Iron overload diseases: the chemical speciation of non-heme iron deposits in iron loaded mammalian tissues

⁵⁷Fe Mössbauer spectra of iron overloaded human spleen, rat spleen and rat liver tissue samples at 78 K were found to consist of a quadrupole doublet (major component) with magnetic sextet (minor component with fractional spectral area F _s). The distributions of F _s for spleen tissue from two different clinically identifiable groups (n = 7 and n = 12) of thalassemic patients were found to be significantly different. The value of F _s for dietary-iron loaded rat liver was found to rise significantly with age/duration (up to 24 months) of iron loading.     

Optical trapping microrheology in cultured human cells

We present the microrheological study of the two close human epithelial cell lines: non-cancerous HCV29 and cancerous T24. The optical tweezers tracking was applied to extract the several seconds long trajectories of endogenous lipid granules at time step of 1μs. They were analyzed using a recently proposed equation for mean square displacement (MSD) in the case of subdiffusion influenced by an optical trap. This equation leads to an explicit form for viscoelastic moduli. The moduli of the two cell lines were found to be the same within the experimental accuracy for frequencies 10(2) - 10(5) Hz. For both cell lines subdiffusion was observed with the exponent close to 3/4, the value predicted by the theory of semiflexible polymers. For times longer than 0.1s the MSD of cancerous cells exceeds the MSD of non-cancerous cells for all values of the trapping force. Such behavior can be interpreted as a signature of the active processes and prevents the extraction of the low-frequency viscoelastic moduli for the living cells by passive microrheology.     

Iron homeostasis in neuronal cells: a role for IREG1

Background  

Iron is necessary for neuronal function but in excess generates neurodegeneration. Although most of the components of the iron homeostasis machinery have been described in neurons, little is known about the particulars of their iron homeostasis. In this work we characterized the response of SH-SY5Y neuroblastoma cells and hippocampal neurons to a model of progressive iron accumulation.     

Fluorescence spectra of cultured normal and malignant lung cells

Fluorescence Spectroscopy has emerged as a new modality to characterize physicochemical properties of biomolecules. The biomolecules have certain photophysical properties based on their molecular structure and these properties have been considered as useful parameters to monitor alterations in the functional, morphological and micro environmental changes in the cells and tissues. In this study the fluorescence emission spectra of normal and malignant lung cells were recorded for different excitation wavelengths: 230, 300, 340, and 450 nm, corresponding to the absorption of tyrosine, tryptophan, collagen or elastin, Nicotinamide adenine dinucleotide (NADH) and flavin adeno dinucleotide (FAD). Similarly excitation spectra were also recorded at 340 nm. The emission profiles showed considerable difference between the malignant and normal cells with the malignant cells having more fluorescence intensity than that of normal cells keeping emission at 340 nm. Our study had shown the discriminating features between normal and carcinoma cells lines because of higher concentration of tryptophan (1.5 times), NADH (3 times), and flavin (4 times) in carcinoma cell lines.     

Fluorescence labeling and interaction of atherogenic lipoproteins with cultured cells

We prepared lipoprotein (a) and LDL covalently labeled with either BODIPY or rhodamine. A dual wavelength method was used for the microscopic observation of both lipoproteins during their interaction with HepG2 cells. Since a large proportion of Lp(a) colocalized with LDL on the cell surface and inside the cells, it was concluded that Lp(a) uptake into cells is mediated by LDL via internalization of LDL.     

Biophysical characteristics of cells cultured on cholesteryl ester liquid crystals

This study aimed at examining the biophysical characteristics of human derived keratinocytes (HaCaT) cultured on cholesteryl ester liquid crystals (CELC). CELC was previously shown to improve sensitivity in sensing cell contractions. Characteristics of the cell integrin expressions and presence of extracellular matrix (ECM) proteins on the liquid crystals were interrogated using various immunocytochemical techniques. The investigation was followed by characterization of the chemical properties of the liquid crystals (LC) after immersion in cell culture media using Fourier transform infrared spectroscopy (FTIR). The surface morphology of cells adhered to the LC was studied using atomic force microscopy (AFM). Consistent with the expressions of the integrins α2, α3 and β1, extracellular matrix proteins (laminin, collagen type IV and fibronectin) were found secreted by the HaCaT onto CELC and these proteins were also secreted by cells cultured on the glass substrates. FTIR analysis of the LC revealed the existence of spectrum assigned to cholesterol and ester moieties that are essential compounds for the metabolizing activities of keratinocytes. The immunostainings indicated that cell adhesion on the LC is mediated by self-secreted ECM proteins. As revealed by the AFM imaging, the constraint in cell membrane spread on the LC leads to the increase in cell surface roughness and thickness of cell membrane. The biophysical expressions of cells on biocompatible CELC suggested that CELC could be a new class of biological relevant material.     

Local stimulation of cultured myocyte cells by femtosecond laser-induced stress wave

When an 800 nm femtosecond laser is tightly focused into cell culture medium a stress wave is generated at the laser focal point. Since the stress wave localizes in a few tens of μm, it is possible to locally stimulate single cells in vitro. In this work, several kinds of cultured mammalian cells, HeLa, PC12, P19CL6, and C2C12, were stimulated by the stress wave and the cell growth after the stress loading with the laser irradiation was investigated. In comparison with the control conditions, cell growth after the laser irradiation was enhanced for the cells of C2C12 and P19CL6, which can differentiate into myocytes, and suppressed for PC12 and HeLa cell lines. These results suggest a possibility of cell growth enhancement due to myogenic cells response to the femtosecond laser-induced stress.     

Nondestructive isolation of single cultured animal cells by femtosecond laser-induced shockwave

A shockwave induced by focusing an intense femtosecond laser onto a culture medium under a microscope was used to manipulate single cultured animal cells. Mouse NIH3T3 fibroblasts were cultured on a collagen matrix and placed on a microscope installation that included an intense femtosecond laser. Cells were detached individually from the matrix by the shockwave, after their filopodia were cut by focusing the laser directly on them. The appearance and locomotion of cells after detachment was monitored with a CCD camera. Filopodia of the detached cells were regenerated, and the cell adhered again to the matrix within 4 hrs after detachment. When the shockwave was induced at a pulse energy of 0.72 J/pulse, 80% of cells were successfully detached from the culture plate in a non-destructive manner. The force required to detach a cell was estimated to be a few N/m², which is larger than the photon force resulting from conventional laser trapping. PACS 42.62.Be; 47.40.Nm; 87.80.Rb     

含铜铁矿石全铁的测定时铜对铁的影响探讨

探讨了含铜铁矿石分析过程中的铜对铁分析的影响.分析过程中采用标准溶液模拟实验、国家标准物质中铁的容量分析、方法比对等实验,得出一个大约的经验系数R,为含铜铁矿石中全铁含量的质量控制提供参考.该系数经过对多个含铜国家标准物质中的铁进行分析验证,样正值与标准值基本相符.     

Ultrasound-assisted extraction of ginseng saponins from ginseng roots and cultured ginseng cells

Ultrasound-assisted extraction was evaluated as a simpler and more effective alternative to conventional extraction methods for the isolation of ginsenosides (saponins) from various types of ginseng. The ginseng samples were extracted with different solvents, under either direct sonication by an ultrasound probe horn or indirect sonication in an ultrasound cleaning bath. The ultrasonic extraction was compared with the conventional method of refluxing boiling solvents in a soxhlet extractor, on the yields of both the total saponin isolated by thin-layer chromatography and the individual ginsenosides by high performance liquid chromatography. It was found that the sonication-assisted extraction of ginseng saponins was about three times faster than the traditional extraction method. The ultrasonic extraction was not only more efficient but also convenient for the recovery and purification of the active ingredients of plant materials. In addition, the sonication-assisted extraction can be carried out at lower temperatures which are favorable for the thermally unstable compounds.     

Polyethyleneimine-mediated transfection of cultured postmitotic neurons from rat sympathetic ganglia and adult human retina

Background  

Chemical methods of transfection that have proven successful with cell lines often do not work with primary cultures of neurons. Recent data, however, suggest that linear polymers of the cation polyethyleneimine (PEI) can facilitate the uptake of nucleic acids by neurons. Consequently, we examined the ability of a commercial PEI preparation to allow the introduction of foreign genes into postmitotic mammalian neurons. Sympathetic neurons were obtained from perinatal rat pups and maintained for 5 days in vitro in the absence of nonneuronal cells. Cultures were then transfected with varying amounts of a plasmid encoding either E. coli β-galactosidase or enhanced green fluorescence protein (EGFP) using PEI.     

Nuclear magnetic resonance study of iron overload in liver tissue

Whole-tissue and homogenized samples of human liver were studied in a NMR spectrometer, T1 and T2 relaxation times were measured as a function of added inorganic or organic iron. When inorganic iron (Fe+3) was added, pronounced T1 and T2 shortening was noted. However, when organic iron, in the form of ferritin, was added, the amount of T1 and T2 relaxation enhancement was much reduced for the same amount of added iron. The in vitro ferritin results model the situation found in clinical studies of hemochromatosis. Only in cases of severe iron overload were significant decreases in relaxation times observed. The T2 relaxation time was the more reliable indicator of excessive levels of iron in the liver. The large range of T1 and T2 values encountered in normal volunteers precludes the use of MR to quantitatively measure iron levels in the liver. The T1 and T2 relaxation times measured at intervals for one individual tend to fluctuate as well, making the use of MR to follow the course of treatment of iron overload disorders unreliable.     

Biological tissue magnetism in the frame of iron overload diseases

The conspicuous magnetic properties of iron, paradoxically, rarely participate in the methods routinely employed in the clinical environment to detect iron containing species in tissues. In the organism iron is just a trace metal and it mostly occurs as part of haemoproteins or ferritin, which show paramagnetic, diamagnetic or antiferromagnetic behaviour, hence resulting in a very low contribution to the tissue susceptibility. Detailed magnetic measurements make it nowadays possible to identify such species in tissues that correspond to individuals with iron overload pathologies. Since, as alternatives to the conventional biopsy, magnetism-based noninvasive techniques to diagnose and manage such diseases are recently under development, the deep knowledge of the magnetic properties of the different forms of iron in tissues is of high applied interest.     

Iron distribution in chevkinite

In this paper Mössbauer spectra of chevkinite at 283K and 83K are presented for the first time. The spectra consist of two Fe²⁺ and two Fe³⁺ doublets, and the assignments were made according to the structure of chevkinite. Iron distribution obtained from the spectrum at 83 K is consistent with that at 283 K.     

Iron in the brain

The results of our studies of iron in three brain structures, substantia nigra (SN), globus pallidus (GP), and hippocampus (Hip), are presented. Mössbauer spectroscopy, electron microscopy and ELISA (enzyme-linked immuno-absorbent assay) were applied. Mössbauer studies show that most of the iron in the brain is ferritin-like. The concentration of iron is similar in SN and GP, but less than half of this in Hip. ELISA studies showed that the H/L ratio of ferritin in SN and GP is also similar, but is about three times higher in Hip. These results suggest that the role of iron in SN and GP may be different from that in Hip. Electron microscopy shows that the diameters of the ferritin iron cores in the brain are smaller that in the liver (3.5 ± 0.5 nm vs. 6.0 ± 0.5 nm). Mössbauer studies yield the ratio between the concentration of iron in control and parkinsonian SN as 1.00 ± 0.13.     

Neurospheres from rat adipose-derived stem cells could be induced into functional Schwann cell-like cells in vitro

Background  

Schwann cells (SC) which are myelin-forming cells in peripheral nervous system are very useful for the treatment of diseases of peripheral nervous system and central nervous system. However, it is difficult to obtain sufficient large number of SC for clinical use, so alternative cell systems are desired.     

Microdisperse Iron Silicide Structures Produced by Implantation of Iron Ions in Silicon

Iron implanted and subsequently annealed n-type Si(111) was studied by conversion electron Mössbauer spectroscopy for phase analysis and Auger electron spectroscopy for sputter depth profiling and element mapping. During implantation (200 keV, 3 × 10¹⁷ cm^?2, 350°C) a mixture of β- and α-FeSi₂ is firmed and after the subsequent annealing (900°C for 18 h and 1150°C for 1 h) a complete transition to the β- and the α-phase can be detected. The as-implanted profile has Gaussian shape and is broadening during annealing at 900°C to a plateau-like profile and shows only a slight broadening and depth depending fluctuations of the iron concentration after the 1150°C annealing. With scanning Auger electron spectroscopy the lateral iron and silicon distribution were investigated and show for the sample annealed at 900°C large separated β-FeSi₂ precipitates which grow due to the process of Ostwald ripening. At 1150°C additionally coalescence of the precipitates occur and a wide extended penetration α-FeSi₂ network structure is formed.     

The biological characteristics of different generation mesenchymal stem cells cultured in vitro and cocultured with vascular scaffold

Mesenchymal stem cells (MSCs) were used widely as seed cells in tissue engineering blood vessel construction. However, the biological characteristics difference of different generation MSCs in vitro culture is unknown, which laid a foundation for appropriate generation seeded cells selection for tissue engineering blood vessel construction. In this report, MSCs were isolated from SD rat bone marrow and identified by flow cytometry; cell growth curve test, cell surface antigen expression rate detection, cryopreservation resuscitation rate test, CD31 expression rate test, cell cycle analysis, and adhesion difference on vascular scaffold test were performed. The research results indicated that the MSCs shape was spindle and uniform with vigorous growth. CD105 and CD90 factor expression rate reached 82.5 and 84.9%, respectively, and the expression rate of CD45 was only 7.3%. The proliferation capacity of the fourth generation MSCs were more exuberant, with proliferation index as 20.3%; the cell proliferation index of the eighth generation decreased to only 9.1%. The cryopreservation resuscitation rate of the second generation and fourth generation MSCs were both higher than 80%, and the cryopreservation resuscitation rate of the eighth generation MSCs was only about 60%. After the induction for 5 days, MSCs had weak CD31 expression, and with the prolonged induction time, expression increased. All generation MSCs expressed CD31 after being induced for 10 days; however, the CD31 positive expression rate of the second generation, fourth generation, and sixth generation MSCs had significant difference with the eighth generation MSCs. Adhesion rate of MSCs before sixth generation was around 40%, but the adhesion rate of eighth generation MSCs was only about 27%. In all, biological characteristics of different generation MSCs existed certain differences, and especially the eighth generation MSCs aged seriously, whose cell activity decreased significantly. The researchers believed that the MSCs before the sixth generation can maintain excellent properties of MSCs, and can be used as seed cells for vascular tissue engineering.     

Preparation and Properties of Iron and Iron Oxide Nanocrystals in MgO Matrix

We have prepared α-iron and magnetite (Fe₃O₄) nanoparticles in MgO matrix from a mixture of nanocrystalline Fe₂O₃ with Mg(H,O) powders calcinated in hydrogen. This procedure yielded spherical magnetic nanoparticles embedded in MgO. Transmission electron microscopy and Mössbauer spectroscopy were used for structure and phase analysis. The measurements of magnetic properties showed increased coercivity of the nanocomposite samples.