Extreme diversity among amacrine cells: implications for function

We report a quantitative survey of the population of amacrine cells present in the retina of the rabbit. The cells' dendritic shape and level of stratification were visualized by a photochemical method in which a fluorescent product was created within an individual cell by focal irradiation of that cell's nucleus. A systematically random sample of 261 amacrine cells was examined. Four previously known amacrine cells were revealed at their correct frequencies. Our central finding is that the heterogeneous collection of other amacrine cells is broadly distributed among at least 22 types: only one type of amacrine cell makes up more than 5% of the total amacrine cell population. With these results, the program of identification and classification of retinal neurons begun by Cajal is nearing completion. The complexity encountered has implications both for the retina and for the many regions of the central nervous system where less is known.     

Induced and synchronized estrus in cattle: dose titration of estradiol benzoate in peripubertal heifers and postpartum cows after treatment with an intravaginal progesterone-releasing insert and prostaglandin F2alpha

Seven cysteine-rich repeats form the ligand-binding region of the low-density lipoprotein (LDL) receptor. Each of these repeats is assumed to bind a calcium ion, which is needed for association of the receptor with its ligands, LDL and beta-VLDL. The effects of metal ions on the folding of the reduced N-terminal cysteine-rich repeat have been examined by using reverse-phase high-performance liquid chromatography to follow the formation of fully oxidized isomers with different disulfide connectivities. In the absence of calcium many of the 15 possible isomers formed on oxidation, whereas in its presence the predominant product at equilibrium had the native disulfide bond connectivities. Other metals were far less effective at directing disulfide bond formation: Mn2+ partly mimicked the action of Ca2+, but Ba2+, Sr2+, and Mg2+ had little effect. This metal-ion specificity was also observed in two-dimensional 1H NMR spectral studies; only Ca2+ induced the native three-dimensional fold. The two paramagnetic ions, Gd3+ and Mn2+, and Cd2+ did not promote adoption of a well-defined structure, and the two paramagnetic ions did not displace calcium ions. The location of calcium ion binding sites in the repeat was also explored by NMR spectroscopy. The absence of chemical shift changes for the side chain proton resonances of Asp26, Asp36, and Glu37 from pH 3.9 to 6.8 in the presence of calcium ions and their proximal location in the NMR structures implicated these side chains as calcium ligands. Deuterium exchange NMR experiments also revealed a network of hydrogen bonds that stabilizes the putative calcium-binding loop.     

Effect of exercise on natural cytotoxicity and pulmonary tumor metastases in mice

Exercise has been demonstrated to reduce experimental tumor formation in rodents when the exercise is present during the tumor initiation or promotion phases. This study evaluated whether exercise influenced the process of tumor metastasis and subsequent growth in a secondary implant site. Male C3H mice remained sedentary, were given free access to running wheels, ran on a treadmill (15 m.min-1, 30 min.d-1), or walked on a treadmill (5 m.min-1, 5 min.d-1)(N = 20/group). Following 9 wk of this protocol, exercise was discontinued. At this time all animals received a tumor cell dose (CIRAS 1, 3 x 10(5)) i.v., and remained sedentary until sacrificed 3 wk later. Splenic natural killer (NK) cell activity was elevated in the wheel running mice compared with sedentary controls 3 wk after cessation of exercise (F(3,74) = 6.266, P < 0.002). Exercised mice displayed lower tumor cell retention in the lungs relative to nonexercised mice (F(1,37) = 6.593, P < 0.02). Tumor incidence was not different across activity groups whereas tumor multiplicity was higher in mice that had been previously exposed to exercise. (However, it should be noted that the significant exercise-tumor effect was due to a small number of exercised mice with extreme multiplicity, > 200 foci/lung.) More extensive tumor colony formation was present in wheel-trained mice that displayed the greatest volumes of daily running. The results from this study suggest that exercise was able to augment natural immune cytotoxic function for up to 3 wk after cessation of activity. However, this augmentation of natural immune function was not associated with reduced tumor incidence in the exercised animals.     

Production of titania nanoparticles by a green process route

The manufacture of heterogeneous catalysts and catalyst supports produces substantial amounts of nitrate containing aqueous effluent. The use of nitrate free precursors and an environmentally friendly process would change the manufacture so that the entire process of catalyst synthesis and use can be considered green. In this work the precipitation of titania acetylacetonate nanoparticles for use as catalytic supports using a supercritical carbon dioxide anti-solvent process was investigated over a range of conditions. The effects of 1) pressure, 2) temperature, 3) solution flowrate, 4) solution concentration of TiO(acac)₂ in methanol, 5) nozzle diameter and 6) CO₂/methanol flow ratio on the mean particle size and morphology were studied. Particle sizes between 27 and 78 nm were obtained and were generally string and branch-like with an amorphous nature. Pressure and temperature had little effect on the mean particle size. A decrease in the velocity of the solution flow rate led to an increase in mean particle size and to particles that exhibited greater interconnectivity. It was also observed that an increase in concentration of TiO(acac)₂ in methanol led to an increase in mean particle size. The process shows promise for the production of catalysts by an environmentally acceptable route.     

Fabrication of Single-Nanocrystal Arrays

To realize the full potential of nanocrystals in nanotechnology, it is necessary to integrate single nanocrystals into addressable structures; for example, arrays and periodic lattices. The current methods for achieving this are reviewed. It is shown that a combination of top-down lithography techniques with directed assembly offers a platform for attaining this goal. The most promising of these directed assembly methods are reviewed: capillary force assembly, electrostatic assembly, optical printing, DNA-based assembly, and electrophoretic deposition. The last of these appears to offer a generic approach to fabrication of single-nanocrystal arrays.     

The "spotted" locus maps to bovine chromosome 6 in a Hereford-Cross population

The PROMISE (prosthetic centres andmetalions in protein activesites) database aims to present comprehensive sequence, structural, functional and bibliographic information on metalloproteins and other complex proteins, with an emphasis on active site structure and function. The database is available on the WorldWide Web at http://bioinf.leeds.ac.uk/promise/     

Cyp1 Inhibition Prevents Doxorubicin-Induced Cardiomyopathy in a Zebrafish Heart-Failure Model

Doxorubicin is a highly effective chemotherapy agent used to treat many common malignancies. However, its use is limited by cardiotoxicity, and cumulative doses exponentially increase the risk of heart failure. To identify novel heart failure treatment targets, a zebrafish model of doxorubicin-induced cardiomyopathy was previously established for small-molecule screening. Using this model, several small molecules that prevent doxorubicin-induced cardiotoxicity both in zebrafish and in mouse models have previously been identified. In this study, exploration of doxorubicin cardiotoxicity is expanded by screening 2271 small molecules from a proprietary, target-annotated tool compound collection. It is found that 120 small molecules can prevent doxorubicin-induced cardiotoxicity, including 7 highly effective compounds. Of these, all seven exhibited inhibitory activity towards cytochrome P450 family 1 (CYP1). These results are consistent with previous findings, in which visnagin, a CYP1 inhibitor, also prevents doxorubicin-induced cardiotoxicity. Importantly, genetic mutation of cyp1a protected zebrafish against doxorubicin-induced cardiotoxicity phenotypes. Together, these results provide strong evidence that CYP1 is an important contributor to doxorubicin-induced cardiotoxicity and highlight the CYP1 pathway as a candidate therapeutic target for clinical cardioprotection.     

Transdermal drug delivery: from micro to nano

Delivery across skin offers many advantages compared to oral or intravenous routes of drug administration. Skin however is highly impermeable to most molecules on the basis of size, hydrophilicity, lipophilicity and charge. For this reason it is often necessary to temporarily alter the barrier properties of skin for effective administration. This can be done by applying chemical enhancers, which alter the lipid structure of the top layer of skin (the stratum corneum, SC), by applying external forces such as electric currents and ultrasounds, by bypassing the stratum corneum via minimally invasive microneedles or by using nano-delivery vehicles that can cross and deliver their payload to the deeper layers of skin. Here we present a critical summary of the latest technologies used to increase transdermal delivery.     

Supercritical carbon dioxide extraction of polyunsaturated fatty acids from Northern shrimp (Pandalus borealis Kreyer) processing by-products

This study investigated the potential of Northern shrimp (Pandelus borealis Kreyer) by-products as a source of omega-3 polyunsaturated fatty acids (ω-3 PUFAs). The by-products (heads, shell and tail) of processing accounted for approximately 50–60% of the catch. Supercritical CO₂ extraction (SFE) of the by-products at 35 MPa and 40 °C generated a deep red oil, rich in ω-3 PUFAs, specifically 7.8 ± 0.06% eicosapentaenoic acid (EPA) and 8.0 ± 0.07 % docosahexaenoic acid (DHA).     

Nanostructured nanoparticles of self-assembled lipid pro-drugs as a route to improved chemotherapeutic agents

We demonstrate that oral delivery of self-assembled nanostructured nanoparticles consisting of 5-fluorouracil (5-FU) lipid prodrugs results in a highly effective, target-activated, chemotherapeutic agent, and offers significantly enhanced efficacy over a commercially available alternative that does not self-assemble. The lipid prodrug nanoparticles have been found to significantly slow the growth of a highly aggressive mouse 4T1 breast tumour, and essentially halt the growth of a human MDA-MB-231 breast tumour in mouse xenografts. Systemic toxicity is avoided as prodrug activation requires a three-step, enzymatic conversion to 5-FU, with the third step occurring preferentially at the tumour site. Additionally, differences in the lipid prodrug chemical structure and internal nanostructure of the nanoparticle dictate the enzymatic conversion rate and can be used to control sustained release profiles. Thus, we have developed novel oral nanomedicines that combine sustained release properties with target-selective activation.