Transcriptome and proteome analysis reveal new insight into proximal and distal responses of wheat to foliar infection by Xanthomonas translucens.

The molecular details of local plant response against Xanthomonas translucens infection is largely unknown. Moreover, there is no knowledge about effects of the pathogen on the root's transcriptome and proteome. Therefore, we investigated the global gene and protein expression changes both in leaves and roots of wheat (Triticum aestivum) 24 h post leaf infection of X. translucens. This simultaneous analysis allowed us to obtain insight into possible metabolic rearrangements in above- and belowground tissues and to identify common responses as well as specific alterations. At the site of infection, we observed the implication of various components of the recognition, signaling, and amplification mechanisms in plant response to the pathogen. Moreover, data indicate a massive down-regulation of photosynthesis and confirm the chloroplast as crucial signaling hub during pathogen attack. Notably, roots responded as well to foliar attack and their response significantly differed from that locally triggered in infected leaves. Data indicate that roots as a site of energy production and synthesis of various secondary metabolites may actively influence the composition and colonisation level of root-associated microbes. Finally, our results emphasize the accumulation of jasmonic acid, pipecolic acid and/or the downstream mediator of hydrogen peroxide as long distal signals from infected leaves to roots.

Proteomic changes and molecular effects associated with Cr(III) and Cr(VI) treatments on germinating kiwifruit pollen.

The present study is aimed at identifying molecular changes elicited by Cr(III) and Cr(VI) on germinating kiwifruit pollen. To address this question, comparative proteomic and DNA laddering analyses were performed. While no genotoxic effect was detected, a number of proteins whose accumulation levels were altered by treatments were identified. In particular, the upregulation of some proteins involved in the scavenging response, cell redox homeostasis and lipid synthesis could be interpreted as an oxidative stress response induced by Cr treatment. The strong reduction of two proteins involved in mitochondrial oxidative phosphorylation and a decline in ATP levels were also observed. The decrease of pollen energy availability could be one of the causes of the severe inhibition of the pollen germination observed upon exposure to both Cr(III) and Cr(VI). Finally, proteomic and biochemical data indicate proteasome impairment: the consequential accumulation of misfolded/damaged proteins could be an important molecular mechanism of Cr(III) toxicity in pollen.

Analytic Hierarchy Process (AHP) for examining healthcare professionals' assessments of risk factors. The relative importance of risk factors for falls in community-dwelling older people.

BACKGROUND: A gap exists between evidence-based medicine and clinical-practice. Every day, healthcare professionals (HCPs) combine empirical evidence and subjective experience in order to maximize the effectiveness of interventions. Consequently, it is important to understand how HCPs interpret the research evidence and apply it in everyday practice. We focused on the prevention of falls, a common cause of injury-related morbidity and mortality in later life, for which there is a wide range of known risk factors. OBJECTIVES: To use the Analytic Hierarchy Process (AHP) to investigate the opinions of HCPs in prioritizing risk factors for preventing falls. METHODS: We used the AHP to develop a hierarchy of risk factors for falls based on the knowledge and experience of experts. We submitted electronic questionnaires via the web, in order to reach a wider number of respondents. With a web service, we pooled the results and weighted the coherence and the experience of respondents. RESULTS: Overall, 232 respondents participated in the study: 32 in the technical pilot study, nine in the scientific pilot study and 191 respondents in the main study. We identified a hierarchy of 35 risk factors, organized in two categories and six sub-categories. CONCLUSIONS: The hierarchy of risk factors provides further insights into clinicians' perceptions of risk factors for falls. This hierarchy helps understand the relative importance that clinicians place on risk factors for falls in older people and why evidence-based guidelines are not always followed. This information may be helpful in improving intervention programs and in understanding how clinicians prioritize multiple risk factors in individual patients. The AHP method allows the opinions of HCPs to be investigated, giving appropriate weight to their coherence, background and experience.

An algorithm for the recovery of fetal heart rate series from CTG data.

Cardiotocography (simultaneous recording of fetal heart rate (FHR) and uterine contractions) is one of the most used diagnostic techniques to evaluate fetal well-being and to investigate the functional state of the fetal autonomic nervous system. Recently, great interest has been paid to the variability of the FHR, and its frequency analysis, as a base for a more objective analysis of the cardiotocographic (CTG) tracings. FHR signals are unevenly sampled series. To obtain evenly sampled series, cardiotocographs often use zero-order interpolation. Such process is simple and fast but results unsuitable for frequency analyses because it introduces alterations in the FHR power spectrum. An algorithm for the recovery of the true FHR series out of the zero-order interpolated CTG data was developed and evaluated.

Foetal heart rate power spectrum response to uterine contraction.

Cardiotocography is the most diffused prenatal diagnostic technique in clinical routine. The simultaneous recording of foetal heart rate (FHR) and uterine contractions (UC) provides useful information about foetal well-being during pregnancy and labour. However, foetal electronic monitoring interpretation still lacks reproducibility and objectivity. New methods of interpretation and new parameters can further support physicians' decisions. Besides common time-domain analysis, study of the variability of FHR can potentially reveal autonomic nervous system activity of the foetus. In particular, it is clinically relevant to investigate foetal reactions to UC to diagnose foetal distress early. Uterine contraction being a strong stimulus for the foetus and its autonomic nervous system, it is worth exploring the FHR variability response. This study aims to analyse modifications of the power spectrum of FHR variability corresponding to UC. Cardiotocographic signal tracts corresponding to 127 UC relative to 30 healthy foetuses were analysed. Results mainly show a general, statistically significant (t test, p<0.01) power increase of the FHR variability in the LF 0.03-0.2 Hz and HF 0.2-1 in correspondence of the contraction with respect to a reference tract set before contraction onset. Time evolution of the power within these bands was computed by means of time-varying spectral estimation to concisely show the FHR response along a uterine contraction. A synchronised grand average of these responses was also computed to verify repeatability, using the contraction apex as time reference. Such modifications of the foetal HRV that follow a contraction can be a sign of ANS reaction and, therefore, additional, objective information about foetal reactivity during labour.

Genomic stability in Arabidopsis thaliana transgenic plants obtained by floral dip.

The occurrence of DNA modification is an undesired phenomenon accompanying plant cell transformation. The event has been correlated with the stress imposed by the presently utilised transformation procedures, all depending on plant differentiation from in vitro cell culture, but other causes have not been excluded. In this work, transgenic Arabidopsis thaliana plants have been produced by an approach that does not require cell dedifferentiation, being based on in planta Agrobacterium-mediated gene transfer by flower infiltration, which is followed by recovery and selection of transgenic progeny. Genomic DNA changes in transgenic and control plants have been investigated by AFLP and RAMP analysis. Results show no statistically relevant genomic modifications in transgenic plants, as compared with control untreated plants. Variations were observed in callus-derived A. thaliana plants, thus supporting the conclusion that somaclonal variation is essentially correlated with the stress imposed by the in vitro cell culture, rather than with the integration of a foreign gene.

AFLP analysis as biomarker of exposure to organic and inorganic genotoxic substances in plants.

In recent years several plant species have been in use as bioindicators and several tests have been developed to evaluate the toxicity of environmental pollutants in vegetal organisms. In the present paper Arabidopsis thaliana (L.) Heynh. (ecotype Wassilewskija) was used as bioindicators of two genotoxic substances: potassium dichromate and dihydrophenanthrene. Inhibition of seed germination was observed with both pollutants. AFLP analysis (i) indicated that both substances are genotoxic, (ii) showed that dihydrophenanthrene induces DNA changes in different target sequences than potassium dichromate, (iii) quantified the genotoxic effect using cluster analysis by comparing DNA from treated plants with that of control plants. On the basis of these considerations we suggest that AFLP method is a powerful tool for measuring qualitative and quantitative genotoxic activity due to environmental pollutants. AFLP method can be applied to a wide range of bioindicator organisms and may become a universal methodology to identify target genes for specific genotoxic agents. This could open up possibilities for designing specifically targeted assays and new approaches to risk assessment.

Efficiency of transient transformation in tobacco protoplasts is independent of plasmid amount.

We describe an optimized protocol for the transient transformation of tobacco protoplasts mediated by polyethylene-glycol (PEG). As expected, the quantitative beta-glucuronidase (Gus) activity driven by pCaMVGus was dependent on the amount of plasmid used. Nevertheless, we demonstrate by an immunodetection method that transformation efficiency did not depend on the amount of plasmid used but on the limitation imposed by cell competence. In fact, we obtained the same percentage of transformed cells (about 60%) using a wide range of plasmid concentrations (0.1-10 microg per test). Finally, we show that, when we used two plasmid types in a mixture at a concentration ranging from 0.1 to 10 microg for each, all transformed cells expressed proteins encoded by both plasmids. Transient expression and co-transformation experiments are routinely used methods and, probably, the major results from this work were assumed by many researchers in this field, but our data experimentally support this assumption.

Automatic recognition of vertebral landmarks in fluoroscopic sequences for analysis of intervertebral kinematics.

Intervertebral kinematics closely relates to the functionality of the spinal segments. Direct measurement of the intervertebral kinematics in vivo is very problematic. The use of a fluoroscopic device can provide continuous screening of the lumbar tract during patient spontaneous motion, with an acceptable, low X-ray dose. The kinematic analysis is intended to be limited to planar motion. Kinematic parameters are computed from vertebral landmarks on each frame of the image sequence. Landmarks are normally selected manually in spite of the fact that this is subjective, tedious to perform and regarded as one of the major contributors to errors in the computed kinematic parameters. The aim of this work is to present an innovative method for the automatic recognition of vertebral landmarks throughout a fluoroscopic image sequence to provide an objective and more precise quantification of intervertebral kinematics. The recognition procedure is based upon comparing vertebral features in two adjacent frames by means of a cross-correlation index, which is also robust despite the low signal-to-noise ratio of the lumbar fluoroscopic images. To provide a quantitative assessment of this method a calibration model was used which consisted of two lumbar vertebrae linked by a universal joint. The reliability and accuracy of the kinematic measurements have been investigated. The errors are of the order of a millimetre for the localisation of the intervertebral centre of rotation and tenths of a degree for the intervertebral angle. Error analysis suggests that this method improves the accuracy of the intervertebral kinematic calculations and has the potential to automate the selection of anatomical landmarks.

Relationship between visual acuity and eye position variability during foveations in congenital nystagmus.

Visual acuity in congenital nystagmus has proven to be primarily related to the duration of foveation periods, during which the image of a target falls onto the fovea and eye velocity slows down. It was found that the longer the foveation time the higher the visual acuity. However, the cycle-to-cycle variability of the eye position and velocity during foveation periods also contribute to visual acuity. A high variability of the eye position during the foveations hinders a stable placement of the target image on the centralmost fovea and consequently decreases visual acuity. To investigate the relationship between different nystagmus features and visual acuity, infrared- oculographic and electro-oculographic eye position recordings of 20 patients affected by congenital nystagmus were analysed in different gaze positions. In several patients' recordings, a high variability of the eye position during foveations (i.e. greater than 0.5 degrees) was detected. Correspondingly, low visual acuity was measured, in spite of sufficiently long foveation periods. The standard deviation of eye positions during foveation periods was used to measure this variability and it was found to be correlated to visual acuity, in conjunction with the mean duration of the foveation periods. On the basis of the data analysis, an exponential relationship is proposed to relate visual acuity and the standard deviation of the eye position during foveations.

Study of the control strategy of the quadriceps muscles in anterior knee pain.

Anterior knee pain (AKP) is a common pathological condition, particularly among young people and athletes, associated to an abnormal motion of the patella during the bending of the knee and possibly dependent on a muscular or structural imbalance. A lack of synergy in the quadriceps muscles results in a dynamic misalignment of the patella, which in turn produces pain. AKP rehabilitative therapy consists of conservative treatment whose main objective is to strengthen the Vastus Medialis. The aim of this article is to study the quadriceps muscle control strategy in AKP patients during an isokinetic exercise. Analysis of the muscle activation strategy is important for an objective measurement of the knee functionality in that it helps to diagnose and monitor the rehabilitative treatment. Surface electromyography (EMG) from the three superficial muscles of the femoral quadriceps during a concentric isokinetic exercise has been analyzed along with the signals of knee joint position and torque. A group of 12 AKP patients has been compared with a group of 30 normal subjects. Analysis of the grand ensemble average of the EMG linear envelopes in AKP patients reveals significant modifications in Vastus Medialis activity compared to the other quadriceps muscles. In order to study the synergy of the muscles, temporal identifiers have been associated to the EMG linear envelopes. To this end, EMG linear envelope decomposition in Gaussian pulses turned out to be effective and the results highlight an appreciable delay in the activation of the Vastus Medialis in AKP patients. This muscular unbalance can explain the abnormal motion of the patella.

The product of the rice myb7 unspliced mRNA dimerizes with the maize leucine zipper Opaque2 and stimulates its activity in a transient expression assay.

myb7 mRNA is present in rice in spliced and unspliced forms, splicing being enhanced by anoxia. The protein (Mybleu) encoded by the unspliced mRNA is composed of an incomplete Myb domain followed by a leucine zipper; however, it lacks canonical sequences for DNA binding, transcriptional activation, and nuclear localization. We show here that in transiently transformed tobacco protoplasts, Mybleu is able to enhance the transcriptional activity of the maize leucine zipper Opaque2 on its target b32 promoter. The Mybleu transactivation effect is strictly dependent on the presence of Opaque2 and is driven by Mybleu-Opaque2 heterodimers. Mybleu is located in the nucleus, both in rice and in transformed tobacco protoplasts. In rice, the protein is expressed in regions corresponding to undifferentiated cells of roots and coleoptiles. Therefore, myb7 mRNA encodes, depending on its splicing, two transcription factors belonging to separate classes. One of them, Mybleu, has novel structural characteristics, suggesting the existence of new mechanisms acting in the activation of transcription.

Quadriceps muscles activation in anterior knee pain during isokinetic exercise.

The aim of this paper is to study the femoral quadriceps muscles activation in patients suffering from anterior knee pain (AKP) during an isokinetic exercise. AKP is a common pathological condition, associated to an abnormal motion of the patella. It possibly depends on a muscular or structural imbalance. A lack of synergy in the quadriceps muscles results in a dynamic misalignment of the patella, which in turn produces pain. A quantitative analysis of the muscle activation strategy is important for an objective measurement of the knee functionality in that it helps to diagnose and monitor the rehabilitative treatment. Surface electromyography (EMG) from the three superficial muscles of the femoral quadriceps during a concentric isokinetic exercise has been analysed. A group of AKP patients has been compared with a control group of healthy subjects. Ensemble average of the EMG linear envelopes reveals significant modifications in Vastus Medialis activity in AKP patients. In order to quantify the synergy of the muscles, different parameters have been associated to EMG linear envelopes significant features. A decomposition in gaussian pulses and an asymmetry coefficient have been utilised. The results relative to these concise parameters highlight an appreciable delay and a modification in the activation of the Vastus Medialis in AKP patients.

Localisation and characterisation of auditory perception through Functional Magnetic Resonance Imaging.

In the last few years, Functional Magnetic Resonance Imaging (fMRI) has been widely accepted as an effective tool for mapping brain activities in both the neurosensorial and the cognitive field. The present work aims to assess the possibility of using fMRI methods to perform a non-invasive evaluation of the human auditory function. To this end the cortical response to different non speech stimuli (pure tones, pulsed tones) was examined for ten subjects with no audiological impairment. Our findings point out some remarkable differences in both the spatial and the temporal features of the primary auditory cortex response to pulsed tones and to pure tones.

AUDIOMAT: an electronic newspaper for visually impaired people.

Electronic newspapers are now becoming available via wide-area computer networks, such as the internet. Unfortunately, the Windows-based graphical user interface of the client programs makes it difficult for visually impaired people to use such services. Furthermore, few visually impaired people have a PC. The AUDIOMAT project was designed to provide a reading system for the visually impaired, thus giving this community in the Campania region of southern Italy the opportunity to read their local newspaper. In a pilot trial, three blind subjects used the AUDIOMAT system daily for two months. Despite some minor technical problems the system performed well and a wider validation phase is now planned.

A quantitative approach in the treatment of hemiplegic patients using an electropneumatic platform.

Exercise is the central feature in the rehabilitative treatment of hemiplegic patients. In cognitive rehabilitation, great importance is attributed to the recovery of a sufficient number of proprioceptive channels before expecting the patient to regain voluntary control. The PERM [Piattaforma (Platform), Elettropneumatica (Electropneumatic) per (for) Riabilitazione (Rehabilitation) Motoria (Motor)] is a new device that can be used in the treatment of the lower limbs to provide the patient with controlled and quantifiable mechanical stimuli, thus enabling the therapist to adopt a more rigorous approach to treatment even in the very early stages of rehabilitation. Alternatively, the system can be used in a different operating mode as a normal biofeedback system capable of displaying on-screen the extent to which voluntary load has been transferred to the lower limb.

Medical technologies in developing countries: a feasibility study on the maintenance of medical equipment in Ethiopia.

The authors report the results of a feasibility study, sponsored by the Italian Foreign Ministry, of setting up a Regional Centre for the Management and Maintenance of Medical Technologies in Ethiopia. After some general considerations regarding the problems of co-operation in this field, the authors draft some concrete proposals for an operative programme.