Reliability of the International Spinal Cord Injury Upper Extremity Basic Data Set.

STUDY DESIGN: It is a psychometrics study. OBJECTIVE: To assess the inter-rater reliability of the International Spinal Cord Injury Upper Extremity Basic Data Set (ISCI-UE). SETTING: Maharaj Nakorn Chiang Mai Hospital, Chiang Mai, Thailand. METHODS: Individuals with subacute and chronic cervical spinal cord injury (SCI) were recruited. One examiner rated five parts of the ISCI-UE, including the ability to reach and grasp, the shoulder function classification, utilization of adaptive devices used to enhance upper-extremity function, complications affecting upper-extremity function, and upper extremity/hand reconstructive surgery. A second blinded examiner repeated the procedures within 1 day. Quadratic weighted kappa was calculated to determine the inter-rater reliability. RESULTS: Sixty participants were included in the study. Fifty-two patients were men, and the mean (SD) age of participants was 42.9 (14.3) years. The median (interquartile range) time since injury was 9.5 (1-53) months. A total of 117 upper limbs were assessed. The inter-rater reliability was substantial, with almost perfect agreement in all items (ability to reach and grasp = 0.98; shoulder function classification = 0.97; use of assistive devices = 0.89; complications = 0.74; and surgery = 1). CONCLUSION: The International Spinal Cord Injury Upper Extremity Basic Data Set (ISCI-UE) has very good inter-rater reliability for evaluating individuals with cervical SCI.

International Spinal Cord Society (ISCoS) database: Asian Spinal Cord Network (ASCoN) Pilot Project [IDAPP].

STUDY DESIGN: Global mapping project of ISCoS for traumatic spinal cord injury (T-SCI) highlighted paucity of data from low and middle income countries (LMICs). Recognizing this gap, IDAPP study of one year duration was proposed as the first step to develop an International SCI database. OBJECTIVES: Primary objective was to assess database variables, processes involved and web platform for their suitability with a view to provide guidance for a large scale global project. Secondary objective was to capture demographic and selected injury/safety data on patients with T-SCI with a view to formulate prevention strategies. SETTING: Nine centers from Asia. METHODS: All patients with T-SCI admitted for first time were included. International SCI Core Data Set and especially compiled Minimal Safety Data Set were used as data elements. Questionnaire was used for feedback from centers. RESULTS: Results showed relevance and appropriateness of processes, data variables and web platform of the study. Ease of entering and retrieval of data from web platform was confirmed. Cost of one year IDAPP study was USD 7780. 975 patients were enrolled. 790 (81%) were males. High falls (n = 513, 52%) as a cause and complete injuries (n = 547, 56%) were more common. There was a higher percentage of thoracic and lumbar injuries (n = 516, 53%). CONCLUSIONS: The study confirms that establishing the SCI database is possible using the variables, processes and web platform of the pilot study. It also provides a low cost solution. Expansion to other centers/regions and including non-traumatic SCI would be the next step forward.

Prevalence of pressure ulcers in Thai wheelchair users with chronic spinal cord injuries.

STUDY DESIGN: A cross-sectional study. OBJECTIVES: To study prevalence of pressure ulcers (PrUs), quality of life (QoL) and effect of wheelchair cushions used by Thai wheelchair users with chronic spinal cord injury (SCI). SETTING: Maharaj Hospital, Chiang Mai, Thailand. METHODS: Thai chronic SCI wheelchair users, aged over 18 years and non-ambulatory with ASIA impairment scale A, B or C were recruited. They completed the PrUs questionnaire and rated the EuroQoL-5D and their health status with a visual analog scale (VAS). Demographic data of each participant were extracted from medical records. The EQ-5D health states were transformed to utility scores by using the Thai algorithm and the prevalence of PrUs was reported. The EQ-5D, the utility scores and the health status VAS were compared between those with and without current PrUs and between those participants using foam and air-filled cushions. RESULTS: Of 129 participants, 26.4% had current PrUs at the time of the study, 27.9% had healed PrUs and 45.7% never had PrUs. The median VAS score for health status was 70 (Q1=50, Q3=80). Based on the EQ-5D, only one dimension (anxiety/depression) was significantly different between those with and those without current PrUs (P=0.015). Those using an air-filled cushions had a mean utility score four times higher than of those using a foam cushion (0.131 vs. 0.032, P=0.089) but not statistically significant. CONCLUSIONS: PrUs are still prevalent among Thai wheelchair users with chronic SCI. Anxiety/depression is associated with current ulcers.

Pulmonary embolism after manual muscle testing in an incomplete paraplegic patient: a case report.

OBJECTIVE: To report and discuss the case of an incomplete paraplegic patient who died of pulmonary embolism (PE) aggravated by manual muscle testing. SETTING: Acute spinal ward, Maharaj Hospital, Chiang Mai, Thailand. CASE REPORT: A 79-year-old man suffering from chest trauma, fractured ribs and a fracture of T11 with incomplete paraplegia, American Spinal Injury Association impairment scale D. Intercostal tubes were inserted at both sides due to haemothorax. Ten days after onset, T9 to L2 posterior instrumentation was successfully completed. A week after the operation, he was allowed to stand on a tilt-table and a rehabilitation specialist was consulted to assess and plan to encourage ambulation. After manual muscle testing of the right hip flexors and knee extensors, the patient suffered from a short period of unconsciousness and breathlessness. Electrocardiography showed right bundle branch block and a drop in oxygen saturation from 98 to 70%. After oxygenation with mask and bag, oxygen saturation increased to 90%. PE or acute myocardial infarction was suspected. After insertion of an endotracheal tube, the patient went into cardiac arrest. Cardiopulmonary resuscitation failed. The autopsy revealed large and small thromboemboli in both lungs, particularly in the pulmonary artery. CONCLUSION: Strong hip and knee muscle contractions during manual muscle testing were suspected of triggering massive pulmonary emboli from the proximal vein of the right leg of a paraplegic patient who had functional motor movements and did not receive any thromboembolic prophylaxis which caused unexpected fatal pulmonary emboli. Screening of venous thromboembolism risks and its symptoms/signs before mobilisation is mandatory.

Identification of radiation-induced expression changes in nonimmortalized human T cells.

In the event of a radiation accident or attack, it will be imperative to quickly assess the amount of radiation exposure to accurately triage victims for appropriate care. RNA-based radiation dosimetry assays offer the potential to rapidly screen thousands of individuals in an efficient and cost-effective manner. However, prior to the development of these assays, it will be critical to identify those genes that will be most useful to delineate different radiation doses. Using global expression profiling, we examined expression changes in nonimmortalized T cells across a wide range of doses (0.15-12 Gy). Because many radiation responses are highly dependent on time, expression changes were examined at three different times (3, 8, and 24 h). Analyses identified 61, 512 and 1310 genes with significant linear dose-dependent expression changes at 3, 8 and 24 h, respectively. Using a stepwise regression procedure, a model was developed to estimate in vitro radiation exposures using the expression of three genes (CDKN1A, PSRC1 and TNFSF4) and validated in an independent test set with 86% accuracy. These findings suggest that RNA-based expression assays for a small subset of genes can be employed to develop clinical biodosimetry assays to be used in assessments of radiation exposure and toxicity.

IL10 and IL10 receptor gene variation and outcomes after unrelated and related hematopoietic cell transplantation.

BACKGROUND: Results of a previous study with human leukocyte antigen (HLA)-identical siblings showed individual and synergistic associations of single nucleotide polymorphisms in the promoter region of the recipient's IL10 gene and the donor's IL10 receptor beta (IL-10RB) gene with development of grades III-IV acute graft-versus-host disease (GVHD) after allogeneic hematopoietic cell transplantation. METHODS: In this study of 936 patients who had unrelated donors, genotypes of single nucleotide polymorphisms in the IL10 gene and the IL-10RB gene were evaluated as correlates with outcomes after transplantation. RESULTS: We found no statistically significant associations of polymorphisms at positions -3575, -2763, -1082, and -592 of the IL10 gene or codon 238 of the IL10RB gene with severe acute GVHD, extensive chronic GVHD or nonrelapse mortality after hematopoietic cell transplantation. Among HLA-matched unrelated pairs, the patient's IL10/-592 genotype and donor's IL10RB/c238 genotype showed trends suggesting individual and combined associations with grades III-IV acute GVHD similar to those observed among patients with HLA-identical sibling donors. CONCLUSIONS: Although genetic variation in IL10 pathway affects risk of acute GVHD and non-relapse mortality in HLA-identical sibling transplants, the current results indicate that genetic variation in the IL10 pathway does not significant affect these outcomes in unrelated donor transplants suggesting that the strength of the alloimmune response in the latter exceeds the anti-inflammatory activity of IL10.

Simultaneous isolation of DNA and RNA from the same cell population obtained by laser capture microdissection for genome and transcriptome profiling.

Laser capture microdissection (LCM) is used extensively for genome and transcriptome profiling. Traditionally, however, DNA and RNA are purified from separate populations of LCM-harvested cells, limiting the strength of inferences about the relationship between gene expression and gene sequence variation. There have been no published protocols for the simultaneous isolation of DNA and RNA from the same cells that are obtained by LCM of patient tissue specimens. Here we report an adaptation of the Qiagen AllPrep method that allows the purification of DNA and RNA from the same LCM-harvested cells. We compared DNA and RNA purified by the QIAamp DNA Micro kit and the PicoPure RNA Isolation kit, respectively, from LCM-collected cells from adjacent tissue sections of the same specimen. The adapted method yields 90% of DNA and 38% of RNA compared with the individual methods. When tested with the GeneChip 250K Nsp Array, the concordance rate of the single nucleotide polymorphism heterozygosity calls was 98%. When tested with the GeneChip U133 Plus 2.0 Array, the correlation coefficient of the raw gene expression was 97%. Thus, we developed a method to obtain both DNA and RNA material from a single population of LCM-harvested cells and herein discuss the strengths and limitations of this methodology.

Genetic variation in the IL-10 pathway modulates severity of acute graft-versus-host disease following hematopoietic cell transplantation: synergism between IL-10 genotype of patient and IL-10 receptor beta genotype of donor.

We have previously shown that the interleukin 10 (IL-10)/-592*A allele of the recipient is associated with less severe acute graft-versus-host disease (GVHD) and a lower risk of nonrelapse mortality after hematopoietic cell transplantation (HCT) from an HLA-identical sibling. In the present study, we examined variation in the IL-10 receptor beta gene as a further test of the hypothesis that the IL-10 pathway regulates the risk of acute GVHD. A single nucleotide polymorphism (A/G) at cDNA position 238 of the IL-10 receptor beta gene (IL10RB/c238) was genotyped in 953 HC transplant recipients and their HLA-identical sibling donors. IL-10/-592 and IL10RB/c238 genotypes were tested for association with GVHD by multivariable analysis. The IL-10/-592*A allele of the recipient and IL10RB/c238*G allele of the donor were significantly associated with a lower risk of grades III-IV acute GVHD (trend P < .001 and P = .02, respectively). The donor IL10RB/c238*G allele provided protection among patients with the IL-10/-592 A/C or A/A genotypes but not among patients with the high-risk IL-10/-592 C/C genotype. These data suggest an interaction of the patient IL-10/-592 and donor IL10RB/c238 genotypes on risk of GVHD, further supporting the hypothesis that the IL-10 pathway plays an important role in controlling the severity of acute GVHD.

Studies on allosteric phenomena in glycogen phosphorylase b.

This article attempts to trace, from a personal point of view, the history of discoveries of allosteric phenomena in phosphorylase b and the later development of systematic attempts to fit the data into comprehensive theoretical models. Work from our own laboratory is emphasized, but we try to integrate this into the results from other investigators and show their contributions to our ideas and experiments. Finally, some recent unpublished data is presented together with some conclusions and predictions from a new hypothesis. The discoveries by Carl and Gerty Cori of the activation of phosphorylase by AMP, the inhibition of glucose and the enzymatic interconversion of two forms fo the enzyme with different control properties helped lay the foundations of our present understanding of allosteric mechanisms. The later discovery of the oligomeric nature of phosphorylase and its relationship to AMP binding served as a basis for many years of research into the structure-function relationships of phosphorylase and other enzymes. Data showing that AMP lowers the entropy of activation is discussed with respect to the role of the nucleotide and its binding close to the active site. The discovery of the control of phosphorylase b by common metabolites and the impetus this gave to the intensive kinetic studies of the last ten years, wherein fitting to theoretical models has been a common feature, is reviewed.

Studies of the regulation and reaction mechanism of the carbamyl phosphate synthetase and aspartate transcarbamylase of bakers' yeast.

Kinetic studies of the carbamyl phosphate synthetase activity (CPSase) of bakers' yeast revealed an absolute requirement for K+ ions ; KM values for two of the substrates, glutamine and bicarbonate, were found to be 5 X 10(-4) M and 3 X 10(-3) M respectively. CPSase activity of the purified enzyme aggregate (M.W. 800,000) was extremely sensitive to UTP with a Ki of 2.4 X 10(-4) M. The purine nucleotide intermediate, XMP, was a strong activator of CPSase, acting at a site different from the regulatory site at which UTP binds ; XMP activation diminished at high concentrations of the substrate Mg-ATP. Studies of the reaction mechanism of CPSase revealed that it involved the sequential addition of the substrates bicarbonate and Mg-ATP, liberation of ADP, addition of glutamine, binding of ATP and then release of ADP and the product carbamyl phosphate. Studies of the reaction mechanism of the aspartate transcarbamylase (ATCase) of the aggregate yielded data which were not compatible with any of the usual models ; whichever reaction mechanism is ultivately found to fit the data, it will probably prove applicable both to the ATCase of the aggregate and to the disaggregated ATCase subunit (MW 138,000).

Kinetics and reaction mechanism of the carbamylphosphate synthetase of a multienzyme aggregate from yeast.

We have studied the kinetics and reaction mechanism of the carbamylphosphate synthetase of an enzyme aggregate functioning in the pyrimidine pathway of yeast. MG--ATP was found to be one of the substrates of the enzyme reaction which was activated by free Mg-2+ and inhibited by free ATP. Feedback inhibition by UTP was non-competitive with respect to both glutamine and bicarbonate. Potassium ions were essential for activity and could not be replaced by sodium. Glutamine could be replaced partially by ammonium ions as nitrogen donor. A bicarbonate-dependent cleavage of ATP was shown to take place in the absence of L-glutamine; L-glutamate was a competitive inhibitor of L-glutamine and the enzyme was shown to synthesize ATP when incubated with ADP and carbamyl phosphate. The reaction mechanism was found to involve sequential addition of the substrates bicarbonate and Mg--ATP and release of ADP, followed by addition of the third substrate glutamine. The purine nucleotide XMP had a pronounced activating effect on the enzyme, acting at a site different from that of UTP. Saturating levels of Mg--ATP eliminated this activation.