The bend in RNA created by the trans-activation response element bulge of human immunodeficiency virus is straightened by arginine and by Tat-derived peptide.

The trans-activation response element (TAR) found near the 5' end of the viral RNA of the human immunodeficiency virus contains a 3-nt bulge that is recognized by the virally encoded trans-activator protein (Tat), an important mediator of transcriptional activation. Insertion of the TAR bulge into double-stranded RNA is known to result in reduced electrophoretic mobility, suggestive of a bulge-induced bend. Furthermore, NMR studies indicate that Arg causes a change in the structure of the TAR bulge, possibly reducing the bulge angle. However, neither of these effects has been quantified, nor have they been compared with the effects of the TAR-Tat interaction. Recently, an approach for the quantification of bulge-induced bends has been described in which hydrodynamic measurements, employing the method of transient electric birefringence, have yielded precise estimates for the angles of a series of RNA bulges, with the angles ranging from 7 degrees to 93 degrees. In the current study, transient electric birefringence measurements indicate that the TAR bulge introduces a bend of 50 degrees +/- 5 degrees in the absence of Mg2+. Addition of Arg leads to essentially complete straightening of the helix (to < 10 degrees) with a transition midpoint in the 1 mM range. This transition demonstrates specificity for the TAR bulge: no comparable transition was observed for U3 or A3 (control) bulges with differing flanking sequences. An essentially identical structural transition is observed for the Tat-derived peptide, although the transition midpoint for the latter is near 1 microM. Finally, low concentrations of Mg2+ alone reduce the bend angle by approximately 50%, consistent with the effects of Mg2+ on other pyrimidine bulges. This last observation is important in view of the fact that most previous structural/binding studies were performed in the absence of Mg2+.

Detection of one slowly exchanging substrate water molecule in the S3 state of photosystem II.

The exchangeability of the substrate water molecules at the catalytic site of water oxidation in photosystem II has been probed by isotope-exchange measurements using mass spectrometric detection of flash-induced oxygen evolution. A stirred sample chamber was constructed to reduce the lag time between injection of H2(18)O and the detecting flash by a factor of more than 1000 compared to the original experiments by R. Radmer and O. Ollinger [(1986) FEBS Lett. 195, 285-289]. Our data show that there is a slow (t1/2 approximately 500 ms, 10 degrees C) and a fast (t1/2 <25 ms, 10 degrees C) exchanging substrate water molecule in the S3 state of photosystem II. The slow exchange is coupled with an activation energy of about 75 kJ/mol and is discussed in terms of a terminal manganese oxo ligand, while the faster exchanging substrate molecule may represent a water molecule not directly bound to the manganese center.

A cyclophilin from the polycentric anaerobic rumen fungus Orpinomyces sp. strain PC-2 is highly homologous to vertebrate cyclophilin B.

A cyclophilin (CyP) purified to homogeneity from the polycentric anaerobic rumen fungus Orpinomyces sp. strain PC-2 had a molecular mass of 20.5 kDa and a pI of 8.1. The protein catalyzed the isomerization of the prolyl peptide bond of N-succinyl-Ala-Ala-(cis,trans)-Pro-Phe p-nitroanilide with a kcat/Km value of 9.3 x 10(6) M-1.s-1 at 10 degrees C and pH 7.8. Cyclosporin A strongly inhibited this peptidylprolyl cis-trans isomerase activity with an IC50 of 19.6 nM. The sequence of the first 30 N-terminal amino acids of this CyP had high homology with the N-terminal sequences of other eukaryotic CyPs. By use of a DNA hybridization probe amplified by PCR with degenerate oligonucleotide primers designed based on the amino acid sequences of the N terminus of this CyP and highly conserved internal regions of other CyPs, a full-length cDNA clone was isolated. It possessed an open reading frame encoding a polypeptide of 203 amino acids with a calculated molecular weight of 21,969, containing a putative hydrophobic signal peptide sequence of 22 amino acids preceding the N terminus of the mature enzyme and a C-terminal sequence, Lys-Ala-Glu-Leu, characteristic of an endoplasmic reticulum retention signal. The Orpinomyces PC-2 CyP is a typical type B CyP. The amino acid sequence of the Orpinomyces CyP exhibits striking degrees of identity with the corresponding human (70%), bovine (69%), mouse (68%), chicken (66%), maize (61%), and yeast (54%) proteins. Phylogenetic analysis based on the CyP sequences indicated that the evolutionary origin of the Orpinomyces CyP was closely related with CyPs of animals.

Kinetics of hydrogen bond breakage in the process of unfolding of ribonuclease A measured by pulsed hydrogen exchange.

A sensitive test for kinetic unfolding intermediates in ribonuclease A (EC 3.1.27.5) is performed under conditions where the enzyme unfolds slowly (10 degrees C, pH 8.0, 4.5 M guanidinium chloride). Exchange of peptide NH protons (2H-1H) is used to monitor structural opening of individual hydrogen bonds during unfolding, and kinetic models are developed for hydrogen exchange during the process of protein unfolding. The analysis indicates that the kinetic process of unfolding can be monitored by EX1 exchange (limited by the rate of opening) for ribonuclease A in these conditions. Of the 49 protons whose unfolding/exchange kinetics was measured, 47 have known hydrogen bond acceptor groups. To test whether exchange during unfolding follows the EX2 (base-catalyzed) or the EX1 (uncatalyzed) mechanism, unfolding/exchange was measured both at pH 8.0 and at pH 9.0. A few faster-exchanging protons were found that undergo exchange by both EX1 and EX2 processes, but the 43 slower-exchanging protons at pH 8 undergo exchange only by the EX1 mechanism, and they have closely similar rates. Thus, it is likely that all 49 protons undergo EX1 exchange at the same rate. The results indicate that a single rate-limiting step in unfolding breaks the entire network of peptide hydrogen bonds and causes the overall unfolding of ribonuclease A. The additional exchange observed for some protons that follows the EX2 mechanism probably results from equilibrium unfolding intermediates and will be discussed elsewhere.

The large scale distribution of galaxies in the Shapley Supercluster

We present new results of our wide-field redshift survey of galaxies in a 182 square degree region of the Shapley Supercluster (SSC) based on observations with the FLAIR-II spectrograph on the UK Schmidt Telescope (UKST). In this paper we present new measurements to give a total sample of redshifts for 710 bright (R less than or equal to 16.6) galaxies, of which 464 are members of the SSC (8000 < υ < 18 000 km s(-1)). Our data reveal that the main plane of the SSC (upsilon approximate to 14 500 km s(-1)) extends further than previously realised, filling the whole extent of our survey region of 10 degrees by 20 degrees on the sky (35 Mpc by 70 Mpc, for H-0 = 75 km s(-1) Mpc(-1)). There is also a significant structure associated with the slightly nearer Abell 3571 cluster complex (upsilon approximate to 12 000 km s(-1)) with a caustic structure evident out to a radius of 6 Mpc. These galaxies seem to link two previously identified sheets of galaxies and establish a connection with a third one at (V) over bar = 15 000 km s(-1) near RA = 13(h). They also tend to fill the gap of galaxies between the foreground Hydra-Centaurus region and the more distant SSC. We calculate galaxy overdensities of 5.0+/-0.1 over the 182 square degree region surveyed and 3.3+.-0.1 in a 159 square degree region excluding rich clusters. Over the large region of our survey the inter-cluster galaxies make up 46 per cent of all galaxies in the SSC region and may contribute a similar amount of mass to the cluster galaxies.

Effect of seasonal variation and storage temperature on leaf chlorophyll fluorescence and vase life of cut roses

Low temperature injury (LTI) of roses (Rosa hybrida L.) is difficult to assess by visual observation. Relative chlorophyll fluorescence (CF; F-v/F-m) is a non-invasive technique that provides an index of stress effects on photosystem 11 (PS 11) activity. This instrumental technique allows determination of the photosynthetic efficiency of plant tissues containing chloroplasts, such as rose leaves. In the present study, pre- and Post-Storage measurements of F-v/F-m were carried out to assess LTI in 'First Red' and 'Akito' roses harvested year round. Relationships between the pre-harvest environment conditions of temperature, relative humidity and photon flux density (PFD), F-v/F-m, and, vase life duration after storage are reported. After harvest, roses were stored at 1, 5 and 10 degrees C for 10 days. Non-stored roses were the control treatment. F-v/F-m ratios were reduced following storage, suggesting LTI of roses. However, reductions in F-v/F-m were not closely correlated with reduced vase life duration and were seasonally dependent. Only during winter experiments was F-v/F-m of roses stored at 1 degrees C significantly (P <= 0.001) lower compared to F-v/F-m of non-stored control roses and roses stored at 5 and 10 degrees C. Thus, the fall of F-v/F-m was due to an interaction of growing season and storage at 1 degrees C. Vase lives of roses grown during winter were significantly (P <= 0.001) shorter compared to roses grown during summer. Length of vase life was intermediate for roses grown during autumn and spring. Because of the lack of correlation between F-v/F-m and post-storage vase life it is concluded that the CF parameter F-v/F-m is nota practical index for assessing LTI in cold-stored roses. Higher PFD and temperature in summer were positively and significantly correlated with maintenance of post-storage FvIF ratios and longer vase life. It is suggested that shorter vase lives and lower post-storage F-v/F-m values after storage at 1 degrees C are consequences of reduced photosynthesis and smaller carbohydrate pools in winter-harvested roses. (c) 2004 Elsevier B.V All rights reserved.

Effect of root zone temperature on oilseed rape (Brassica napus) response to boron

Oilseed rape (Brassica napus) is sensitive to low boron (B) supply, and its growth response to B may be influenced by soil temperature. To test the relationship between B and temperature, oilseed rape (cv. Hyola 42) seedlings were grown at 10 degrees C (low) root zone temperature (RZT) with B supply from deficient to adequate B levels until growth of low B plants just began to slow down. Half of the pots were then transferred to 20 degrees C (warm) RZT for 11 days before they were moved back to 10 degrees C RZT for the final 4 days. Both plant dry mass and B uptake increased after plants were exposed to warm RZT. However, plant B deficiency was exacerbated by warm RZT in low B plants because of increased relative growth rate and shoot-root ratio without a commensurate increase in B uptake rate. It is concluded that RZT above the critical threshold for chilling injury in oilseed rape can nevertheless affect the incidence of B deficiency by altering shoot-root ratio and hence the balance between shoot B demand and B uptake.

Rotational and centration stability of an aspheric intraocular lens with a simulated toric design

Purpose: To assess the stability of the Akreos AO intraocular lens (IOL) platform with a simulated toric design using objective image analysis. Setting: Six hospital eye clinics across Europe. Methods: After implantation in 1 eye of patients, IOLs with orientation marks were imaged at 1 to 2 days, 7 to 14 days, 30 to 60 days, and 120 to 180 days. The axis of rotation and IOL centration were objectively assessed using validated image analysis. Results: The study enrolled 107 patients with a mean age of 69.9 years ± 7.7 (SD). The image quality was sufficient for IOL rotation analysis in 91% of eyes. The mean rotation between the first day postoperatively and 120 to 180 days was 1.93 ± 2.33 degrees, with 96% of IOLs rotating fewer than 5 degrees and 99% rotating fewer than 10 degrees. There was no significant rotation between visits and no clear bias in the direction of rotation. In 71% of eyes, the dilation and image quality was sufficient for image analysis of centration. The mean change in centration between 1 day and 120 to 180 days was 0.21 ± 0.11 mm, with all IOLs decentering less than 0.5 mm. There was no significant decentration between visits and no clear bias in the direction of the decentration. Conclusion: Objective analysis of digital retroillumination images taken at different postoperative periods shows the aspheric IOL platform was stable in the eye and is therefore suitable for the application of a toric surface to correct corneal astigmatism.

Does rebound tonometry probe misalignment modify intraocular pressure measurements in human eyes?

Purpose. To examine the influence of positional misalignments on intraocular pressure (IOP) measurement with a rebound tonometer. Methods. Using the iCare rebound tonometer, IOP readings were taken from the right eye of 36 healthy subjects at the central corneal apex (CC) and compared to IOP measures using the Goldmann applanation tonometer (GAT). Using a bespoke rig, iCare IOP readings were also taken 2 mm laterally from CC, both nasally and temporally, along with angular deviations of 5 and 10 degrees, both nasally and temporally to the visual axis. Results. Mean IOP ± SD, as measured by GAT, was 14.7±2.5 mmHg versus iCare tonometer readings of 17.4±3.6 mmHg at CC, representing an iCare IOP overestimation of 2.7±2.8 mmHg (P<0.001), which increased at higher average IOPs. IOP at CC using the iCare tonometer was not significantly different to values at lateral displacements. IOP was marginally underestimated with angular deviation of the probe but only reaching significance at 10 degrees nasally. Conclusions. As shown previously, the iCare tonometer overestimates IOP compared to GAT. However, IOP measurement in normal, healthy subjects using the iCare rebound tonometer appears insensitive to misalignments. An IOP underestimation of <1 mmHg with the probe deviated 10 degrees nasally reached statistical but not clinical significance levels. © 2013 Ian G. Beasley et al.

Distribution, density and abundance of Antarctic ice seals off Queen Maud Land and the eastern Weddell Sea

The Antarctic Pack Ice Seal (APIS) Program was initiated in 1994 to estimate the abundance of four species of Antarctic phocids: the crabeater seal Lobodon carcinophaga, Weddell seal Leptonychotes weddellii, Ross seal Ommatophoca rossii and leopard seal Hydrurga leptonyx and to identify ecological relationships and habitat use patterns. The Atlantic sector of the Southern Ocean (the eastern sector of the Weddell Sea) was surveyed by research teams from Germany, Norway and South Africa using a range of aerial methods over five austral summers between 1996-1997 and 2000-2001. We used these observations to model densities of seals in the area, taking into account haul-out probabilities, survey-specific sighting probabilities and covariates derived from satellite-based ice concentrations and bathymetry. These models predicted the total abundance over the area bounded by the surveys (30°W and 10°E). In this sector of the coast, we estimated seal abundances of: 514 (95 % CI 337-886) x 10**3 crabeater seals, 60.0 (43.2-94.4) x 10**3 Weddell seals and 13.2 (5.50-39.7) x 10**3 leopard seals. The crabeater seal densities, approximately 14,000 seals per degree longitude, are similar to estimates obtained by surveys in the Pacific and Indian sectors by other APIS researchers. Very few Ross seals were observed (24 total), leading to a conservative estimate of 830 (119-2894) individuals over the study area. These results provide an important baseline against which to compare future changes in seal distribution and abundance.

On the Movements of the North Atlantic Subpolar Front in the Preinstrumental Past*

Three sediment records of sea surface temperature (SST) are analyzed that originate from distant locations in the North Atlantic, have centennial-to-multicentennial resolution, are based on the same reconstruction method and chronological assumptions, and span the past 15 000 yr. Using recursive least squares techniques, an estimate of the time-dependent North Atlantic SST field over the last 15 kyr is sought that is consistent with both the SST records and a surface ocean circulation model, given estimates of their respective error (co)variances. Under the authors' assumptions about data and model errors, it is found that the 10 degrees C mixed layer isotherm, which approximately traces the modern Subpolar Front, would have moved by ~15 degrees of latitude southward (northward) in the eastern North Atlantic at the onset (termination) of the Younger Dryas cold interval (YD), a result significant at the level of two standard deviations in the isotherm position. In contrast, meridional movements of the isotherm in the Newfoundland basin are estimated to be small and not significant. Thus, the isotherm would have pivoted twice around a region southeast of the Grand Banks, with a southwest-northeast orientation during the warm intervals of the Bolling-Allerod and the Holocene and a more zonal orientation and southerly position during the cold interval of the YD. This study provides an assessment of the significance of similar previous inferences and illustrates the potential of recursive least squares in paleoceanography.

Differential effects of low-temperature inhibition on the propylene induced autocatalysis of ethylene production, respiration and ripening of 'Hayward' kiwifruit

Previous studies (Stavroulakis and Sfakiotakis, 1993) have shown an inhibition of propylene-induced ethylene production in kiwifruit below a critical temperature range of 11-14.8 degrees C. The aim of this research was to identify the biochemical basis of this inhibition in kiwifruit below 11-14.8 degrees C. 'Hayward' kiwifruit were treated with increasing propylene concentrations at 10 and 20 degrees C. Ethylene biosynthesis pathways and fruit ripening were investigated. Kiwifruit at 20 degrees C in air started autocatalysis of ethylene production and ripened after 19 d with a concomitant increase in respiration. Ethylene production and the respiration rise appeared earlier with increased propylene concentrations. Ripening proceeded immediately after propylene treatment, while ethylene autocatalysis needed a lag period of 24-72 h. The latter event was attributed to the delay found in the induction of 1-aminocyclopropane-1-carboxylate synthase (ACC synthase) activity and consequently to the delayed increase of l-aminocyclopropane l-carboxylic acid (ACC) content. In contrast propylene treatment induced 1-aminocyclopropane-1-carboxylate oxidase (ACC oxidase) activity with no lag period. Moreover, transcription of ACC synthase and ACC oxidase genes was active only in ethylene-producing kiwifruit at 20 degrees C. In contrast, treatment at 10 degrees C with propylene strongly inhibited ethylene production, which was attributed to the low activities of both ACC synthase and ACC oxidase as well as the low initial ACC level. Interestingly, fruit treated with propylene at 10 degrees C appeared to be able to transcribe the ACC oxidase but not the ACC synthase gene. However, propylene induced ripening of that fruit almost as rapidly as in the propylene-treated fruit at 20 degrees C. Respiration rate was increased together with propylene concentration. It is concluded that kiwifruit stored at 20 degrees C behaves as a typical climacteric fruit, while at 10 degrees C behaves like a non-climacteric fruit. We propose that the main reasons for the inhibition of the propylene induced (autocatalytic) ethylene production in kiwifruit at low temperature (less than or equal to 10 degrees C), are primarily the suppression of the propylene-induced ACC synthase gene expression and the possible post-transcriptional modification of ACC oxidase.

Differential effects of low-temperature inhibition on the propylene induced autocatalysis of ethylene production, respiration and ripening of 'Hayward' kiwifruit

Previous studies (Stavroulakis and Sfakiotakis, 1993) have shown an inhibition of propylene-induced ethylene production in kiwifruit below a critical temperature range of 11-14.8 degrees C. The aim of this research was to identify the biochemical basis of this inhibition in kiwifruit below 11-14.8 degrees C. 'Hayward' kiwifruit were treated with increasing propylene concentrations at 10 and 20 degrees C. Ethylene biosynthesis pathways and fruit ripening were investigated. Kiwifruit at 20 degrees C in air started autocatalysis of ethylene production and ripened after 19 d with a concomitant increase in respiration. Ethylene production and the respiration rise appeared earlier with increased propylene concentrations. Ripening proceeded immediately after propylene treatment, while ethylene autocatalysis needed a lag period of 24-72 h. The latter event was attributed to the delay found in the induction of 1-aminocyclopropane-1-carboxylate synthase (ACC synthase) activity and consequently to the delayed increase of l-aminocyclopropane l-carboxylic acid (ACC) content. In contrast propylene treatment induced 1-aminocyclopropane-1-carboxylate oxidase (ACC oxidase) activity with no lag period. Moreover, transcription of ACC synthase and ACC oxidase genes was active only in ethylene-producing kiwifruit at 20 degrees C. In contrast, treatment at 10 degrees C with propylene strongly inhibited ethylene production, which was attributed to the low activities of both ACC synthase and ACC oxidase as well as the low initial ACC level. Interestingly, fruit treated with propylene at 10 degrees C appeared to be able to transcribe the ACC oxidase but not the ACC synthase gene. However, propylene induced ripening of that fruit almost as rapidly as in the propylene-treated fruit at 20 degrees C. Respiration rate was increased together with propylene concentration. It is concluded that kiwifruit stored at 20 degrees C behaves as a typical climacteric fruit, while at 10 degrees C behaves like a non-climacteric fruit. We propose that the main reasons for the inhibition of the propylene induced (autocatalytic) ethylene production in kiwifruit at low temperature (less than or equal to 10 degrees C), are primarily the suppression of the propylene-induced ACC synthase gene expression and the possible post-transcriptional modification of ACC oxidase.

Complicaciones perioperatorias en el manejo de corrección de escoliosis en un hospital de tercer nivel en la ciudad de Bogotá entre el 2010-2014

Introducción: la escoliosis, definida como una deformidad de la columna vertebral en más de 10 grados, se agrupa en 4 orígenes distintos: idiopática, congénita, neuromuscular y sindromática. Cada una de ellas con diferente riesgo de progresión en severidad, lo que determina la necesidad de corrección quirúrgica para cada caso en su tratamiento. Conocer las probabilidades de complicación en la etapa peri operatoria, abre la posibilidad de dar asesoría integral que mida la relación riesgo - beneficio de la medida terapéutica. Métodos: se realiza un estudio retrospectivo de corte transversal. La información se obtiene de los registros de las historias clínicas desde el año 2010 al 2014, de pacientes intervenidos quirúrgicamente para la corrección de escoliosis. Resultados: Se obtuvieron 318 registros de procedimientos en 230 pacientes. El tipo de escoliosis presentado con mayor frecuencia es de origen idiopático 108 (47%); en los 4 tipos de escoliosis se observa mayor número de mujeres 169 (73,4%). La edad donde se concentran la mayor cantidad de cirugías para corrección de escoliosis está entre 10 - 14 años. De 13 complicaciones seleccionadas, aquellas de origen respiratorio son las de mayor probabilidad de ocurrencia (OR 30 - sig 0,000). La característica sociodemográfica “edad” logra predecir el 46% de las complicaciones perioperatorias. Discusión: La corrección de escoliosis va acompañada de comorbilidades, datos sociodemográficos y diagnósticos que en conjunto determinan el grado de complicación peri operatoria. Se necesitan registros clínicos muy completos para poder determinar la asociación entre la etiología de la escoliosis con las complicaciones más comunes. Este trabajo propone y evidencia los datos de los registros clínicos como predictores de complicaciones quirúrgicas de escoliosis. Esto exige un trabajo institucional interno que garantice la calidad en los registros de datos clínicos.