The influence of lead on the biomechanical properties of bone tissue in rats.

INTRODUCTION AND OBJECTIVE: Environmental lead (Pb) is a serious public health problem. At high levels, Pb is devastating to almost all organs. On the other hand, it is difficult to determine a safe level of exposure to Pb. More than 90% of the Pb in the adult human body and 70% in a child's body is stored in the bones. In the presented study, the effects of lead exposure on bones were studied for rats treated orally with Pb acetate in drinking water for 14 days. The hypothesis was tested that lead exposure negatively affects bone structure. MATERIALS AND METHODS: Femur strength was measured in a three-point bending test, whereas infrared spectroscopy (FTIR) was used to measure molecular structural changes. RESULTS: Lead significantly decreased the ratio of area of two types of vibrational transitions, which are highly specific to mineral to matrix ratio. The results of the biomechanical study show that femurs of rats treated by Pb-acetate appeared to be weaker than bones of the control group, and may produce a condition for the development of higher risk of fractures. Additionally, a great difference in body mass was observed between control and the Pb acetate-treated groups. CONCLUSIONS: The lower bone mineral content and the weaker mechanical properties of bones from Pb-treated rats are associated with the pathologic state dependent of the exposure of lead.

Biomechanical parameters of the BP-enriched bone cement.

Bisphosphonates (BPs) are well-known substances with very efficient antiresorptive properties. Their beneficial actions are useful not only in achieving better bone mineral density but also in improving bone microarchitecture, strength and, consequently, its quality. Surgical cement, being a polymer composite, is required to be highly biocompatible and biotolerant. The goal of the presented study was to assess whether the enrichment of cement with pamidronate has changed its biomechanical properties. We compared the biomechanical parameters of clean bone cement and BP-enriched bone cement, which were both used formerly in our rat models. Biomechanical properties of BP-enriched bone cement are defined by two basic terms: stress and strain, which are caused by the influence of external force. In the investigatory process of the bone's biomechanical parameters, the compressive test and the three-point flexural tests were used. During the three-point flexural investigation, the sample was supported at both ends and loaded in the middle, resulting in a flexure. After a specific range of flexure, the sample was fractured. In obtained results, there were no significant differences in the values of the stress determined at the point of maximal load and the energy stored in the samples for proportional stress-strain limit (elastic region). There were also no significant differences in the density of the samples. The study shows that the enrichment of bisphosphonates causes yielding of the bone cement material. In the presented data, we conclude that use of pamidronate implanted in bone cement did not have a detrimental effect on its biomechanical properties. Therefore, the obtained results encouraged us to perform further in vivo experiments which assess the biomechanical properties of bones implanted with BP-enriched bone cement.

Light-induced isomerization of the LHCII-bound xanthophyll neoxanthin: possible implications for photoprotection in plants.

Light-harvesting pigment-protein complex of Photosystem II (LHCII) is the largest photosynthetic antenna complex of plants and the most abundant membrane protein in the biosphere. Plant fitness and productivity depend directly on a balance between excitations in the photosynthetic apparatus, generated by captured light quanta, and the rate of photochemical processes. Excess excitation energy leads to oxidative damage of the photosynthetic apparatus and entire organism and therefore the balance between the excitation density and photosynthesis requires precise and efficient regulation, operating also at the level of antenna complexes. We show that illumination of the isolated LHCII leads to isomerization of the protein-bound neoxanthin from conformation 9'-cis to 9',13- and 9',13'-dicis forms. At the same time light-driven excitation quenching is observed, manifested by a decrease in chlorophyll a fluorescence intensity and shortened fluorescence lifetimes. Both processes, the neoxanthin isomerization and the chlorophyll excitation quenching, are reversible in dim light. The results of the 77K florescence measurements of LHCII show that illumination is associated with appearance of the low-energy states, which can serve as energy traps in the pigment-protein complex subjected to excess excitation. Possible sequence of the molecular events is proposed, leading to a protective excess excitation energy quenching: neoxanthin photo-isomerization→formation of LHCII supramolecular structures which potentiate creation of energy traps→excitation quenching.

Investigation of the molecular mechanism of the blue-light-specific excitation energy quenching in the plant antenna complex LHCII.

Excitation of the major photosynthetic antenna complex of plants, LHCII, with blue light (470nm) provides an advantage to plants, as it gives rise to chlorophyll a fluorescence lifetimes shorter than with excitation with red light (635nm). This difference is particularly pronounced in fluorescence emission wavelengths longer than 715nm. Illumination of LHCII preparation with blue light additionally induces fluorescence quenching, which develops on a minute timescale. This effect is much less efficient when induced by red light, despite the equalized energy absorbed in both the spectral regions. Simultaneous analysis of the fluorescence and photoacoustic signals in LHCII demonstrated that the light-driven fluorescence quenching is not associated with an increase in heat emission. Instead, a reversible light-induced conformational transformation of the protein takes place, as demonstrated by the FTIR technique. These findings are discussed in terms of the blue-light-specific excitation energy quenching in LHCII, which may have photoprotective applications.

Blue-light-controlled photoprotection in plants at the level of the photosynthetic antenna complex LHCII.

Plants have developed several adaptive regulatory mechanisms, operating at all the organization levels, to optimize utilization of light energy and to protect themselves against over-excitation-related damage. We report activity of a previously unknown possible regulatory mechanism that operates at the molecular level of the major photosynthetic pigment-protein complexes of plants, LHCII. This mechanism is driven exclusively by blue light, operates in the trimeric but not in the monomeric complex, and results in singlet excitation quenching leading to thermal energy dissipation. The conclusions are based on single molecule fluorescence lifetime analysis, direct measurements of thermal energy dissipation by photo-thermal spectroscopy, and on fluorescence spectroscopy. Possible molecular mechanisms involved in the blue-light-induced photoprotective effect are discussed, including xanthophyll photo-isomerization and the thermo-optic effect.

Light-induced change of configuration of the LHCII-bound xanthophyll (tentatively assigned to violaxanthin): a resonance Raman study.

Raman scattering spectra of light-harvesting complex LHCII isolated from spinach were recorded with an argon laser, tuned to excite the most red-absorbing LHCII-bound xanthophylls (514.5 nm). The intensity of the nu(4) band (at ca. 950 cm-1) corresponding to the out-of-plane wagging modes of the C-H groups in the resonance Raman spectra of carotenoids appears to be inversely dependent on the probing laser power density. This observation can be interpreted in terms of excitation-induced change of configuration of the protein-bound xanthophyll owing to the fact that the intensity of this particular band is diagnostic of a chromophore twisting resulting from its binding to the protein environment. The comparison of the shape of the nu(4) band of a xanthophyll involved in the light-induced spectral changes with the shape of the nu(4) band of the xanthophylls present in LHCII, reported in the literature, lets us conclude that, most probably, violaxanthin is a pigment that undergoes light-driven changes of molecular configuration but also the involvement of lutein may not be excluded. Possible physical mechanisms responsible for the configuration changes and physiological importance of the effect observed are discussed.

Xanthophyll-induced aggregation of LHCII as a switch between light-harvesting and energy dissipation systems.

The xanthophyll cycle pigments, violaxanthin and zeaxanthin, present outside the light-harvesting pigment-protein complexes of Photosystem II (LHCII) considerably enhance specific aggregation of proteins as revealed by analysis of the 77 K chlorophyll a fluorescence emission spectra. Analysis of the infrared absorption spectra in the Amide I region shows that the aggregation is associated with formation of intermolecular hydrogen bonding between the alpha helices of neighboring complexes. The aggregation gives rise to new electronic energy levels, in the Soret region (530 nm) and corresponding to the Q spectral region (691 nm), as revealed by analysis of the resonance light scattering spectra. New electronic energy levels are interpreted in terms of exciton coupling of protein-bound photosynthetic pigments. The energy of the Q excitonic level of chlorophyll is not high enough to drive the light reactions of Photosystem II but better suited to transfer excitation energy to Photosystem I, which creates favourable energetic conditions for the state I-state II transition. The lack of fluorescence emission from this energy level, at physiological temperatures, is indicative of either very high thermal energy conversion rate or efficient excitation quenching by carotenoids. Chlorophyll a fluorescence was quenched up to 61% and 34% in the zeaxanthin- and violaxanthin-containing samples, respectively, as compared to pure LHCII. Enhanced aggregation of LHCII, observed in the presence of the xanthophyll cycle pigments, is discussed in terms of the switch between light-harvesting and energy dissipation systems.