalasemia in the children of the patients and in 1 of their brothers, while the other members of the family had normal blood catalase activity. The biochemical characterization (catalase activity, electrophoretic migration, isoelectric point and enzyme stability) of the blood as well as tissue catalase of the acatalasemic patients yielded a catalase form which did not differ from normal.

Acatalasia↗
PubMed2005

Improved yields for baculovirus-mediated expression of human His(6)-PDK1 and His(6)-PKBbeta/Akt2 and characterization of phospho-specific isoforms for design of inhibitors that stabilize inactive conformations.

PDK1 and PKB/Akt have a pleckstrin homology (PH) domain at the C-terminus and N-terminus, respectively, which stabilizes an unphosphorylated, autoinhibited conformation. Binding of the PH domain to a phospholipid second messenger causes relief of autoinhibition, which results in kinase phosphorylation and activation. Baculovirus-mediated expression in Sf9 insect cells of both His(6)-PDK1 and His(6)-PKBbeta/Akt2 were optimized, which significantly improved the yields (5-fold) of the affinity purified enzymes over previously reported values. Isoelectric focusing (IEF) and Western analyses indicated that the apparent V(max)=192+/-13 U/mg and K(m) (PDK-Tide)=55+/-10 microM of purified His(6)-PDK1 results from a mixture of at least three different phospho-specific isoforms (pI values of 6.8, 6.5, and 6.4). A purely unphosphorylated isoform of His(6)-PDK1 (pI=6.8) was generated by treatment with lambda protein phosphatase (lambdaPP), which decreased V(max) to 2.4+/-0.4 U/mg and increased K(m) (PDK-Tide) to 217+/-61 microM. Isoelectric focusing and Western analyses indicated that the apparent V(max)=0.21+/-0.03 U/mg and K(m) (Crosstide)=87+/-30 microM of purified His(6)-PKBbeta/Akt2 results from a mixture of the enzyme monophosphorylated either at Ser-474 ( approximately 90%) or at Thr-309 ( approximately 10%). A purely unphosphorylated isoform of His(6)-PKBbeta/Akt2 (pI=6.4) was generated by treatment with lambdaPP, which decreased V(max) approximately 2-fold. The optimization of high-level production and detailed characterization of purified and lambdaPP-treated His(6)-PDK1 and His(6)-PKBbeta/Akt2 will facilitate detailed structural and kinetic studies aimed at understanding the mechanism of second messenger-induced activation.

3-Phosphoinositide-Dependent Protein Kinases↗
PubMed2000

Synthesis of methyl beta-D-fructoside catalyzed by levansucrase from Rahnella aquatilis.

Methyl beta-D-fructoside(MF) was formed from sucrose and methanol by a transfructosylation reaction using recombinant levansucrase from Rahnella aquatilis. The increase in the yield of MF formation was achieved by increasing methanol concentration. The enzyme stability at higher concentrations of methanol was maintained by lowering the reaction temperature. The optimum temperature and sucrose concentration for MF formation was 10 degrees C and 50 gL(-1) respectively and the yield of MF was 70%.

Journal Article↗
PubMed2006

A test of the transition-metal nanocluster formation and stabilization ability of the most common polymeric stabilizer, poly(vinylpyrrolidone), as well as four other polymeric protectants.

Following an introduction to the nanocluster stabilization literature and DLVO (Derjaugin-Landau-Verwey-Overbeek) theory of colloidal stability, the most common steric stabilizer of transition-metal nanoclusters, poly(vinylpyrrolidone) (PVP), has been examined for its efficacy in the formation, stabilization, and subsequent catalytic activity of prototype, test case Ir(0)n nanoclusters. First, the five criteria established previously for ranking nanocluster protectants for their nanocluster formation and stabilization ability were evaluated for 1 monomer equiv of 10000 average molecular weight (MWav) PVP in the absence, and then presence, of the traditionally weakly coordinating anion BF4- as well as the absence and presence of the strongly coordinating, superior anionic stabilizer P2W15Nb3O62(9-), all in propylene carbonate solvent. It is found that neither 1 equiv of BF4- in propylene carbonate nor 1 monomer equiv of (undried) PVP alone allows for isolable and redissolvable nanoclusters without bulk Ir(0)n metal formation. Careful predrying of the PVP, and by implication other polymers, is shown to be necessary for the formation and stabilization of the nanoclusters. Next, 40 monomer equiv of 10000 MWav PVP and 1 equiv of BF4- in propylene carbonate are shown to allow isolable, redissolvable nanoclusters. Control experiments reveal little difference on nanocluster stabilization by 3500 or 55000 (i.e., vs 10,000) MWav PVP, but yield interesting effects on nanocluster nucleation by the 3500 MWav PVP, as well as by the polymer poly(bis(ethoxy)phosphazene) (PBEP). Four other key polymers reported in the literature to be nanocluster stabilizers are tested by the five criteria method for their efficacy in the formation and stabilization of Ir0n nanoclusters (now in acetone due to the polymers' solubility) and in comparison to each other, specifically, poly(methyl methacrylate) (PMMA), poly(styrene) (PS), poly(methylhydrosilane) (PMHS), and PBEP. Only 40 monomer equiv dried PMMA allows isolable and redissolvable nanoclusters in acetone. Control/reference point experiments show that the electrostatic stabilizer P2W15Nb3O62(9-) is superior to each of the five polymeric stabilizers studied herein in both acetone and propylene carbonate, at least for the test case of Ir(0)n nanoclusters. Further controls show that 40 monomer equiv of PVP added to P2W15Nb3O(62)9--stabilized nanoclusters has no discernible effect on the five criteria other than to reduce by approximately 50% the nanocluster catalytic activity and total catalytic lifetime for cyclohexene hydrogenation. The main finding of this work is that DLVO theory as applied to nanocluster stabilization is fully supported; that is, surface-bound anions in high dielectric constant solvents provide superior stabilization. The importance of even traditionally weakly coordinating anions such as BF4- in nanocluster stabilization is a second, important finding of this work. The fact that HPO4(2-) has been shown to be a simple, cheap, commercially available, thermally robust, and 31P-NMR-handle-containing analogue of the more esoteric P2W15Nb3O62(9-) stabilizer is also discussed in the 14 total Conclusions from this first study ranking polymeric stabilizers of modern transition-metal nanoclusters.

Journal Article↗
PubMed1995

Proteolysis of fusion proteins: stabilization and destabilization of staphylococcal protein A and Escherichia coli beta-galactosidase.

The product yield of staphylococcal Protein A reached only 1.8% of the cell dry weight, while the corresponding value was 14% for a fusion protein composed of Protein A and Escherichia coli beta-galactosidase [1], when produced in the same E. coli host strain, with the same promoter and under identical process conditions. Measurement of the stability of Protein A in vivo showed that it was quickly degraded in the cell with a half-life of 30 min when the protein was expressed alone, but after fusion to beta-galactosidase, the Protein A part became considerably stabilized. In spite of the fast intracellular proteolysis of Protein A, few degradation products could be identified on Coomassie Brilliant Blue-stained SDS/PAGE gels after IgG purification, indicating an even faster degradation of the Protein A fragments. Such degradation products, however, accumulated during incubation of the disintegrated cells. Intracellular degradation intermediates could be demonstrated with the more sensitive Western-blot technique. This technique also revealed that a slow degradation took place not only in the Protein A moiety of the fusion protein, but also in the beta-galactosidase moiety. A control with native beta-galactosidase also showed a weak in vivo proteolysis of this molecule, but it was more stable in free form than in the fused form. This means that the proteolytically very sensitive Protein A was stabilized by fusion with beta-galactosidase, but the originally rather stable beta-galactosidase became slightly more susceptible to proteolysis after the fusion.

Amino Acid Sequence↗
PubMed2003

Dynamics of the anaerobic process: effects of volatile fatty acids.

A complex and fast dynamic response of the anaerobic biogas system was observed when the system was subjected to pulses of volatile fatty acids (VFAs). It was shown that a pulse of specific VFAs into a well-functioning continuous stirred tank reactor (CSTR) system operating on cow manure affected both CH(4) yield, pH, and gas production and that a unique reaction pattern was seen for the higher VFAs as a result of these pulses. In this study, two thermophilic laboratory reactors were equipped with a novel VFA-sensor for monitoring specific VFAs online. Pulses of VFAs were shown to have a positive effect on process yield and the levels of all VFA were shown to stabilize at a lower level after the biomass had been subjected to several pulses. The response to pulses of propionate or acetate was different from the response to butyrate, iso-butyrate, valerate, or iso-valerate. High concentrations of propionate affected the degradation of all VFAs, while a pulse of acetate affected primarily the degradation of iso-valerate or 2-methylbutyrate. Pulses of n-butyrate, iso-butyrate, and iso-valerate yielded only acetate, while degradation of n-valerate gave both propionate and acetate. Product sensitivity or inhibition was shown for the degradation of all VFAs tested. Based on the results, it was concluded that measurements of all specific VFAs are important for control purposes and increase and decrease in a specific VFA should always be evaluated in close relationship to the conversion of other VFAs and the history of the reactor process. It should be pointed out that the observed dynamics of VFA responses were based on hourly measurements, meaning that the response duration was much lower than the hydraulic retention time, which exceeds several days in anaerobic CSTR systems.

Acetates↗
PubMedDate not supplied

[Exchange plasmapheresis in combined treatment of severe uveitis].

Exchange plasmapheresis (EP) has been employed in multiple-modality therapy of 15 patients suffering from severe uveitis. The traditional therapy has yielded but a short-term effect and failed to stabilize the process in these patients. EP has resulted in improvement in all the patients: the injection has reduced, as well as the number of precipitates and oracities in the vitreous body; hemorrhages in the fundus oculi have resolved, and the vision acuity has considerably improved in 11 of the 15 patients; no cases of the deterioration of the vision acuity have been recorded. The relapses have ceased in 10 patients. EP has been well tolerated and yielded no complications; it has normalized the levels of the blood serum immunoglobulins and circulating immune complexes and activized phagocytosis.

Adult↗
PubMed2005

Conversion analysis for mutation detection in MLH1 and MSH2 in patients with colorectal cancer.

CONTEXT: The accurate identification and interpretation of germline mutations in mismatch repair genes in colorectal cancer cases is critical for clinical management. Current data suggest that mismatch repair mutations are highly heterogeneous and that many mutations are not detected when conventional DNA sequencing alone is used. OBJECTIVE: To evaluate the potential of conversion analysis compared with DNA sequencing alone to detect heterogeneous germline mutations in MLH1, MSH2, and MSH6 in colorectal cancer patients. DESIGN, SETTING, AND PARTICIPANTS: Multicenter study with patients who participate in the Colon Cancer Family Registry. Mutation analyses were performed in participant samples determined to have a high probability of carrying mismatch repair germline mutations. Samples from a total of 64 hereditary nonpolyposis colorectal cancer cases, 8 hereditary nonpolyposis colorectal cancer-like cases, and 17 cases diagnosed prior to age 50 years were analyzed from June 2002 to June 2003. MAIN OUTCOME MEASURES: Classification of family members as carriers or noncarriers of germline mutations in MLH1, MSH2, or MSH6; mutation data from conversion analysis compared with genomic DNA sequencing. RESULTS: Genomic DNA sequencing identified 28 likely deleterious exon mutations, 4 in-frame deletion mutations, 16 missense changes, and 22 putative splice site mutations. Conversion analysis identified all mutations detected by genomic DNA sequencing--plus an additional exon mutation, 12 large genomic deletions, and 1 exon duplication mutation--yielding an increase of 33% (14/42) in diagnostic yield of deleterious mutations. Conversion analysis also showed that 4 of 16 missense changes resulted in exon skipping in transcripts and that 17 of 22 putative splice site mutations affected splicing or mRNA transcript stability. Conversion analysis provided an increase of 56% (35/63) in the diagnostic yield of genetic testing compared with genomic DNA sequencing alone. CONCLUSIONS: The data confirm the heterogeneity of mismatch repair mutations and reveal that many mutations in colorectal cancer cases would be missed using conventional genomic DNA sequencing alone. Conversion analysis substantially increases the diagnostic yield of genetic testing for mismatch repair mutations in patients diagnosed as having colorectal cancer.

Adaptor Proteins, Signal Transducing↗
PubMed1994

Characterization of a D1-D2-cyt b-559 complex containing 4 chlorophyll a/2 pheophytin a isolated with the use of MgSO4.

A D1-D2-cyt b-559 complex containing 4 chlorophyll alpha, 1 beta-carotene and 1 cytochrome b-559 per 2 pheophytin a has been isolated from spinach with 30% yield using a Q-Sepharose Fast-Flow anion-exchange column equilibrated with 0.1% Triton X-100, 10 mM MgSO4 and 50 mM Tris-HCl (pH 7.2). The preparation was then stabilized with 0.1% dodecyl-beta-D-maltoside. This method gave a yield 10 times higher than that using a Fractogel TSK-DEAE 650(S) column equilibrated with 0.1% Triton X-100, 30 mM NaCl and 50 mM Tris-HCl (pH 7.2). The PS II RC complex was characterized using absorption and fluorescence spectroscopy at 277 and 77 K. A selective reversible bleaching under reducing conditions with maximum at 682 nm, associated with pheophytin a reduction, and light-induced absorption differences with a lifetime of 1.0 ms, ascribed to the triplet state of P680 were measured and indicated that the isolated D1-D2-cyt b-559 complex is active in charge separation. The results are compared with the data obtained for a PS II RC preparation containing 6 chlorophyll alpha, 2 beta-carotene and 1 cyt b-559 per 2 pheophytin a.

Chlorophyll↗
PubMed2003

Temporal stability of high-frequency brain oscillations in the human EEG.

Temporal stability of a given measurement within individual participants is a desirable property of psychophysiological measures. The present study aims to examine the reliability of high-frequency oscillatory activity in the human electroencephalogram (EEG) across 4 sessions. Convolution of the EEG time series with Morlet wavelets yielded time-frequency representations of the signal for each session. Stability of both topography and time course of gamma-band activity (GBA) was determined for two participants performing a feature-based selective attention task in four separate sessions, spaced at weekly intervals. We found high temporal stability of non-phase-locked GBA typically occurring in time ranges between 200 and 500 ms following presentation of a stimulus, both in terms of topography and time course. Early phase-locked GBA (80-120 ms) showed higher variability with respect to topography, but was consistent in terms of time course. We conclude that measures of high-frequency oscillatory activity as used in the cognitive neurosciences meet stability requirements necessary for meaningful interpretation of this parameter of brain function.

Adult↗