Comparative Characterization and Identification of Poly-3-Hydroxybutyrate Producing Bacteria with Subsequent Optimization of Polymer Yield

In this work, the strains Bacillus megaterium BM 1, Azotobacter chrocococcumAz 3, Bacillus araybhattay RA 5 were used as an effective producer of poly-3-hydroxybutyrate P(3HB). The purpose of the study was to isolate and obtain an effective producer of P(3HB) isolated from regional chestnut soils of northern Kazakhstan. This study demonstrates the possibility of combining the protective system of cells to physical stress as a way to optimize the synthesis of PHA by strains. Molecular identification of strains and amplification of the phbC gene, transmission electron microscope (TEM), extracted and dried PHB were subjected to Fourier infrared transmission spectroscopy (FTIR). The melting point of the isolated P(3HB) was determined. The optimal concentration of bean broth for the synthesis of P(3HB) for the modified type of Bacillus megaterium RAZ 3 was 20 g/L, at which the dry weight of cells was 25.7 g/L−1 and P(3HB) yield of 13.83 g/L−1, while the percentage yield of P(3HB) was 53.75%. The FTIR spectra of the extracted polymer showed noticeable peaks at long wavelengths. Based on a proof of concept, this study demonstrates encouraging results.

Green preparation and characterization of silver nanocolloids used as antibacterial material in soap

This study aimed to assess the characteristics, including morphology, physicochemical properties, and antibacterial properties, of silver nanocolloids obtained by D-glucose reduction. Silver nanoparticles were synthesized in accordance with the principles of green chemistry using D-glucose as a reductor. The obtained nanostructures were characterized by UV–vis spectroscopy, transmission electron microscopy, and dynamic light scattering. Stability tests performed after 1 month of storage revealed that the colloids prepared with and without polyvinylpyrrolidone as a stabilizer had the same properties. Distribution of the nanoparticles was tested using inductively coupled plasma mass spectrometry by doping the silver colloids into a natural soap mass. The antibacterial activity of the soap containing silver nanoparticles was tested on dirty hands. The antibacterial activity test demonstrated that the novel green soap materials improved with D-glucose-reduced silver nanoparticles possessed better antibacterial properties than a pure soap, and thus, they could be recommended for quotidian use by dermatological patients.

Dynamics of optical vortices in 2D materials

Optical vortices in planar geometries are a universal wave phenomenon, where electromagnetic waves possess topologically protected integer values of orbital angular momentum (OAM). The conservation of OAM governs their dynamics, including their rules of creation and annihilation. However, such dynamics remained so far beyond experimental reach. Here, we present a first observation of creation and annihilation of optical vortex pairs. The vortices conserve their combined OAM during pair creation/annihilation events and determine the field profile throughout their motion between these events. We utilize free electrons in an ultrafast transmission electron microscope to probe the vortices, which appear in the form of phonon polaritons in the 2D material hexagonal boron nitride. These results provide the first observation of optical vortices in any 2D material, which were predicted but never observed. Our findings promote future investigation of vortices in 2D materials and their use for chiral plasmonics, toward the control of selection rules in light-matter interactions and the creation of optical simulators of phase transitions in condensed matter physics.

Collagen-binding integrin alpha11beta1 contributes to joint destruction in arthritic hTNFtg mice

Background: In rheumatoid arthritis (RA), fibroblast-like synoviocytes (FLS) undergo a tumor-like transformation, wherein they develop an aggressive phenotype that is characterized by increased adhesion to components of cartilage extracellular matrix (ECM) and that contributes extensively to joint destruction. The collagen-binding integrin alpha11beta1 was previously shown to be involved in similar processes in cancer-associated fibroblasts mediating tumorigenicity and metastasis in certain tumors. Therefore, this study aimed to study the role of integrin alpha11beta1 in RA and to characterize the effects of alpha11beta1 deficiency on the disease course and severity in arthritic hTNFtg mice. Methods: The expression levels of integrin alpha11beta1 were analyzed by immunohistochemistry, immunofluorescence, and western blot analysis in synovial samples and FLS of patients with RA and osteoarthritis (OA) as well as in samples from wild type (wt) and arthritic hTNFtg mice. Furthermore, the subcellular expression of integrin alpha11beta1 was investigated in co-culture experiments with cartilage explants and analyzed by transmission electron microscopy. To investigate the effects of integrin alpha11beta1 deficiency, itga11-/- mice were interbred with hTNFtg mice and disease severity was assessed by clinical scoring of grip strength and paw swelling over the disease course. Hind paws of 12-weeks-old mice of all genotypes were analyzed by uCT imaging followed by stainings of paraffin-embedded tissue sections with Toluidine-blue and tartrate-resistant acid phosphatase (TRAP) to evaluate established parameters of joint destruction such as inflammation area, cartilage destaining, FLS attachment to the cartilage surface, and bone damage. Results: Expression levels of integrin alpha11beta1 were clearly elevated in synovial tissues and FLS from RA patients and hTNFtg mice, compared to the controls derived from OA patients and wt mice. Interestingly, this expression was shown to be particularly localized in focal adhesions of the FLS. As revealed by transmission electron microscopy, integrin alpha11beta1 expression was particularly evident in areas of direct cellular contact with the ECM of cartilage. Evaluations of clinical scorings and histomorphological analyses demonstrated that itga11-/-hTNFtg displayed alleviated clinical symptoms, higher bone volume, less cartilage destruction, and reduced FLS attachment to the cartilage in comparison to hTNFtg mice. Conclusions: The collagen-binding integrin alpha11beta1 is upregulated in the context of RA and its deficiency in mice with an inflammatory hTNFtg background leads to a significant reduction in the arthritic phenotype which makes integrin alpha11beta1 an interesting target for therapeutical intervention.

Mechanical Activation of Smectite-Based Nanocomposites for Creation of Smart Fertilizers

This research presents the mechanical creation of smart fertilizers from a mixture of smectite and urea in a 3:2 ratio by using the planetary milling technique. The smectite–urea composites show intercalation between urea and mineral, which increases steadily with increasing activation time. A shift of X-Ray Diffraction basal reflections, intensities of Fourier transform infrared spectroscopy (FTIR) peaks, and weight losses in thermogravimetric analysis (TG) document the systematic crystallo-chemical changes of the composites related to nitrogen interaction with activation. Observations of the nanocomposites by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) corroborate the inference. Nitrogen intercalates with smectite in the interlayer space and remains absorbed either within micro-aggregates or on the surface of activated smectites. Soil leaching tests reveal a slower rate of nitrogen than that of traditional urea fertilizers. Different forms of nitrogen within the composites cause their differential release rates to the soil. The formulated nanocomposite fertilizer enhances the quality and quantity of oat yield.

Novel Data About Unusual Location of Toxoplasma Gondii Within Active Microbial Community in the Anterior Chamber of the Eye of Egyptian Rural Children

In Egypt, many cases of granulomatous anterior uveitis consisting of single or multiple gelatinous nodules were detected in children living in rural areas. These lesions are believed to be waterborne and were previously attributed to flatworms ‘stage, showing some improvement after antiparasitic treatment. In a trial to explore the nature of these ocular lesions among rural Egyptian children, twenty surgically excised ocular lesions were subjected to transmission electron microscopy (TEM) examination. TEM results were combined with previous results of the metagenomic analysis performed for four cases out of the twenty samples, revealing the presence of Toxoplasma gondii (T. gondii), besides, a wide range of microbial communities, including variable species of fungi, bacteria, and archaea. The excised lesions ranged from 1 to 5 mm in size and demonstrated an extensive inflammatory cellular infiltrate. Using TEM, five out of twenty samples revealed active eukaryotic organisms with intact energetic cellular organelles, besides, numerous nuclei encircled within a syncytial layer and enclosed by a hyaline layer rich in mitochondria. Six samples showed inactivity in the cellular and the covering portions, while just inflammatory reaction was seen in the remaining nine samples. Toxoplasma gondii was found free within the distal part of the syncytium while, the proximal part showed the active synthesis of possibly extra polymeric substance, perhaps secreted by the microbial community. In a conclusion, Toxoplasma gondii has been detected among a microbial community in an atypical lesion in the eye. Further studies need to be sustained on genotype characterization, proteomic analysis, besides, the aquatic transmission of these mixed microbial species to the ocular tissues to clarify the reason behind such ocular illness.

Bisphenol-A Exposure Induced Neurotoxicity and Synapse and Cytoskeleton Dysfunction in Neuro-2a Cells

Bisphenol A (BPA) is one of the typical environmental endocrine disruptors. BPA was leached from polycarbonate containers into food and water, and it has been detected in collective samples from humans. Microtubule-associated protein 2 (MAP2) and Tau maintain microtubule normal function and promote the normal development of the nervous system. Synaptophysin (SYP) and drebrin (Dbn) proteins are involved in regulating synaptic plasticity. This study aimed to determine the adverse effects of BPA on Neuro-2a cells by investigating the synaptic and cytoskeletal damage. Cells were exposed to 0 (Minimum Essential Medium, MEM), 0.01% (v/v) DMSO and 150 µM BPA for 12, 24, or 36 h. Morphological analysis revealed that the cells in the BPA-treated groups shrank, collapsed, and had a reduced number of synapses compared with those in the control groups. CCK-8 and LDH assays showed that the mortality of Neuro-2a cells increased as the BPA treatment time was prolonged. Transmission electron microscopic analysis further revealed that cells demonstrated nucleolar swelling and nuclear membrane and partial mitochondrial dissolution or condensation following BPA exposure. BPA also significantly decreased the relative protein expression levels of MAP2, Tau, and Dbn (P < 0.01). Interestingly, the relative protein expression levels of SYP increased (P < 0.01). These results indicated that BPA damaged the development and proliferation of Neuro-2a cells by disrupting cytoskeleton and synaptic integrity.

Synthesis, Characterization, and Biodistribution of GdF3:Tb3+@RB Nanocomposites

Herein we report the development of a nanocomposite for X-ray-induced photodynamic therapy (X-PDT) and computed tomography (CT) based on PEG-capped GdF3:Tb3+ scintillating nanoparticles conjugated with Rose Bengal photosensitizer via electrostatic interactions. Scintillating GdF3:Tb3+ nanoparticles were synthesized by a facile and cost-effective wet chemical precipitation method. All synthesized nanoparticles had an elongated “spindle-like” clustered morphology with an orthorhombic structure. The structure, particle size, and morphology were determined by transmission electron microscopy (TEM), X-ray diffraction (XRD), and dynamic light scattering (DLS) analysis. The presence of a polyethylene glycol (PEG) coating and Rose Bengal conjugates was proved by Fourier-transform infrared spectroscopy (FTIR), thermogravimetric analysis (TG), and ultraviolet–visible (UV-vis) analysis. Upon X-ray irradiation of the colloidal PEG-capped GdF3:Tb3+–Rose Bengal nanocomposite solution, an efficient fluorescent resonant energy transfer between scintillating nanoparticles and Rose Bengal was detected. The biodistribution of the synthesized nanoparticles in mice after intravenous administration was studied by in vivo CT imaging.

Mechanism of a Volatile Organic Compound (6-Methyl-2-Heptanone) Emitted From Bacillus subtilis ZD01 Against Alternaria solani in Potato

The antagonistic mechanisms of soluble non-volatile bioactive compounds, such as proteins and lipopeptides emitted from Bacillus have been widely studied. However, there are limited studies on the antifungal mechanisms of volatile organic compounds (VOCs) produced by Bacillus against plant fungal diseases. In this study, the antagonistic mechanisms of one specific VOC, 6-methyl-2-heptanone, against Alternaria solani were investigated. To optimize the extraction conditions of headspace solid-phase microextraction, a 50/30-μm divinylbenzene/carboxen/polydimethylsiloxane fiber at 50°C for 40 min was used. For gas chromatography-mass spectrometry using a free fatty acid phase capillary column, 6-methyl-2-heptanone accounted for the highest content, at 22.27%, of the total VOCs from Bacillus subtilis ZD01, which inhibited A. solani mycelial growth strongly in vitro. Therefore, 6-methyl-2-heptanone was selected as the main active chemical to elucidate the action mechanisms against A. solani. Scanning and transmission electron microscopy analyses revealed that after exposure to an EC50 dose of 6-methyl-2-heptanone, A. solani hyphal cells had a wide range of abnormalities. 6-Methyl-2-heptanone also caused the capture of cellular fluorescent green label and the release of adenosine triphosphate (ATP) from outer membranes A. solani cells, which may enhance 6-methyl-2-heptanone ability to reach the cytoplasmic membrane. In addition, 6-methyl-2-heptanone showed strong inhibitory effect on A. solani conidial germination. It also damaged conidial internal structures, with the treated group having collapsed shrunken small vesicles as observed by transmission electron microscopy. Because 6-methyl-2-heptanone showed strong effects on mycelial integrity and conidial structure, the expression levels of related pathogenic genes in A. solani treated with 6-methyl-2-heptanone were investigated. The qRT-PCR results showed that transcriptional expression levels of slt2 and wetA genes were strongly down-regulated after exposure to 6-methyl-2-heptanone. Finally, because identifying the functions of pathogenic genes will be important for the biological control of A. solani, the wetA gene was identified as a conidia-associated gene that plays roles in regulating sporulation yield and conidial maturation. These findings provide further insights into the mechanisms of VOCs secreted by Bacillus against A. solani.