Blood testing in sport: hematological profiling.

Hemoglobin concentration and percent reticulocytes (%retics) were analyzed in blood samples taken pre-competition, post-competition, and during out of competition testing in elite speed skaters. Percent reticulocytes during screening was not different from the values obtained post-race, and no significant gender difference was found. Mean hemoglobin concentration both in males and females was slightly higher at 1 425 m altitude compared to <750 m altitude (0.23 g/dl increase in males and 0.48 g/dl increase in females; p<0.05 and p<0.01, respectively). Mean %retics at 1 425 m altitude is higher (0.24% in males and 0.27% in females, respectively, p<0.01) compared to blood sampled <750 m altitude. The distribution of percent reticulocytes shows 11 out of 11 500 samples with %reticulocytes below 0.4%. From the 171 samples with a values >2.4% in 52 skaters at least two consecutive samples yielded a percent reticulocytes above 2.4%. In 50 individuals with generally normal values but at least in two consecutive samples values above 2.4% the pattern required additional testing. In conclusion, percent reticulocytes are a robust hematological parameter, including acute exercise.

Hemoglobin level in elite speed skaters from 2000 up to 2005, and its relationship with competitive results.

We studied the mean hemoglobin (Hb) concentration in elite male and female long track junior and senior speed skaters from 2000 to 2005. In addition, the number of abnormal hematological findings observed over this period of time was investigated. We also studied whether there were differences in Hb concentration between top ranked and lower ranked skaters, and whether a relationship between Hb concentration and competitive results could be observed. The present study shows that the mean Hb level in male and female junior and senior long track speed skaters remained fairly stable and did not change from 2000 through 2005. The number and percentage of abnormal hematological findings were found to vary between 0 and 2 %, and failed to show a clear pattern or trend over the years. There was no difference in mean Hb levels between top ranked and lower ranked skaters, and no meaningful relationship between Hb concentration and ranking could be found.

Molecular nodes in memory processing: insights from Aplysia.

Recent research in a variety of systems indicates that memory formation can involve the activation of a wide range of molecular cascades. In assessing this recent work it is clear that no single cascade is uniquely important for all forms of memory, nor is a single form of memory uniquely dependent on a single cascade. Rather, it appears that molecular networks are differentially engaged in the induction of various forms of memory. Despite this highly interactive array of possible cascades, specific 'molecular nodes' have emerged as critical regulatory points in memory formation. Functionally, these nodes can operate in two sequential steps, beginning with a convergence of inputs which coordinately influence the activation state of the node, in which the nature of stimulation determines the dynamics of nodal activity, followed by a divergence of substrate selection, in which the node serves as a gateway that activates specific downstream effectors. Finally, specific nodes can be differentially engaged (i.e. have different 'weights') depending upon the nature and pattern of the activating stimulus. The marine mollusk Aplysia has proven useful for a molecular analysis of memory formation. We will use this system to highlight some of the molecular strategies employed by the nervous system in the formation of memory for sensitization, and we will focus on extracellular signal-related kinase as a candidate node integral to these processes.

Hemoglobin and hematocrit values after saline infusion and tourniquet.

This study attempted to contribute to standardization of blood testing in sport, and to investigate the effect of artificial dilution with saline. In 10 healthy, physically active males and 3 healthy physically active females hemoglobin (Hb), hematocrit (Ht), and % reticulocytes (%retics) were measured at different time points to look for possible fluctuations during day time, while the subjects had regular coffee breaks and lunch. In 7 of the subjects in a separate experiment 500 ml of saline were infused around 8 am and Hb, Ht, and %retics were measured before and every hour thereafter until 7 hours after infusion. In addition Ht was measured on a hematological analyzer as well as with a centrifuge. In a separate experiment the effect of tourniquet duration on Hb and Ht was studied in 9 of the subjects. The results show that Hb, Ht, and %retics are stable from 8 am to 4 pm, but that infusion of 500 ml of saline induces an acute decrease in Hb and Ht within one hour (Hb decreased from 15.2+/-0.9 g/dl to 14.5+/-1.0 g/dl, and Ht from 45.6+/-2.8 % to 44.0+/-2.5 %). The decline in Hb and Ht was maintained during the 7-hour observation period. Ht values of the same samples measured with a hematological analyzer and a centrifuge were not different. Application of the tourniquet did significantly affect Hb and Ht values only from two minutes, and thereafter Hb and Ht remained stable during the rest of the 5-minute tourniquet. With blood testing in sport these results have to be taken into consideration.

Comparison of dosimetry calibration factors at the NRCC and the NIST. National Research Council of Canada. National Institute of Standards and Technology.

In early 1998, three transfer ionization chambers were used to compare the air-kerma and absorbed-dose-to-water calibration factors measured by the National Research Council of Canada (NRCC) and the National Institute of Standards and Technology (NIST). The ratios between the NRCC and NIST calibration factors are 0.9950 and 1.0061 in the case of the absorbed-dose-to-water and air-kerma standards, respectively. In the case of the standard of absorbed dose to water, the combined uncertainty of the ratio between the standards of the two laboratories is about 0.6% and consequently, the observed difference of 0.5% is not significant at the one sigma level. In the case of the standard of air kerma, the combined uncertainty of the ratio between the standards of the two laboratories is about 0.4%, and so the observed difference of 0.61% is significant at the one sigma level. However, this discrepancy is due to the known differences in the methods of assessing the wall correction factor at the two laboratories. Taking into account changes implemented in the standards that form the basis of the calibrations, the present results are consistent with those of the previous comparison done in 1990/91. As a direct result of these differences in the calibration factors, changing from an air-kerma based protocol following TG-21 to an absorbed-dose-to-water based protocol following TG-51, would alter the relationship between clinical dosimetry in Canada and the United States by about 1%. For clinical reference dosimetry, the change from TG-21 to TG-51 could result in an increase of up to 2% depending upon the ion chamber used, the details of the protocol followed and the source of traceability, either NRCC or NIST.

Environmental dosimetry performance criteria: pilot test of proposed ANSI draft standard N13.29. American National Standards Institute.

American National Standards Institute (ANSI) standards are a source of technical guidance on the application of passive radiation detectors in personnel and environmental monitoring programs. The proposed new ANSI Draft N13.29 describes performance tests for environmental dosimetry providers that are analogous to those now required in personnel dosimetry. The objective of this study was to pilot test the procedural specifications before Draft N13.29 is submitted for final balloting. The results of the performance tests are relevant to environmental surveillance programs, which may be affected if Draft N13.29 is approved. Seven environmental dosimetry providers submitted dosimeters for the pilot test, which included two phases. The first phase involved exposing dosimeters to laboratory gamma, beta, and x-ray sources at routine and accident dose levels. In the second phase, dosimeters were subjected to 90 d of simulated environmental conditions in a chamber that cycled through prescribed temperature and humidity parameters and lighting conditions. Two participants passed all categories of the laboratory test phase, and all seven passed the environmental test phase. We report here on the results of the performance tests; additional results relevant to finalizing Draft N13.29 are detailed elsewhere.

Reoperation after primary posterior instrumentation and fusion for idiopathic scoliosis. Toward defining late operative site pain of unknown cause.

STUDY DESIGN: Retrospective case series. OBJECTIVES: To determine the frequency and categorize the indications for reoperation in three implant systems (Harrington, Cotrel-Dubousset [C-D], and Isola). To define late operative site pain (LOSP) of no apparent cause as an indication for implant removal and determine the success of implant removal in relieving LOSP. SUMMARY OF BACKGROUND DATA: Late operative site pain of no apparent cause has been discussed briefly in the literature but has not been investigated as a major indication for implant removal. METHODS: One hundred eighty-two of 190 consecutive patients with idiopathic scoliosis (96%) who underwent primary surgery between January 1, 1981, and December 31, 1992, by one surgeon in one hospital, with use of Harrington, C-D, or Isola instrumentation were studied an average of 9 years after surgery to determine the indications for and frequency of reoperation. RESULTS: The overall frequency of reoperation for all instrumentation types combined was 19%: Harrington, 19%; C-D, 24%; and Isola, 14%. By 6 years' follow-up the cumulative risk of reoperation by Kaplan-Meier analysis was Harrington, 14%; C-D, 21%; and Isola, 14%. (statistically nonsignificant difference). The most frequent indication for reoperation was LOSP of no apparent cause: 8% (14 patients) for all instrumentation types combined. The average interval between the initial operation and reoperation for LOSP was 46 months (range, 20-97 months). The frequency of each implant type was Harrington, 6%; C-D, 12%; and Isola, 6%. By 6 years' follow-up, the cumulative Kaplan-Meier risk for reoperation due to LOSP was Harrington, 5%; C-D, 13%; and Isola, 8% (statistically non-significant difference). Of the 14 patients who had instrumentation removal for LOSP, 10 (71%) had successful relief of pain after implant removal. CONCLUSIONS: Occurring regardless of implant type, LOSP of no apparent cause after posterior instrumentation of scoliosis is a distinct clinical entity and is relieved by implant removal in most patients.

Scatter fractions from linear accelerators with x-ray energies from 6 to 24 MV.

Computation of shielding requirements for a linear accelerator must take into account the amount of radiation scattered from the patient to areas outside the primary beam. Currently, the most frequently used data are from NCRP 49 that only includes data for x-ray energies up to 6 MV and angles from 30 degrees to 135 degrees. In this work we have determined by Monte Carlo simulation the scattered fractions of dose for a wide range of energies and angles of clinical significance including 6, 10, 18, and 24 MV and scattering angles from 10 degrees to 150 degrees. Calculations were made for a 400 cm2 circular field size impinging onto a spherical phantom. Scattered fractions of dose were determined at 1 m from the phantom. Angles from 10 degrees to 30 degrees are of concern for higher energies where the scatter is primarily in the forward direction. An error in scatter fraction may result in too little secondary shielding near the junction with the primary barrier. The Monte Carlo code ITS (Version 3.0) developed at Sandia National Laboratory and NIST was used to simulate scatter from the patient to the barrier. Of significance was the variation of calculated scattered dose with depth of measurement within the barrier indicating that accurate values may be difficult to obtain. Mean energies of scatter x-ray spectra are presented.

Macromolecular substrate affinity for the tissue factor-factor VIIa complex is independent of scissile bond docking.

The upstream coagulation enzymes are homologous trypsin-like serine proteases that typically function in enzyme-cofactor complexes, exemplified by coagulation factor VIIa (VIIa), which is allosterically activated upon binding to its cell surface receptor tissue factor (TF). TF cooperates with VIIa to create a bimolecular recognition surface that serves as an exosite for factor X binding. This study analyzes to what extent scissile bond docking to the catalytic cleft contributes to macromolecular substrate affinity. Mutation of the P1 Arg residue in factor X to Gln prevented activation by the TF.VIIa complex but did not reduce macromolecular substrate affinity for TF.VIIa. Similarly, mutations of the S and S' subsites in the catalytic cleft of the enzyme VIIa failed to reduce affinity for factor X, although the affinity for small chromogenic substrates and the efficiency of factor X scissile bond cleavage were reduced. Thus, docking of the activation peptide bond to the catalytic cleft of this enzyme-cofactor complex does not significantly contribute to affinity for macromolecular substrate. Rather, it appears that the creation of an extended macromolecular substrate recognition surface involving enzyme and cofactor is utilized to generate substrate specificity between the highly homologous, regulatory proteases of the coagulation cascade.

Regulation of the catalytic function of coagulation factor VIIa by a conformational linkage of surface residue Glu 154 to the active site.

Coagulation factor VIIa is an allosterically regulated trypsin-like serine protease that initiates the coagulation pathways upon complex formation with its cellular receptor and cofactor tissue factor (TF). The analysis of a conformation-sensitive monoclonal antibody directed to the macromolecular substrate exosite in the VIIa protease domain demonstrated a conformational link from this exosite to the catalytic cleft that is independent of cofactor-induced allosteric changes. In this study, we identify Glu 154 as a critical surface-exposed exosite residue side chain that undergoes conformational changes upon active site inhibitor binding. The Glu 154 side chain is important for hydrolysis of scissile bond mimicking peptidyl p-nitroanilide substrates, and for inhibition of VIIa's amidolytic function upon antibody binding. This exosite residue is not linked to the catalytic cleft residue Lys 192 which plays an important role in thrombin's allosteric coupling to exosite I. Allosteric linkages between VIIa's active site and the cofactor binding site or between the cofactor binding site and the macromolecular substrate exosite were not influenced by mutation of Glu 154. Glu 154 thus only influences the linkage of the macromolecular substrate binding exosite to the catalytic center. These data provide novel evidence that allosteric regulation of VIIa's catalytic function involves discrete and independent conformational linkages and that allosteric transitions in the VIIa protease domain are not globally coupled.

Importance of factor VIIa Gla-domain residue Arg-36 for recognition of the macromolecular substrate factor X Gla-domain.

Macromolecular substrate docking with coagulation enzyme-cofactor complexes involves multiple contacts distant from the enzyme's catalytic cleft. Here we characterize the binding of the Gla-domain of macromolecular substrate coagulation factor X to the complex of tissue factor (TF) and VIIa. Site-directed mutagenesis of charged residue side chains in the VIIa Gla-domain identified Arg-36 as being important for macromolecular substrate docking. Ala substitution for Arg-36 resulted in an increased KM and a decreased rate of X activation. X with a truncated Gla-domain was activated by mutant and wild-type VIIa at indistinguishable rates, demonstrating that Arg-36 interactions require a properly folded Gla-domain of the macromolecular substrate. VIIa Arg-36 was also required for effective docking of the X Gla-domain in the absence of phospholipid, demonstrating that the Gla-domain of VIIa participates in protein-protein interactions with X. In the absence of TF, the mutant VIIa had essentially normal function, indicating that the cofactor positions VIIa's Gla-domain for optimal macromolecular substrate docking. Computational docking suggests multiple charge complementary contacts of the X Gla-domain with TF.VIIa. A prominent interaction is made by the functionally important X residue Gla-14 with the center of the extended docking site created by residues in the carboxyl module of TF and the contiguous VIIa Gla-domain. These data demonstrate the functional importance of interactions of the Gla-domains of enzyme and substrate, and begin to elucidate the molecular details of the ternary TF.VIIa.X complex.

Scattered fractions of dose from 6,10,18, and 25 MV linear accelerator x rays in radiotherapy facilities.

Data for computation of primary and secondary shielding barriers in radiotherapy facilities are presented in the National Council on Radiation Protection and Measurements Report 49, which covers primary barriers for accelerating voltages up to 10 MV and secondary barriers up to 6 MV. Inconsistencies in reported scattered fraction values found in the literature for megavoltage accelerators are due, in part, to differences in measurement techniques. A consistent approach was used to measure the scattered fractions of 6, 10, 18, and 25 MV photon dose. A reference ion chamber was placed on the symmetry axis of a cylindrical water phantom, approximating the human torso, 25.6 cm diameter, at the gantry isocenter. Measurements were made with a second ion chamber at 2m from the isocenter at angles ranging from 10 degrees through 150 degrees from the beam central axis with a field size of 20 cm x 20 cm at the isocenter. The dose measured at each angle is reported as a fraction of the reference chamber dose at the center of the phantom and as calculated to a reference chamber depth of d(max), also at the isocenter. The results of these measurements were compared with Monte Carlo calculations and show reasonable agreement. A comparison with previously published data was not as good due to the differences in measurement techniques. However, after some adjustments to correct for the differences, the comparison is very good. Scattered fractions beyond 30 degrees are highest for the lowest energy beam investigated. This indicates that the scattered fraction values reported in NCRP 49 for 6 MV are sufficient for use in conservative shielding calculations for the higher energy x rays beyond 30 degrees. For angles smaller than 30 degrees, the new data are more appropriate and show the scatter fractions are larger for higher MV x rays.

Influence of cofactor binding and active site occupancy on the conformation of the macromolecular substrate exosite of factor VIIa.

The catalytic activity of the trypsin-like serine protease coagulation factor VIIa is allosterically regulated. In this work, we employed monoclonal antibodies as probes to analyze conformational changes in the VII protease domain that are induced by zymogen activation, cofactor tissue factor (TF) binding, and active site occupancy. The epitopes of three monoclonal antibodies were mapped using a panel of 57 individual alanine replacement mutants in the protease domain. Two of the antibodies had typical "hot spot" epitopes in a basic cluster above the active site cleft and antibody binding to these epitopes was not affected by zymogen activation, TF binding, or active site occupancy. In contrast, the binding kinetics of VII/VIIa to a monoclonal antibody that mapped to an extended epitope overlapping with the macromolecular substrate exosite was affected by each of the conformational transitions of the VIIa protease domain. The changes in antibody affinity are consistent with a transition from zymogen VII to the TF.VIIa complex, with free enzyme VIIa as an intermediate that retains some zymogen-like features responsible for its low catalytic activity. In contrast, active site occupancy resulted in effects that were qualitatively different from the effects of zymogen activation on the antibody epitope. This provides novel insight into the conformational interdependence between the active site, the region for macromolecular substrate recognition, and the cofactor binding exosite of this allosterically regulated serine protease.

Nitrogen balance in patients receiving either fat or carbohydrate for total intravenous nutrition.

Eighteen pre and/or postoperative patients underwent TIN using either fat or carbohydrate as a caloric substrate source for a similar period. Positive nitrogen balance was achieved with either solution for an equal number of days when balance studies were complete. Both groups demonstrated weight gain but it was more marked in the carbohydrate TIN group. One patient in the carbohydrate TIN group was changed to Intralipid because of a catheter related complication. This patient continued to show positive nitrogen balance while on Intralipid. There were 4 deaths in the 18 patients unrelated to TIN. It is concluded that Intralipid, when given through a peripheral vein with a nitrogen source, can produce positive nitrogen balance and is a safe and effective means for doing this.

Prevention of peritoneal adhesions in the rat. The effects of dexamethasone, methylprednisolone, promethazine, and human fibrinolysin.

Peritoneal adhesions were created in rats by brisk scrubbing of the terminal part of the ileum. Adhesions were graded by total number and the presence of small bowel obstruction. Adhesion prophylaxis was evaluated using dexamethasone, methylprednisolone sodium succinate, promethazine hydrochloride, and human fibrinolysin (Thrombolysin) in various combinations, doses, and routes of administration. Methylprednisolone and dexamethasone, depending on the route of administration, modified the total number of adhesions but did not modify their severity when compared to control animals. Promethazine by itself modified peritoneal adhesions in the rat. Used together, methylprednisolone and promethazine also modified adhesions, but were not substantially better than the combination of dexamethasone and promethazine. Methylprednisolone, promethazine, and human fibrinolyzin, when used in combination intraperitoneally, virtually eliminated adhesion formation.