Current and emerging commercial optical biosensors.

The field of commercial optical biosensors is rapidly evolving, with new systems and detection methods being developed each year. This review outlines the currently available biosensor hardware and highlights unique features of each platform. Affinity-based biosensor technology, with its high sensitivity, wide versatility and high throughput, is playing a significant role in basic research, pharmaceutical development, and the food and environmental sciences. Likewise, the increasing popularity of biosensors is prompting manufacturers to develop new instrumentation for dedicated applications. We provide a preview of some of the emerging commercial systems that are dedicated to drug discovery, proteomics, clinical diagnostics and routine biomolecular interaction analysis.     

A novel array-based assay of in situ tissue transglutaminase activity in human umbilical vein endothelial cells

Transglutaminases (TGs), a family of calcium-dependent transamidating enzymes, are involved in functions such as apoptosis and inflammation and play a role in autoimmune diseases and neurodegenerative disorders. In this study, we describe a novel array-based approach to rapidly determine in situ TG activity in human umbilical vein endothelial cells and J82 human bladder carcinoma cells. Amine arrays were fabricated by immobilizing 3-aminopropyltrimethoxysilane on glass slides. The assay was specific and highly reproducible. The average coefficient of variation betweens spots was 2.6% (n = 3 arrays), and the average correlation coefficients between arrays and between arrays/reactions were 0.998 and 0.976, respectively (n = 3 arrays). The assay was successfully applied to detect changes in TG activity induced by maitotoxin and to analyze inhibition of the TG activation with cystamine and monodansyl cadaverine. In addition, the assay demonstrated that intracellular reactive oxygen species regulate the maitotoxin-induced activation of TG. Thus, the array-based in situ TG activity assay constitutes a rapid and high-throughput approach to investigating the roles of TGs in cell signaling.     

Prenatal diagnosis of Pallister-Killian syndrome and literature review

Pallister-Killian syndrome (PKS) is a rare sporadic genetic disorder usually caused by mosaicism of an extra isochromosome of 12p (i(12p)). This retrospective study analysed the prenatal ultrasound manifestations and molecular and cytogenetic results of five PKS foetuses. Samples of amniotic fluid and/or cord blood, skin biopsy and placenta were collected. Conventional karyotyping and single nucleotide polymorphism array (SNP array) were performed on all the amniotic fluid or cord blood samples. Copy number variants sequencing (CNV-seq) and fluorescence in situ hybridization (FISH) were also used for the validation for one foetus. All the five foetuses were from pregnancies with advanced parental age. Two foetuses involved structural abnormalities and one foetus had only soft markers, all of which included increased nuchal translucency. The rest two foetuses had normal ultrasounds in the second trimester, which has rarely been reported before. The karyotype revealed typical i(12p) in four cases and a small supernumerary marker chromosome consisting of 12p and 20p in the remaining one case. The proportion of cells with i(12p) ranged from 0 to 100% in cultural cells, while SNP array results suggested 2−4 copies of 12p. For one foetus, metaphase FISH showed normal results, but the interphase FISH suggested cell lines with two, three and four copies of 12p in the amniotic fluid. Advanced parental age may be an important risk factor for PKS, and there were no typical ultrasound manifestations related to PKS. A combination of karyotype analysis and molecular diagnosis is an effective method for the diagnosis of PKS.     

On non-metric multidimensional scaling ordination and interpretation of the matorral vegetation in lowland Murcía

The plant-environment relationships in a range of matorral communities, having different rainfall conditions in semi-arid lowland habitats in Murcía, S.E. Spain, were examined using the non-metric multidimensional scaling ordination technique. Hypotheses on floristic variations were derived based on an interpretative strategy which involved a site configuration rotation, followed by stepwise multiple regression analysis. An environmental data set was used to isolate variables associated with site co-ordinate trends. Results showed that floristic variation in these communities was mainly determined by aspect induced radiation receipt. Besides, most environmental trends in the study areas were found to be oblique to original site ordination axes and configuration rotation seemed to be a prerequisite in quantitative interpretation. The interpretative strategy introduced in this study was effective. It enhanced straightforward, quantitative and objective interpretation whereby inductive inferences on environmental trends could be readily formulated.Abbreviations ARAD radiation receipt surrogate - DEPTH soil depth - DISTURB disturbance index - NMDS non-metric multidimensional scaling - NUT nutrient scalar - SLOPE slope angle - TEXTIN texture index     

Consistent handedness of microtubule helical arrays in maize and Arabidopsis primary roots

Summary The orientation of cortical microtubules in plant cells has been extensively studied, in part because of their influence on the expansion of most plant cell types. Cortical microtubules are often arranged in helical arrays, which are well known to occur with a specific pitch as a function of development or experimental treatment; however, it is not known if the handedness of helical arrays can also be specified. We have studied the handedness of helical arrays by using Vibratome sectioning of maize primary roots and confocal microscopy of Arabidopsis primary roots. In cortical cells of maize roots, the helical array was found to have the same handedness at a given position, not only for the cells of a single root, but also for the cells of more than one hundred roots examined. Quantification of angular distribution of apparent individual microtubules showed that defined regions of the root were composed of cells with highly uniform microtubule orientation. In the region between transverse and longitudinal microtubules (5–10.5 mm from the tip), the array formed a right-handed helix, and basal of cells with longitudinal microtubules (11.5–15 mm from the tip), the array formed a left-handed helix. Similarly, in epidermal cells of Arabidopsis roots right-handed helical arrays were found in the region between transverse and longitudinal microtubules. These results suggest that, in addition to the orientation of microtubules, the handedness of helical microtubule arrays is under cellular control.Abbreviations Cy3 indocarbocyanine - PBS phosphate-buffered saline - PIPES piperazine-N,N-bis-[2-ethanesulfonic acid]     

Sulfide-quinone and sulfide-cytochrome reduction in Rhodobacter capsulatus

The reduction by sulfide of exogenous ubiquinone is compared to the reduction of cytochromes in chromatophores of Rhodobacter capsulatus. From titrations with sulfide values for V_max of 300 and 10 moles reduced/mg bacteriochlorophyll a·h, and for K_m of 5 and 3 M were estimated, for decyl-ubiquinone-and cytochrome c-reduction, respectively. Both reactions are sensitive to KCN, as has been found for sulfide-quinone reductase (SQR) in Oscillatoria limnetica, which is a flavoprotein. Effects of inhibitors interfering with quinone binding sites suggest that at least part of the electron transport from sulfide in R. capsulatus employs the cytochrome bc ₁-complex via the ubiquinone pool.Abbreviations BChl a bacteriochlorophyll a - DAD diaminodurene - decyl-UQ decyl-ubiquinone - LED light emitting diode - NQNO 2-n-nonyl-4-hydroxyquinoline-N-oxide - PQ-1 plastoquinone 1 - SQR sulfide-quinone reductase (E.C. 1.8.5.'.) - UQ ubiquinone 10 - Q_c the quinone reduction site on the cytochrome b ₆ f/bc ₁, complex (also termed Q_i or Q_r or Q_n) - Q_s the quinone reduction site on SQR - Q_z quinol oxidation site on the b ₆ f/bc ₁, complex (also termed Q_o or Q_p)     

Cell wall synthesis during growth and maturation of Nitella internodal cells

An improved ¹³C-density-labeling method was used to study cell wall synthesis in rapidly expanding, slowly expanding and recently mature internodes of Nitella translucens var axillaris (A.Br.) R.D.W. As cells matured, the rate of wall synthesis slowed and the deposition of cellulose microfibrils changed from a predominantly transverse direction in the primary wall of rapidly expanding internodes to a helicoidal array in the secondary wall of mature internodes. The secondary wall was characterized by relatively higher rates of cellulose synthesis and lower rates of pectin synthesis than the primary wall. The synthesis of xyloglucan also decreased markedly at the transition to secondary wall synthesis, while the synthesis of mannose-rich hemicellulose increased. Even though structural differences were striking between the primary and secondary walls of Nitella, compositional differences between the two types of wall were quantitative rather than qualitative. The authors appreciate the assistance of Martin Yousef with the electron microscopy.     

Action potential velocity measurements in the upper trapezius muscle

The feasibility of action potential velocity (APV) measurements in the upper trapezius muscle with surface electrodes has been investigated. A four-bar electrode array connected to a double differential amplifier system was used. APV was estimated by a polarity correlation algorithm implemented on a PC computer. Six females and six males participated in the investigation. Attempts to get acceptable APV estimates were made in five electrode locations, 5 mm interspaced along the upper rim, beginning in the most distal part. Data were collected while holding out the arm horizontally in the sagittal plane. The results indicate that the method worked in five out of six males while it was difficult to get reliable estimates in the female group. Furthermore, the two most distal electrode locations gave the best results. In these two locations, the average APV for males was 4.8 m s⁻¹, 0.9. The difficulties in the female group were possibly due to small muscle dimensions and subcutaneous fat. Use of the double differential technique seems to be essential; attempts with the single differential technique were fruitless.     

The Structure and Characterization of the Surface Protein Layer of a Comamonas-Like Organism

The regular surface layer of a strain of a Comamonas-like organism was examined by electron microscopy. The surface layer protein was easily extracted from the cell surface by a 2.5 M solution of lithium chloride. The protein subunit has a molecular size of 32,000 daltons, but usually forms a large aggregate of more than 1,200,000 daltons. In the extract it formed a regular array of p4 symmetry and was observed to be intimately associated with fragments of lipopolysaccharide. The size of a subunit determined by the negative staining method and the image processing method measured 5.2 × 6.4 nm (width and length), was arranged in a cobblestone-like pattern, and was located in a lattice space measuring 13.0 nm square.     

On the integrated interpretation of indirect site ordinations: a case study using semi-arid vegetation in southeastern Spain

The need for rigorous methods of interpreting indirect site ordinations is discussed and the problem of oblique habitat and phytosociological trends is emphasized. A sequential approach, termed integrated interpretation, is introduced to overcome this problem. It first involves a technique termed rotational correlation a linear form of trend surface analysis, which locates the best fit between a habitat variable and the coordinates of a pluridimensional ordination. The coordinates at positions of best fit are then used to produce two-way site-species tables (seriation arrays) which summarize floristic variation in relation to each major habitat trend. The seriation arrays can also be used to identify groups of differential species. Integrated interpretation is demonstrated using semi-arid vegetation from Murcia Province, SE Spain. A two-axis site ordination of vegetation data by non-metric multidimensional scaling is shown to have two oblique trends related to aspect-induced topoclimates and types of past anthropogenic disturbance. Rotation of the configuration to maximum linear correlation achieves very high levels of explanation for each factor. Rotated ordination coordinates from the correlation analysis are used to obtain two seriation arrays and floristic noda which relate directly to the two underlying habitat trends. A floristic discontinuity is revealed on the topoclimate continuum and it is hypothesized that temperature conditions are a major determinant of floristic composition, but that moisture partly determines vegetational cover and phytomass. Several patterns of past cultivation are detected and associated with five main kinds of geomorphological unit, together with some evidence that the rate of succession is partly determined by topoclimate. The integrated approach to interpretation is compared with visual methods and other multivariate techniques. A case is made for increased interpretational rigour in site coordinate studies, especially the discontinuation of trend-seeking using individual unrotated ordination axes.Nomenclature of vascular plants follows Tutin et al. (1964–1980) Flora Europaea 5 Vols. Cambridge University Press, Cambridge. Nomenclature of syntaxa follows Rivas-Goday & Rivas-Martinez (1967).I am indebted to J. A. Michel, Dr M. A. Abdelrahman and Monica M. Dargie for field assistance. Field discussion with Dr R. L. Wright, Dr M. P. Austin and Professor F. Esteve Chueca was most helpful; the latter assisted with confirmation of vascular plant voucher specimens. Non-vascular taxonomy was determined by Dr C. Humphries (mosses and one alga) and Dr O. L. Gilbert (lichens). An anonymous referee suggested an approximation of the analytical solution to rotational correlation, and Dr N. R. J. Fieller assisted with its confirmation. The project was assisted by a grant from the University of Sheffield Research Fund.