RNA tertiary structure mediation by adenosine platforms

The crystal structure of a group I intron domain reveals an unexpected motif that mediates both intra- and intermolecular interactions. At three separate locations in the 160-nucleotide domain, adjacent adenosines in the sequence lie side-by-side and form a pseudo-base pair within a helix. This adenosine platform opens the minor groove for base stacking or base pairing with nucleotides from a noncontiguous RNA strand. The platform motif has a distinctive chemical modification signature that may enable its detection in other structured RNAs. The ability of this motif to facilitate higher order folding provides one explanation for the abundance of adenosine residues in internal loops of many RNAs.     

Electrodeposited polytyramine as an immobilisation matrix for enzyme biosensors

The application of an electrodeposited polytyramine film as an immobilisation matrix for the construction of enzyme biosensors is described. Glucose oxidase (used as a model enzyme) is covalently attached to free amine groups on the polytyramine film using the coupling reagents 1-ethyl-3(3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide. The resultant recognition interface consisted of multilayers of GOx immobilised onto the polymer surface. This method of constructing enzyme biosensors is shown to produce a highly reproducible and stable device. The biosensor showed no loss in electrode response after four months of dry storage and exhibited only minor loss in response after 20 days of repeated use. The resultant biosensor had a linear range of 0.1-28 mM glucose and a detection limit of 0.01 mM.     

Antisaccade task performance in questionnaire-identified schizotypes

Individuals who scored high on Perceptual Aberration-Magical Ideation Scales (Per-Mag; n = 90), the Social Anhedonia Scale (SocAnh; n = 39), and control participants (n = 89) were administered saccadic refixation (prosaccade) and saccadic suppression (antisaccade) tasks. Eye movements were scored in terms of error rates and latency. None of the groups differed in terms of their performance on the prosaccade task. Both the Per-Mag (p < 0.01) and SocAnh (p < 0.05) groups exceeded the controls in terms of mean antisaccade errors. The high-risk groups did not differ from each other. Eighteen of the Per-Mag individuals and 10 of the SocAnh individuals displayed deviant antisaccade performance. These findings are particularly interesting in light of suggestive evidence that antisaccade task deficits may serve as a marker of susceptibility to schizophrenia. It is hypothesized that the individuals who scored aberrantly on the Chapman scales and displayed antisaccade performance deficits are most likely to be at risk for the development of psychosis.     

Role of an inhibitory pyrimidine element and polypyrimidine tract binding protein in repression of a regulated alpha-tropomyosin exon

The lipogenic enzyme fatty acid synthase (FAS) is elevated in various human primary cancers and certain human cancer cell lines. FAS overexpression in human neoplasia has clinical relevance because of its association with tumor aggression and potential chemotherapeutic intervention. Here, we surveyed FAS in cell lines established from normal murine mammary epithelium (NMuMG) and from mammary tumors induced by either rodent polyoma (Py) virus or murine mammary tumor virus (MMTV). Western blotting revealed greater content of FAS in Py-transformed A1-1 and T1 than NMuMG or MMTV-transformed Mm5MT, RIIIMT and MMT060562. These data suggest that signaling events mediated by Py transformation may increase cellular amounts of FAS. Although FAS content was elevated to similar levels in A1-1 and T1, specific activities were significantly different as enzyme activity in T1 was 3-fold higher than A1-1. Likewise, FAS activity in NMuMG was about 0.5-fold higher than the MMTV-transformed lines, even though enzyme content was similar. Immunoprecipitation studies employing anti-phosphoamino acid antibodies followed by immunoblot analysis with anti-FAS antisera (and vice versa) were used to characterize the constitutive phosphorylation state of the enzyme. Phosphoserine and phosphothreonine residues were detected in the more active FAS from T1 and NMuMG, but not in the less active FAS from Mm5MT or A1-1. Discovery of phosphorylated FAS suggests that the enzyme may have more immediate control over lipogenesis than previously thought. High-dose (10-4 M) dexamethasone induced FAS content and activity in NMuMG and MMTV-transformed lines but not Py-transformed cells. Lower concentrations (10-8, 10-6 M) of dexamethasone also activated FAS but without concomitant elevation of its protein content, which was consistent with a phosphorylated form of FAS. Finally, cell lines were treated with the FAS inhibitor cerulenin: almost all breast cancer lines were growth inhibited at significantly lower amounts of drug than normal cell lineages, suggesting that FAS plays a greater role in viability of tumor cells than normal cells. Pretreatment with palmitate (a primary end-product of FAS) prior to cerulenin rescued A1-1 cells only slightly from growth inhibition, whereas pretreatment with oleate (a monounsaturated fatty acid synthesized from palmitate) synergized cerulenin's cytotoxic effects.     

Preserved pattern completion priming for novel, abstract geometric shapes in amnesics of several aetiologies

Two experiments were conducted using a paradigm developed by Gabrieli et al., Neuropsychologia 28, 417-427, 1990, which assessed both indirect and direct memory performance in a completion task for novel abstract geometric patterns. The preferred method of scoring was the lines method, based on the number of correct and incorrect lines produced for each item. It was chosen because it is both the simplest and the most informative measure. Two methods of scoring were used in previous work, namely, the strict whole figure method and the lenient whole figure method (Gabrielli et al., 1990; Verfaelie et al., Brain Cognit. 18, 34-45, 1992). Therefore to facilitate comparisons between studies and to determine the characteristics of different scoring methods, results with all three measures were included. In Experiment 1, two different encoding strategies of naming and copying were used in order to explore the relationship between indirect and direct memory performance. Indirect memory performance in the naming condition was at baseline whereas in the copying condition it was significantly above baseline. Cued recall did not differ across encoding conditions but recognition was higher in the naming condition than the copying condition. In Experiment 2, an attempt was made to extend the findings of two studies, one with H.M. (Gabrielli et al., 1990) and one with nine Korsakoffs (Verfaelie et al., 1992), to a larger group of 14 amnesics of several aetiologies. Indirect memory performance was found to be equivalent for the amnesics and their matched controls, only when the lenient and the lines methods of scoring were used. Recognition and cued recall performance was impaired for the amnesics compared to the controls.     

Charge transfer through DNA: A selective electrochemical DNA biosensor

The charge-transfer properties of DNA duplexes are exploited to produce a fast, simple, sensitive, and selective DNA biosensor by exposing the DNA recognition interface to a sample containing target DNA and the redox-active intercalator, anthraquinonemonosulfonic acid (AQMS). Electrochemistry from electron transfer through the DNA to AQMS intercalated into DNA duplexes can be differentiated from electrochemistry due to direct access of the AQMS to the electrode surface due to the difference in the environment of the AQMS giving a shift in the potential at which the molecule is reduced. The ability to distinguish between the two electrochemical signals enables DNA hybridization to be monitored in real time. This in situ detection scheme has good selectivity, being able to differentiate between a complementary target DNA sequence and one containing either C-A or G-A single-base mismatches. The concentration detection limit of the biosensor is 0.5 nM (1 pmol) with an assay time of 1 h. The fact that the end user is only required to simultaneously add the sample containing the target DNA and AQMS gives a DNA biosensor that is highly compatible with PCR on chip technologies.     

A Fill-and-Flow Biosensor

An alternative design for an amperometric biosensor for lactate is described in which the analyte flows through a rectangular duct without the aid of external pumping or a flow system. With this deviation from the traditional "stacked layers" of the biosensor geometry, the biorecognition matrix is located upstream of the detector electrode rather than directly over the electrode; this general design is reminiscent of the "dipstick" technology of the home test kits, but in the latter case the "flow" is a function of the nature of the wicking material rather than the dimensions of the channel. A droplet of analyte, l-lactate, is placed in the inlet well of the channel which flows through the cell and reacts with an immobilized lactate oxidase surface matrix zone, and the coproduct, hydrogen peroxide, is swept downstream to the monitoring electrode, where it is detected. The flow through the channel biosensor is shown to be laminar, and the biosensor response varies with the rate of flow of the analyte, which could be controlled by the outlet porosity; the sensitivity was observed to increase, but the dynamic range decreased with an increase in flow rate until a threshold loading, where the biosensor response became independent of the enzyme loading.     

Advances in the Application of Magnetic Nanoparticles for Sensing

Magnetic nanoparticles (MNPs) are of high significance in sensing as they provide viable solutions to the enduring challenges related to lower detection limits and nonspecific effects. The rapid expansion in the applications of MNPs creates a need to overview the current state of the field of MNPs for sensing applications. In this review, the trends and concepts in the literature are critically appraised in terms of the opportunities and limitations of MNPs used for the most advanced sensing applications. The latest progress in MNP sensor technologies is overviewed with a focus on MNP structures and properties, as well as the strategies of incorporating these MNPs into devices. By looking at recent synthetic advancements, and the key challenges that face nanoparticle‐based sensors, this review aims to outline how to design, synthesize, and use MNPs to make the most effective and sensitive sensors.     

Can the Shape of Nanoparticles Enable the Targeting to Cancer Cells over Healthy Cells?

Macropinocytosis is a consequence of oncogenic alterations of cancer cells while most healthy cells are non-macropinocytic. It is currently unclear whether macropinocytic cancer cells can be targeted rather than healthy cells, by adjusting the shape and size of nanoparticles. Herein, the endocytosis of two differently shaped nanoparticles; nanorods and nanospheres are compared in cancer and healthy cells. The cells are breast epithelial cancer cells (MCF7) and breast epithelial healthy cells (MCF10A) and pancreas cancer cells (PANC-1 cells) and non-tumourogenic patient-derived cancer-associated fibroblasts (CAFs). The endocytosis pathway is quantified by a combination of pair correlation microscopy and endocytosis inhibitors. MCF7 cells use clathrin-mediated endocytosis and macropinocytosis to take up the nanorods while MCF10A cells use predominantly clathrin-mediated endocytosis. Based on the comparison of endocytic behavior of cancer and healthy cells, MCF7 cells can be induced to take up more nanorods and suppress the metabolism and endocytosis of nanorods in MCF10A cells. The nanorods allow targeting to breast cancer MCF7 cells and pancreas cancer cells over the healthy cells. This study opens exciting possibilities for shape to target the cancer cells over healthy cells, by adjusting nanoparticle shape.