Hair diameter distribution in sheep interspecies hybrids

Hair diameter distribution was studied in wild sheep species (argali—Ovis ammon, mouflon—O. Musimon; domestic sheep—O. aries) and their hybrids. In wild species and their hybrids, rather distinctly separated subpopulations of thin (underwool) hairs (10–22 μm diameter) and thick (kemp) hairs (85–302 μm) were detected, and intermediate diameters were rare. In domestic sheep, the kemp subpopulation was very poor, not distinctly separated from the underwool, while maximal hair diameter was <150 μm. In wild × domestic crosses, the kemp subpopulation was clearly expressed, shifted to lower diameter range, and maximal diameters was <200 μm. The same restriction was found in two-month-old wild species lambs compared with adults.     

Influence of glass microbeads on growth,activity and morphological changes of Bacillus megaterium

Cells of Bacillus megaterium growing in the presence of glass microbeads with average diameters of 29 and 53 μm were frequently filamentous and sometimes reached lengths of 600 μm. Some of the filaments were nonseptate. The formation of filaments was prevented by magnesium but not by several other cations. In media with supplemental magnesium, the time required before active proliferation commenced was inversely related to the diameter of the particles. B. megaterium growing in media with the smaller size beads consumed oxygen and utilized glucose at greater rates than bacteria in media with the larger spheres or in bead-free solutions, and the uptake of oxygen was maintained for a longer period.     

Synthesis of cubic ZnS microspheres exhibiting broad visible emission for bioimaging applications

Biocompatible ZnS microspheres with an average diameter of 3.85 µm were grown by solvo‐hydrothermal (S‐H) method using water–acetonitrile–ethylenediamine (EDA) solution combination. ZnS microspheres were characterized by X‐ray diffraction (XRD), scanning electron microscopy (SEM), high‐resolution transmission electron microscopy (HRTEM), Fourier transform (FT)‐Raman spectroscopy and Fourier transform infrared spectroscopy (FTIR) techniques. The broad photoluminescence (PL) emissions from 380–580 nm that were seen from the ZnS microspheres attributed to the increase in carrier concentration, as understood from the observed intense Raman band at 257 cm^–1. Cytotoxicity and haemocompatibility investigations of these ZnS microspheres revealed its biocompatibility. ZnS microspheres, along with biological cell lines, were giving visible light emission and could be used for bioimaging applications. Copyright © 2015 John Wiley & Sons, Ltd.     

In vitro delivery of curcumin with cholesterol-based cationic liposomes

A new cholesterol-based cationic lipid was synthesized; liposomes prepared on its basis were evaluated as drug delivery vehicles for curcumin. Free and liposome-encapsulated curcumin cytotoxicity against HeLa, A549, HepG2, K562 and 1301 cell lines was assessed. Liposomal curcumin with ED₅₀ values ranging from 2.5–10 μM exhibited 2–8 times higher cytotoxicity than free curcumin. The synthetic cholesterol-based cationic lipid also enhanced cellular uptake of curcumin into tested cells. Cationic liposome alone showed low cytotoxicity at high doses with ED₅₀ values of 90–210 μM.     

Defect-related luminescent bur-like hydroxyapatite microspheres induced apoptosis of MC3T3-E1 cells by lysosomal and mitochondrial pathways

When orthopedic joints coated by hydroxyapatite(HA) were implanted in the human body, they release wear debris into the surrounding tissues. The generation and accumulation of wear particles will induce aseptic loosening. However, the potential bioeffect and mechanism of HA-coated orthopedic implants on bone cells are poorly understood. In this study, defect-related luminescent bur-like hydroxyapatite(BHA) microspheres with the average diameter of 7–9 μm which are comparable to that of the wear-debris particles from aseptically loosened HA implants or HA debris have been synthesized by hydrothermal synthesis and the MC3 T3-E1 cells were set as a cells model to study the potential bioeffect and mechanism of BHA microspheres. The studies demonstrated that BHA microspheres could be taken into MC3 T3-E1 cells via endocytosis involved in micropinocytosisand clathrin-mediated endocytosis process, and exert cytotoxicity effect. BHA microspheres could induce the cell apoptosis by intracellular production of reactive oxygen species(ROS), which led to not only an increase in the permeability of lysosome and release of cathepsins B, but also mitochondrial dysfunction and DNA damage. Our results provide novel evidence to elucidate their toxicity mechanisms and might be helpful for more reasonable applications of HA-based orthopaedic implants in the future.     

Synthesis and toxicity characterization of carbon coated iron oxide nanoparticles with highly defined size distributions

Background

Iron oxide nanoparticles hold great promise for future biomedical applications. To this end numerous studies on iron oxide nanoparticles have been conducted. One aspect these studies reveal is that nanoparticle size and shape can trigger different cellular responses through endocytic pathways, cell viability and early apoptosis. However, systematic studies investigating the size dependence of iron oxide nanoparticles with highly defined diameters across multiple cells lines are not available yet.

Methods

Iron oxide nanoparticles with well-defined size distributions were prepared. All samples were thoroughly characterized and the cytotoxicity for four standard cell lines (HeLa Kyoto, human osteosarcoma (U2OS), mouse fibroblasts (NIH 3T3) and mouse macrophages (J7442)) where investigated.

Results

Our findings show that small differences in size distribution (ca. 10 nm) of iron oxide nanoparticles do not influence cytotoxicity, while uptake is size dependent. Cytotoxicity is dose-dependent. Broad distributions of nanoparticles are more easily internalized as compared to the narrow distributions for two of the cell lines tested (HeLa Kyoto and mouse macrophages (J7442)).

Conclusion

The data indicate that it is not feasible to probe changes in cytotoxicity within a small size range (10 nm). However, TEM investigations of the nanoparticles indicate that cellular uptake is size dependent.

General significance

The present work compares narrow and broad distributions for various samples of carbon-coated iron oxide nanoparticles. The data highlights that cells differentiate between nanoparticle sizes as indicated by differences in cellular uptake. This information provides valuable knowledge to better understand the interaction of nanoparticles and cells.     

Diterpenoids from Euphorbia neriifolia and Their Related Anti‐HIV and Cytotoxic Activity

Fifteen diterpenoids ( 1 – 15 ), including three undescribed ones with ent‐atisane skeleton, eupnerias G–I ( 1 – 3 ), were obtained from Euphorbia neriifolia. Compounds 1 – 3 were established through comprehensive spectroscopic analysis. Compounds 4 and 5 exhibited obvious anti‐HIV‐1 effect, and their EC₅₀ were 6.6±3.2 and 6.4±2.5 μg mL^?1, respectively. Compound 6 exhibited moderate cytotoxicity on HepG2 and HepG2/Adr cells with IC₅₀ at 13.70 and 15.57 μm , respectively. In addition, compound 15 exhibited significant cytotoxicity on HepG2 cell lines (IC₅₀=0.01 μm ), while it did not show any cytotoxicity against HepG2/Adr cell lines.     

On the Polarization and Innervation of the Pars Inferior Sensory Epithelia of the Herring Labyrinth

The macula sacculi and the macula lagenae of the herring, Clupea harengus L., were examined by light microscopy, the macula lagenae is large compared to what is normal among non-ostariophysan fishes, the morphological polarization of the hair cells in the inferior maculae shows a pattern which is similar to that usually seen in teleost fishes. The fibres in the nerves supplying the macula sacculi and the macula lagenae were counted and their diameters measured. The ramulus saccularis is divided in two separate ramuli innervating populations of hair cells with different morphological polarization. The saccular rostral nerve trunk contains 1800–2300 fibres, with 1300–1800 fibres in the caudal nerve trunk. The lagenar nerve is composed of 2100–4000 fibres. The fibre diameters are 1–14 μm in all ramuli. Silver staining of the nerve axoplasm reveals a unique differentiation of the maculae, which can be divided into a central area surrounded by a peripheral part. The hair cells in the central area are innervated by thick nerve fibres (5–14 μm diameter) as well as a few thin nerve fibres (about 1 μm diameter), while the receptor cells in the peripheral area are exclusively innervated by thin fibres having diameters of 2 μm or less.     

Molecular simulation of size-selective gas adsorption in idealised carbon nanotubes

Molecular simulations were used to examine the adsorption of diatomic molecules (nitrogen and oxygen) and similarly sized gases (argon and methane) in pores with van der Waals diameters similar in size to the gas diameters. Idealised carbon nanotubes were used to model generic pores, to better understand the effect of pore diameter on guest adsorption in the absence of defects, specific adsorption sites, or variations in pore diameter that often complicate studies of gas adsorption in other porous materials. Molecular dynamics simulations of open nanotubes show that argon and methane are able to enter tubes whose diameters are slightly smaller than the gas diameters. Diatomic gases are able to enter tubes that are significantly smaller than their kinetic diameters with the molecular axis aligned parallel to the nanotube. The results indicate that size-selective adsorption of these gases is theoretically possible, although differences in pore diameters of only a few tenths of an Angstrom are required. Grand canonical Monte Carlo simulations of a 3.38 Å nanotube indicate significant uptake by argon and oxygen, but not nitrogen or methane. The adsorption of nitrogen and methane gradually increases as the nanotube diameter approaches 4.07 Å, and all gases fully saturate a 4.54 Å nanotube. Of the nanotubes studied, the largest adsorption enthalpy for any gas corresponds to the 4.54 Å nanotube, with significantly lower enthalpies seen in the 5.07 Å nanotube. These results suggest an ideal pore diameter for each gas based on the gas–pore van der Waals interaction energies. Trends in the ideal diameter correlate with the minimum tube diameter accessible to each gas.     

Characteristics of microspheres formed in PCR with bacterial genomic DNA or plasmid DNA as templates

Earlier, we discovered that, along with linear DNA fragments, nano- and microparticles of DNA and their aggregates are formed in the PCR with yeast genomic DNA used as a template and gene-specific or partially complementary primers. The size of the microparticles (microspheres) varied in the range of 0.5 to 3–4 μm. Only thermostable KlenTaq polymerase but not Taq polymerase could effectively generate microspheres. In this work, we demonstrate that KlenTaq polymerase can produce microspheres of variable size (1 to 7 μm in diameter) if genomic DNA of the bacterium Acidithiobacillus ferrooxidans and partially complementary primers are present in the PCR mixture. Conditions for generation of DNA microparticles in PCR with Taq-polymerase and bacterial genomic DNA as template were also elaborated. It was also found that mainly large microspheres of up to 7 μm accumulated in PCR with plasmid DNAs used as templates and gene-specific primers in the presence of KlenTaq polymerase or mixtures of KlenTaq and Pfu polymerases. Besides, small aggregates, as well as linear branched structures and three-dimensional conglomerates of fused microspheres, were also revealed in the PCR mixtures. UV absorption spectra of native DNA microspheres and microspheres that had undergone heating at 93°C were registered. The key role of Mg²⁺ cations in the formation and stabilization of the microsphere structure was established.     

Combination of microwell structures and direct oxygenation enables efficient and size‐regulated aggregate formation of an insulin‐secreting pancreatic β‐cell line

Spherical three‐dimensional (3D) cellular aggregates are valuable for various applications such as regenerative medicine or cell‐based assays due to their stable and high functionality. However, previous methods to form aggregates have shown drawbacks, being labor‐intensive, showing low productivity per unit area or volume and difficulty to form homogeneous aggregates. We proposed a novel strategy based on oxygen‐permeable polydimethylsiloxane (PDMS) honeycomb microwell sheets, which can theoretically supply about 80 times as much oxygen as conventional polystyrene culture dishes, to produce recoverable aggregates in controllable sizes using mouse insulinoma cells (MIN6‐m9). In 48 hours of culture, the PDMS sheets produced aggregates whose diameters were strictly controlled (?32, 60, 90, 150 and 280 mm) even at an inoculum density eight times higher (8.0×105 cells/cm²) than that of normal confluent monolayers (1.0×105 cells/cm²). Measurement of the oxygen tension near the cell layer and glucose/lactate analysis clearly showed that cells exhibit aerobic respiration on the PDMS‐based culture system. Glucose‐responsive insulin secretion of the recovered aggregates showed that the aggregates around 90 mm in diameter secreted the largest amounts of insulin. This confirmed the advantages of 3D cellular organization and the existence of a suitable aggregate size, above which excess organization leads to a decreased metabolic response. These results demonstrated that this microwell‐based PDMS culture system provides a promising method to form size‐regulated and better functioning 3D cellular aggregates of various kinds of cells with a high yield per surface area. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 30:178–187, 2014     

Preparation of liposomes of defined size distribution by extrusion through polycarbonate membranes

Liposomes of defined size and homogeneity have been prepared by sequential extrusion of the usual multilamellar vesicles through polycarbonate membranes. The process is easy, reproducible, produces no detectable degradation of the phospholipids, and can double the encapsulation efficiency of the liposome preparation. Multilamellar vesicles extruded by this technique are shown by both negative stain and freeze-fracture electron microscopy to have mean diameters approaching the pore diameter of the polycarbonate membrane through which they were extruded. When sequentially extruded down through a 0.2 μm membrane, the resulting vesicles exhibit a very homogeneous size distribution with a mean diameter of 0.27 μm while maintaining an acceptable level of encapsulation of the aqueous phase.     

What is the upper size limit for cosmopolitan distribution in free‐living microorganisms?

A comparison of testate amoebae assemblages from the Arctic and Antarctic (areas of similar habitat a maximum distance apart) is used to try and answer the question ‘What is the upper size limit for cosmopolitan distribution in free‐living microbes?’ Species restricted to either the Arctic or Antarctic exhibited sizes up to 230 μm while the largest cosmopolitan species was 135 μm in size. Comparison of the testate assemblages using a multivariate classificatory technique (TWINSPAN ) also suggested more restricted distribution for the larger species. There was a negative relationship between species size and number of sites at which it was recorded (r_s=?0.261, P < 0.05), with all the more widespread species having a size of below 100 μm. It is suggested that for testate amoebae cosmopolitan distributions become common below 100–150 μm. This suggests that most species of testate (indeed most free‐living microbes) have low species richness because of lack of opportunities for allopatric speciation as most are below 100 μm and so geographical isolation is unlikely. It is suggested that if this is correct, only the largest free‐living microbes (> 150 μm) are likely to be of conservation concern because of their smaller ranges. However, I point out that currently different studies are giving very different answers to the question, how ubiquitous and species rich are free‐living microbes? The subject requires further work to try and reconcile these different results.     

Size Effects on the Uptake of Particles by Populations of Tetrahymena pyriformis Cells

Flow cytometry has been used to make direct measurements of rates of uptake of latex microspheres from dilute, monodisperse suspensions by Tetrahymena pyriformis. Measurements were made for five different sizes of microspheres, ranging from 1.09 to 6.17 μm diameter. Fractions of cells in the population that did not ingest the microspheres offered were also determined. In addition, the size distributions, as indicated by the forward angle light scattering intensity which is measured by the instrument, were determined for the whole population and for the subpopulations of cells that did and did not ingest the particles, for each particle size used. It was found that the fraction of cells that did not ingest the particles was small and independent of particle size when this was less than about 2.7 μm, but increased with particle size when particle size was increased above this value. The so-called maximum clearance rate, which can be calculated from the data, was found to increase monotonically with particle size if it were based only on those cells which actually ingested the particles offered. However, a plot of maximum clearance rate vs. particle size exhibited a maximum if the clearance rate were based on all cells present in the population.     

Combined retrograde tracing and immunohistochemistry of trigeminal ganglion neurons projecting to gingiva or tooth pulps in the lower jaw of the cichlid Tilapia mariae

Rat trigeminal ganglion neurons projecting to the oral mucosa or to tooth pulps have different cell diameters and contain different chemical markers. In the present paper we examine whether trigeminal ganglion neurons sending axons to gingiva or tooth pulps in the lower jaw of the cichlid Tilapia mariae differ in a similar way. Retrograde tracing with fluorescent latex microspheres revealed labelled gingival and pulpal neurons in the caudal part of the trigeminal ganglion. The gingival neurons had a unimodal size distribution (peak 11 μm; range 8–14 μm) and the pulpal neurons exhibited a bimodal size distribution (peaks 12 and 25 μm; range 10–40 μm). Immunohistochemistry revealed a calcitonin gene-related peptide-like immunoreactivity in some 40% of the gingival neurons and a substance P-like immunoreactivity in 30%. Of the small pulpal neurons about 60% exhibited a calcitonin gene-related peptide-like immunoreactivity and 15% showed a substance P-like immunoreactivity. Of the large pulpal neurons some 70% exhibited a calcitonin gene-related peptide-like immunoreactivity. These neurons did not show a substance P-like immunoreactivity. In some animals a few trigeminal ganglion neurons showed a neuropeptide Y- or a vasoactive intestinal polypeptide-like immunoreactivity. Perikarya with a tyrosine hydroxylase- or a choline acetyl transferase-like immunoreactivity were not observed. We conclude that gingiva and tooth pulps in the lower jaw of T. mariae are innervated by trigeminal ganglion neurons, the cell diameters and neuropeptide contents of which differ in a pattern similar to that in the rat. Hence, this seems to represent a conserved evolutionary pattern.     

Process and formulation variables in the preparation of injectable and biodegradable magnetic microspheres

The aim of this study was to prepare biodegradable sustained release magnetite microspheres sized between 1 to 2 μm. The microspheres with or without magnetic materials were prepared by a W/O/W double emulsion solvent evaporation technique using poly(lactide-co-glycolide) (PLGA) as the biodegradable matrix forming polymer. Effects of manufacturing and formulation variables on particle size were investigated with non-magnetic microspheres. Microsphere size could be controlled by modification of homogenization speed, PLGA concentration in the oil phase, oil phase volume, solvent composition, and polyvinyl alcohol (PVA) concentration in the outer water phase. Most influential were the agitation velocity and all parameters that influence the kinematic viscosity of oil and outer water phase, specifically the type and concentration of the oil phase. The magnetic component yielding homogeneous magnetic microspheres consisted of magnetite nanoparticles of 8 nm diameter stabilized with a polyethylene glycole/polyacrylic acid (PEG/PAA) coating and a saturation magnetization of 47.8 emu/g. Non-magnetic and magnetic microspheres had very similar size, morphology, and size distribution, as shown by scanning electron microscopy. The optimized conditions yielded microspheres with 13.7 weight% of magnetite and an average diameter of 1.37 μm. Such biodegradable magnetic microspheres seem appropriate for vascular administration followed by magnetic drug targeting.     

Cuceolatins A–D: New Bioactive Diterpenoids from the Leaves of Cunninghamia lanceolata

Four new diterpenoids named cuceolatins A–D, including three labdane‐type ( 1 – 3 ) and one abietane‐type ( 4 ) as well as three known labdane analogs ( 5 – 7 ), were reported from the leaves of Cunninghamia lanceolata. Structural assignments for these compounds were conducted by analyses of spectroscopic data, and their absolute configurations were determined by time‐dependent density functional theory (TD‐DFT) based electronic circular dichroism (ECD) calculations. Among them, the abietane‐type diterpenoid (11‐hydroxy‐12‐methoxyabieta‐8,11,13‐trien‐3‐one ( 4 )) showed significant cytotoxicity against human MDA‐MB‐231, MCF‐7, and HeLa tumor cell lines with IC₅₀ measurements of 4.3, 2.8 and 4.5 μm , respectively, while the labdane‐type diterpenoids with a 4α‐carboxy group ( 1 – 3 and 5 ) exhibited moderate antibacterial activity towards Bacillus subtilis and Staphylococcus aureus with IC₅₀ values all below 25 μm .     

Microparticle‐mediated gene delivery for the enhanced expression of a 19‐kDa fragment of merozoite surface protein 1 of Plasmodium falciparum

The 19 kDa carboxyl‐terminal fragment of merozoite surface protein 1 (MSP1₁₉) is a major component of the invasion‐inhibitory response in individual immunity to malaria. A novel ultrasonic atomization approach for the formulation of biodegradable poly(lactic‐co‐glycolic acid) (PLGA) microparticles of malaria DNA vaccines encoding MSP1₁₉ is presented here. After condensing the plasmid DNA (pDNA) molecules with a cationic polymer polyethylenimine (PEI), a 40 kHz ultrasonic atomization frequency was used to formulate PLGA microparticles at a flow rate of 18 mL h^?1. High levels of gene expression and moderate cytotoxicity in COS‐7 cells were achieved with the condensed pDNA at a nitrogen to phosphate (N/P) ratio of 20, thus demonstrating enhanced cellular uptake and expression of the transgene. The ability of the microparticles to convey pDNA was examined by characterizing the formulated microparticles. The microparticles displayed Z‐average hydrodynamic diameters of 1.50–2.10 μm and zeta potentials of 17.8–23.2 mV. The encapsulation efficiencies were between 78 and 83%, and 76 and 85% of the embedded malaria pDNA molecules were released under physiological conditions in vitro. These results indicate that PLGA‐mediated microparticles can be employed as potential gene delivery systems to antigen‐presenting cells in the prevention of malaria. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2010