Undecalcified Bone Preparation for Histology,Histomorphometry and Fluorochrome Analysis

Undecalcified bone histology demonstrates the micro-architecture of bone. It shows both the mineralised and cellular components of bone, which provides vital information on bone turnover or bone formation and resorption. This has tremendous importance in a variety of clinical and research applications. It yields beautiful images¹ and allows for techniques such as fluorochrome assessment and histomorphometry². Fluorochrome analysis is a technique where fluorescent dyes that bind to calcium are injected at a particular time point, which allows for quantification of the amount of mineralisation at that given time. Histomorphometry is a process of bone quantification at the microscopic level. Performing undecalcified bone histology is technically challenging, particularly with large size specimens. It requires variations in technique from those used in standard paraffin embedded histology. This video illustrates the process of producing good quality sections and demonstrates the technical difficulties and methods with which to overcome them. Specimen preparation, fixation and processing are achieved with a manner similar to other soft tissues, however due to the density and lower permeability of bone considerably longer fixation and processing times are required, often taking several weeks. Embedding is achieved using a supporting medium with similar or equal hardness and density to the bone such as methacrylate- based resins, but unlike paraffin infiltration and embedding, this is an irreversible step. Sectioning can be achieved by grinding which produces a thicker section, which is optimal for studies such as fluorochrome analysis. This is best achieved using a diamond blade on a macrotome. Alternatively, thinner sections can be produced for light microscopy and this is achieved using a sledge microtome with a very sharp blade. The sledge microtome provides the additional strength and stability required for large, hard blocks. Resin embedded sections can be stained with a variety of stains, which are demonstrated.Download video file.^{(88M, mp4)}     

Be<Superscript>2+ </Superscript>V<Superscript>-</Superscript> Dipole and Adsorptivity of Atomic H on LiH(001) Surface: ab initio Study

An attempt has been made to examine the properties of the title divalent cation impurity - cation vacancy dipole as well as the adsorptivity and diffusion of atomic hydrogen over the LiH (001) surface using ab initio methods of molecular electronic structure calculations. The LiH crystal surface was simulated and quantum clusters of variable size were surrounded by point charges to examine the I-V dipole orientation, cation vacancy migration and overlap effects. The effects of introducing an I-V dipole on the nature of adsorbate-substrate interactions and diffusion of an H atom over the surface were also examined. The results confirm that the (100) orientation of the I-V dipole is energetically more favorable than the (110) orientation and that the cation vacancy migrates without activation energy barriers. The I-V dipole enhances the adsorptivity of atomic hydrogen by ca. 4.96 eV, changes the nature of adsorption from physisorption to chemisorption and introduces an activation energy barrier to H diffusion over the surface. As the I-V dipole is introduced, the HOMO and LUMO levels of the substrate shift to higher energies, the band gap becomes narrower and the atomic charges on the adsorbed H increases. This change in the electronic structure makes the adsorptivity process, through the charge transfer from the H 1s singly occupied AO to the conduction band and from the valence band to the H 1s singly occupied AO more facile.     

Fluoride,Some Selected Elements,Lipids, and Protein in the Muscle and Liver Tissues of Five Fish Species along the Egyptian Mediterranean Sea Coast

Fish is a vital, healthy source of animal protein and vitamins. This study was oriented to estimate fluoride, some selected elements, lipids, and protein concentrations in fish. Five fish species were collected from the Egyptian Mediterranean Sea coast during April 2007. Variable content of fluoride, calcium, magnesium, sulphate, phosphorus, lipids, and protein were determined in muscle and liver of fish samples and yielded average values in fish muscle of 11.0 ± 2.0 μg/g, 6.5 ± 3.8 mg/g, 100.2 ± 39.3 mg/g, 4.4 ± 6.3 mg/g, 2.4 ± 0.5 mg/g, 6.0 ± 2.2 mg/l, and 2.6 ± 1.0 g% wet wt, respectively. Special attention was given to fluoride because of its hazardous classification. Interestingly, the hazard index values of fluoride were less than 1 for all collected fish species. Additionally, the daily fluoride exposure was generally below the nutrient reference values for Australian and New Zealand populations. Accordingly, there is little human health risk from the selected fish species due to consumption of fluoride-contaminated fish.     

Oxidative stress and DNA damage in relation to transition metals overload in Abu-Qir Bay,Egypt

The aim of the present study is to evaluate the transition metals overload in Abu-Qir Bay in Egypt, as compared to a less polluted area (reference area) through some biomarkers of oxidative stress. Catalase enzyme activity, malondialdehyde (MDA) concentration and DNA damage (number of apurinic/apyrimidinic sites) were the tested biomarkers. The levels of iron and copper in Mugil cephalus liver tissues were significantly higher in samples from the polluted area as compared to the reference area: Fe: 407 ± 38 vs. 216 ± 21 μg/g wet wt; p = 0.008, Cu: 54 ± 6 vs. 17.7 ± 4 μg/g wet wt; p = 0.0001. This could account for the observed increase in MDA concentration (15.7 ± 5.7 vs. 2.5 ± 0.5 U/g; p = 0.035), and the elevated number of AP sites (13.9 ± 2.6 vs. 0.37 ± 0.2 AP site/1 × 10⁵ bp; p = 0.0001). Similarly, the activity of catalase enzyme responsible for the cellular defense was significantly high (58.3 ± 12.2 vs. 28.4 ± 4.0 U/mg; p = 0.032). The present data indicated a clear relationship between the pollution degree of the above marine environment and both biochemical and molecular responses of the piscine system.     

Antimicrobial Activity of Two Novel Venoms from Saudi Arabian Scorpions (Leiurus quinquestriatus and Androctonus crassicauda)

International Journal of Peptide Research and Therapeutics - Many recent studies have shown the benefits of scorpion venom, as it components may be used as potential candidates for drug...     

Effects of Silver Nanoparticles on Burn Wound Healing in a Mouse Model

Biological Trace Element Research - Healing of injuries caused by exposure to heat has been discussed in many studies, although a few drugs have been shown to produce satisfactory results. In this...     

Cysteine supplementation reverses methionine restriction effects on rat adiposity: significance of stearoyl-coenzyme A desaturase

Stearoyl-CoA desaturase-1 (SCD1) is a key enzyme in fatty acid and energy metabolism, but little is known about its nutritional regulation. Dietary methionine restriction in rats decreases hepatic Scd1 mRNA and protein, increases energy expenditure, and decreases fat-pad mass/body-weight% (FM/BW%). In humans, plasma concentrations of the methionine product, cysteine, are associated with obesity. To determine which consequences of methionine-restriction are mediated by decreased cysteine availability, we monitored obesity-related variables in 4 dietary groups for 12 weeks: control-fed (CF), methionine-restricted (MR), MR supplemented with 0.5% l-cysteine (MR+Cys) and CF+Cys rats. MR lowered weight gain and FM/BW% despite higher food intake/weight than CF, and lowered serum cysteine. Hepatic Scd1 expression was decreased, with decreased serum SCD1 activity indices (calculated from serum fatty acid profile), decreased serum insulin, leptin and triglycerides, and higher adiponectin. Cysteine supplementation (MR+Cys) essentially reversed all these phenotypes and raised serum cysteine but not methionine to CF levels. Adding extra cysteine to control diet (CF+Cys) increased serum taurine but did not affect serum cysteine, lipids, proteins, or total weight gain. FM/BW% and serum leptin were modestly decreased. Our results indicate that anti-obesity effects of MR are caused by low cysteine and that dietary sulfur amino acid composition contributes to SCD1 regulation.