Impacts of a word-picture training on reading in youth with mixed intellectual disabilities. A waiting-list control group comparison

Individuals with intellectual disabilities (ID) often struggle with learning how to read. Reading difficulties seem to be the most common secondary condition of ID. Only one in five children with mild or moderate ID achieves even minimal literacy skills. However, literacy education for children and adolescents with ID has been largely overlooked by researchers and educators. While there is little research on reading of children with ID, many training studies have been conducted with other populations with reading difficulties. The most common approach of acquiring literacy skills consists of sophisticated programs that train phonological skills and auditory perception. Only few studies investigated the influence of implicit learning on literacy skills. Implicit learning processes seem to be largely independent of age and IQ. Children are sensitive to the statistics of their learning environment. By frequent word reading they acquire implicit knowledge about the frequency of single letters and letter patterns in written words. Additionally, semantic connections not only improve the word understanding, but also facilitate storage of words in memory. Advances in communication technology have introduced new possibilities for remediating literacy skills. Computers can provide training material in attractive ways, for example through animations and immediate feedback .These opportunities can scaffold and support attention processes central to learning. Thus, the aim of this intervention study was to develop and implement a computer based word-picture training, which is based on statistical and semantic learning, and to examine the training effects on reading, spelling and attention in children and adolescents (9-16 years) diagnosed with mental retardation (general IQ  74). Fifty children participated in four to five weekly training sessions of 15-20 minutes over 4 weeks, and completed assessments of attention, reading, spelling, short-term memory and fluid intelligence before and after training. After a first assessment (T1), the entire sample was divided in a training group (group A) and a waiting control group (group B). After 4 weeks of training with group A, a second assessment (T2) was administered with both training groups. Afterwards, group B was trained for 4 weeks, before a last assessment (T3) was carried out in both groups. Overall, the results showed that the word-picture training led to substantial gains on word decoding and attention for both training groups. These effects were preserved six weeks later (group A). There was also a clear tendency of improvement in spelling after training for both groups, although the effect did not reach significance. These findings highlight the fact that an implicit statistical learning training in a playful way by motivating computer programs can not only promote reading development, but also attention in children with intellectual disabilities.

Feasibility of cardiopulmonary exercise testing and training using a robotics-assisted tilt table in dependent-ambulatory stroke patients

BACKGROUND: We evaluated the feasibility of an augmented robotics-assisted tilt table (RATT) for incremental cardiopulmonary exercise testing (CPET) and exercise training in dependent-ambulatory stroke patients. METHODS: Stroke patients (Functional Ambulation Category ≤ 3) underwent familiarization, an incremental exercise test (IET) and a constant load test (CLT) on separate days. A RATT equipped with force sensors in the thigh cuffs, a work rate estimation algorithm and real-time visual feedback to guide the exercise work rate was used. Feasibility assessment considered technical feasibility, patient tolerability, and cardiopulmonary responsiveness. RESULTS: Eight patients (4 female) aged 58.3 ± 9.2 years (mean ± SD) were recruited and all completed the study. For IETs, peak oxygen uptake (V'O2peak), peak heart rate (HRpeak) and peak work rate (WRpeak) were 11.9 ± 4.0 ml/kg/min (45 % of predicted V'O2max), 117 ± 32 beats/min (72 % of predicted HRmax) and 22.5 ± 13.0 W, respectively. Peak ratings of perceived exertion (RPE) were on the range "hard" to "very hard". All 8 patients reached their limit of functional capacity in terms of either their cardiopulmonary or neuromuscular performance. A ventilatory threshold (VT) was identified in 7 patients and a respiratory compensation point (RCP) in 6 patients: mean V'O2 at VT and RCP was 8.9 and 10.7 ml/kg/min, respectively, which represent 75 % (VT) and 85 % (RCP) of mean V'O2peak. Incremental CPET provided sufficient information to satisfy the responsiveness criteria and identification of key outcomes in all 8 patients. For CLTs, mean steady-state V'O2 was 6.9 ml/kg/min (49 % of V'O2 reserve), mean HR was 90 beats/min (56 % of HRmax), RPEs were > 2, and all patients maintained the active work rate for 10 min: these values meet recommended intensity levels for bouts of training. CONCLUSIONS: The augmented RATT is deemed feasible for incremental cardiopulmonary exercise testing and exercise training in dependent-ambulatory stroke patients: the approach was found to be technically implementable, acceptable to the patients, and it showed substantial cardiopulmonary responsiveness. This work has clinical implications for patients with severe disability who otherwise are not able to be tested.

Selective Transport of Zinc, Manganese, Nickel, Cobalt and Cadmium in the Root System and Transfer to the Leaves in Young Wheat Plants

• Background and Aims The uptake, translocation and redistribution of the heavy metals zinc, manganese, nickel, cobalt and cadmium are relevant for plant nutrition as well as for the quality of harvested plant products. The long-distance transport of these heavy metals within the root system and the release to the shoot in young wheat (Triticum aestivum ‘Arina’) plants were investigated. • Methods After the application of 65Zn, 54Mn, 63Ni, 57Co and 109Cd for 24 h to one seminal root (the other seminal roots being excised) of 54-h-old wheat seedlings, the labelled plants were incubated for several days in hydroponic culture on a medium without radionuclides. • Key Results The content of 65Zn decreased quickly in the labelled part of the root. After the transfer of 65Zn from the roots to the shoot, a further redistribution in the phloem from older to younger leaves was observed. In contrast to 65Zn, 109Cd was released more slowly from the roots to the leaves and was subsequently redistributed in the phloem to the youngest leaves only at trace levels. The content of 63Ni decreased quickly in the labelled part of the root, moving to the newly formed parts of the root system and also accumulating transiently in the expanding leaves. The 54Mn content decreased quickly in the labelled part of the root and increased simultaneously in leaf 1. A strong retention in the labelled part of the root was observed after supplying 57Co. • Conclusions The dynamics of redistribution of 65Zn, 54Mn, 63Ni, 57Co and 109Cd differed considerably. The rapid redistribution of 63Ni from older to younger leaves throughout the experiment indicated a high mobility in the phloem, while 54Mn was mobile only in the xylem and 57Co was retained in the labelled root without being loaded into the xylem.

The Cell Agglutination Agent, Phytohemagglutinin-L, Improves the Efficiency of Somatic Nuclear Transfer Cloning in Cattle (Bos taurus)

One of the several factors that contribute to the low efficiency of mammalian somatic cloning is poor fusion between the small somatic donor cell and the large recipient oocyte. This study was designed to test phytohemagglutinin (PHA) agglutination activity on fusion rate, and subsequent developmental potential of cloned bovine embryos. The toxicity of PHA was established by examining its effects on the development of parthenogenetic bovine oocytes treated with different doses (Experiment 1), and for different durations (Experiment 2). The effective dose and duration of PHA treatment (150 microg/mL, 20 min incubation) was selected and used to compare membrane fusion efficiency and embryo development following somatic cell nuclear transfer (Experiment 3). Cloning with somatic donor fibroblasts versus cumulus cells was also compared, both with and without PHA treatment (150 microg/mL, 20 min). Fusion rate of nuclear donor fibroblasts, after phytohemagglutinin treatment, was increased from 33 to 61% (P < 0.05), and from 59 to 88% (P < 0.05) with cumulus cell nuclear donors. The nuclear transfer (NT) efficiency per oocyte used was improved following PHA treatment, for both fibroblast (13% versus 22%) as well as cumulus cells (17% versus 34%; P < 0.05). The cloned embryos, both with and without PHA treatment, were subjected to vitrification and embryo transfer testing, and resulted in similar survival (approximately 90% hatching) and pregnancy rates (17-25%). Three calves were born following vitrification and embryo transfer of these embryos; two from the PHA-treated group, and one from non-PHA control group. We concluded that PHA treatment significantly improved the fusion efficiency of somatic NT in cattle, and therefore, increased the development of cloned blastocysts. Furthermore, within a determined range of dose and duration, PHA had no detrimental effect on embryo survival post-vitrification, nor on pregnancy or calving rates following embryo transfer.

The assessment of standardized training programs in public health preparedness

The Texas Bioterrorism Continuing Education Consortium (BCE) provided National Disaster Life Support (NDLS) training courses throughout the state of Texas in 2005, to help improve knowledge and skills pertaining to bioterrorism and other public health emergencies. The NDLS training courses include curriculum in Basic Disaster Life Support (BDLS) and Core Disaster Life Support (CDLS). A course evaluation which included items assessing ability and willingness of training participants, role of responders, and other variables was mailed to all NDLS participants who provided contact information. An analysis was conducted to determine whether the survey respondents participated in the Hurricanes Katrina and/or Rita relief efforts, as well as to evaluate the impact of the NDLS training courses on the participant's ability and willingness to respond during a disaster. The study population (n = 2150) consisted mostly of nurses (50%) (n=1074). A chi-square test of analysis indicated the following results. Among the survey respondents who took the CDLS course, there was no statically significant difference by occupation pertaining to ability or willingness to respond (x2 [df = 5] = 4.02, p= 0.546); (x2 [df = 5] = 2.45, p = .783). However, there was a statistically significant difference among those respondents who took the BDLS course with respect to ability, and a slightly significant difference with respect to willingness (x2 [df = 5] = 13.35, p = .020 and (x2 = [df = 5] = 10.299, p = .067). These findings are similar to previous studies assessing willingness to respond to a disaster.^ A second analysis was conducted with these survey data to evaluate the implications for disaster response training for the NDLS courses. Results indicated that the majority of disaster responders served in the role for which they were professionally trained (Physicians=68%; Nurses = 50.4%). Nurses, EMT, and Fire professionals served in multiple roles. These results suggest the importance of developing training programs that will prepare professionals to serve in multiple roles. The development of standardized evaluation methods would fill an important gap in assessing impact of national training programs. ^

Competence Training in Higher Education: The case of the Technological Master "Prestación de Servicios Profesionales" from the Colegio de Postgraduados (México).

The Competency-Based Education in the context of training is intended as a comprehensive approach that seeks to link education with the productive sector and increase the potential of individuals, in the face of social, economic, political and cultural transformations that suffers the world and the contemporary society; this is how educational services associated to the rural area takes part of the global revalorization of the role of learning and knowledge. Under the competence approach and taking into account the CONOCER model, we design a Technological Master from the “Colegio de Postgraduados” identifying the competences needed so that the students, professional from different areas of knowledge, managed to develop them, but mainly to achieve the goal of developing the capacities of producers in Mexican rural area.

The application of biomechanics to penalty corner drag-flick training: a case of Study

The penalty corner is one of the most important game situations in field hockey with one third of all goals resulting from this tactical situation. The aim of this study was to develop and apply a training method, based on previous studies, to improve the drag- flick skill on a young top-class field hockey player. A young top-class player exercised three times per week using specific drills over a four week period. A VICON optoelectronic system (Oxford Metrics, Oxford, UK) was employed to capture twenty drag-flicks, with six cameras sampling at 250 Hz, prior and after the training period. In order to analyze pre- and post-test differences a dependent t-test was carried out. Angular velocities and the kinematic sequence were similar to previous studies. The player improved (albeit not significantly) the angular velocity of the stick. The player increased front foot to the ball at T1 (p < 0.01) and the drag-flick distances. The range of motion from the front leg decreased from T1 to T6 after the training period (p < 0.01). The specific training sessions conducted with the player improved some features of this particular skill. This article shows how technical knowledge can help with the design of training programs and whether some drills are more effective than others.

Effect of endurance, speed and strength training on skin temperature measured by infrared thermography = Efecto del entrenamiento de resistencia, velocidad y fuerza en la temperatura de la piel a través de la termografía infrarroja

La termografía infrarroja (TI) es una técnica no invasiva y de bajo coste que permite, con el simple acto de tomar una fotografía, el registro sin contacto de la energía que irradia el cuerpo humano (Akimov & Son’kin, 2011, Merla et al., 2005, Ng et al., 2009, Costello et al., 2012, Hildebrandt et al., 2010). Esta técnica comenzó a utilizarse en el ámbito médico en los años 60, pero debido a los malos resultados como herramienta diagnóstica y la falta de protocolos estandarizados (Head & Elliot, 2002), ésta se dejó de utilizar en detrimento de otras técnicas más precisas a nivel diagnóstico. No obstante, las mejoras tecnológicas de la TI en los últimos años han hecho posible un resurgimiento de la misma (Jiang et al., 2005, Vainer et al., 2005, Cheng et al., 2009, Spalding et al., 2011, Skala et al., 2012), abriendo el camino a nuevas aplicaciones no sólo centradas en el uso diagnóstico. Entre las nuevas aplicaciones, destacamos las que se desarrollan en el ámbito de la actividad física y el deporte, donde recientemente se ha demostrado que los nuevos avances con imágenes de alta resolución pueden proporcionar información muy interesante sobre el complejo sistema de termorregulación humana (Hildebrandt et al., 2010). Entre las nuevas aplicaciones destacan: la cuantificación de la asimilación de la carga de trabajo físico (Čoh & Širok, 2007), la valoración de la condición física (Chudecka et al., 2010, 2012, Akimov et al., 2009, 2011, Merla et al., 2010), la prevención y seguimiento de lesiones (Hildebrandt et al., 2010, 2012, Badža et al., 2012, Gómez Carmona, 2012) e incluso la detección de agujetas (Al-Nakhli et al., 2012). Bajo estas circunstancias, se acusa cada vez más la necesidad de ampliar el conocimiento sobre los factores que influyen en la aplicación de la TI en los seres humanos, así como la descripción de la respuesta de la temperatura de la piel (TP) en condiciones normales, y bajo la influencia de los diferentes tipos de ejercicio. Por consiguiente, este estudio presenta en una primera parte una revisión bibliográfica sobre los factores que afectan al uso de la TI en los seres humanos y una propuesta de clasificación de los mismos. Hemos analizado la fiabilidad del software Termotracker, así como su reproducibilidad de la temperatura de la piel en sujetos jóvenes, sanos y con normopeso. Finalmente, se analizó la respuesta térmica de la piel antes de un entrenamiento de resistencia, velocidad y fuerza, inmediatamente después y durante un período de recuperación de 8 horas. En cuanto a la revisión bibliográfica, hemos propuesto una clasificación para organizar los factores en tres grupos principales: los factores ambientales, individuales y técnicos. El análisis y descripción de estas influencias deben representar la base de nuevas investigaciones con el fin de utilizar la TI en las mejores condiciones. En cuanto a la reproducibilidad, los resultados mostraron valores excelentes para imágenes consecutivas, aunque la reproducibilidad de la TP disminuyó ligeramente con imágenes separadas por 24 horas, sobre todo en las zonas con valores más fríos (es decir, zonas distales y articulaciones). Las asimetrías térmicas (que normalmente se utilizan para seguir la evolución de zonas sobrecargadas o lesionadas) también mostraron excelentes resultados pero, en este caso, con mejores valores para las articulaciones y el zonas centrales (es decir, rodillas, tobillos, dorsales y pectorales) que las Zonas de Interés (ZDI) con valores medios más calientes (como los muslos e isquiotibiales). Los resultados de fiabilidad del software Termotracker fueron excelentes en todas las condiciones y parámetros. En el caso del estudio sobre los efectos de los entrenamientos de la velocidad resistencia y fuerza en la TP, los resultados muestran respuestas específicas según el tipo de entrenamiento, zona de interés, el momento de la evaluación y la función de las zonas analizadas. Los resultados mostraron que la mayoría de las ZDI musculares se mantuvieron significativamente más calientes 8 horas después del entrenamiento, lo que indica que el efecto del ejercicio sobre la TP perdura por lo menos 8 horas en la mayoría de zonas analizadas. La TI podría ser útil para cuantificar la asimilación y recuperación física después de una carga física de trabajo. Estos resultados podrían ser muy útiles para entender mejor el complejo sistema de termorregulación humano, y por lo tanto, para utilizar la TI de una manera más objetiva, precisa y profesional con visos a mejorar las nuevas aplicaciones termográficas en el sector de la actividad física y el deporte Infrared Thermography (IRT) is a safe, non-invasive and low-cost technique that allows the rapid and non-contact recording of the irradiated energy released from the body (Akimov & Son’kin, 2011; Merla et al., 2005; Ng et al., 2009; Costello et al., 2012; Hildebrandt et al., 2010). It has been used since the early 1960’s, but due to poor results as diagnostic tool and a lack of methodological standards and quality assurance (Head et al., 2002), it was rejected from the medical field. Nevertheless, the technological improvements of IRT in the last years have made possible a resurgence of this technique (Jiang et al., 2005; Vainer et al., 2005; Cheng et al., 2009; Spalding et al., 2011; Skala et al., 2012), paving the way to new applications not only focused on the diagnose usages. Among the new applications, we highlighted those in physical activity and sport fields, where it has been recently proven that a high resolution thermal images can provide us with interesting information about the complex thermoregulation system of the body (Hildebrandt et al., 2010), information than can be used as: training workload quantification (Čoh & Širok, 2007), fitness and performance conditions (Chudecka et al., 2010, 2012; Akimov et al., 2009, 2011; Merla et al., 2010; Arfaoui et al., 2012), prevention and monitoring of injuries (Hildebrandt et al., 2010, 2012; Badža et al., 2012, Gómez Carmona, 2012) and even detection of Delayed Onset Muscle Soreness – DOMS- (Al-Nakhli et al., 2012). Under this context, there is a relevant necessity to broaden the knowledge about factors influencing the application of IRT on humans, and to better explore and describe the thermal response of Skin Temperature (Tsk) in normal conditions, and under the influence of different types of exercise. Consequently, this study presents a literature review about factors affecting the application of IRT on human beings and a classification proposal about them. We analysed the reliability of the software Termotracker®, and also its reproducibility of Tsk on young, healthy and normal weight subjects. Finally, we examined the Tsk thermal response before an endurance, speed and strength training, immediately after and during an 8-hour recovery period. Concerning the literature review, we proposed a classification to organise the factors into three main groups: environmental, individual and technical factors. Thus, better exploring and describing these influence factors should represent the basis of further investigations in order to use IRT in the best and optimal conditions to improve its accuracy and results. Regarding the reproducibility results, the outcomes showed excellent values for consecutive images, but the reproducibility of Tsk slightly decreased with time, above all in the colder Regions of Interest (ROI) (i.e. distal and joint areas). The side-to-side differences (ΔT) (normally used to follow the evolution of some injured or overloaded ROI) also showed highly accurate results, but in this case with better values for joints and central ROI (i.e. Knee, Ankles, Dorsal and Pectoral) than the hottest muscle ROI (as Thigh or Hamstrings). The reliability results of the IRT software Termotracker® were excellent in all conditions and parameters. In the part of the study about the effects on Tsk of aerobic, speed and strength training, the results of Tsk demonstrated specific responses depending on the type of training, ROI, moment of the assessment and the function of the considered ROI. The results showed that most of muscular ROI maintained warmer significant Tsk 8 hours after the training, indicating that the effect of exercise on Tsk last at least 8 hours in most of ROI, as well as IRT could help to quantify the recovery status of the athlete as workload assimilation indicator. Those results could be very useful to better understand the complex skin thermoregulation behaviour, and therefore, to use IRT in a more objective, accurate and professional way to improve the new IRT applications for the physical activity and sport sector.

Nutrition and physical activity programs for obesity treatment (PRONAF study). methodological approach of the project

Este trabajo aborda la metodología seguida para llevar a cabo el proyecto de investigación PRONAF (Clinical Trials Gov.: number NCT01116856.) Background: At present, scientific consensus exists on the multifactorial etiopatogenia of obesity. Both professionals and researchers agree that treatment must also have a multifactorial approach, including diet, physical activity, pharmacology and/or surgical treatment. These two last ones should be reserved for those cases of morbid obesities or in case of failure of the previous ones. The aim of the PRONAF study is to determine what type of exercise combined with caloric restriction is the most appropriate to be included in overweigth and obesity intervention programs, and the aim of this paper is to describe the design and the evaluation methods used to carry out the PRONAF study. Methods/design: One-hundred nineteen overweight (46 males) and 120 obese (61 males) subjects aged 18–50 years were randomly assigned to a strength training group, an endurance training group, a combined strength + endurance training group or a diet and physical activity recommendations group. The intervention period was 22 weeks (in all cases 3 times/wk of training for 22 weeks and 2 weeks for pre and post evaluation). All subjects followed a hypocaloric diet (25-30% less energy intake than the daily energy expenditure estimated by accelerometry). 29–34% of the total energy intake came from fat, 14–20% from protein, and 50–55% from carbohydrates. The mayor outcome variables assesed were, biochemical and inflamatory markers, body composition, energy balance, physical fitness, nutritional habits, genetic profile and quality of life. 180 (75.3%) subjects finished the study, with a dropout rate of 24.7%. Dropout reasons included: personal reasons 17 (28.8%), low adherence to exercise 3 (5.1%), low adherence to diet 6 (10.2%), job change 6 (10.2%), and lost interest 27 (45.8%). Discussion: Feasibility of the study has been proven, with a low dropout rate which corresponds to the estimated sample size. Transfer of knowledge is foreseen as a spin-off, in order that overweight and obese subjects can benefit from the results. The aim is to transfer it to sports centres. Effectiveness on individual health-related parameter in order to determine the most effective training programme will be analysed in forthcoming publications.

Design of a training tool for improving the use of hand-held detectors in humanitarian demining

Purpose – The purpose of this paper is to introduce the design of a training tool intended to improve deminers' technique during close-in detection tasks. Design/methodology/approach – Following an introduction that highlights the impact of mines and improvised explosive devices (IEDs), and the importance of training for enhancing the safety and the efficiency of the deminers, this paper considers the utilization of a sensory tracking system to study the skill of the hand-held detector expert operators. With the compiled information, some critical performance variables can be extracted, assessed, and quantified, so that they can be used afterwards as reference values for the training task. In a second stage, the sensory tracking system is used for analysing the trainee skills. The experimentation phase aims to test the effectiveness of the elements that compose the sensory system to track the hand-held detector during the training sessions. Findings – The proposed training tool will be able to evaluate the deminers' efficiency during the scanning tasks and will provide important information for improving their competences. Originality/value – This paper highlights the need of introducing emerging technologies for enhancing the current training techniques for deminers and proposes a sensory tracking system that can be successfully utilised for evaluating trainees' performance with hand-held detectors.

The chair of food Banks UPM as a tool of raising awareness and promoting a culture of rational food comsumption

The Chair of Food Banks UPM arises from a cooperation agreement between the Spanish Federation of Food Banks (FESBAL) and the Technical University of Madrid (UPM), with the aim of raising awareness and promoting rational food consumption to avoid food waste, through activities of training, transfer of knowledge and promotion of I+D+i. The aim of this paper is to reflect on the activities carried out during the first year in order to obtain learning lessons and improve the management of activities and resources.

Restoration of β-adrenergic signaling in failing cardiac ventricular myocytes via adenoviral-mediated gene transfer

Cardiovascular gene therapy is a novel approach to the treatment of diseases such as congestive heart failure (CHF). Gene transfer to the heart would allow for the replacement of defective or missing cellular proteins that may improve cardiac performance. Our laboratory has been focusing on the feasibility of restoring β-adrenergic signaling deficiencies that are a characteristic of chronic CHF. We have now studied isolated ventricular myocytes from rabbits that have been chronically paced to produce hemodynamic failure. We document molecular β-adrenergic signaling defects including down-regulation of myocardial β-adrenergic receptors (β-ARs), functional β-AR uncoupling, and an up-regulation of the β-AR kinase (βARK1). Adenoviral-mediated gene transfer of the human β2-AR or an inhibitor of βARK1 to these failing myocytes led to the restoration of β-AR signaling. These results demonstrate that defects present in this critical myocardial signaling pathway can be corrected in vitro using genetic modification and raise the possibility of novel inotropic therapies for CHF including the inhibition of βARK1 activity in the heart.

Horizontal gene transfer and mutation: Ngrol genes in the genome of Nicotiana glauca

Ngrol genes (NgrolB, NgrolC, NgORF13, and NgORF14) that are similar in sequence to genes in the left transferred DNA (TL-DNA) of Agrobacterium rhizogenes have been found in the genome of untransformed plants of Nicotiana glauca. It has been suggested that a bacterial infection resulted in transformation of Ngrol genes early in the evolution of the genus Nicotiana. Although the corresponding four rol genes in TL-DNA provoked hairy-root syndrome in plants, present-day N. glauca and plants transformed with Ngrol genes did not exhibit this phenotype. Sequenced complementation analysis revealed that the NgrolB gene did not induce adventitious roots because it contained two point mutations. Single-base site-directed mutagenesis at these two positions restored the capacity for root induction to the NgrolB gene. When the NgrolB, with these two base substitutions, was positioned under the control of the cauliflower mosaic virus 35S promoter (P35S), transgenic tobacco plants exhibited morphological abnormalities that were not observed in P35s-RirolB plants. In contrast, the activity of the NgrolC gene may have been conserved after an ancient infection by bacteria. Discussed is the effect of the horizontal gene transfer of the Ngrol genes and mutations in the NgrolB gene on the phenotype of ancient plants during the evolution of N. glauca.

Supernatant protein factor, which stimulates the conversion of squalene to lanosterol, is a cytosolic squalene transfer protein and enhances cholesterol biosynthesis

Squalene epoxidase, a membrane-associated enzyme that converts squalene to squalene 2,3-oxide, plays an important role in the maintenance of cholesterol homeostasis. In 1957, Bloch and colleagues identified a factor from rat liver cytosol termed “supernatant protein factor (SPF),” which promotes the squalene epoxidation catalyzed by rat liver microsomes with oxygen, NADPH, FAD, and phospholipid [Tchen, T. T. & Bloch, K. (1957) J. Biol. Chem. 226, 921–930]. Although purification of SPF by 11,000-fold was reported, no information is so far available on the primary structure or biological function of SPF. Here we report the cDNA cloning and expression of SPF from rat and human. The encoded protein of 403 amino acids belongs to a family of cytosolic lipid-binding/transfer proteins such as α-tocopherol transfer protein, cellular retinal binding protein, yeast phosphatidylinositol transfer protein (Sec14p), and squid retinal binding protein. Recombinant SPF produced in Escherichia coli enhances microsomal squalene epoxidase activity and promotes intermembrane transfer of squalene in vitro. SPF mRNA is expressed abundantly in the liver and small intestine, both of which are important sites of cholesterol biosynthesis. SPF is expressed significantly in isolated hepatocytes, but the expression level was markedly decreased after 48 h of in vitro culture. Moreover, SPF was not detectable in most of the cell lines tested, including HepG2 and McARH7777 hepatomas. Transfection of SPF cDNA in McARH7777 significantly stimulated de novo cholesterol biosynthesis. These data suggest that SPF is a cytosolic squalene transfer protein capable of regulating cholesterol biosynthesis.

On the role of the K-proton transfer pathway in cytochrome c oxidase

Cytochrome c oxidase is a membrane-bound enzyme that catalyzes the four-electron reduction of oxygen to water. This highly exergonic reaction drives proton pumping across the membrane. One of the key questions associated with the function of cytochrome c oxidase is how the transfer of electrons and protons is coupled and how proton transfer is controlled by the enzyme. In this study we focus on the function of one of the proton transfer pathways of the R. sphaeroides enzyme, the so-called K-proton transfer pathway (containing a highly conserved Lys(I-362) residue), leading from the protein surface to the catalytic site. We have investigated the kinetics of the reaction of the reduced enzyme with oxygen in mutants of the enzyme in which a residue [Ser(I-299)] near the entry point of the pathway was modified with the use of site-directed mutagenesis. The results show that during the initial steps of oxygen reduction, electron transfer to the catalytic site (to form the “peroxy” state, Pr) requires charge compensation through the proton pathway, but no proton uptake from the bulk solution. The charge compensation is proposed to involve a movement of the K(I-362) side chain toward the binuclear center. Thus, in contrast to what has been assumed previously, the results indicate that the K-pathway is used during oxygen reduction and that K(I-362) is charged at pH ≈ 7.5. The movement of the Lys is proposed to regulate proton transfer by “shutting off” the protonic connectivity through the K-pathway after initiation of the O2 reduction chemistry. This “shutoff” prevents a short-circuit of the proton-pumping machinery of the enzyme during the subsequent reaction steps.