Diatoms are considered unicellular eukaryotic organisms exclusively depositing biogenic silica. Heretofore there has been no report of calcification by these algae. Here it is shown that calcium ...carbonate within the stalks of Didymosphenia geminata, a nuisance species that has prolifically colonized streams and rivers globally, is biogenic in origin and occurs as a network of calcite nanofibers. The nanofibrous framework in the mineralized polysaccharide matrix imparts mechanical support to the stalks, providing stability in variable flow conditions. The results demonstrate that D. geminata possesses cellular and periplasmic carbonic‐anhydrases that contribute to carbon fixation and biomineralization, respectively. The activity of external carbonic‐anhydrase was more than 50% of the total activity, which points to its role in anchoring this bioeroding diatom on hard surfaces. The first evidence of multiphase biomineralization by diatoms that deposit both biogenic silica and crystalline biogenic calcite which are imparting distinct functional advantage to the organism is provided.
Calcification by diatoms is demonstrated for the first time. The polysaccharide‐based adhesive stalks of Didymosphenia geminata represent a unique biocomposite containing amorphous silica and nanocrystalline calcite fibers. These are produced as a result of the activity of periplasmic carbonic anhydrases which also alters the pH of the cell‐substrate interface leading to bioerosion and biomineralization.
Evolution of bacteria to selective chemical pressure (e.g. antibiotics) is well studied in contrast to the influence of physical stressors. Here we show that instantaneous physical stress in a ...homogeneous environment (without concentration gradient) induces fast adaptation of Escherichia coli. We exposed E. coli to a large number of collisions of around 10
per bacterium per second with sharp ZnO nanorods. The pressure exerted on the bacterial cell wall was up to 10 GPa and induced phenotype changes. The bacteria's shape became more spherical, the density of their periplasm increased by around 15% and the average thickness of the cell wall by 30%. Such E. coli cells appeared almost as Gram-positive bacteria in the standard Gram staining. Additionally, we observed a combination of changes occurring at the genomic level (mutations identified in form of single nucleotide polymorphisms) and down-regulation of expression of 61 genes encoding proteins involved in β-oxidation of fatty acids, glycolysis, the citric acid cycle, as well as uptake of amino acids and enzyme cofactors. Thus, we show that bacteria undergo phenotypic changes upon instantaneous, acute physical stress without any obviously available time for gradual adaptation.
Plants have developed a range of strategies for resisting environmental stresses. One of the most common is the synthesis and deposition of callose, which functions as a barrier against stress factor ...penetration. The aim of our study was to examine whether callose forms an efficient barrier against Pb penetration in the roots of Lemna minor L. exposed to this metal. The obtained results showed that Pb induced callose synthesis in L. minor roots, but it was not deposited regularly in all tissues and cells. Callose occurred mainly in the protoderm and in the centre of the root tip (procambial central cylinder). Moreover, continuous callose bands, which could form an efficient barrier for Pb penetration, were formed only in the newly formed and anticlinal cell walls (CWs); while in other CWs, callose formed only small clusters or incomplete bands. Such an arrangement of callose within root CWs inefficiently protected the protoplast from Pb penetration. As a result, Pb was commonly present inside the root cells. In the light of the results, the barrier role of callose against metal ion penetration appears to be less obvious than previously believed. It was indicated that induction of callose synthesis is not enough for a successful blockade of the stress factor penetration. Furthermore, it would appear that the pattern of callose distribution has an important role in this defence strategy.
The cell wall emerged as one of the important structures in plant stress responses. To investigate the effect of cold on the cell wall properties, the content and localization of pectins and pectin ...methylesterase (PME) activity, were studied in two maize inbred lines characterized by different sensitivity to cold. Low temperature (14/12 °C) caused a reduction of pectin content and PME activity in leaves of chilling-sensitive maize line, especially after prolonged treatment (28 h and 7 days). Furthermore, immunocytohistological studies, using JIM5 and JIM7 antibodies, revealed a decrease of labeling of both low- and high-methylesterified pectins in this maize line. The osmotic potential, quantified by means of incipient plasmolysis was lower in several types of cells of chilling-sensitive maize line which was correlated with the accumulation of sucrose. These studies present new finding on the effect of cold stress on the cell wall properties in conjunction with changes in the osmotic potential of maize leaf cells.
The hypothesis that lead (Pb) can be uptake or remobilized from the cell wall (CW) by internalization withlow-esterified pectins (up to 40% – JIM5-P), was studied in tip-growing apical cell of
...Funaria hygrometrica protonemata. Treatment 4
h with 1
mM PbCl
2 caused marked vesicular traffic intensification and the common internalization of JIM5-P from the CW. Lead bound to JIM5-P was internalized from the CW, together with this compound and entered the protoplast. It showed that Pb deposited in CW is not as safe for plant cell as previously believed.
However, pulse-chase experiments (recovering 4
h and 24
h) indicated that CW and its thickenings can function as the final sequestration compartments. In Pb deposition sites, a callose layer occurred. It was localized from the protoplast site, next to Pb deposits separating sequestrated to CW and its thickenings Pb from plasma membrane almost certainly protecting the plant cell from its returning into the protoplast.
Lead bound to low-esterified pectins in cell wall can be uptake or remobilized by endocytosis together with this pectin epitope.
Low-methylesterified homogalacturonans have been suggested to play a role in the binding and immobilization of Pb in CW. Using root apices of hybrid aspen, a plant with a high phytoremediation ...potential, as a model, we demonstrated that the in situ distribution pattern of low-methylesterified homogalacturonan, pectin epitope (JIM5-P), reflects the pattern of Pb occurrence. The region which indicated high JIM5-P level corresponded with “Pb accumulation zone”. Moreover, JIM5-P was especially abundant in cell junctions, CWs lining the intercellular spaces and the corners of intercellular spaces indicating the highest accumulation of Pb. Furthermore, JIM5-P and Pb commonly co-localized.
The observations indicate that low-methylesterified homogalacturonan is the CW polymer that determines the capacity of CW for Pb sequestration. Our results suggest a promising directions for CW modification for enhancing the efficiency of plant roots in Pb accumulation, an important aspect in the phytoremediation of soils contaminated with trace metals.
•Co-localization of low-methylesterified pectins and Pb was analysed in situ.•The pattern of Pb accumulation matched low-methylesterified pectins distribution.•Low-methylesterified pectins and Pb commonly co-localized in cell walls.•Low-methylesterified pectins revealed an important compound in Pb sequestration.•We suggest a new direction in enhancing plant efficiency for phytoremediation.
The distribution of lead in developing tissues of aspen root tips exposed to short-term lead treatment mimics the distribution of low-methylesterified pectin epitope.
A composite in which gold nanoparticles (AuNPs) approximately 10 nm in size are embedded in amorphous transparent silica matrix has been produced. The synthetic protocol uses HAuCl4 as the Au ion ...source, tetraethoxysilane (TEOS) as the SiO2 precursor, and l‐ascorbic acid (AA) as the reducing agent. AA is employed before the sol‐gel process in an amount sufficient only for reduction of Au3+ ions to Au+. By using a cationic surfactant, benzylcetyldimethylammonium chloride hydrate (BDAC) and/or cetyltrimethylammonium bromide (CTAB), the Au+ ions are encapsulated within metalomicelles, which prevents them from being reduced to Au0 and enables their homogeneous distribution in the gel. Reduction of Au+ to Au0 and the growth of the AuNPs occurs at room temperature during the gelation, and arises from the release of EtOH during the hydrolysis of TEOS. The composites contain 0.027 wt % of Au. They exhibit nonlinear optical behavior characterized by the third‐order nonlinear refraction index, n2, in the range 3.6–5.7×10−16 cm2 W−1 at λ=1.030 μm. The composites are capable of effective third‐harmonic generation of ultrashort near‐IR (210 fs, 1.030 μm) laser pulse through a direct third‐order mechanism.
Sequential gold reduction: A synthetic protocol employing the two‐step reduction of gold ions enables in situ sol‐gel fabrication of vitreous silica composites containing uniformly distributed gold nanoparticles. First, Au3+ ions that form metallomicelles with cationic surfactant are reduced to Au+ by l‐ascorbic acid. Then, EtOH released during hydrolysis of tetraethoxysilane reduces Au+ to Au0. The composite exhibits nonlinear optical properties, and is capable of effective third‐harmonic generation in the near‐IR.
Rab7 GTPases are involved in membrane trafficking in the late endosomal/lysosomal pathway. In Paramecium octaurelia Rab7a and Rab7b are encoded by paralogous genes. Antipeptide antibodies generated ...against divergent C-termini recognize Rab7a of 22.5 kDa and Rab7b of 25 kDa, respectively. In 2D gel electrophoresis two immunoreactive spots were identified for Rab7b at pI about 6.34 and about 6.18 and only one spot for Rab7a of pI about 6.34 suggesting post-translational modification of Rab7b. Mass spectrometry revealed eight identical phosphorylated residues in the both proteins. ProQ Emerald staining and ConA overlay of immunoprecipitated Rab7b indicated its putative glycosylation that was further supported by a faster electrophoretic mobility of this protein upon deglycosylation. Such a post-translational modification and substitution of Ala(140) in Rab7a for Ser(140) in Rab7b may result in distinct targeting to the oral apparatus where Rab7b associates with the microtubular structures as revealed by STED confocal and electron microscopy. Rab7a was mapped to phagosomal compartment. Absolute qReal-Time PCR analysis revealed that expression of Rab7a was 2.6-fold higher than that of Rab7b. Upon latex internalization it was further 2-fold increased for Rab7a and only slightly for Rab7b. Post-transcriptional gene silencing of rab7a suppressed phagosome formation by 70 % and impaired their acidification. Ultrastructural analysis with double immunogold labeling revealed that this effect was due to the lack of V-ATPase recruitment to phagolysosomes. No significant phenotype changes were noticed in cells upon rab7b silencing. In conclusion, Rab7b acquired a new function, whereas Rab7a can be assigned to the phagolysosomal pathway.
Objective: Mechanism of denervation atrophy remains poorly understood. In particular, the question about irreversibility of the late atrophy is still open. Therefore, in the present study, we ...investigated whether and how a passive movement can affect a progress of atrophy in rat soleus muscle. To address this issue, a locomotor training on a treadmill was applied to rats with their right hindlimb muscles denervated.
Methods: The hindlimb muscles were denervated by cutting the sciatic nerve. Starting either 7 days or 1 month after the surgery, the animals were trained on a treadmill. Two months after denervation, the soleus muscle was investigated using light and electron microscopy and biochemical methods. Control soleus muscles were obtained from non-trained animals: the untreated and the 2-month denervated.
Results: Locomotor training caused slight increase in denervated rat soleus muscle weight and significant increase in its fiber diameter. The training positively affected some of the factors that were believed to be the reasons of atrophy irreversibility, because of significant increase in the number of capillary blood vessels and muscle fiber nuclei with the concomitant decrease in the number of severely damaged muscle fibers and amount of collagen. Morphology of the contractile apparatus was also improved as more regular organization of sarcomeres and the hexagonal arrangement of myosin filaments was evident. Moreover, the amount of myosin heavy chains (MHC) significantly increased after training. The effects were more evident in the animals with longer training.
Conclusion: Passive movement seems to attenuate some of the pathologic processes within the denervated muscle.
Background. Tinnitus is defined as a phantom auditory perception, i.e., sound experience despite the lack of acoustic stimuli in the environment. The aim of this study was to assess the state of the ...natural antioxidant barrier of a body in patients complaining about the presence of tinnitus. Material and Methods. The study included a total of 51 patients aged from 20 to 62 years with diagnosed idiopathic tinnitus and 19 healthy subjects as a control group. All patients underwent the audiometric tone test, speech audiometry, distortion otoacoustic emission product testing, study of evoked auditory potentials of short latency, and biochemical analysis of venous blood concerning values of activity or concentration of glutathione, glutathione peroxidase, S-transferase, glutathione reductase superoxide dismutase, malondialdehyde, and ceruloplasmin as the selected parameters of oxidative stress. Results. Disorders of the auditory pathway were not only limited to the cochlea but also covered its further episodes. Mean values of activity or concentration of the selected parameters of oxidative stress in the study and control groups showed reduced effectiveness of the body’s natural antioxidant barrier. Discussion. Patients complaining about the presence of tinnitus showed reduced effectiveness of the body’s natural antioxidant barrier compared to the control group. Conclusions. The main indication to undertake further research on the functioning of the antioxidant barrier in people suffering from ailments in the form of tinnitus is to determine a suitable therapy aimed at improving the quality of life of these patients, which might be the administration of antioxidant medications.
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