Water-soluble chlorophyll-binding proteins from Brassica oleracea allow for stable photobiocatalytic oxidation of cellulose by a lytic polysaccharide monooxygenase.

BACKGROUND: Lytic polysaccharide monooxygenases (LPMOs) are indispensable redox enzymes used in industry for the saccharification of plant biomass. LPMO-driven cellulose oxidation can be enhanced considerably through photobiocatalysis using chlorophyll derivatives and light. Water soluble chlorophyll binding proteins (WSCPs) make it is possible to stabilize and solubilize chlorophyll in aqueous solution, allowing for in vitro studies on photostability and ROS production. Here we aim to apply WSCP-Chl a as a photosensitizing complex for photobiocatalysis with the LPMO, TtAA9. RESULTS: We have in this study demonstrated how WSCP reconstituted with chlorophyll a (WSCP-Chl a) can create a stable photosensitizing complex which produces controlled amounts of H2O2 in the presence of ascorbic acid and light. WSCP-Chl a is highly reactive and allows for tightly controlled formation of H2O2 by regulating light intensity. TtAA9 together with WSCP-Chl a shows increased cellulose oxidation under low light conditions, and the WSCP-Chl a complex remains stable after 24 h of light exposure. Additionally, the WSCP-Chl a complex demonstrates stability over a range of temperatures and pH conditions relevant for enzyme activity in industrial settings. CONCLUSION: With WSCP-Chl a as the photosensitizer, the need to replenish Chl is greatly reduced, enhancing the catalytic lifetime of light-driven LPMOs and increasing the efficiency of cellulose depolymerization. WSCP-Chl a allows for stable photobiocatalysis providing a sustainable solution for biomass processing.

Current Espghan Guidelines for Celiac Disease in Pediatric Age, Tertiary Care Center Experience: A Proposal for Further Simplification.

According to the 2012 ESPGHAN criteria for diagnosis of celiac disease (CD), duodenal biopsy (DB) can be avoided in children with a clear malabsorption syndrome, anti-tissue transglutaminase IgA (tTG2) ≥ 10x the cut-off, anti-endomysium IgA (EMA) and HLA DQ2/DQ8 genes. The aim of this study is to report our experience and evaluate the accuracy of the actual guidelines. PATIENTS AND METHODS: This is a retrospective study conducted on all patients diagnosed CD from 2012 to 2018 in our Center. For all patients enrolled were analyzed: data of family history, symptoms, serology, genetics, Marsh grade and follow-up. RESULTS: A total of 481 children [mean age 6,4 yrs; F:M= 1.8:1] were included in the study. The mean age of patients who were not subject to DB was lower (4.51 yrs) comparing with patients that received DB (6.48 yrs). Out of the 256 patients with anti-tTG2 ≥ 10 fold, 121 underwent DB because of mild symptoms (84/121) or no symptoms (37/121). In all cases Marsh type 3 was found and HLA haplotypes was compatible with CD diagnosis. CONCLUSIONS: Our study confirms that the serology has a primary importance to diagnose CD, regardless of the symptoms. These data suggest that biopsy and HLA haplotypes search, in presence of anti-tTG2 IgA ≥ 10x the cut-off, are wasteful and unhelpful for the patients.

Nanophotonics of higher-plant photosynthetic membranes.

The thylakoid membrane inside chloroplasts hosts the light-dependent reactions of photosynthesis. Its embedded protein complexes are responsible for light harvesting, excitation energy transfer, charge separation, and transport. In higher plants, when the illumination conditions vary, the membrane adapts its composition and nanoscale morphology, which is characterized by appressed and non-appressed regions known as grana and stroma lamellae, respectively. Here we investigate the nanophotonic regime of light propagation in chloroplasts of higher plants and identify novel mechanisms in the optical response of the thylakoid membrane. Our results indicate that the relative contributions of light scattering and absorption to the overall optical response of grana strongly depend on the concentration of the light-harvesting complexes. For the pigment concentrations typically found in chloroplasts, the two mechanisms have comparable strengths, and their relative value can be tuned by variations in the protein composition or in the granal diameter. Furthermore, we find that collective modes in ensembles of grana significantly increase light absorption at selected wavelengths, even in the presence of moderate biological disorder. Small variations in the granal separation or a large disorder can dismantle this collective response. We propose that chloroplasts use this mechanism as a strategy against dangerously high illumination conditions, triggering a transition to low-absorbing states. We conclude that the morphological separation of the thylakoid membrane in higher plants supports strong nanophotonic effects, which may be used by chloroplasts to regulate light absorption. This adaptive self-organization capability is of interest as a model for novel bioinspired optical materials for artificial photosynthesis, imaging, and sensing.

Thermal unfolding and refolding of a lytic polysaccharide monooxygenase from Thermoascus aurantiacus.

Lytic polysaccharide monooxygenases (LPMOs) are copper-containing enzymes which promote the degradation of recalcitrant polysaccharides like cellulose or chitin. Here, we have investigated the thermostability of an LPMO from Thermoascus aurantiacus (TaLPMO9A). TaLPMO9A was found to retain most of its initial activity after incubating at 100 °C while its apparent melting temperature (T m) is 69 °C at neutral pH. Interestingly, our studies show that holoTaLPMO9A, apoTaLPMO9A and deglycosylated TaLPMO9A can fold back to their original conformation upon lowering the temperature. In the presence of ß-mercaptoethanol the protein does not refold. Activity of TaLPMO9A and refolded TaLPMO9A was studied by an Amplex® Red assay as well as by TaLPMO9A catalysed oxidation of phosphoric acid swollen cellulose (PASC). These studies confirm the functional regain of TaLPMO9A activity upon going through one cycle of unfolding and refolding. The thermal unfolding and refolding of TaLPMO9A was measured spectroscopically. Utilizing the two-state model, detailed thermodynamic parameters were obtained for holoTaLPMO. Furthermore, we have investigated the kinetics of TaLPMO9A unfolding and refolding. Our results have implications in understanding LPMO stability, which is crucial for the efficient application of LPMOs as biocatalysts during biomass degradation.

The influence of dual-task conditions on movement in young adults with and without Down syndrome.

This investigation compared spatial and temporal movement parameters of a sample of young adults with Down syndrome (DS) (N=12) and individuals without disabilities (IWD) (N=12) under dual-task conditions. Subjects performed a walking task at a preferred speed in isolation and again while holding a plate and cup, carrying tray and cups, talking on a phone, or buttoning a shirt. Spatial and temporal values were compared using a 2 (group) × 5 (conditions) repeated measures analysis of variance. Analysis of spatial components separately indicated that step length, step width, stride length and stride width revealed significant group and condition interactions (p ≤.01). Temporal components yielded significance in velocity and single-leg support time (p ≤.01). The current results support the notion that along with impairments to qualitative motor skills, individuals with DS are also impaired in higher order executive functioning (EF), as measured by a dual-task paradigm. It was concluded that movements are less efficient and functional in individuals with DS when an additional task is encountered while walking. We theorized that the motor program was sufficient for general locomotion but was not sufficiently developed to allow individuals with DS to modify or alter their movements to changing cognitive conditions that increasingly taxed EF. As gait and balance are trainable in this population, we recommend developing appropriate exercise and motor skill interventions during childhood and adolescents to increase strength, stability, and more "robust" ambulatory motor schema.

Spatial and temporal variability of movement parameters in individuals with Down syndrome.

This investigation compared spatial and temporal gait movement parameters of a sample of individuals with Down syndrome (n=12) and one of individuals without disabilities (n=12). All participants were evaluated on responses to a preferred pace and fast walk with the GAITRite Electronic Walkway. Spatial outcomes included step and stride length, step and stride width, toe-in/toe-out, and base of support. Temporal outcomes included step time, velocity, single and double leg support time, stance, and swing time. There were significant group differences for step length, step width, stride length, and velocity in the preferred walk condition. Significant group differences for step length, step width, and stride length were observed in the fast walk condition. Percentage differences also indicated lower scores for all spatial and temporal variables in relation to the control group. The ability to control gait movements appears to reflect earlier movement experiences, so it may be possible to use variable sensory feedback and specific training to modify and adjust movement responses and improve gait performance in Down syndrome.

The number of pregnancies is a risk factor for Alzheimer's disease.

Epidemiological data show a higher prevalence of late-onset Alzheimer's disease (AD) in women. The estrogenic deficiency in the post-menopausal period is suspected to be the cause of the gender-related risk of the disease, but studies on the estrogenic therapy and occurrence of AD were not consistent and sometimes contradicting. The aim of this study is to investigate whether a higher exposure to endogenous estrogens is associated with lower risk of dementia or not. Two hundred and four AD patients and 201 control women were considered. By interviews, we evaluated different variables, indirectly correlated to estrogenic natural exposure, as well as educational level and head trauma. These data were correlated in the AD group with the disease progression, as well as with the age at onset. Unexpectedly, we found a significant higher number of pregnancies in the AD than in the control group. Within the AD cases, the number of lifetime pregnancies is related to an earlier onset of the disease. As previously reported, we confirmed that the educational level is a protective factor and that major head trauma represents a risk factor in developing AD. The higher number of pregnancies and a less frequency of nulliparous women, indirectly relate the AD group to a higher estro-progestinic exposure. These findings suggest that it is the increase of progesterone or estrogens level--and not the estrogens decrease, as previously indicated by other authors--that could play a role in the Alzheimer's pathology.

Angle- and velocity-specific alterations in torque and semg activity of the quadriceps and hamstrings during isokinetic extension-flexion movements.

The purpose of this study was to investigate the effect of movement velocity (100 degrees, 200 degrees , 300 degrees s(-1), and 400 degrees s(-1)) and joint position (0 degrees - 15 degrees [L0], 25 degrees - 40 degees [L25], 55 degrees - 70 degrees [L55], and 75 degrees - 90 degrees [L75]) on peak torque (PT) parameters and surface electromyography (SEMG) of the knee-joint muscles during reciprocal isokinetic extension and flexion movements. Thirteen subjects (age = 22.7 +/- 2.1 years, mean height = 161.1 +/- 6.6 cm, mean weight = 63.5 +/- 5.8 kg) participated in the study. Bipolar surface electrodes were placed over the vastus medialis, vastus lateralis, biceps femoris, and medial hamstrings for determination of the root mean square (SEMGrms) and median frequency (SEMGmf) of the SEMG. Peak torque, angle of peak torque (PTang), percentage of peak torque (PTper), SEMGrms, and SEMGmf were analyzed using separate repeated measures analysis of variance (ANOVA). The following main results, significant at p < or = 0.05 or better, were found: The PTang was influenced by movement velocity (in extension there was a decrease in PTang moving from 300 degrees x s(-1) to 400 degrees x s(-1) and inflexion there was an increase in PTang moving from 300 degrees x s(-1) to 400 degrees x s(-1)). Secondly, a greater percentage of peak torque (PTper) was maintained during knee flexion than knee extension. And thirdly, both the quadriceps and hamstrings exhibited changing amplitudes and spectral frequencies based on joint position and movement velocity. There was a trend of decreasing SEMGrms for the quadriceps as the knee moved into extension, and a lower SEMGmf during early (L75) and end stages of knee extension (L0). For the hamstrings, SEMGrms was lowest at the more shortened position (L75) and highest near the mid-position (L25); the lowest SEMGmf occurred at the more lengthened position (L0) and the highest occurred at the more shortened position (L75). Finally, velocity influenced hamstrings and quadriceps muscle amplitude such that SEMGrms was highest at the slower velocities and lowest at the higher velocities. Velocity had no impact on quadriceps spectral properties (p > 0.05), but had a cyclic effect on hamstrings spectral properties. Changes in amplitude and frequency spectrum in tested muscles could be explained, in part, by neural drive to these muscles. Data support the hypothesis of lower activation levels of the quadriceps muscle in the extended position espoused by several authors as a way to protect the knee-joint in the knee-extended position.

Coactivation patterns of the medial and lateral hamstrings based on joint position and movement velocity during isokinetic movements.

The purpose of this investigation was to determine the effect of movement velocity (100 degrees x s(-1), 200 degrees x s(-1), 300 degees x s(-1), and 400 degrees x s(-1)) and joint position (0 degrees - 20 degrees [L0] 30 degrees - 50 degrees [L30], and 70 degrees - 90 degrees [L70] knee flexion) on reciprocal coactivation patterns of the medial and lateral hamstrings as determined by the amplitude and frequency spectrum of surface electromyography (SEMG). Thirteen female subjects (age = 22.7 +/- 2.1 years, mean height = 161.1 +/- 6.6 cm, mean weight = 63.5 +/- 5.8 kg) participated in the study. Bipolar surface electrodes were placed over the biceps femoris (BF) and medial hamstrings (MH) for determination of the root mean square (SEMGrms) and median frequency (SEMGmf) of the SEMG. Normalized SEMGrms values for the MH and BF were determined as a percentage of agonist SEMGrms activity for the same muscle during its agonist phase. Data were analyzed using separate 2 x 3 x 4 (muscle x position x angular velocity) repeated measures analysis of variance (ANOVA). For SEMGrms, there were significant muscle (p < 0.01) and position (p < or = 0.0001) main effects. Post-hoc analyses indicated the BF displayed greater muscle amplitude than the MH and that there was greater muscle amplitude at the L0 position (as the knee approached terminal extension). No velocity effect was noted (p > 0.05). For SEMGmf there were muscle x position (p < or = 0.05) and muscle x position x velocity (p < or = 0.01) interaction effects. Post-hoc analyses indicated the BF displayed a higher frequency spectrum than the MH at the L0 position. Secondly, velocity affected the BF and MH frequency spectrum such that values for both the MH and BF were lowest at 200 degrees x s(-1) and highest at 300 degrees x s(-1) (BF) and 400 degrees x s(-1) (MH). Velocity had little impact on the frequency spectrum in the midrange of the ROM (L30 position). Higher SEMGrms and SEMGmf values for the BF could be explained by the locking or screw home mechanism of the knee, and a way in which the human motor control system provides the limb with a dynamic braking system to control both extension and lateral rotational forces during the final stage of knee extension. It would appear that the way in which the body performs this function is not only to increase the amplitude of BF muscle firing but also to shift toward the recruitment of more fast-twitch motor units.

Excitation energy transfer pathways in Lhca4.

EET in reconstituted Lhca4, a peripheral light-harvesting complex from Photosystem I of Arabidopsis thaliana, containing 10 chlorophylls and 2 carotenoids, was studied at room temperature by femtosecond transient absorption spectroscopy. Two spectral forms of Lut were observed in the sites L1 and L2, characterized by significantly different interactions with nearby chlorophyll a molecules. A favorable interpretation of these differences is that the efficiency of EET to Chls is about two times lower from the "blue" Lut in the site L1 than from the "red" Lut in the site L2 due to fast IC in the former case. A major part of the energy absorbed by the "red" Lut, approximately 60%-70%, is transferred to Chls on a sub-100-fs timescale from the state S(2) but, in addition, minor EET from the hot S(1) state within 400-500 fs is also observed. EET from the S(1) state to chlorophylls occurs also within 2-3 ps and is ascribed to Vio and/or "blue" Lut. EET from Chl b to Chl a is biphasic and characterized by time constants of approximately 300 fs and 3.0 ps. These rates are ascribed to EET from Chl b spectral forms absorbing at approximately 644 nm and approximately 650 nm, respectively. About 25% of the excited Chls a decays very fast-within approximately 15 ps. This decay is proposed to be related to the presence of the interacting Chls A5 and B5 located next to the carotenoid in the site L2 and may imply some photoprotective role for Lhca4 in the photosystem I super-complex.

Knee muscular response strategies differ by developmental level but not gender during jump landing.

The purpose of this investigation was to determine differences between pre- and post-pubescent males and females in quadriceps (vastus medialis; VM) and hamstrings (medial hamstrings and biceps femoris; HAMS) muscular activation patterns via the root mean square of surface electromyography (SEMG) during self-initiated vertical jump landing. Fifty-eight subjects, divided into age and gender groupings, were compared on kinematic variables during pre-landing (100 msec preceeding initial ground contact), post-landing (100 msec following initial ground contact), and initial-contact-to-maximum-knee-flexion stages. Kinematic variables investigated were (1) SEMG values during each stage of the vertical-jump landing; (2) Co-contraction ratios (CCR), which represented the ratio of normalized hamstrings' activity to normalized quadriceps' activity; and, (3) knee angle at initial contact. Results indicated (1) no significant gender differences in variables measured; and, (2) significant developmental level differences. Post-pubescent subjects displayed greater HAMS acitivity and CCR values in the pre-landing stage relative to post-landing stages, indicating that post-pubescent subjects had a greater level of hamstrings co-contraction prior to landing than pre-pubescent subjects. Conversely, pre-pubescent subjects displayed greater post-landing and initial-contact-to-maximum-knee-flexion ratios, indicating a greater level of hamstrings' co-contraction during post-landing stages than post-pubescent subjects. There were no significant differences in knee angle at initial contact. The greater level of hamstrings' co-activation prior to landing by post-pubescent subjects indicated that they used a strategy of pre-tuning the hamstrings prior to landing (more CNS pre-activation) to control the ground reaction forces and anterior tibial displacement experienced by the knee during landing. On the other hand, pre-pubescent subjects controlled these forces by having a greater level of hamstrings' co-activation during landing, which represents more of a reflexive activation in response to ground impact.

Rejection properties of stochastic-resonance-based detectors of weak harmonic signals.

In [Phys. Rev. E 57, 6470 (1998)]] a thorough characterization in terms of receiver operating characteristics of stochastic-resonance detectors of weak harmonic signals of known frequency in additive Gaussian noise was given. It was shown that strobed sign-counting based strategies can be used to achieve a nice trade-off between performance and cost, by comparison with noncoherent correlators. Here we discuss the more realistic case where besides the sought signal (whose frequency is assumed known) further unwanted spectrally nearby signals with comparable amplitude are present. Rejection properties are discussed in terms of suitably defined false-alarm and false-dismissal probabilities for various values of interfering signal(s) strength and spectral separation.

The effect of movement velocity and movement pattern on the reciprocal co-activation of the hamstrings.

The effect of velocity and movement pattern (reciprocal vs. non-reciprocal) on the reciprocal co-activation of the hamstrings was investigated through analysis of the root mean square (RMS) and the median frequency (MDF) of surface electromyography (SEMG). Fourteen subjects performed six continuous repetitions of a reciprocal isokinetic movement pattern (maximal extension followed by maximal flexion), and six continuous repetitions of a non-reciprocal movement pattern (maximal extension only) at 100 degrees, 200 degrees, 300 degrees s-1, and 400 degrees s-1. Data were analyzed using separate 2 x 4 (movement pattern x angular velocity) repeated measures analysis of variance (ANOVA). No significant differences (p > 0.05) were noted between reciprocal and non-reciprocal movement patterns for RMS. However, results did reveal a velocity effect for RMS (F = 5.0, p < 0.01), with significant differences observed between 100 degrees s-1 and 400 degrees s-1 (F = 9.4, p < 0.01), 200 degrees s-1 and 400 degrees s-1 (F = 9.5, p < 0.01), and 300 degrees s-1 and 400 degrees s-1 (F = 11.0, p < 0.001), with RMS values at 400 degrees s-1 being the highest. There was also a velocity effect for MDF (F = 8.03, p < 0.001), with significant differences observed between 100 degrees s-1 and 300 degrees s-1 (F = 4.2, p < 0.05), 100 degrees s-1 and 400 degrees s-1 (F = 20.2, p < 0.0001), 200 degrees s-1 and 400 degrees s-1 (F = 15.221, p < 0.001), and 300 degrees s-1 and 400 degrees s-1 (F = 5.9, p < 0.01). In all cases the highest MDF values were exhibited at the lower velocities. Lastly, there was an interaction effect when comparing movement patterns at 400 degrees s-1, with MDF values being significantly higher during the non-reciprocal movement pattern than during the reciprocal movement pattern (F = 10.9, p < 0.01). Results indicated that during isokinetic movements, RMS and MDF activity of the hamstrings are altered as velocity changes. More specifically, as velocity increases overall hamstrings' co-activation increases and there is a shift in the power spectrum toward the recruitment of slower-twitch muscle fibers. Results also indicate that movement pattern (reciprocal vs. non reciprocal) does not effect appreciably SEMG activity of hamstrings' co-contraction.

Reliability and concurrent validity of the movement assessment battery for children.

Reliability and concurrent validity of the Movement Assessment Battery for Children were evaluated with a sample of 106 boys and girls, distributed into four age groups: 5-6 yr. (n = 20), 7-8 yr. (n = 20), 9-10 yr. (n = 46), and 11-12 yr. (n = 20). Test-retest reliability of the Movement Assessment Battery for Children, estimated using intraclass correlation coefficients, was high across all age groups, and concurrent validity yielded moderate Pearson correlation coefficients between the Move ment battery and long and short forms of the Bruininks-Oseretsky Test of Motor Proficiency. These results support the use of the Movement battery as a measure of motor ability in children, ages 5 to 12 years.

The major antenna complex of photosystem II has a xanthophyll binding site not involved in light harvesting.

We have characterized a xanthophyll binding site, called V1, in the major light harvesting complex of photosystem II, distinct from the three tightly binding sites previously described as L1, L2, and N1. Xanthophyll binding to the V1 site can be preserved upon solubilization of the chloroplast membranes with the mild detergent dodecyl-alpha-d-maltoside, while an IEF purification step completely removes the ligand. Surprisingly, spectroscopic analysis showed that when bound in this site, xanthophylls are unable to transfer absorbed light energy to chlorophyll a. Pigments bound to sites L1, L2, and N1, in contrast, readily transfer energy to chlorophyll a. This result suggests that this binding site is not directly involved in light harvesting function. When violaxanthin, which in normal conditions is the main carotenoid in this site, is depleted by the de-epoxidation in strong light, the site binds other xanthophyll species, including newly synthesized zeaxanthin, which does not induce detectable changes in the properties of the complex. It is proposed that this xanthophyll binding site represents a reservoir of readily available violaxanthin for the operation of the xanthophyll cycle in excess light conditions.

Carotenoid-to-chlorophyll energy transfer in recombinant major light-harvesting complex (LHCII) of higher plants. I. Femtosecond transient absorption measurements.

The energy transfer kinetics from carotenoids to chlorophylls and among chlorophylls has been measured by femtosecond transient absorption kinetics in a monomeric unit of the major light-harvesting complex (LHCII) from higher plants. The samples were reconstituted complexes with different carotenoid contents. The kinetics was measured both in the carotenoid absorption region and in the chlorophyll Q(y) region using two different excitation wavelengths suitable for selective excitation of the carotenoids. Analysis of the data shows that the overwhelming part of the energy transfer from the carotenoids occurs directly from the initially excited S(2) state of the carotenoids. Only a small part (<20%) may possibly take an S(1) pathway. All the S(2) energy transfer from carotenoids to chlorophylls occurs with time constants <100 fs. We have been able to differentiate among the three carotenoids, two luteins and neoxanthin, which have transfer times of approximately 50 and 75 fs for the two luteins, and approximately 90 fs for neoxanthin. About 50% of the energy absorbed by carotenoids is initially transferred directly to chlorophyll b (Chl b), while the rest is transferred to Chl a. Neoxanthin almost exclusively transfers to Chl b. Due to various complex effects discussed in the paper, such as a specific coupling of Chl b and Chl a excited states, the percentage of direct Chl b transfer thus is somewhat lower than estimated by us previously for LHCII from Arabidopsis thaliana. (Connelly, J. P., M. G. Müller, R. Bassi, R. Croce, and A. R. Holzwarth. 1997. Biochemistry. 36:281). We can distinguish three different Chls b receiving energy directly from carotenoids. We propose as a new mechanism that the carotenoid-to-Chl b transfer occurs to a large part via the B(x) state of Chl b and to the Q(x) state, while the transfer to Chl a occurs only via the Q(x) state. We find no compelling evidence in favor of a substantial S(1) transfer path of the carotenoids, although some transfer via the S(1) state of neoxanthin can not be entirely excluded. The S(1) lifetimes of the two luteins were determined to be 15 ps and 3.9 ps. A detailed quantitative analysis and kinetic model of the processes described here will be presented in a separate paper.

Photochemical behavior of xanthophylls in the recombinant photosystem II antenna complex, CP26.

The steady state absorption and fluorescence spectroscopic properties of the xanthophylls, violaxanthin, zeaxanthin, and lutein, and the efficiencies of singlet energy transfer from the individual xanthophylls to chlorophyll have been investigated in recombinant CP26 protein overexpressed in Escherichia coli and then refolded in vitro with purified pigments. Also, the effect of the different xanthophylls on the extents of static and dynamic quenching of chlorophyll fluorescence has been investigated. Absorption, fluorescence, and fluorescence excitation demonstrate that the efficiency of light harvesting from the xanthophylls to chlorophyll a is relatively high and insensitive to the particular xanthophyll that is present. A small effect of the different xanthophylls is observed on the extent of quenching of Chl fluorescence. The data provide the precise wavelengths of the absorption and fluorescence features of the bound pigments in the highly congested spectral profiles from these light-harvesting complexes. This information is important in assessing the mechanisms by which higher plants dissipate excess energy in light-harvesting proteins.

Energy transfer among CP29 chlorophylls: calculated Förster rates and experimental transient absorption at room temperature.

The energy transfer rates between chlorophylls in the light harvesting complex CP29 of higher plants at room temperature were calculated ab initio according to the Förster mechanism (Förster T. 1948, Ann. Physik. 2:55-67). Recently, the transition moment orientation of CP29 chlorophylls was determined by differential linear dichroism and absorption spectroscopy of wild-type versus mutant proteins in which single chromophores were missing (Simonetto R., Crimi M., Sandonà D., Croce R., Cinque G., Breton J., and Bassi R. 1999. Biochemistry. 38:12974-12983). In this way the Q(y) transition energy and chlorophyll a/b affinity of each binding site was obtained and their characteristics supported by reconstruction of steady-state linear dichroism and absorption spectra at room temperature. In this study, the spectral form of individual chlorophyll a and b ligands within the protein environment was experimentally determined, and their extinction coefficients were also used to evaluate the absolute overlap integral between donors and acceptors employing the Stepanov relation for both the emission spectrum and the Stokes shift. This information was used to calculate the time-dependent excitation redistribution among CP29 chlorophylls on solving numerically the Pauli master equation of the complex: transient absorption measurements in the (sub)picosecond time scale were simulated and compared to pump-and-probe experimental data in the Q(y) region on the native CP29 at room temperature upon selective excitation of chlorophylls b at 640 or 650 nm. The kinetic model indicates a bidirectional excitation transfer over all CP29 chlorophylls a species, which is particularly rapid between the pure sites A1-A2 and A4-A5. Chlorophylls b in mixed sites act mostly as energy donors for chlorophylls a, whereas site B5 shows high and bidirectional coupling independent of the pigment hosted.

Effect of ankle position fixation on peak torque and electromyographic activity of the knee flexors and extensors.

The purpose of this study was to examine the effect of ankle position fixation on peak torque (PT) and electromyographic (EMG) activity of knee-joint muscles during isokinetic testing. Twelve female athletes performed isokinetic knee flexion and extension at 60 degrees and 180 degrees/s under two conditions: with the ankle fixed in a position of plantarflexion and with the ankle fixed in a position of dorsiflexion. Bipolar surface electrodes were placed on the vastus lateralis, vastus medialis, biceps femoris, medial hamstrings, and the lateral head of the gastrocnemius for determination of the root mean square of the EMG (rmsEMG) and the median frequency of the EMG (mfEMG). No significant differences in knee extensor PT were noted in either ankle position for each velocity tested. Significant differences were noted, however, in knee flexor PT (p < 0.05) at both 60 degrees and 180 degrees/s, with the greatest PT observed with the ankle fixed in dorsiflexion. Neither quadriceps, hamstrings, nor gastrocnemius rmsEMG activity was affected by ankle position; however, there was a significant difference in mfEMG for the gastrocnemius, with higher frequencies observed with the ankle fixed in plantarflexion (p < 0.01). These results suggest that ankle position effects knee flexor PT during open chain isokinetic movements. The reason for decreased knee flexor PT with the ankle fixed in plantarflexion is probably due to the gastrocnemius muscle being in a too shortened position, thereby preventing it from effectively producing force at the knee joint.

Reciprocal coactivation patterns of the medial and lateral quadriceps and hamstrings during slow, medium and high speed isokinetic movements.

The effect of movement velocity and fatigue on the reciprocal coactivation of the quadriceps and hamstrings was investigated through analysis of the root mean square (RMS) and the median frequency (MDF) of surface electromyography for the vastus medialis (VM), vastus lateralis (VL), medial hamstrings (MH) and biceps femoris (BF). Fourteen subjects performed six continuous isokinetic knee extension and flexion movements at 60 degrees, 180 degrees and 300 degrees s(-1), and 30 continuous movements at 300 degrees s(-1) to examine muscular fatigue patterns. Statistical analyses revealed that the RMS activity of the VM displayed greater coactivation than the VL (P<0.01) and the BF displayed greater coactivation than the MH (P<0.0001). There was no effect of velocity on the coactivation levels of the VM, the VL, or the MH; however, there was an effect of velocity on the coactivation levels of the BF (P<0.0001). Relative to MDF activity, the MH shifted upward as velocity increased (P<0. 01) while the BF decreased between 180 and 300 degrees s(-1) (P<0. 01). Results of the muscular fatigue test indicated that the RMS activity of the VM showed a higher degree of coactivation than the VL (P<0.01) and the BF showed approximately three times the coactivation level of the MH (P<0.001). The MDF of the VL and MH shifted downward as the repetitions progressed (P<0.01) with no changes for the VM or for the BF. Results of this study suggest that during isokinetic testing, both the VM and BF have significantly greater reciprocal coactivation levels when compared to the VL and MH, respectively. In addition, these results suggest that motor unit recruitment patterns of the VM and VL and the MH and BF differ with regard to the effects of velocity and fatigue.