Concepts of aging with paralysis: implications for recovery and treatment.

This chapter deals with the impact of age on the occurrence, clinical presentation, outcome, and course of a spinal cord injury (SCI). This is of importance in a society where the population of elderly people continuously increases. The chapter is focused first, on the actual problems of a SCI in elderly subjects and second, on age-specific sequelae after a SCI. The etiology and clinical presentation of a SCI differs in elderly subjects compared to young subjects. With advanced age, incomplete cervical lesions following falls or due to spondylotic degeneration of the cervical spine and non-traumatic SCI occur more frequently. Research pertaining to the comparison of different age groups is prone to bias due to survival and treatment cohort effects. There is an increased risk of complications and mortality after a complete SCI in elderly people. Surprisingly, the recovery of the neurological deficit does not depend on age. However, elderly subjects with SCI have more problems in transferring an improvement in motor score into a functional improvement in their ability to carry out the activities of daily living. With increasing age after a SCI the completeness and level of injury determine the occurrence of complications and outcome restrictions. In addition, problems in general health (e.g., in circulation, kidney function, diabetes mellitus) may affect the functional independence of elderly subjects with SCI.

Long-term follow-up of patients with spinal cord injury with a new ICF-based tool.

STUDY DESIGN: To develop a computer program that supports the overview of a follow-up care process in people with spinal cord injury (SCI) in daily clinical practice. OBJECTIVES: To create a new electronic tool based on the International Classification of Functioning, Disability and Health (ICF) that enables information to be registered and visualized, including the use of a net-diagram ('spider') to show a patient's long-term development. This diagram helps the clinician to recognize predispositions over time, as well as making information accessible to the patient, so as to involve him as a participant in defining current and future treatment options. Furthermore, guidelines for the prevention of common diseases, based on the recommendations of internal medicine, rehabilitation medicine and findings in the SCI literature, were implemented to provide enhanced health coaching in the area of preventative care. METHODS: In an outpatient setting, four perspectives were assessed: patient, physician, occupational therapist and physiotherapist for a comprehensive bio-psycho-social consideration. All categories were assessed and graphically visualized with the electronic tool, on the basis of the ICF domains: body function, activities/participation and environmental factors. RESULTS: The assessed data were summarized and graphically represented using three spider charts. CONCLUSION: The tool facilitates the patient counselling and the interdisciplinary work in daily clinical practice. Such a visual report helps to recognize predispositions over time. Furthermore, it helps to explain the clinical and patient-related findings accessible to the patients, to involve them as participants in defining the goals and the treatment plan.

External quality assessment programmes for detection of HCV RNA, HIV RNA and HBV DNA in plasma: improved proficiency of the participants observed over a 2-year period.

BACKGROUND AND OBJECTIVES: Two External Quality Assessment Programmes (EQAPs) were run in 2008 and 2009 to evaluate the proficiency of blood centres in detecting, by nucleic acid amplification techniques (NAT), the possible contamination of plasma with hepatitis C virus (HCV), human immunodeficiency virus (HIV) and hepatitis B virus (HBV). MATERIALS AND METHODS: In the EQAP-2008, three customized panels were designed; each containing positive samples with a viral nominal concentration for the three viruses of about three times the 95% DL of the respective commercial NAT assay. In the EQAP-2009, the proficiency of the participants was evaluated with a single panel, independently on the NAT method used. RESULTS: While 84% (102/122) of the participants in the EQAP-2008 correctly identified the positive and negative samples of the panels, in the EQAP-2009 the percentage of proficient laboratories increased to 97% (118/122). Most importantly, in this 2-year experience, we observed a decrease in the number of pre-/postanalytical errors, from 14 in 2008 to two in 2009. CONCLUSIONS: The design of these two EQAPs allowed participants to assess the performance of the NAT methods applied in their routine screening of blood donations, not only with respect to analytical errors but also to human errors that, despite the high level of automation reached by NAT methods, can still occur.

Outcome after incomplete spinal cord injury: central cord versus Brown-Sequard syndrome.

STUDY DESIGN: A retrospective analysis of prospectively collected data. OBJECTIVE: A hemisection of the spinal cord is a frequently used animal model for spinal cord injury (SCI), the corresponding human condition, that is, the Brown-Sequard syndrome (BS), is relatively rare as compared with the central cord syndrome (CC). The time course of neurological deficit, functional recovery, impulse conductivity and rehabilitation length of stay in BS and CC subjects were compared. SETTING: Nine European Spinal Cord Injury Rehabilitation Centers. METHODS: Motor score, walking function, daily life activities, somatosensory evoked potentials and length of stay were evaluated 1 and 6 months after SCI, and were compared between age-matched groups of tetraparetic BS and CC subjects. RESULTS: For all analyzed measures no difference in the time course of improvement was found in 15 matched pairs. CONCLUSION: In contrast to the assumption of a better outcome of subjects with BS, no difference was found between the two incomplete SCI groups. This is of interest with respect to the different potential mechanisms leading to a recovery of functions in these two SCI subgroups.

External quality assessment for the detection of HCV RNA, HIV RNA and HBV DNA in plasma by nucleic acid amplification technology: a novel approach.

BACKGROUND AND OBJECTIVES: In this EQA study a novel approach was used to assess the performance of blood centres and blood product manufacturers in detecting the possible contamination of plasma with HCV, HIV and HBV by NAT. MATERIALS AND METHODS: A panel of 12 samples, three negative and three positive for each virus, was distributed to the EQA participants. The positive samples were prepared, using the respective WHO standards, in order to obtain a viral concentration of about three times the 95% DL of the methods most commonly used by laboratories involved in blood screening by NAT. Participants were requested to test each sample of the panel on different days, possibly by different operators using their routine NAT assay. RESULTS: Overall, the participants' performance was satisfactory. In particular, 49 of the 59 participants (83%) were able to correctly identify all samples. Regarding the remaining 10 laboratories, in three cases a deviation from the laboratory's procedure that could be attributed to an operator's mistake was observed, in two cases a possible cross-contamination occurred while in the remaining five cases the failure to detect the positive samples couldn't be ascribed to any relevant deviation in the laboratory's procedure. CONCLUSIONS: The novel design of this EQA study allowed participants to verify their day by day activity as the study was carried out in the context of their routine testing. Under these conditions, it was demonstrated that, despite the high level of automation reached by NAT assays, human errors can still occur.

Standardized assessment of walking capacity after spinal cord injury: the European network approach.

OBJECTIVES: After a spinal cord injury (SCI), walking function is an important outcome measure for rehabilitation and new treatment interventions. The current status of four walking capacity tests that are applied to SCI subjects is presented: the revised walking index for spinal cord injury (WISCI II), the 6 minute walk test (6MinWT), 10 meter walk test (10MWT) and the timed up and go (TUG) test. Then, we investigated which categories of the WISCI II apply to SCI subjects who participated in the European Multicenter Study of Human Spinal Cord Injury (EM-SCI), and the relationship between the 10MWT and the TUG. METHODS: In the EM-SCI, the walking tests were applied 2 weeks and 1, 3, 6 and 12 months after SCI. We identified the WISCI II categories that applied to the EM-SCI subjects at each time point and quantified the relationship between the 10MWT and the TUG using Spearman's correlation coefficients (rho) and linear regression. RESULTS: Five WISCI II categories applied to 71% of the EM-SCI subjects with walking ability, while 11 items applied to 11% of the subjects. The 10MWT correlated excellently with the TUG at each time point (rho>0.80). However, this relationship changed over time. One year after SCI, the time needed to accomplish the TUG was 1.25 times greater than the 10MWT time. DISCUSSION: Some categories of the WISCI II appear to be redundant, while some discriminate to an insufficient degree. In addition, there appear to be ceiling effects, which limit its usefulness. The relationship between the 10MWT and TUG is high, but changes over time. We suggest that, at present, the 10MWT appears to be the best tool to assess walking capacity in SCI subjects. Additional valuable information is provided by assessing the needs for walking aids or personal assistance. To ensure comparability of study results, proposals for standardized instructions are presented.

Improving walking assessment in subjects with an incomplete spinal cord injury: responsiveness.

STUDY DESIGN: Prospective longitudinal study. OBJECTIVES: To investigate the responsiveness of the Walking Index for Spinal Cord Injury II (WISCI II), 6-Min Walk (6MWT) and 10-Meter Walk Tests (10MWT) for the assessment of walking capacity in incomplete spinal cord injury (iSCI) and to validate these tests with the lower extremity motor score (LEMS). SETTING: European Multicenter Study of Human Spinal Cord Injury. METHODS: The walking tests of 22 iSCI subjects who achieved functional ambulation and could stand or walk within 1 month after iSCI were analyzed at 3, 6 and 12 months after injury. Responsiveness was assessed by determining differences between the time intervals, and Spearman's correlation coefficient was calculated to quantify validity. RESULTS: All walking tests were able to assess the improvement of walking capacity within the first 3 months after injury. Between 3 and 6 months, only the 10MWT and 6MWT were responsive to the ongoing improvement in locomotor capacity. Overall, correlations between the tests were good within the first month, but became poorer over time. CONCLUSION: The 6MWT and 10MWT were more responsive in demonstrating an improvement in walking capacity compared to the WISCI II. The testing of functional outcome after iSCI as provided by the ordinal ASIA motor score can be improved by interval scaled measures. This allows increasing the responsiveness of functional outcome measures and should be advantageous in assessing therapeutical approaches in iSCI subjects. In iSCI subjects with walking ability, we recommend the additional use of timed tests to monitor improvement in locomotor capacity.

External quality assessment for the detection of blood-borne viruses in plasma by nucleic acid amplification technology: the first human immunodeficiency virus and hepatitis B virus studies (HIV EQA/1 and HBV EQA/1) and the fifth hepatitis C virus study (HCV EQA/5).

BACKGROUND AND OBJECTIVES: This External Quality Assessment (EQA) study was aimed at assessing the proficiency of blood centres and blood product manufacturers in detecting, by nucleic acid amplification technology (NAT), the possible contamination of plasma with hepatitis C virus (HCV), human immunodeficiency virus (HIV) and hepatitis B virus (HBV). MATERIALS AND METHODS: Three independent panels, one for each virus, were prepared at the Istituto Superiore di Sanità (ISS) by diluting the respective reference preparations. NAT methods used by the EQA participants included polymerase chain reaction (PCR) assays by Roche, transcription-mediated amplification (TMA) assays by Chiron and in-house PCR assays. RESULTS: Forty-three of the 45 participants (95.6%) in the HCV EQA/5 who used a validated method were consistently able to detect a nominal concentration of 100 IU/ml for all six major genotypes. In the case of the HIV EQA/1, all 35 participants detected the samples containing 1000 IU/ml HIV, while five (14.3%) did not identify the samples containing 100 IU/ml HIV. With respect to the HBV EQA/1, all 16 participants correctly identified the positive samples containing either 1000 IU/ml or 100 IU/ml HBV. No false-positive results were observed with any of the three panels. CONCLUSIONS: The HCV EQA/5 showed an improved proficiency of laboratories as compared with the HCV EQA/4. In fact, HCV genotypes 1, 2, 3 and 5 were correctly identified in 100% of the assays and genotypes 4 and 6 in 97.8% of the assays. While most of the participants in the HIV EQA/1 showed a good level of proficiency, an excellent performance was shown by all participants in the HBV EQA/1.

High proficiency in detecting the six major hepatitis C virus genotypes of laboratories involved in testing plasma by nucleic acid amplification technology.

Since the introduction of mandatory HCV RNA testing of plasma pools for fractionation by nucleic acid amplification technology, we have organised External Quality Assessment studies (EQAs) addressed to blood products manufacturers and blood centres. Here we report the results of a new EQA, the first one to include all six major HCV genotypes. The results, reported by laboratories worldwide, showed that genotypes 1, 2 and 3 were correctly identified in 100% of the tests, genotype 4 in 96.7% and genotypes 5 and 6 in 98.3% of the assays. As detection of all HCV genotypes is critical for laboratories involved in testing plasma for HCV, all six genotypes should continue to be included in the next EQA studies.

Perceived horizontal body position in healthy and paraplegic subjects: effect of centrifugation.

The perception of body position is mainly mediated by otolith information and visual cues. It has been shown, however, that proprioceptive sources are also involved. To distinguish between the contributions of the vestibular and nonvisual extra-vestibular information to graviception, we tested the effects of a stimulus that leaves the vestibular input unchanged but modifies the information from sense organs located more caudal along the trunk. This was achieved by bringing subjects into a horizontal ear-down position and rotating them around an earth-vertical axis that coincided with the interaural axis. In this paradigm, through centrifugal force, the stimulation of the vestibular and the putative extravestibular graviceptive organs in the body becomes dissociated. Healthy subjects (n = 14) and paraplegic patients with lesions between T4 and T8 (n = 7) adjusted themselves to the perceived horizontal right-ear down body position under two conditions: one with constant velocity rotation (ROT, velocity =120 degrees /s) around the earth-vertical axis of the turntable, and one without rotation (BASE). Among healthy subjects, the individual differences between BASE and ROT varied widely in both the feet-up or feet-down direction. In contrast, adjustments in paraplegic patients during ROT were always in the feet-down direction compared with BASE. A model with two extravestibular graviceptive sensors could explain our results: one sensor is located rostral to T4, and the other is caudal to T8. A load on the rostral graviceptor is interpreted as a tilt of the body in the feet-up direction and shifts the adjustments of perceived body position feet-down; a load on the caudal receptor is interpreted as a tilt in the feet-down direction and shifts the perceived body position feet-up. During ROT, healthy subjects solve the discrepant inputs of both extravestibular graviceptors in a highly variable manner, while paraplegic subjects show less variability because they are restricted to only the rostral graviceptor.

Hepatitis C virus testing of plasma pools by nucleic acid amplification technology: external quality assessment.

BACKGROUND AND OBJECTIVES: Since 1 July 1999, in accordance with European regulations, only batches of blood products obtained from plasma pools tested and found to be non-reactive for hepatitis C virus (HCV) RNA are being released. As monitoring the performance of manufacturers involved in plasma pool testing is important to ensure reliable amplification techniques, the Istituto Superiore di Sanità, as the Italian regulatory authority, organized an external quality assessment study. MATERIALS AND METHODS: A reference HCV RNA panel calibrated in international units (IU) was sent to each participant to be tested according to the validated procedure they routinely used in plasma pool testing. The panel consisted of 20 coded samples, four of which were obtained from a negative plasma pool. The remaining 16 samples, prepared by diluting the national reference preparation (ISS HCV RNA 0498), represented four half-log dilution series, each consisting of four samples containing 100, 32, 10 and 3.2 IU/ml of HCV RNA. RESULTS: The overall performance of the laboratories was very satisfactory. All laboratories correctly identified the negative samples. The 100- and 32-IU/ml samples were both detected in 98.4% of the assays, while the 10- and 3.2-IU/ml samples were detected in 73.4 and 50.0% of the assays, respectively. No substantial differences were observed between in-house procedures and commercial kits. CONCLUSION: This external quality assessment study showed that manufacturers of blood products have reached a high level of proficiency that fully complies with the European Pharmacopoeia requirements. This finding is reassuring in the context of the safety of blood products.

Driven gait orthosis for improvement of locomotor training in paraplegic patients.

DESIGN: Single cases. OBJECTIVE: To compare the effects of manually assisted locomotor training in paraplegic patients with the automated training by a driven gait orthosis. SETTING: ParaCare, University Hospital Balgrist in Zurich, Switzerland. METHODS: Treadmill training with manual assistance and by a driven gait orthosis was applied to two spinal cord injured patients. The first patient had an incomplete lesion at C3, the second a complete lesion at C5. They were selected by convenience sample. The EMG activity of the leg muscles rectus femoris, biceps femoris, gastrocnemius medials (GM) and tibialis anterior (TA) was visually compared for the two training methods. GM and TA activity was also quantified by calculating the variation ratio between the EMG of the patients and a set of healthy subjects. RESULTS: No significant difference between the two training methods was found according to the leg muscle EMG activity. CONCLUSION: Neuronal centers in the spinal cord become activated in a similar way by the manually assisted and the automated locomotor training. With the driven gait orthosis training sessions can be prolonged and workload of therapists can be reduced, and therefore, the automated training represents an alternative to the conventional therapy.

Long term effects of locomotor training in spinal humans.

The long term effects of locomotor training in patients with spinal cord injury (SCI) were studied. In patients with complete or incomplete SCI coordinated stepping movements were induced and trained by bodyweight support and standing on a moving treadmill. The leg extensor muscle EMG activity in both groups of patients increased significantly over the training period, associated with improved locomotor ability in those with incomplete SCI. During a period of more than 3 years after training, the level of leg extensor EMG remained about constant in incomplete SCI in those who regularly maintained locomotor activity. By contrast the EMG significantly fell in those with complete SCI. The results suggest a training induced plasticity of neuronal centres in the isolated spinal cord which may be of relevance for future interventional therapies.

Sensitivity of Mycosphaerella fijiensis from Banana to Trifloxystrobin.

An ascospore germination method was developed and validated to assess the sensitivity of bulk samples of Mycosphaerella fijiensis to trifloxystrobin. Using this method, the sensitivity of 142 ascospore samples from banana plantations not treated with strobilurins was analyzed to establish a baseline of pathogen sensitivity. A bulk method was utilized for monitoring purposes because it avoids potential complications due to the isolation and propagation of single-spore isolates and enables the testing of larger samples. Following intensive use of strobilurins (6 to 11 applications per year) over 4 years, under conditions of high disease pressure and the absence of sanitary measures at a development site in Costa Rica, bulk samples with 50% effective concentration (EC50) resistance factors (RFs) in excess of 500 compared with the mean baseline sensitivity were detected. Single-ascospore isolates derived from spores germinating at the discriminatory dose of 3 µg/ml were also resistant, suggesting that the frequency of resistant individuals in bulk samples could be estimated from the relative numbers of ascospores growing at this dose. The resistance of selected isolates was confirmed in planta. In vitro tests with four resistant and two sensitive single-ascospore isolates collected from different locations and times indicated possible cross-resistance of trifloxystrobin to azoxystrobin, famoxadone, and fenamidone, but not to propiconazole.

Prevalence of TT viral DNA in italian blood donors with and without elevated serum ALT levels: molecular characterization of viral DNA isolates.

BACKGROUND AND OBJECTIVE: A novel non-enveloped DNA virus, called TT virus (TTV), has been reported to be associated with post-transfusion hepatitis of unknown etiology. Although its clinical role still remains obscure, its presence in blood donations might cause problems. It, therefore, appeared of interest to investigate TTV prevalence in voluntary blood donors. DESIGN AND METHODS: A total of 595 Italian blood donors with and without elevated serum alanine aminotransferase (ALT) levels were tested by polymerase chain reaction using two sets of semi-nested primers that amplify the well-known region in the N22 clone. The amplified products were then sequenced to assess the genotype by phylogenetic and restriction fragment length polymorphism analyses. RESULTS: The prevalence of TTV in blood donors was 5+/-1.9% (25 out of 500) with a 95% confidence limit. A similar prevalence was found in 95 selected blood donors with increased ALT levels. A viral load of 10(3)-10(4) viral DNA molecules/mL was found, thus indicating a rather narrow range of variability. A phylogenetic tree built up on the basis of 210 base sequences of ORF1 allowed isolates to be classified into 2 groups corresponding, at least, to two of the putatives TTV genotypes, group 1 and group 2 of Okamoto's classification. A similar classification was also obtained by site restriction enzyme analysis. INTERPRETATION AND CONCLUSIONS: The results show that TTV infection is present among Italian blood donors. No significant difference in prevalence of TTV infection was found between patients with normal and increased ALT, making the association between TTV infection and human hepatitis questionable.