The level of agreement between the numerical rating scale and visual analogue scale for assessing pain intensity in adults with chronic pain.

The numerical rating scale and visual analogue scale are used to quantify pain intensity. However, it has not yet been explored whether these scores are interchangeable in adults with chronic pain. Data from the prospective multicentre cross-sectional INTERVAL study were used to evaluate the one-dimensionality and agreement between numerical rating scale scores and visual analogue scale scores in adults with chronic pain. Pain intensity scores using the numerical rating scale and visual analogue scale were provided by 366 patients with chronic pain for current, average, minimal and maximal pain. To evaluate whether pain intensity scales are completed in accordance with each other, the proportion of patients who satisfied the following condition was calculated: minimal pain intensity ≤ maximal pain intensity. A factor analysis confirmed the one-dimensionality of the pain measures. A significant difference was found between numerical rating scale and visual analogue scale scores for average, current, minimum and maximum pain. Intra-class correlation coefficient estimates ranged from 0.739 to 0.858 and all measures failed to show sufficient and acceptable agreement at the 95% level. The strength of agreement between pain severity categories was classified as 'moderate' for average and minimal pain and 'substantial' for current and maximal pain. The proportion of patients who scored minimal pain ≤ maximal pain was 97.5% for the numerical rating scale and 89.5% for the visual analogue scale. This study failed to show an acceptable agreement between the numerical rating scale and visual analogue scale when pain intensity was rated by adults with chronic pain, despite showing both scales measure the same information.

The sarcopenia and physical frailty in older people: multi-component treatment strategies (SPRINTT) project: description and feasibility of a nutrition intervention in community-dwelling older Europeans.

BACKGROUND: The "Sarcopenia and Physical Frailty in Older People: Multicomponent Treatment Strategies" (SPRINTT) project sponsored a multi-center randomized controlled trial (RCT) with the objective to determine the effect of physical activity and nutrition intervention for prevention of mobility disability in community-dwelling frail older Europeans. We describe here the design and feasibility of the SPRINTT nutrition intervention, including techniques used by nutrition interventionists to identify those at risk of malnutrition and to carry out the nutrition intervention. METHODS: SPRINTT RCT recruited older adults (≥ 70 years) from 11 European countries. Eligible participants (n = 1517) had functional limitations measured with Short Physical Performance Battery (SPPB score 3-9) and low muscle mass as determined by DXA scans, but were able to walk 400 m without assistance within 15 min. Participants were followed up for up to 3 years. The nutrition intervention was carried out mainly by individual nutrition counseling. Nutrition goals included achieving a daily protein intake of 1.0-1.2 g/kg body weight, energy intake of 25-30 kcal/kg of body weight/day, and serum vitamin D concentration ≥ 75 mmol/L. Survey on the method strategies and feasibility of the nutrition intervention was sent to all nutrition interventionists of the 16 SPRINTT study sites. RESULTS: Nutrition interventionists from all study sites responded to the survey. All responders found that the SPRINTT nutrition intervention was feasible for the target population, and it was well received by the majority. The identification of participants at nutritional risk was accomplished by combining information from interviews, questionnaires, clinical and laboratory data. Although the nutrition intervention was mainly carried out using individual nutritional counselling, other assisting methods were used as appropriate. CONCLUSION: The SPRINTT nutrition intervention was feasible and able to adapt flexibly to varying needs of this heterogeneous population. The procedures adopted to identify older adults at risk of malnutrition and to design the appropriate intervention may serve as a model to deliver nutrition intervention for community-dwelling older people with mobility limitations.

Cortical motor output decreases after neuromuscular fatigue induced by electrical stimulation of the plantar flexor muscles.

AIM: Neuromuscular electrical stimulation (NMES) causes early onset of neuromuscular fatigue. Peripheral electrophysiological explorations suggest that supra-spinal alterations are involved through sensitive afferent pathways. As sensory input is projected over the primary somatosensory cortex (S1), S1 area involvement in inhibiting the central motor drive can be hypothesized. This study assessed cortical activity under a fatiguing NMES protocol at low frequency. METHODS: Twenty healthy males performed five NMES sequences of 17 trains over the plantar flexors (30 Hz, 4 s on/6 s off). Before and after each sequence, neuromuscular tests composed of maximal voluntary contractions (MVCs) were carried out. Cortical activity was assessed during MVCs with functional near-infrared spectroscopy over S1 and primary motor (M1) areas, through oxy- [HbO] and deoxy-haemoglobin [HbR] variation. Electrophysiological data (H-reflex during MVC, EMG activity and level of voluntary activation) were also recorded. RESULTS: MVC torque significantly decreased after the first 17 NMES trains (P < 0.001). The electrophysiological data were consistent with supra-spinal alterations. In addition, [HbO] declined significantly during the protocol over the S1 and M1 areas from the first 17 NMES trains (P < 0.01 and P < 0.001 respectively), while [HbR] increased (P < 0.05 and P < 0.01 respectively), indicating early decline in cortical activity over both primary cortical areas. CONCLUSIONS: The declining cortical activity over the M1 area is highly consistent with the electrophysiological findings and supports motor cortex involvement in the loss of force after a fatiguing NMES protocol. In addition, the declining cortical activity over the S1 area indicates that the decreased motor output from M1 is not due to increased S1 inhibitory activity.

How the ankle joint angle alters the antagonist and agonist torques during maximal efforts in dorsi- and plantar flexion.

The aim of this study was to assess, via an EMG bio-feedback method, the ankle joint angle effect on the agonist and antagonist torques in plantar- (PF) and dorsi-flexion (DF). The isometric PF and DF maximal voluntary contractions (MVCs) torques were measured simultaneously with surface EMG activity of triceps surae (TS) and tibialis anterior (TA) muscles in 12 young adults (mean age 27) at five different ankle joint angles. Our results showed that: (i) The coactivation level does not properly reflect the mechanical effect of the antagonist muscle, (ii) TS antagonist torque significantly altered the DF MVC-angle relationship, whereas TA antagonist torque did not influence this MVC-angle relationship in PF. The alteration of the MVC with angular position was due, in part, to the coactivation phenomenon in DF, but not in PF. Thenceforth, when investigating the torque at the ankle joint, it is necessary to take into account both agonist and antagonist torque modifications with ankle joint angle.

High serum vascular endothelial growth factor correlates with disease activity of spondylarthropathies.

Angiogenesis is involved in chronic inflammatory joint diseases such as rheumatoid arthritis (RA). Vascular endothelial growth factor (VEGF) plays a crucial role in angiogenesis. The spondylarthropathies (SpA) are characterized by enthesitis and synovitis, in which blood vessels participate. The objective of this study was to investigate serum VEGF levels and their potential associations with disease activity markers for SpA. Sera were collected from 105 patients with SpA (72 with ankylosing spondylitis (AS), four with psoriatic arthritis (PsA), six with reactive arthritis (ReA), eight with enteropathic arthropathy and 15 with undifferentiated SpA), 50 patients with rheumatoid arthritis (RA) and 64 healthy controls. Disease activity in SpA patients was assessed using the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and laboratory parameters of inflammation [erythrocyte sedimentation rate (ESR) and C-reactive protein level (CRP)]. Serum VEGF levels were significantly higher in SpA patients (316.4 +/- 215.6 pg/ml) and RA patients (405.2 +/- 366.5) than in controls (217.3 +/- 145.2) (P = 0.003). In SpA patients, serum VEGF levels correlated with disease activity indices (BASDAI: r = 0.22, P = 0.04; ESR: r = 0.3, P = 0.003; and CRP: r = 0.23, P = 0.02). Serum VEGF levels were not associated with presence of extra-articular manifestations or syndesmophytes or with the grade of sacroiliitis. These results suggest that VEGF and therefore angiogenesis may play a role in SpA pathogenesis and may serve as a disease activity marker in SpAs.

TWEAK stimulation of astrocytes and the proinflammatory consequences.

Astrocytes exert many active roles in brain homeostasis, potentially including the regulation of immune reactions. They possess a substantial aptitude for plasticity and, indeed, functional and phenotypic changes are frequently encountered in reactive gliosis observed in brain injuries. The significance of reactive astrocytes is still poorly defined, but it is clear that these cells are an important source of cytokines in inflamed brain. How tumor necrosis factor (TNF) and TNF-receptor family members contribute to this reaction is an interesting issue that is currently being explored. It was previously shown that reactive astrocytes express high levels of Fas (CD95) and respond to Fas ligand (CD95L) by apoptosis or IL-8 production. TWEAK (Apo-3 ligand) is a recently identified member of the TNF family that is produced mainly by leukocytes that can infiltrate the inflamed brain and thus influence astrocyte behavior. Here we show that human astrocytes derived from different regions of the brain specifically bind TWEAK and are totally resistant to TWEAK mediated apoptosis. In addition, high amounts of IL-8 and IL-6 were secreted by astrocytes after TWEAK exposure. Finally, expression of cell surface molecules involved in the propagation and/or maintenance of brain inflammation was determined. TWEAK significantly increased ICAM-1 expression on astrocytes, whereas no modification was detected in the expression of Fas, TNFRI, B7-1, or MHC molecules. In conclusion, the proinflammatory effects induced by TWEAK on astrocytes in culture recapitulate many characteristics of reactive astrocytes observed in vivo, suggesting that TWEAK could play a significant role in brain inflammation.

Detection of intracellular cytokines in citrated whole blood or marrow samples by flow cytometry.

We have used three-color flow cytometric analysis for the detection of intracellular cytokines (IFN-gamma and IL-4) in CD3(+) cells, after stimulation for 4 h with phorbol 12-myristate 13-acetate (PMA) and ionomycin in the presence of monensin. We report in the present paper a validation study for analysing IFN-gamma and IL-4 production by bone marrow (BM)-derived T cells and peripheral blood T cells after BM transplantation. Using citrate as anticoagulant for blood and marrow sampling interfered with PMA+ionomycin-based cell stimulation. Indeed, removing this anticoagulant by two washes with 10% pooled human AB serum-supplemented RPMI 1640 before cell stimulation improved the percentages of IL-4(+) (0.02+/-0.01% to 0. 47+/-0.17% without and with washes, respectively; p<0.01) and IFN-gamma(+) (6.8+/-2.75% to 39.33+/-4.6%; p<0.01) cells to levels similar to those observed in heparin-based whole blood cultures (0.38+/-0.17% IL-4(+) and 34.27+/-4.96% IFN-gamma(+) cells; p>0.05). Delaying the cell cultures for 24 h did not significantly modify the detection of IFN-gamma in washed whole blood, but significantly altered IFN-gamma secretion in culture supernatants, as assessed by ELISA. Moreover, the percentage of IFN-gamma-producing cells within the CD3(+) lymphocyte population was stable, since similar results were obtained in two or three different independent experiments performed with the same healthy donors. This method was shown to be applicable for different kinds of citrated samples, such as blood or BM-derived cells. Overall, our data suggest that in addition to allowing for the identification of cytokine-producing cell phenotype, intracellular cytokines staining using flow cytometry is more reliable than ELISA for the biological follow-up of clinical samples.

Comparative analysis of class I, II and III epitope-detecting CD34 monoclonal antibodies by quantitative flow cytometry.

UNLABELLED: The aim of this study was to compare different CD34 monoclonal antibodies (MAbs) belonging to three different classes: MY10 class I, QBend10 class II, a mixture of three selected MAbs class I and II designated as CD34 Pool, and 8G12 class III. Bone marrow (BM) samples from 13 healthy donors were analyzed for: 1) percentage of CD34+ cells, 2) quantitative expression of CD34 epitopes (antigen's density - AgD) using a quantitative indirect immunofluorescence (QIFI) test, 3) study of CD34+ cell subsets defined by CD34 and CD38 coexpression. 8G12 MAb showed the highest reactivity with regard to the percentage of detected CD34+ cells and AgD on these cells. A nearly identical percentage of CD34+ cells was detected with CD34 Pool, but with a lower AgD. With QBend10, the percentage of CD34 expressing cells was insignificantly decreased and the AgD was slightly lower. The expression of the MY10 epitope was the lowest and was detected on the lowest number of CD34+ cells. Concerning CD34 and CD38 coexpressing subset, we observed that 8G12 class III MAb detected CD34loCD38dim cells with comparable efficiency with MY10 class I MAb, but with significantly higher level than QBend10 class II and CD34 Pool class I+II MAbs. The CD34hiCD38dim subset was detected with the same efficiency by QBend10, CD34 Pool or 8G12 MAbs but with significantly higher frequency than MY10 MAb. IN CONCLUSION: class II and III MAbs appear preferable for flow cytometric quantification of CD34+ cells; for CD34+ cell subsets determination class III MAbs should be suitable.

Comparison of T cell depletion strategies from bone marrow, umbilical cord and peripheral blood using five separation systems.

The development of bone marrow transplantation in mismatched or matched unrelated donor situations, the recent use of peripheral blood stem cells for allogeneic transplants, the standardization and respect of good methodology practices highlight the need to evaluate new safe methods of T cell depletion (TCD). We have performed 79 in vitro TCD using five techniques: rabbit complement cytotoxicity, CD2-CD7 immunomagnetic depletion, CD5-CD8 panning system, CD34 positive purging and counterflow centrifugation elutriation (CCE). We analyzed these different approaches with regard to the degree of T and B depletion, recovery of progenitors and NK cells. In our hands, the 5 systems evaluated showed a TCD of between 1.3 and 3 log. The CCE, immunomagnetic, complement and panning methods all give similar a TCD of around 2 log. In contrast, we obtained a TCD of approximately 3 log with CD34 positive purging. The progenitor yield was around 50% regardless of the technique used. However, the degree of B and NK cell depletion was dependent on the method: specific TCD resulted in low BCD (under 0.5 log), whereas CCE or CD34 positive purging gave a BCD of greater than 1 log. Moreover, CD34 positive selection resulted in a virtually complete elimination of NK cells. CCE was the only technique allowing isolation of the small lymphocyte population which can be useful for adoptive therapy. To obtain TCD over three logarithms, double purging techniques are necessary. Because specific roles of T cells subsets in engraftment, graft versus host disease, Epstein Barr virus associated B cell lymphoproliferative disorders and disease relapse have not yet been completely elucidated, new techniques such as CD34 positive purging and double purging methods (positive and negative purging) need to be clinically evaluated, especially with respect to peripheral blood stem cells.

Variability of the alloreactive T-cell response to human leukemic blasts.

Clinical and experimental data suggest a role for the immune response in preventing leukemic relapses after allogeneic bone marrow transplantation (BMT): the graft-versus-leukemia (GVL) effect. In this report, we have evaluated the response of normal donor lymphocytes against allogeneic leukemic cells as an in vitro model of the GVL effect. We used a limiting dilution technique in order to determine the frequency of cytotoxic T-lymphocyte precursors (pre-CTL) against allogeneic leukemic blasts among normal donor lymphocytes. We demonstrate a considerable variability of CTL precursor frequency. This variability depended on leukemic populations since, for a given leukemia, the pre-CTL frequency was comparable among our tested normal allogeneic donors. Moreover, when HLA-DR negative leukemias were used as allostimulators, the pre-CTL frequencies were extremely low. In order to verify the impact of leukemic DR expression on the stimulatory capacity of leukemic cells, we selected and analyzed in mixed lymphocyte tumor cell culture (MLTC), a panel of myelogenous and lymphoblastic leukemias with variable levels of DR expression, each against different allogeneic responders. Our results demonstrated a close correlation (r = 0.953, p < 0.0001) between the proliferative response of alloactivated lymphocytes and the percentage of stimulatory leukemic cells expressing HLA-DR molecules. Anti-MHC class II monoclonal antibodies inhibited the lymphocyte proliferation in the MLTC, confirming the preponderant role of DR in the generation of this response. Overall, our results demonstrate the extreme variability of leukemic cells in their allostimulatory capacity and the central role of DR expression in determining leukemic allo-recognition. In the setting of a clinical protocol, our data suggest that the infusion of allogeneic T lymphocytes in a DR negative leukemia will not lead to an alloreactive T-cell anti-tumor effect.

Immunological characteristics of umbilical cord blood cells: phenotypic and functional analysis.

The purpose of this study was to phenotype and assay immunological function on cord blood from 70 samples. Immunophenotyping indicated similar numbers of CD2, 3, and 8-positive cells as in adult bone marrow. CD4-positive cells were increased and CD19, 20, and CD56-positive cells were decreased in numbers. Proliferative responses to nonspecific mitogens were lower in cord blood cells than in adult cells, as was NK activity and spontaneous secretion of IL-6 and TNF-alpha. This study confirms the relative immaturity of cord blood cells even when some functional assays appear normal.

Interleukin-2-induced increase of a monoclonal B-cell lymphocytosis. A novel in vivo interleukin-2 effect?

A 56-year-old man with refractory B-cell lymphocytic non-Hodgkin's lymphoma was treated in a Phase II study with interleukin-2 (IL-2) (Roussel-Uclaf, Romainville, France). The patient had involvement of multiple lymph nodes and medullary and peripheral blood (3.6 x 10(9) monoclonal CD19-positive [CD19+] B-lymphocytes/l). After a 5-day cycle of IL-2 treatment, an eightfold increase of the monoclonal CD19+ population was observed (27 x 10(9) monoclonal CD19+ cells). The lymphocytosis decreased dramatically during the second cycle (days 15 to 19) of IL-2 treatment, resulting in 6 x 10(9)/l peripheral lymphocytes, with 5.5 x 10(9) B-lymphocytes. As soon as day 20, peripheral B-cells again increased considerably, with 32 x 10(9) CD19+ cells/l at day 27. The CD19+ population remained monoclonal as assessed by kappa/lambda cell-surface phenotyping and kappa gene rearrangement evaluation. Kinetics of the monoclonal B-lymphocyte response to IL-2 paralleled the natural killer/lymphokine-activated killer and T-cell response, with a 4-day latency period, suggesting an indirect enhancing effect of IL-2. Before and during IL-2 treatment, peripheral B-lymphocytes never expressed detectable levels of the p55 IL-2 receptor. However, the p75 IL-2 receptor was expressed significantly in the IL-2-responsive monoclonal B-cell population. Tumor necrosis factor alpha, a known (in vitro) B-cell tumor growth factor, reached high serum levels during IL-2 treatment. Response evaluation at day 45 showed stability of the lymph node involvement and the marrow lymphocyte infiltrate. At day 45, peripheral B-cell lymphocytosis was 7.5 x 10(9)/l. To the knowledge of the authors, this is the first report of an in vivo IL-2-induced reversible increase of peripheral monoclonal B-cell lymphocytosis.

[Immunomodulation by in vivo monoclonal antibodies during bone marrow grafts: parameters of biological monitoring]. / Immunomodulation par anticorps monoclonaux in vivo au cours des greffes de moelle osseuse: éléments de la surveillance biologique.

Fifteen patients undergoing allogeneic BMT were given monoclonal antibodies (MAb). One patient received OKT3 and 5 patients an anti-LFA1 MAb to prevent the graft rejection, 7 patients received a CD5-CD8 combination for treatment of acute GvHD and 2 patients received a CD8 MAb to overcome graft rejection. One autografted patient was administered with a CD8 MAb for an hypoplasia associated with a CD8+ lymphocytosis. Biologic monitoring associated the evaluation of serum MAb level, the analysis of target cell depletion and the detection of anti MAb antibodies. Except for OKT3, the MAb dosage had to be adjusted so that the recipients remained in antibody excess as determined by in vitro monitoring. Although the patients treated with CD5-CD8 or CD8 alone exhibited complete coating of reactive cells we did not observe a complete depletion of these populations.