Electron-dense Core Research Articles

Three-Dimensional Ultrastructure of Flower-Spray Nerve Endings in the Rat Carotid Sinus.

The flower-spray nerve endings are afferent nerve terminals in the carotid sinus that arise from carotid sinus nerve of glossopharyngeal nerve. However, the three-dimensional ultrastructural characteristics of flower-spray nerve endings and spatial relationships between the terminal parts and other cellular elements have not been fully understood. To elucidate their detailed relationship, backscattered electron imaging of serial sections was performed with a scanning electron microscope to produce a three-dimensional reconstruction of the flower-spray endings. The terminal parts of flower-spray endings were distributed horizontally approximately 5µm outside the external elastic membrane in the tunica adventitia of the internal carotid artery. The three-dimensional reconstruction showed that the terminal parts of flower-spray endings were flat with irregular contours and were partially covered by the thin cytoplasmic processes of Schwann cells. The complex consisting of the nerve terminals and associated Schwann cells was surrounded by a multilayered basement membrane. The terminal parts of the endings were also surrounded by fibroblasts with elastic fibers and collagen fibrils. Secretory vesicles without an electron-dense core were observed in the terminal parts of the endings. The accumulation of vesicles just below the axonal membrane was observed in terminal parts not covered by Schwann cell cytoplasmic processes on both the luminal and basal sides. Swollen mitochondria, concentric membranous structures, and glycogen granule-like electron-dense materials were often noted in some of the terminal parts of the endings and the parent axon. Collectively, the present results suggest that flower-spray endings are baroreceptors because their morphology was similar to other mechanoreceptors. Furthermore, flower-spray endings may be affected by glutamate secreted in an autocrine manner.

Free zinc determines the formability of the vesicular dense core in diabetic beta cells.

During the progression of type 2 diabetes, total body zinc deficiency disrupts the formability of the electron-dense core in beta-cell vesicles, but the mechanism is unclear. Using fluorescence imaging, transmission electron microscopy and pharmacokinetics assays, we established a strong link between an increasing concentration of free zinc and the formability enhancement of the dense core electron density. Thus, our results highlight a mechanism by which zinc supplementation enhances the maturation of dense cores and restores the secretion of insulin in two diabetic mouse models both in vitro and in vivo. This study provides a potential research direction for investigating the etiology and nutrition of zinc in the management of type 2 diabetes.

A novel herpes-like virus inducing branchial lesions in a tiger shark (Galeocerdo cuvier).

A juvenile, male tiger shark (Galeocerdo cuvier) developed illness after capture in Florida waters and was euthanized. Gross lesions included mild skin abrasions, hepatic atrophy, and coelomic fluid. Histologically, gills contained multifocal lamellar epithelial cell necrosis and thromboses. Scattered gill and esophageal epithelial cells had large, basophilic, intracytoplasmic, and intranuclear inclusions. Ultrastructurally, lamellar epithelial cells contained arrays of intracytoplasmic viral particles and scattered intranuclear nucleocapsids. Capsulated virions were 148 ± 11 nm with an 84 ± 8 nm icosahedral nucleocapsid and an electron-dense core. Next-generation sequencing, quantitative polymerase chain reaction, and in situ hybridization performed on formalin-fixed tissue confirmed a herpes-like viral infection. The viral polymerase shared 24% to 31% protein homology with other alloherpesviruses of fish, indicating a divergent virus. This report documents the pathologic findings associated with a molecularly confirmed novel herpes-like virus in an elasmobranch.

Structural heterogeneity of cellular K5/K14 filaments as revealed by cryo-electron microscopy.

Keratin intermediate filaments are an essential and major component of the cytoskeleton in epithelial cells. They form a stable yet dynamic filamentous network extending from the nucleus to the cell periphery, which provides resistance to mechanical stresses. Mutations in keratin genes are related to a variety of epithelial tissue diseases. Despite their importance, the molecular structure of keratin filaments remains largely unknown. In this study, we analyzed the structure of keratin 5/keratin 14 filaments within ghost mouse keratinocytes by cryo-electron microscopy and cryo-electron tomography. By averaging a large number of keratin segments, we have gained insights into the helical architecture of the filaments. Two-dimensional classification revealed profound variations in the diameter of keratin filaments and their subunit organization. Computational reconstitution of filaments of substantial length uncovered a high degree of internal heterogeneity along single filaments, which can contain regions of helical symmetry, regions with less symmetry and regions with significant diameter fluctuations. Cross-section views of filaments revealed that keratins form hollow cylinders consisting of multiple protofilaments, with an electron dense core located in the center of the filament. These findings shed light on the complex and remarkable heterogenic architecture of keratin filaments, suggesting that they are highly flexible, dynamic cytoskeletal structures.

The Alpha-Granule Release Reaction Is Mediated By SNARE-Dependent Solubilization of the Granule Matrix

Within their electron dense granule matrix, alpha-granules package a wide range of proteins important to hemostasis, thrombosis, inflammation, and angiogenesis. During the platelet release reaction, these proteins are secreted in a kinetically diverse manner that is proportional to agonist stimulation. The release reaction is accompanied by a large expansion in granule volume, matrix decondensation, and the solubilization of the included proteins. We have previously posited that some combination of differential protein proximity to granule exit sites and protein solubilization could contribute to the diversity of protein release.Here we have used thrombin stimulation and SNARE knockout mice to test two possible mechanisms for granule decondensation/matrix solubilization. In the first, granule swelling and decondensation precedes fusion with the plasma membrane and is regulated by aquaporin-mediated water influx presumably driving the volume change. In the second, matrix solubilization is plasma membrane fusion dependent with the extracellular influx of water through the fusion pore driving volume change and granule decondensation.In wild-type mouse platelets, we found a precursor-product relationship between the incidence of condensed and decondensed granules with the incidence of decondensed granules peaking at 90 sec post-stimulus (0.1 U/mL thrombin). The subsequent decrease in decondensed granules was due to an increasing incidence of compound granule fusion. By electron tomography, granules were connected to the plasma membrane by “necks” or short “pipes”. The granule membrane remained physically distinct during the entire granule decondensation process. Granule decondensation was marked by the conversion of the granule matrix into a diffuse fibrous structure that decreased progressively in electron density over the 5 min stimulation period. A residual electron-dense core was most obvious at 90 sec post stimulation. At the plasma membrane fused granule neck/pipe, solubilized granule matrix fibers were observed to be extruded extracellularly.Overall, the data suggest a progressive solubilization of granule matrix contents and their accompanying extrusion through the granule neck/pipe. In SNARE knockout mouse platelets, SNARE-selective inhibition of matrix solubilization/granule decondensation was observed. Both qualitatively and quantitatively, knockout of VAMP2/3 had little-to-no effect on 1) alpha-granule fusion with the plasma membrane, 2) granule decondensation, 3) matrix solubilization, or 4) compound granule fusion. In striking contrast, VAMP8 knockout initially produced a substantial inhibition of the incidence of all four steps in the granule releasate reaction while a VAMP2/3/8 knockout almost totally inhibited all four steps. In sum, our data strongly indicate that alpha-granule decondensation and matrix protein solubilization/extrusion are highly dependent on SNARE-mediated granule fusion with the plasma membrane. These data provide no support for the alternative mechanism that regulated aquaporin-dependent water influx mediates granule decondensation prior to granule fusion with the plasma membrane or canalicular system.This work was supported in part by NIH grant R01 HL 119393 to BS, NIH grant R01 HL56652 to SWW, and AHA grant 15PRE2555020 to SJ. DisclosuresNo relevant conflicts of interest to declare.

Snapshots into carbon dots formation through a combined spectroscopic approach

The design of novel carbon dots with ad hoc properties requires a comprehensive understanding of their formation mechanism, which is a complex task considering the number of variables involved, such as reaction time, structure of precursors or synthetic protocol employed. Herein, we systematically investigated the formation of carbon nanodots by tracking structural, chemical and photophysical features during the hydrothermal synthesis. We demonstrate that the formation of carbon nanodots consists of 4 consecutive steps: (i) aggregation of small organic molecules, (ii) formation of a dense core with an extended shell, (iii) collapse of the shell and (iv) aromatization of the core. In addition, we provide examples of routes towards tuning the core-shell design, synthesizing five novel carbon dots that all consist of an electron-dense core covered by an amine rich ligand shell.

Light and electron microscopy imaging unveils new aspects of the antiviral capacity of silver nanoparticles in bunyavirus-infected cells

Drug repurposing is an important source of new antivirals because many compounds used to treat a variety of pathologies also hamper viral infections. Habitually, silver nanoparticles (AgNPs) have been used to treat bacterial and fungal infections and their antiviral properties have been also reported. In this work, we have studied the antiviral capacity of AgNPs in cells infected with Bunyamwera virus (BUNV), the prototype of the Bunyavirales order. This group of viruses contains important pathogens for humans, animals and plants. Incubation of BUNV-infected Vero cells with non-toxic concentrations of AgNPs, reduced the production of extracellular infectious viruses in up to three orders of magnitude. With a combination of imaging techniques, we have visualized the intracellular distribution of AgNPs in mock- and BUNV-infected cells and studied their effects on intracellular organelles. In mock-infected cells and at short times post-incubation, AgNPs were detected inside nuclei and mitochondria by transmission electron microscopy (TEM). At long times post-treatment, they accumulated inside lysosome-like organelles. Cell compartments did not exhibit any appreciable ultrastructural alterations after incubation with AgNPs. In BUNV-infected cells, AgNPs attached to extracellular virions, that showed a disrupted morphology. Inside cells, they were detected inside the nucleus, in mitochondria and around characteristic Golgi-associated, single-membrane spherules. These membranous structures are the replication organelles (ROs) of bunyaviruses and contain active viral replication complexes (VRCs). Compared to normal spherules that are round, compact and have an electron-dense core, spherules in AgNPs-treated cells were deformed and their core was electron-lucent. Interestingly, in BUNV-infected cells treated with the typical antiviral ribavirin (RBV), spherules with VRCs exhibit also an anomalous morphology and an electron-lucent core. Both AgNPs and RBV might interfere with BUNV-induced dismantling of cell nucleoli and with the intercellular propagation of large groups of virions, a mechanism of BUNV transmission observed for the first time in cultured cells. Our results point to silver nanoparticles as good candidates for antiviral therapy, either alone or in combination with other antiviral drugs, such as RBV-related compounds.

Morphologic, cytochemical, and ultrastructural features of gray eosinophils in nine cats.

Gray eosinophils, resembling those in sighthound dog breeds, have not previously been reported in cats. We aimed to provide a morphologic, cytochemical, and ultrastructural description of gray eosinophils in cats. Blood films examined as part of routine hematology profiles in cats from May 2015 to July 2018 were evaluated for the presence of gray eosinophils. When identified with modified Wright stain, cells were morphologically assessed and additionally stained with Diff-Quik, ALP, Luna, and Luxol fast blue stains and compared with feline controls. Two cases were prepared for transmission electron microscopy (TEM) and compared with a feline control. Gray eosinophils were identified in 9 of 2641 cats during the study period. Compared with typical feline eosinophils, these cells contained abundant round granules instead of the characteristic rod-shaped specific granules. These granules lacked the characteristic intense pink/red staining with Romanowsky stains and did not stain with ALP, Luna, or Luxol fast blue stains. On TEM, the classical electron-dense core of these granules was replaced by a core with fragmented or amorphous internal material. Typical eosinophils were not identified in any cat in which gray eosinophils were identified. The distinct morphologic, cytochemical, and ultrastructural changes in gray feline eosinophils might be associated with a reduction or lack of major basic protein (MBP) in specific granule cores. Similar to canine gray eosinophils, accurate recognition of these cells is essential to prevent their misclassification as toxic neutrophils.

Fluorescent platinum nanoclusters as correlative light electron microscopy probes.

Correlative Imaging (CI) visualizes a single sample/region of interest with two or more imaging modalities. The technique seeks to elucidate information that may not be discernible by using either of the constituent techniques in isolation. Correlative Light Electron Microscopy (CLEM) can be employed to streamline workflows, i.e., using fluorescent signals in the light microscope (LM) to inform the user of regions which should be imaged with electron microscopy (EM). The efficacy of correlative techniques requires high spatial resolution of signals from both imaging modalities. Ideally, a single point should originate from both the fluorescence and electron density. However, many of the ubiquitously used probes have a significant distance between their fluorescence and electron dense portions. Furthermore, electron dense metal nanoparticles used for EM visualization readily quench any proximal adjacent fluorophores. Therefore, accurate registration of both signals becomes difficult. Here we describe fluorescent nanoclusters in the context of a CLEM probe as they are composed of several atoms of a noble metal, in this case platinum, providing electron density. In addition, their structure confers them with fluorescence via a mechanism analogous to quantum dots. The electron dense core gives rise to the fluorescence which enables highly accurate signal registration between epifluorescence and electron imaging modalities. We provide a protocol for the synthesis of the nanoclusters with some additional techniques for their characterization and finally show how they can be used in a CLEM set up.

First Evidence of Carp Edema Virus Infection of Koi Cyprinus carpio in Chiang Mai Province, Thailand.

The presence of carp edema virus (CEV) was confirmed in imported ornamental koi in Chiang Mai province, Thailand. The koi showed lethargy, loss of swimming activity, were lying at the bottom of the pond, and gasping at the water’s surface. Some clinical signs such as skin hemorrhages and ulcers, swelling of the primary gill lamella, and necrosis of gill tissue, presented. Clinical examination showed co-infection by opportunistic pathogens including Dactylogyrus sp., Gyrodactylus sp. and Saprolegnia sp. on the skin and gills. Histopathologically, the gill of infected fish showed severe necrosis of epithelial cells and infiltrating of eosinophilic granular cells. Electron microscope examination detected few numbers of virions were present in the cytoplasm of gill tissue which showed an electron dense core with surface membranes worn by surface globular units. Molecular detection of CEV DNA from gill samples of fish was performed by polymerase chain reaction (PCR) and confirmed by nested-PCR. Phylogenetic analyses revealed that CEV isolate had 99.8% homology with the CEV isolated from South Korea (KY946715) and Germany (KY550420), and was assigned to genogroup IIa. In conclusion, this report confirmed the presence of CEV infection of koi Cyprinus carpio in Chiang Mai province, Thailand using pathological and molecular approaches.

Identification of putative viroplasms within banana cells infected by banana streak MY virus.

The badnavirus replication cycle is poorly understood and most knowledge is based on extrapolations from model viruses such as Cauliflower mosaic virus (CaMV). However, in contrast to CaMV, badnaviruses are thought not to produce viroplasms and therefore it has been a mystery as to where virion assembly occurs. In this study, ultrathin sections of a banana leaf infected with a badnavirus, banana streak MY virus (BSMYV), were examined by transmission electron microscopy. Electron-dense inclusion bodies (EDIBs) were sporadically distributed in parenchymatous tissues of the leaf, most commonly in the palisade and spongy mesophyll cells. These EDIBs had a characteristic structure, comprising an electron-dense core, a single, encircling lacuna and an outer ring of electron-dense material. However, much less frequently, EDIBs with two or three lacunae were observed. In the outer ring, densely packed virions were visible with a shape and size consistent with that expected for badnaviruses. Immunogold labelling was done with primary antibodies that detected the N-terminus of the capsid protein and strong labelling of the outer ring but not the central core or lacuna was observed. It is concluded that the EDIBs that were observed are equivalent in function to the viroplasms of CaMV, although obviously different in composition as there is not a paralogue of the transactivation/viroplasm protein in the badnavirus genome. It is postulated that production of a viroplasm could be a conserved characteristic of all members of the Caulimoviridae.

Structural and bio-functional assessment of the postaxillary gland in Nidirana pleuraden (Amphibia: Anura: Ranidae)

BackgroundOwing to their incomplete adaptation to the terrestrial environment, amphibians possess complex cutaneous glandular systems. The skin glands not only regulate water loss and respiratory gas and salt exchange, but are also involved in defense against predators and microorganisms, social communication, and reproduction. These glands are distributed throughout the integument, but can accumulate in specific regions, forming visible outgrowths known as macroglands. Some macroglands are sexually dimorphic and mediate intersexual communication and reproductive success. The postaxillary gland is a sexually dimorphic macrogland in Nidirana pleuraden. Its biological function and its morphological and histochemical characteristics are unclear. In the present study, we describe the structure and ultrastructure of the postaxillary gland, and explore its main function.ResultsThe postaxillary gland has a thinner epidermis than the dorsal region of N. pleuraden. In addition to ordinary serous glands (OSG), type I and II mucous gland (I MG & II MG), a type of specialized mucous gland (SMG) is also found to constitute the postaxillary gland. The SMG is larger than other gland types, and consists of high columnar mucocytes with basal nuclei arranged radially toward a lumen. SMGs are positive to periodic acid-Schiff stain and stained blue in Masson’s trichrome stain. A discontinuous myoepithelial sheath lacking innervation encircles SMG mucocytes, and the outlets of such glands are X- or Y-shaped. Transmission electron microscopy reveals abundant secretory granules in SMG, which are biphasic, composed of an electron-opaque outer ring and a less electron-dense core. Lipid droplets, and organelles, such as rough endoplasmic reticulum and Golgi stacks, are located in the supranuclear cytoplasm of the mucocytes in SMG. Female N. pleuraden exhibits chemotaxis toward homogenate of the postaxillary gland, but male does not. On treatment with trypsin, this sexual attraction disappears.ConclusionsThe postaxillary gland of N. pleuraden is a male-specific macrogland that consists primarily of SMGs, together with OSGs, I MGs and II MGs. Other than their extremely large size, SMGs structurally and histochemically resemble many reported specialized gland types in amphibian sexually dimorphic skin glands. Secretions of the postaxillary gland are proteinaceous sexual pheromones, which are believed to attract females at male calling intermissions.

Seasonal emergence of benign epidermal tumor in farm-reared adult grass carp (Ctenopharyngodon idella) caused by lymphocystis disease virus at Uttarakhand, India

From 2017‐2019, during the winter months (November–January), epidermal papilloma like benign tumor was observed in farm-reared adult grass carp, Ctenopharyngodon idella at the northern part of India. An investigation was undertaken to establish the causative agent of this epizootiology. Based on the clinical signs and ineffectiveness of antibiotics to control this disease, viral infection was suspected. A primer was designed to PCR amplify the partial DNA polymerase gene of lymphocystis disease virus (LCDV) from the tumor of moribund grass carp, which yielded 460 bp amplicon. After sequencing this 460 bp amplicon, the viral agent from the tumor of grass carp was identified as lymphocystis disease virus and given strain designation LRI-18 (Genbank accession number-MK347473). Phylogenetic analysis based on the nucleotide sequence of DNA polymerase gene of LCDV revealed the close association of LCDV LRI-18 to LCDV strain from Israel with 99.0% and 96.5% nucleotide and amino acid sequence identity, respectively. LCDV LRI-18 form single (type A; 0.5–2.0 μm) or multiple (type B; 0.5–2.0 μm) intracytoplasmic inclusion bodies on lymphocystis and erythrocyte cells of infected grass carp. Transmission Electron Microscopy (TEM) of blood revealed the presence of enveloped icosahedral virions of approximately 280 nm in diameter with electron dense core and an outer capsid. Histopathological study of various organs of LCDV infected grass carp revealed extensive hypertrophy and fusion of secondary gill lamellae, focal hemorrhage in the eye, pyknotic nuclei in the liver, inflammation in the myocardium, fused villi of intestine and degeneration of the glomeruli in the kidney. Experimental infection by a dip to grass carp fingerlings (average body weight 4.4 ± 0.9 g) using 0.45 μm filtered LCDV crude suspension did not reproduce the disease, and also no mortality were observed. This is the first report of the occurrence of lymphocystis disease in grass carp in India.

Avian coxiellosis in nine psittacine birds, one black-browed barbet, and one paradise tanager

ABSTRACTInfection with a novel species of the genus Coxiella was first described in three Swainson’s blue mountain rainbow lorikeets from a zoological collection, and days later in a group of seven other psittacine birds and a toucan. We provide an update on coxiellosis in nine additional psittacines, and two non-psittacines. Psittacines originated in New England, the mid Atlantic, the Midwest, the South, and the Northwest. Psittacines most commonly had lesions in the brain, spleen, liver, and lungs, consisting of meningoencephalitis, hepatosplenomegaly, and interstitial pneumonia. Lesions contained histiocytic infiltrate, with intracytoplasmic, Gimenez-positive coccobacilli. Transmission electron microscopy revealed bacteria with trilaminar cell walls, electron dense cores, and spore-like forms. PCR revealed homology to the organism in index cases. In addition, one black-browed barbet and one paradise tanager were found with systemic coxiellosis; sequencing identified the same pathogen. These are the second piciforme and the first passerine affected by this pathogen, indicating expanded infectivity and pathogenicity.RESEARCH HIGHLIGHTSReport of coxiellosis in nine psittacines; lesions often in brain, spleen, liver, lung.Second piciforme with coxiellosis, a black-browed barbet.First case of avian coxiellosis described in a passerine, a paradise tanager.

Core-shell nanospheres behind the blue eyes of the bay scallop Argopecten irradians.

The bay scallop Argopecten irradians (Mollusca: Bivalvia) has dozens of iridescent blue eyes that focus light using mirror-based optics. Here, we test the hypothesis that these eyes appear blue because of photonic nanostructures that preferentially scatter short-wavelength light. Using transmission electron microscopy, we found that the epithelial cells covering the eyes of A. irradians have three distinct layers: an outer layer of microvilli, a middle layer of random close-packed nanospheres and an inner layer of pigment granules. The nanospheres are approximately 180 nm in diameter and consist of electron-dense cores approximately 140 nm in diameter surrounded by less electron-dense shells 20 nm thick. They are packed at a volume density of approximately 60% and energy-dispersive X-ray spectroscopy indicates that they are not mineralized. Optical modelling revealed that the nanospheres are an ideal size for producing angle-weighted scattering that is bright and blue. A comparative perspective supports our hypothesis: epithelial cells from the black eyes of the sea scallop Placopecten magellanicus have an outer layer of microvilli and an inner layer of pigment granules but lack a layer of nanospheres between them. We speculate that light-scattering nanospheres help to prevent UV wavelengths from damaging the internal structures of the eyes of A. irradians and other blue-eyed scallops.

New data on spermiogenesis and trepaxonematan axoneme in basal tapeworms (Cestoda, Caryophyllidea, Lytocestidae) parasitizing cyprinid fishes

Monozoic caryophyllidean cestodes, intestinal parasites of cyprinid fishes, represent a group of tapeworms with an unclear evolutionary history. As spermatology may provide phylogenetically important data, the spermiogenesis and ultrastructure of the mature spermatozoon have been investigated using an integrative approach combining transmission electron microscopy, cytochemistry and electron tomography in Khawia rossittensis (Szidat, 1937). The process of spermatid formation is accompanied by the presence of ultrastructural characters not described in traditional models of spermiogenesis, e.g., apical electron-dense material, the two striated roots situated unusually opposite each other, branching of typical striated roots, an intercentriolar body comprising five electron-dense and four electron-lucent layers, rotation of both free flagella and flagellar buds to the median cytoplasmic process at 90°, and a complete proximodistal fusion. The synchronous rotation of both flagellar buds and growing free flagella is an evolutionarily linked pattern favouring the hypothesis that the Caryophyllidea are not ancestral but are secondarily derived from polyzoic forms. Electron tomography analysis has revealed a unique feature of two helicoidal tubular structures in the central electron-dense core of the axoneme of mature spermatozoon. These data provide new insights into the architecture of the 9 + ‘1’ axoneme, which is shared by male gametes of all trepaxonematan Platyhelminthes.

Targeted Derivation of Organotypic Glucose- and GLP-1-Responsive β Cells Prior to Transplantation into Diabetic Recipients.

SummaryGeneration of functional β cells from pluripotent sources would accelerate diagnostic and therapeutic applications for diabetes research and therapy. However, it has been challenging to generate competent β cells with dynamic insulin-secretory capacity to glucose and incretin stimulations. We introduced transcription factors, critical for β-cell development and function, in differentiating human induced pluripotent stem cells (PSCs) and assessed the impact on the functionality of derived β-cell (psBC) progeny. A perifusion system revealed stepwise transduction of the PDX1, NEUROG3, and MAFA triad (PNM) enabled in vitro generation of psBCs with glucose and GLP-1 responsiveness within 3 weeks. PNM transduction upregulated genes associated with glucose sensing, insulin secretion, and β-cell maturation. In recipient diabetic mice, PNM-transduced psBCs showed glucose-responsive insulin secretion as early as 1 week post transplantation. Thus, enhanced pre-emptive β-cell specification of PSCs by PNM drives generation of glucose- and incretin-responsive psBCs in vitro, offering a competent tissue-primed biotherapy.

A green chemistry to produce iron oxide – Chitosan nanocomposite (CS-IONC) for the upgraded bio-restorative and pharmacotherapeutic activities - Supra molecular nanoformulation against drug-resistant pathogens and malignant growth

The logical research on fundamentally adjusted iron oxide nanoparticles has turned out to expanded in biomedicine because of the improved activity and best biocompatibility. In this present work upgraded bio-restorative and pharmacotherapeutic property of chitosan‑iron oxide nanocomposite, which was set up by eco-friendly in situ substance technique. Characterisation of the synthesised nanocomposite by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), x-ray diffraction,(XRD) and Vibrating test magnetometer (VSM) studies reveals that highly stable spherical, electron-dense core shelled rough particles of 50-60 nm. Particle morphology of the synthesised nanocomposite utilising scanning electron microscopy (SEM) uncovers spherical; thick electron centre shelled harsh particles with the size scope of 50-60 nm. FTIR studies show that the specific interaction of practical gatherings of chitosan with iron oxide nanoparticles. Crystalline phase and magnetisation impact of the composite resolved from XRD and VSM studies. Anti-bacterial activity of the nanocomposite examined against human bacterial pathogens which suggest that the readied nanocomposite successfully restrained the development of the tried bacterial strains by recording maximum zone of inhibition, least minimum inhibition concentration (MIC) and biofilm damage against the both tested strains. 100 μg dosages of nanocomposites recorded 20.0 and 21.0 mm of the zone of inhibition against E. coli and S. aureus respectively. Biofilm restraint was additionally observed to be high in nanocomposite treatment by recording lower optical density of ethanol solubilised biofilm of both tested strains. Anticancer activity was examined against the A549 cell line by the assurance of cell feasibility as opposed to oxidative proteins, articulation example of TNF-α, Bax, PARP qualities and apoptosis. Composite prompted 50% of cytotoxicity at 80 μg/mL unmistakably uncovers cytotoxicity against A549 cells. Nanocomposite treatment revealed a high decrease of cell feasibility at all the fixation and most extreme impact seen in 100 μg. Nanocomposite treated cells demonstrated striking changes in cell morphology, the build-up of atomic material related to trademark changes in against oxidative enzymes, quality articulation design which brought about apoptosis-like necrotic cell death. The present findings would propose the conceivable usage of chitosan‑iron oxide nanocomposite as a viable remedial against safe medication pathogens and malignant growth cells.

Cadmium induces reactive oxygen species-dependent pexophagy in Arabidopsis leaves.

Cadmium treatment induces transient peroxisome proliferation in Arabidopsis leaves. To determine whether this process is regulated by pexophagy and to identify the mechanisms involved, we analysed time course-dependent changes in ATG8, an autophagy marker, and the accumulation of peroxisomal marker PEX14a. After 3hr of Cd exposure, the transcript levels of ATG8h, ATG8c, a, and i were slightly up-regulated and then returned to normal. ATG8 protein levels also increased after 3hr of Cd treatment, although an opposite pattern was observed in PEX14. Arabidopsis lines expressing GFP-ATG8a and CFP-SKL enabled us to demonstrate the presence of pexophagic processes in leaves. The Cd-dependent induction of pexophagy was demonstrated by the accumulation of peroxisomes in autophagy gene (ATG)-related Arabidopsis knockout mutants atg5 and atg7. We show that ATG8a colocalizes with catalase and NBR1 in the electron-dense peroxisomal core, thus suggesting that NBR1 may be an autophagic receptor for peroxisomes, with catalase being possibly involved in targeting pexophagy. Protein carbonylation and peroxisomal redox state suggest that protein oxidation may trigger pexophagy. Cathepsine B, legumain, and caspase 6 may also be involved in the regulation of pexophagy. Our results suggest that pexophagy could be an important step in rapid cell responses to cadmium.

FAM81A protein, a novel component of the postsynaptic density in adult brain

Analysis of affinity-purified PSD-95 complexes had previously identified a ‘hypothetical protein’, product of the gene FAM81A [1]. The present study examined the tissue and subcellular distribution of FAM81A protein and its expression levels during development. Comparison of different organs indicates selective expression of FAM81A protein in brain. FAM81A is expressed late in development, with a post-natal gradual increase in brain levels that parallels the expression of PSD-95. Comparison of subcellular fractions from adult brain shows that the distribution of FAM81A protein is similar to that of PSD-95, with a drastic enrichment in the postsynaptic density fraction. Immuno-electron microscopy of adult brain tissue reveals specific immunogold labeling for FAM81A protein at postsynaptic densities in the forebrain. The label for FAM81A protein is concentrated at the cytoplasmic edge of the electron-dense core of the postsynaptic density, with a mean distance of ∼33 nm from the postsynaptic membrane. These observations firmly establish FAM81A protein as a component of the postsynaptic density in the adult brain, suggesting a role in synaptic function.