Host and cell type affect the mode of degradation by Meripilus giganteus

Wood degradation by the white-rot basidiomycete Meripilus giganteus (Pers.: Pers.) Karst. was studied in naturally infected and artificially inoculated wood of beech ( Fagus sylvatica L.) and large-leaved lime ( Tilia platyphyllos Scop.). Semi-thin sections revealed that the secondary walls of most fibres contained internal cavities. Three distinct types of cavity formation, which differed not only between hosts, but also between cell type and location in the annual ring, were identified.
Within discoloured wood of naturally infected beech, the structure of the cavities and their formation by the associated hyphae were reminiscent of a soft-rot. By contrast, cavity formation in artificially inoculated beech and large-leaved lime wood differed from a soft-rot mode of attack as extensive delignification always preceded cavity formation, and neither T-branching, L-bending, nor hyphal growth were found within cell walls. The formation of half-moon shaped cavities in beech wood was present only in tension-wood fibres. From large diameter hyphae, growing within the fibre lumen, numerous fine perforation hyphae extended transversely via helical cracks into the cell wall. Subsequent degradation of cellulose within concentric layers of the tension-wood fibres commenced from the apices of perforation hyphae.
Sections stained with ruthenium red and hydroxlamine-ferric chloride, revealed that M. giganteus preferentially degrades pectin-rich regions of the middle lamellae in xylary ray cells. In large-leaved lime, such regions were uniformly located in the middle lamellae of axial and ray parenchyma. In beech wood, degradation of pectin-rich middle lamellae regions commenced after the delignification of secondary walls and resulted in a conspicuous hollowing of multiseriate xylem rays. Plasticity in wood degradation modes by M. giganteus in large-leaved lime and beech wood reflects variations in cell wall structure and/or prevailing wood conditions.     

Qualitative and quantitative changes of beech wood degraded by wood-rotting basidiomycetes monitored by Fourier transform infrared spectroscopic methods and multivariate data analysis

Beech wood (Fagus sylvatica L.) veneers were cultivated with white and brown rot fungi for up to 10 weeks. Fungal wood modification was traced with Fourier transform near infrared (FT-NIR) and Fourier transform mid infrared (FT-MIR) methods. Partial least square regression (PLSR) models to predict the total lignin content before and after fungal decay in the range between 17.0% and 26.6% were developed for FT-MIR transmission spectra as well as for FT-NIR reflectance spectra. Weight loss of the decayed samples between 0% and 38.2% could be estimated from the wood surface using individual PLSR models for white rot and brown rot fungi, and from a model including samples subjected to both degradation types.     

Quantitative studies on the distribution of myofilaments in intrafusal muscle fibres

Summary Muscle spindles contain two types of intrafusal muscle fibre, nuclear bag fibres and nuclear chain fibres. The intrafusal fibres of rabbit and guinea pig spindles have been studied using quantitative stereological techniques at the ultrastructural level. The crosssectional areas occupied by myofilaments have been measured in the polar and equatorial regions of both types of intrafusal fibre. There are considerably fewer myofilaments in the equatorial regions of both types of fibre compared with their polar regions.This work was carried out with the aid of grants from the Medical Research Council and the Science Research Council of Great Britain.     

Electron microscopic immunocytochemical demonstration of separate neurophysin-vasopressinergic and neurophysin-oxytocinergic nerve fibres in the neural lobe of the rat hypophysis

Summary By means of the unlabeled antibody peroxidase-antiperoxidase (PAP) technique at the electron microscopic level, it was demonstrated that the hormones of the neural lobe of the rat hypophysis are located in separate neurophysin-vasopressinergic and neurophysin-oxytocinergic nerve fibres. These observations confirm the results of our previous immunocytochemical studies at the light microscopic level.This investigation was supported by a grant from the Belgian Nationaal Fonds voor Geneeskundig Wetenschappelijk Onderzoek     

Morphology and organization of muscle fibres in the thoracic coxal muscle receptor organ and the associated promotor muscle, in a crayfish, Cherax destructor, and mud crab, Scylla serrata

Using confocal laser scanning and conventional light microscopy, the morphology and organization of the muscle fibres in a proprioceptor, the thoracic coxal muscle receptor organ (TCMRO), and the associated 'extrafusal' promotor muscle were investigated in two species of decapod crustacea, the crayfish Cherax destructor and the mud crab Scylla serrata . The diameter of the TCMROs was shown to increase distally, with an increase up to 350% recorded for the crayfish. The tapered shape of the crayfish TCMRO was demonstrated to amplify movements mechanically at the transducer region where the afferent nerves attach. Serial sectioning of the TCMROs, showed that the fibre number increased in the proximal to distal direction from 14 to 30 fibres in the crayfish and from 7 to 20 in the crab. Optical sectioning with the laser scanning confocal microscope revealed that the increase in fibre numbers was the result of muscle fibres branching in the distal third section of the TCMRO. The percentage of muscle tissue in the cross-sectional area in the TCMRO was found to be only 35.2% and 64.6% in the crayfish and crab, respectively. Longitudinal sectioning using laser scanning confocal microscopy revealed the average sarcomere length of the TCMRO muscle fibres of both species to be in the intermediate range for crustacean muscle fibres (4.1 ± 0.1 µm and 4.55 ± 0.34 µm for the crayfish and crab) compared with the long sarcomere muscle fibres in the associated promotor muscles (7.87 ± 0.2 and 10.6 ± 0.6 µm). The distinct morphology of the TCMRO muscle fibres – smaller diameter, intermediate sarcomere length and branching of fibres compared to the larger, long sarcomere promotor fibre muscle fibres – suggest that the TCMRO muscle fibres are specialized in their role of proprioception.     

Loss of a DNA binding site within the tail of prelamin A contributes to altered heterochromatin anchorage by progerin

Mutations in the lamin A/C (LMNA) gene that cause Hutchinson-Gilford progeria syndrome (HGPS) lead to expression of a protein called progerin with 50 amino acids deleted from the tail of prelamin A. In cells from patients with HGPS, both the amount and distribution of heterochromatin are altered. We designed in vitro assays to ask whether such alterations might reflect changes in chromatin, DNA and/or histone binding properties of progerin compared to wild-type lamin C-terminal tails. We show that progerin tail has a reduced DNA/chromatin binding capacity and modified trimethylated H3K27 binding pattern, offering a molecular mechanism for heterochromatin alterations related to HGPS.

Structured summary

MINT-7893924, MINT-7893941, MINT-7893990, MINT-7894005, MINT-7894023, MINT-7894038: H3 (uniprotkb:Q71DI3) binds (MI:0407) to LaminA (uniprotkb:P02545) by surface plasmon resonance (MI:0107)MINT-7893957, MINT-7893974, MINT-7894055: H3 (uniprotkb:Q71DI3) binds (MI:0407) to progerin (uniprotkb:Q6UYC3) by surface plasmon resonance (MI:0107)     

Dynamics of electron transfer within and between PS II reaction center complexes indicated by the light-saturation curve of in vivo variable chlorophyll fluorescence emission

The dynamics of light-induced closure of the PS II reaction centers was studied in intact, dark-adapted leaves by measuring the light-irradiance (I) dependence of the relative variable chlorophyll fluorescence V which is the ratio between the amplitude of the variable fluorescence induced by a pulse of actinic light and the maximal variable fluorescence amplitude obtained with an intense, supersaturating light pulse. It is shown that the light-saturation curve of V is a hyperbola of order n. The experimental values of n ranged from around 0.75 to around 2, depending on the plant material and the environmental conditions. A simple theoretical analysis confirmed this hyperbolic relationship between V and I and suggested that n could represent the apparent number of photons necessary to close one reaction center. Thus, experimental conditions leading to n values higher than 1 could indicate that, from a macroscopic viewpoint, more than one photon is necessary to close one PS II center, possibly due to changes in the relative concentrations of the different redox states of the PS II reaction center complexes at the quasi-steady state induced by the actinic light. On the other hand, the existence of environmental conditions resulting in n noticeably lower than 1 suggests the possibility of an electron flow between PS II reaction center complexes.Abbreviations F₀ and F_m minimal and maximal levels of chlorophyll fluorescence emission, respectively - F_p peak fluorescence induced by a pulse of actinic light - I incident light irradiance (in W m^-2) - PS II Photosystem II - P₆₈₀ PS II reaction center - Q_A and Q_B primary and secondary (stable) electron acceptors of PS II - V relative variable chlorophyll fluorescence % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0Jf9crFfpeea0xh9v8qiW7rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaGaaiikaiaadA% facqGH9aqpcaGGOaGaaeOramaaBaaaleaacaqGWbaabeaakiabgkHi% TiaabAeadaWgaaWcbaGaaeimaaqabaGccaGGPaGaai4laiaacIcaca% qGgbWaaSbaaSqaaiaab2gaaeqaaOGaeyOeI0IaaeOramaaBaaaleaa% caqGWaaabeaakiaacMcacaGGPaaaaa!47BD!\[(V = ({\text{F}}_{\text{p}} - {\text{F}}_{\text{0}} )/({\text{F}}_{\text{m}} - {\text{F}}_{\text{0}} ))\]     

Induction of the acrosome reaction in the spermatozoa of the goat by secretions of the male accessory glands and milk

The balancing effects of bulbourethral gland secretion (BUS) and of seminal vesicle secretion (SVS) on goat semen quality were previously demonstrated. In the present study, electron microscope observations revealed a high frequency of spermatozoa with a reacted acrosome among spermatozoa from cauda epididymis exposed to BUS in the presence of milk. This frequency was significantly reduced when SVS had been added either before or after BUS. No reacted acrosome was observed in the absence of milk. All mount spermatozoa were incubated with milk or SVS or BUS or combinations of the three materials labeled with colloidal gold. SVS attached specifically on the plasma membrane covering the anterior part of the acrosome, whereas BUS spread all over the sperm head. Milk attached on the anterior half of the sperm head only when BUS was present in the sperm environment. It is concluded that BUS plays an active role in the induction of the acrosome reaction in the presence of milk and that SVS counteracts this role.     

Orientational information of troponin C within the thin filaments obtained by neutron fiber diffraction

In striated muscles contraction is regulated by the thin filament-based proteins, troponin consisting of three subunits (TnC, TnI, and TnT), and tropomyosin. Knowledge of in situ structures of these proteins is indispensable for elucidating this Ca(2+)-sensitive regulatory mechanism. We employed neutron scattering to investigate the structure of TnC within the thin filament, and found that TnC assumes extended dumbbell-like structures and moves toward the filament axis by binding of Ca(2+). Here, in order to obtain more detailed in situ structural information of TnC, neutron fiber diffraction measurements were performed. Sols of native thin filaments and the thin filaments containing deuterated TnC were prepared in (2)H(2)O. The oriented samples were obtained by placing these sols sealed in quartz capillaries with a diameter of 3 mm in a magnetic field of 18 Tesla. Neutron fiber diffraction patterns were obtained from these oriented samples in the absence and presence of Ca(2+). The patterns obtained showed strong equatorial diffraction due to the thin filaments, 59 A and 51 A layer-lines due to actin, and meridional reflections due to Tn-complex. Analysis of the meridional reflections due to Tn-complex with aid of model calculation showed that the angle between the thin filament axis and the long axis of TnC was estimated to be 67(+/-7) degrees and 49(+/-17) degrees , in the absence and presence of Ca(2+), respectively, suggesting that TnC, which assumes orientations rather perpendicular to the filament axis in the absence of Ca(2+), tilts toward the filament axis and the orientational and positional disorder increases by binding Ca(2+). It also showed that the relative position of the TnC moved by about 22 A by binding Ca(2+), and this apparent movement was concomitant with the movements of other Tn-subunits. This implies that by binding Ca(2+), significant structural rearrangements of Tn-subunits occur.     

Deciphering the Reaction Mechanism of Lithium–Sulfur Batteries by In Situ/Operando Synchrotron‐Based Characterization Techniques

Lithium–sulfur (Li–S) batteries have received extensive attention as one of the most promising next‐generation energy storage systems, mainly because of their high theoretical energy density and low cost. However, the practical application of Li–S batteries has been hindered by technical obstacles arising from the polysulfide shuttle effect and poor electronic conductivity of sulfur and discharge products. Therefore, it is of profound significance for understanding the underlying reaction mechanism of Li–S batteries to circumvent these problems and improve the overall battery performance. Advanced characterization techniques, especially synchrotron‐based X‐ray techniques, have been widely applied to the mechanistic understanding of Li–S batteries. Specifically, in situ/operando synchrotron‐based techniques allows chemical and structural evolution to be directly observed under real operation conditions. Here, recent progress in the understanding of the operating principles of Li–S batteries based on in situ/operando synchrotron‐based techniques, including X‐ray absorption spectroscopy, X‐ray diffraction, and X‐ray microscopy, is reviewed. The aim of this progress report is to provide a comprehensive treatise on in situ/operando synchrotron‐based techniques for mechanism understanding of Li–S batteries, and thereby provide guidance for optimizing their overall electrochemical performances.     

Cryptic organisation within an apparently irregular rostrocaudal distribution of interneurons in the embryonic zebrafish spinal cord

The molecules and mechanisms involved in patterning the dorsoventral axis of the developing vertebrate spinal cord have been investigated extensively and many are well known. Conversely, knowledge of mechanisms patterning cellular distributions along the rostrocaudal axis is relatively more restricted. Much is known about the rostrocaudal distribution of motoneurons and spinal cord cells derived from neural crest but there is little known about the rostrocaudal patterning of most of the other spinal cord neurons. Here we report data from our analyses of the distribution of dorsal longitudinal ascending (DoLA) interneurons in the developing zebrafish spinal cord. We show that, although apparently distributed irregularly, these cells have cryptic organisation. We present a novel cell-labelling technique that reveals that DoLA interneurons migrate rostrally along the dorsal longitudinal fasciculus of the spinal cord during development. This cell-labelling strategy may be useful for in vivo analysis of factors controlling neuron migration in the central nervous system. Additionally, we show that DoLA interneurons persist in the developing spinal cord for longer than previously reported. These findings illustrate the need to investigate factors and mechanisms that determine “irregular” patterns of cell distribution, particularly in the central nervous system but also in other tissues of developing embryos.     

Dimerisation of the Rhodobacter sphaeroides RC-LH1 photosynthetic complex is not facilitated by a GxxxG motif in the PufX polypeptide

In purple photosynthetic bacteria the initial steps of light energy transduction take place in an RC-LH1 complex formed by the photochemical reaction centre (RC) and the LH1 light harvesting pigment-protein. In Rhodobacter sphaeroides, the RC-LH1 complex assembles in a dimeric form in which two RCs are surrounded by an S-shaped LH1 antenna. There is currently debate over the detailed architecture of this dimeric RC-LH1 complex, with particular emphasis on the location and precise function of a minor polypeptide component termed PufX. It has been hypothesised that the membrane-spanning helical region of PufX contains a GxxxG dimerisation motif that facilitates the formation of a dimer of PufX at the interface of the RC-LH1 dimer, and more specifically that the formation of this PufX dimer seeds assembly of the remaining RC-LH1 dimer (J. Busselez et al., 2007). In the present work this hypothesis was tested by site directed mutagenesis of the glycine residues proposed to form the GxxxG motif. Mutation of these glycines to leucine did not decrease the propensity of the RC-LH1 complex to assemble in a dimeric form, as would be expected from experimental studies of the effect of mutation on GxxxG motifs in other membrane proteins. Indeed increased yields of dimer were seen in two of the glycine-to-leucine mutants constructed. It is concluded that the PufX from Rhodobacter sphaeroides does not contain a genuine GxxxG helix dimerisation motif.     

Design and synthesis of new antitumor agents with the 1,7-epoxycyclononane framework. Study of their anticancer action mechanism by a model compound

This article describes the design, synthesis and biological evaluation of a new family of antitumor agents having the 1,7-epoxycyclononane framework. We have developed a versatile synthetic methodology that allows the preparation of a chemical library with structural diversity and in good yield. The synthetic methodology has been scaled up to the multigram level and can be developed in an enantioselective fashion. The study in vitro of a model compound, in front of the cancer cell lines HL-60 and MCF-7, showed a growth inhibitory effect better than that of cisplatin. The observation of cancer cells by fluorescence microscopy showed the presence of apoptotic bodies and a degradation of microtubules. The study of cell cycle and mechanism of death of cancer cells by flow cytometry indicates that the cell cycle arrested at the G₀/G₁ phase and that the cells died by apoptosis preferably over necrosis. A high percentage of apoptotic cells at the subG₀/G₁ level was observed. This indicates that our model compound does not behave as an antimitotic agent like nocodazole, used as a reference, which arrests the cell cycle at G₂/M phase. The interaction of anticancer agents with DNA molecules was evaluated by atomic force microscopy, circular dichroism and electrophoresis on agarose gel. The results indicate that the model compound has not DNA as a target molecule. The in silico study of the model compound showed a potential good oral bioavailability.