Sputum basophils are increased in eosinophilic asthma compared with non-eosinophilic asthma phenotypes.

Sputum basophil numbers are increased in allergic asthmatics, but it is unclear what role airway basophils play in "TH2-low" asthma phenotypes. Using flow cytometry, we found that basophils were significantly increased in all asthmatics (n=26) compared with healthy controls (n=8) (P=0.007) with highest levels observed in eosinophilic asthma (EA); median 0.22%, IQR 0.11%-0.47%; n=14) compared with non-EA (NEA) (0.06%, 0.00%-0.20%; n=12; P<0.05). In asthmatics, basophils were positively correlated with sputum eosinophils (r=0.54; P<0.005) and inversely with sputum neutrophils (r=-0.46: P<0.05), but not with FEV1 (% predicted), FEV1 /FVC or bronchodilator reversibility. In a subgroup initially identified as inadequately controlled asthma (n=7), there was a trend (P=0.08) towards a reduction in sputum basophils following increased inhaled corticosteroid (ICS) treatment. Our findings suggest that basophils may be particularly important in eosinophilic asthma and that sputum basophil assessment could be a useful additional indicator of "TH2-high" asthma.

Cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) can regulate dendritic cell-induced activation and cytotoxicity of CD8(+) T cells independently of CD4(+) T cell help.

The mechanisms that regulate the strength and duration of CD8(+) cytotoxic T cell activity determine the effectiveness of an antitumor immune response. To better understand the antitumor effects of anti-cytotoxic T lymphocyte-associated antigen 4 (CTLA-4) antibody treatment, we analyzed the effect of CTLA-4 signaling on CD8(+) T cells in vitro and in vivo. In vitro, cross-linking of CTLA-4 on purified CD8(+) T cells caused decreased proliferative responses to anti-CD3 stimulation and rapid loss of activation marker expression. In vivo, blockade of CTLA-4 by neutralizing anti-CTLA-4 mAb greatly enhanced the accumulation, activation, and cytotoxic activity of CD8(+) T cells induced by immunization with Ag on dendritic cells (DC). This enhanced response did not require the expression of MHC class II molecules on DC or the presence of CD4(+) T cells. These results demonstrate that CTLA-4 blockade is able to directly enhance the proliferation and activation of specific CD8(+) T cells, indicating its potential for tumor immunotherapy even in situations in which CD4(+) T cell help is limited or absent.

Glycolipid-peptide conjugate vaccines enhance CD8+ T cell responses against human viral proteins.

An important goal of vaccination against viruses and virus-driven cancers is to elicit cytotoxic CD8+ T cells specific for virus-derived peptides. CD8+ T cell responses can be enhanced by engaging help from natural killer T (NKT) cells. We have produced synthetic vaccines that induce strong peptide-specific CD8+ T cell responses in vivo by incorporating an NKT cell-activating glycolipid. Here we examine the effect of a glycolipid-peptide conjugate vaccine incorporating an NKT cell-activating glycolipid linked to an MHC class I-restricted peptide from a viral antigen in human peripheral blood mononuclear cells. The vaccine induces CD1d-dependent activation of human NKT cells following enzymatic cleavage, activates human dendritic cells in an NKT-cell dependent manner, and generates a pool of activated antigen-specific CD8+ T cells with cytotoxic potential. Compared to unconjugated peptide, the vaccine upregulates expression of genes encoding interferon-γ, CD137 and granzyme B. A similar vaccine incorporating a peptide from the clinically-relevant human papilloma virus (HPV) 16 E7 oncoprotein induces cytotoxicity against peptide-expressing targets in vivo, and elicits a better antitumor response in a model of E7-expressing lung cancer than its unconjugated components. Glycolipid-peptide conjugate vaccines may prove useful for the prevention or treatment of viral infections and tumors that express viral antigens.

Antigen expressed on tumor cells fails to elicit an immune response, even in the presence of increased numbers of tumor-specific cytotoxic T lymphocyte precursors.

We have used T-cell receptor (TCR) transgenic mice to analyze the interaction of tumors with the immune system. We show that the tumor cell line Lewis lung-lymphocytic choriomeningitis virus (LL-LCMV), genetically manipulated to express an H-2 Db-restricted epitope of the lymphocytic choriomeningitis virus glycoprotein (LCMV33-41), can grow progressively in TCR transgenic mice, where approximately 50% of CD8+ T cells are specific for LCMV33-41. TCR transgenic T cells were not deleted in tumor-bearing mice, and their surface phenotype and cytokine secretion patterns remained typical of naive T cells. Also, TCR transgenic T cells from tumor-bearing mice had undiminished capacity to proliferate to antigen in vitro. Tumor-protective immune responses could be elicited in TCR transgenic mice by immunization with LCMV33-41 peptide-loaded dendritic cells. Tumor resistance correlated with the switch of TCR transgenic T cells from a CD44low to a CD44high phenotype and increased capacity to produce IFNgamma in vitro. Results similar to those obtained in TCR transgenic mice were also obtained using an adoptive transfer system, where small numbers of TCR transgenic T cells were injected into normal C57BL/6 hosts. These data indicate that even large tumors may not induce specific immunization, tolerance, or anergy to tumor antigens, and that high numbers of tumor-specific CTL precursors are not sufficient to provide tumor resistance.

Immunotherapy with dendritic cells and tumor major histocompatibility complex class I-derived peptides requires a high density of antigen on tumor cells.

Immunization with dendritic cells and unfractionated MHC class I-binding peptides derived from autologous tumor cells has been shown to induce effective antitumor immunity. However, the importance of the relative abundance of tumor peptides on the surface of tumor cells is not known. We have addressed this question using peptides isolated from three tumor cell lines transfected with a minigene encoding amino acids 33-41 of the lymphocytic choriomeningitis virus glycoprotein (LCMV(33-41)). The three cell lines expressed different levels of MHC class I molecules and had different abilities to stimulate proliferation of LCMV(33-41)-specific T cells in vitro. We found that antitumor immune responses were best elicited by immunizing mice with dendritic cells and synthetic LCMV(33-41) peptide. Peptide preparations from a given tumor cell line conferred protection against challenge with the same tumor cell line. However, protective immunity to a different tumor could be induced only if the cell line used for peptide preparation presented a high relative proportion of LCMV(33-41) in association with MHC class I. Our results suggest that multiple peptide epitopes are required for the induction of an effective antitumor immune response using MHC class I-binding peptides from tumor cells. Also, the ability to induce an antitumor immune response appears to correlate with the proportion, rather than the absolute amount, of tumor-specific peptide in the mixture used for immunization.

Selective oxidative dehydrogenation of propane to propene using boron nitride catalysts.

The exothermic oxidative dehydrogenation of propane reaction to generate propene has the potential to be a game-changing technology in the chemical industry. However, even after decades of research, selectivity to propene remains too low to be commercially attractive because of overoxidation of propene to thermodynamically favored CO2 Here, we report that hexagonal boron nitride and boron nitride nanotubes exhibit unique and hitherto unanticipated catalytic properties, resulting in great selectivity to olefins. As an example, at 14% propane conversion, we obtain selectivity of 79% propene and 12% ethene, another desired alkene. Based on catalytic experiments, spectroscopic insights, and ab initio modeling, we put forward a mechanistic hypothesis in which oxygen-terminated armchair boron nitride edges are proposed to be the catalytic active sites.

Dendritic cell elimination as an assay of cytotoxic T lymphocyte activity in vivo.

We show in this paper that the survival of antigen-loaded dendritic cells in vivo may be used as a sensitive readout of CTL activity. We have previously shown that dendritic cells labeled with the fluorescent dye CFSE and injected sub-cutaneously into mice migrate spontaneously to the draining lymph node where they persist for several days. In the presence of effector CTL responses, dendritic cells loaded with specific antigen rapidly disappear from the draining lymph node. In this paper we extend the above observations and set up a simple and sensitive method to reveal CTL activity in individual mice in vivo. Dendritic cells were labeled with two different fluorochromes, loaded with antigen or left untreated, and mixed together before injection into mice. We show that only the dendritic cells loaded with specific antigen were cleared from the draining lymph node, while dendritic cells not loaded with antigen remained unaffected. Cytotoxic responses generated by immunization with peptide-loaded dendritic cells, or by infection with influenza virus, could be revealed using this method. Comparison of the differential survival of dendritic cells populations mixed together also allowed us to accurately evaluate the disappearance of dendritic cells, irrespective of variability in the injection site and other parameters. Given the ability of dendritic cells to efficiently take up and present complex antigens, nucleic acids and apoptotic bodies, this method may also allow the evaluation of cytotoxic activity against antigens that are not characterized in terms of peptide epitopes.

Amineptine, response timing, and time discrimination in the albino rat.

Experiment 1 recorded the effects of single (doses of 1, 5, 10, and 20 mg/kg) and repeated intraperitoneal injections (10 mg/kg) of amineptine (a tricyclic antidepressant drug) on the performance of albino rats in differential reinforcement of low rate (DRL) of 30 s, fixed-interval (FI) of 60 s, and signalled continuous reinforcement (CRF-SD) schedules. In the second experiment, the effects of repeated (10 mg/kg) and single injections (20 mg/kg) were assessed on the discrimination of the duration of auditory stimuli (2 and 8 s). A dose-related increase in response rates was observed in FI and DRL, correlating with a dose-related impairment in the temporal regulation of performance. However, the drug remained without effect on duration discrimination. In other respects, decreases in response latency in CRF-SD or duration discrimination tended to indicate that the drug improved vigilance and reactivity to extraneous significant stimuli. Interpretations in terms of sensitization, tolerance, or dependency could be discarded. Our data support the hypothesis that drug effects on temporal regulation in FI and DRL are secondary to a nonspecific activation of motor activity. They question the plausibility of an antidepressant effect of the drug in humans via modulation of a timing mechanism.

Educating patient educators: enhancing instructional effectiveness in physical therapy for low back pain patients.

The objective of this research project was to study the effectiveness of a training program for the enhancement of patient education skills in physical therapy. In this paper the improvement of five of these skills is tested. These skills are aimed at a better monitoring of adherence problems during the treatment and at enhancing self-efficacy of the patient after treatment. In order to test the effectiveness of the program, complete treatments of 19 physiotherapists have been assessed before (1142 sessions, 130 patients) and after (775 sessions, 88 patients) the training program. Information on the instructions and solutions given to the patients was obtained with a registration form, completed after each session by the physiotherapist. The patient's perception of the effectiveness and feasibility of instructions was obtained from questionnaires, completed by the patient on three occasions. After the training only a minority of the trained skills appeared to be improved. All in all, the training program was not very effective. More effort is needed to develop training programs aimed at promoting patients' self-efficacy as well as measurement instruments to assess the effects of such programs.

Back care instructions in physical therapy: a trend analysis of individualized back care programs.

BACKGROUND AND PURPOSE: The treatment of people with low back pain often includes giving a variety of instructions about back care. The objective of our study was to explore the content and sequence of these instructions. SUBJECTS: Our database contained information on 1,151 therapy sessions for 132 patients who were treated by 21 therapists. METHODS: Hierarchical linear modeling was used to establish trends in instructions during the course of treatment. Instructions were measured by means of a registration form. RESULTS: Pain management instructions were given at the start of treatment and then decreased in later sessions. Instructions about taking care of the back in daily activities followed the same course. Exercise instructions were introduced after the start of treatment and were spread evenly across the visits. The number of recommendations about general fitness decreased during treatment. CONCLUSION AND DISCUSSION: The majority of back care instructions were spread evenly across therapy visits. Relatively little variation in instructions among patients was seen, which may indicate a lack of individualization of the back care programs.

The emerging role of the dendritic cell in novel cancer therapies.

Research over the last two years has explored a number of possible approaches to applying dendritic cell immunotherapy to the treatment of human cancers. The chosen strategy in clinical situations will vary for individual patients and will depend on the type of tumour, availability of tissue samples and potential source of dendritic cells. We believe that the isolation of fractionated tumour peptide from individual patients' tumours for use with autologous stem cell-derived dendritic cells may provide, in at least some cases, an effective and practical approach to cancer immunotherapy. This approach will provide a 'closed' system of tumour immunotherapy with all components (dendritic cells, antigen and cytotoxic T lymphocytes) being provided by the patient and applied in a tailor-made fashion to individual patients as an adjuvant to current anti-tumour therapies.

Refractory ocular hypertension secondary to intravitreal injection of triamcinolone acetonide.

PURPOSE: to report a case of severe ocular hypertension occurring as a complication after a single intravitreal injection of triamcinolone acetonide for the treatment of a diabetic cystoid macular edema. METHODS: interventional case report. RESULTS: a 63-year-old pseudophakic diabetic woman developed a severe and relatively sudden IOP increase to 50 mm Hg one month after receiving a single 4-mg intravitreal injection of triamcinolone acetonide for a chronic progressive macular cystoid edema. Previously the patient who did not develop corticosteroid-induced glaucoma secondary to her cataract surgery was treated with topical beta-blockers for a mild chronic bilateral ocular hypertension. A deep sclerectomy had to be performed in emergency to avoid optic nerve damage and allowed to successfully control the IOP with a 5 month follow-up. Concomitantly visual acuity could be increased from 0.05 before the intravitreal injection to 0.4. CONCLUSIONS: Although unfrequent in the literature, this observation confirms the risk of occurrence of a severe ocular hypertension after intravitreal injection of triamcinolone. A close monitoring of IOP is mandatory after intravitreal injection, especially in patients with altered trabecular function. This potentially devastating complication has to be weighed up with the benefices of intravitreal injection of triamcinolone for improving visual acuity in patients with clinically significant diabetic macular edema.

[Structural adjustment, cultural adjustment?]. / Ajustement structurel, ajustement culturel?

Over the last two decades, multiple studies have been conducted and many articles published about Structural Adjustment Programmes (SAPs). These studies mainly describe the characteristics of SAPs and analyse their economic consequences as well as their effects upon a variety of sectors: health, education, agriculture and environment. However, very few focus on the sociological and cultural effects of SAPs. Following a summary of SAP's content and characteristics, the paper briefly discusses the historical course of SAPs and the different critiques which have been made. The cultural consequences of SAPs are introduced and are described on four different levels: political, community, familial, and individual. These levels are analysed through examples from the literature and individual testimonies from people in the Southern Hemisphere. The paper concludes that SAPs, alongside economic globalisation processes, are responsible for an acute breakdown of social and cultural structures in societies in the South. It should be a priority, not only to better understand the situation and its determining factors, but also to intervene and act with strategies that support and reinvest in the social and cultural sectors, which is vital in order to allow for individuals and communities in the South to strengthen their autonomy and identify.

Beta-D-glucosidase in fractions from rat brain.

Beta-Glucosidase activity has been determined in homogenate and in centrifugation fractions of 7-day-old and adult rat brain; maximum activity was found at pH 4 and pH 5. Of the adult brain, more than 50% of the activity was concentrated in the 800-g sediment fraction (P1), while in the brain of 7-day-old rat about 20% was found in the corresponding fraction. The activity maximum in all fractions after a 2% Triton X-100 treatment occurs at pH 5. Addition of Triton to adult brain homogenate enhances the activity, but this stimulation is less than the sum of the activities observed at pH 4 and pH 5 in the absence of Triton. Triton addition to brain homogenate of 7-day-old rat results in a fall in activity at pH 4 and in a maximum at pH 5. In rat brain homogenate subjected to sonication, a loss of activity is observed at pH 4, scarcely at pH 5; the activity loss is completely abolished and turned into an increase under the influence of Triton. This increase equals the level obtained when Triton is added to an untreated brain homogenate. Sonication of rat brain homogenate leads to changes in the distribution pattern; about 25% of the activity of the adult brain is found in the P1 fraction compared to 50% in the corresponding fraction of the untreated brain. Fractionation of a sonicated brain homogenate from adult rat reveals that at pH 4 most activity (52%) is concentrated in the 20,000-g pellet (P2), 23% in supernatant fluid (S2); at pH 5 the opposite is observed; most activity (49%) is found in the 20,000-g supernatant (S2) and 23% in the 20,000-g pellet (P2). In the presence of Triton the activity of the sonicated brain homogenate of adult rat increases; this stimulation roughly equals the sum of the corresponding activities measured at pH 4 and pH 5 in the absence of Triton.

Analytical performance of three commercially available nephelometers compared for quantifying proteins in serum and cerebrospinal fluid.

We investigated the analytical performance and imprecision of three commercially available nephelometers for the quantification of various proteins in pooled serum and cerebrospinal fluid. Albumin, transferrin, IgG, IgA, and IgM in serum were determined nephelometrically (BNA, Behring; Array, Beckman) and turbidimetrically (Cobas Bio, Hoffmann-La Roche). All these proteins in cerebrospinal fluid except transferrin were determined nephelometrically with all three systems. CVs and lower limits of detection were compared for all instruments, and stability of calibration curves was evaluated.

CD8+ T cell-dependent elimination of dendritic cells in vivo limits the induction of antitumor immunity.

The fate of dendritic cells (DC) after they have initiated a T cell immune response is still undefined. We have monitored the migration of DC labeled with a fluorescent tracer and injected s.c. into naive mice or into mice with an ongoing immune response. DC not loaded with Ag were detected in the draining lymph node in excess of 7 days after injection with maximum numbers detectable approximately 40 h after transfer. In contrast, DC that had been loaded with an MHC class I-binding peptide disappeared from the lymph node with kinetics that parallel the known kinetics of activation of CD8+ T cells to effector function. In the presence of high numbers of specific CTL precursors, as in TCR transgenic mice, DC numbers were significantly decreased by 72 h after injection. The rate of DC disappearance was extremely rapid and efficient in recently immunized mice and was slower in "memory" mice in which memory CD8+ cells needed to reacquire effector function before mediating DC elimination. We also show that CTL-mediated clearance of Ag-loaded DC has a notable effect on immune responses in vivo. Ag-specific CD8+ T cells failed to divide in response to Ag presented on a DC if the DC were targets of a pre-existing CTL response. The induction of antitumor immunity by tumor Ag-loaded DC was also impaired. Therefore, CTL-mediated clearance of Ag-loaded DC may serve as a negative feedback mechanism to limit the activity of DC within the lymph node.

Three cases of disputed paternity in dogs resolved by the use of DNA fingerprinting.

DNA fingerprinting with the probes 33.15 and alpha-globin 3'HVR has been used to resolve three cases of disputed paternity in dogs. For each pedigree it was necessary to establish which bands in the DNA fingerprints of the offspring were of paternal origin, and then establish which putative sire carried all these bands. In the first case, a litter of Rhodesian Ridgebacks, twelve DNA bands were informative in establishing paternity. In the second case, a litter of Afghan hounds, five DNA bands established paternity, Lastly, in a litter of Border collies, five DNA bands established paternity. In each case a single dog only sired the entire litter.

Tumor-peptide-pulsed dendritic cells isolated from spleen or cultured in vitro from bone marrow precursors can provide protection against tumor challenge.

Dendritic cells (DC) purified from murine spleen or generated in vitro from bone marrow precursors were compared for their respective abilities to stimulate T cell responses and provide tumor protection in vivo. In vitro incubation with synthetic tumor peptide conferred on both DC populations the ability to induce proliferation of tumor-peptide-specific T cells in vitro. Spleen DC were reproducibly about twofold more effective than bone-marrow-derived DC in this assay. Both DC populations could also induce cytotoxic activity in vivo. In vitro cytoxicity assays showed that, while cytotoxic activity induced by immunization with spleen DC was clearly peptide-specific, a high non-specific cytotoxic activity was consistently observed after immunization with bone-marrow-derived DC, whether peptide-pulsed or not. Regardless of such high non-specific activity in vitro, only tumor-peptide-pulsed DC could provide protection against subsequent inoculation of tumor cells. DC not pulsed with tumor peptide were ineffective. We conclude that DC isolated from spleen or generated in vitro from bone marrow precursors are suitable reagents for use in tumor vaccination studies.