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Realizar la adaptación cultural de la versión española de la Voice Symptom Scale (VoiSS) y su evaluación métrica inicial mediante un estudio piloto.

ABSTRACTHere we describe the development and validation of an intracellular high-throughput screening assay for finding new antituberculosis compounds active in human macrophages. The assay consists of a luciferase-based primary identification assay, followed by a green fluorescent protein-based secondary profiling assay. Standard tuberculosis drugs and 158 previously recognized active antimycobacterial compounds were used to evaluate assay robustness. Data show that the assay developed is a short and valuable tool for the discovery of new antimycobacterial compounds.

ABSTRACTMycobacterium tuberculosis18b, a streptomycin (STR)-dependent mutant that enters a viable but nonreplicating state in the absence of STR, has been developed as a simple model for drug testing against dormant bacilli. Here, we further evaluated the STR-starved 18b (SS18b) model both in vitroand in vivoby comparing the behavior of 22 approved and experimental tuberculosis drugs. Using the resazurin reduction microplate assay (REMA), rifampin (RIF), rifapentine (RPT), TMC207, clofazimine (CFM), and linezolid (LIN) were found to be active against SS18b in vitro, and their bactericidal activity was confirmed by determining the number of CFU. A latent 18b infection was established in mice, and some of the above-mentioned drugs were used for treatment, either alone or in combination with RIF. RIF, RPT, TMC207, CFM, and pyrazinamide (PZA) were all active in vivo, while cell wall inhibitors were not. A comparative kinetic study of rifamycin efficacy was then undertaken, and the results indicated that RPT clears latent 18b infection in mice faster than RIF. Intrigued by the opposing responses of live and dormant 18b cells to cell wall inhibitors, we conducted a systematic analysis of 14 such inhibitors using REMA. This uncovered an SS18b signature (CWPRED) that accurately predicted the activities of cell wall inhibitors and performed well in a blind study. CWPRED will be useful for establishing the mode of action of compounds with unknown targets, while the SS18b system should facilitate the discovery of drugs for treating latent tuberculosis.

The ability of insulin-like growth factor-1 (IGF-1) to regulate surface expression of the interferon-γ receptor 2 (IFN-γR2) transducing chain and activation of IFN-γ–induced signal transducer and activator of transcription-1 (STAT-1) in human T cells was analyzed. We show that, especially in the absence of serum (which contains IGF-1), IGF-1 down-regulated surface expression of the IFN-γR2 chain and inhibited both IFN-γ–dependent STAT-1 activation and apoptosis in T-cell lines ST4, Jurkat, and Molt-4. IFN-γR2 down-regulation resulted from its enhanced internalization since IGF-1 completely restored the uptake of anti–IFN-γR2 monoclonal antibody (mAb) in serum-deprived T-cell lines. When the interaction between IGF-1 and its receptor was blocked by anti–IGF-1R mAb, enhancement of IFN-γR2 surface expression, STAT-1 activation, and reinstatement of IFN-γ–induced apoptosis were observed. Enhanced expression of IFN-γR2 was also observed in phytohemagglutinin (PHA)–activated T lymphoblasts cultured in the presence of anti–IGF-1R mAb, whereas IGF-1 or anti–IGF-1R mAb did not modify the high IFN-γR2 expression in B and myeloid cell lines. Both IGF-1 and anti–IGF-1R mAb did not modify the constitutive expression of IFN-γR2 mRNA in T cells as well as the high IFN-γR1 binding chain surface expression in T, B, and myeloid cells. These data indicate that IGF-1 plays a critical role in the desensitization of IFN-γ/STAT-1 signaling in T lymphocytes by delivering a signal for IFN-γR2 internalization.

Interleukin (IL)‐12, especially in the presence of neutralizing anti‐IL‐4 monoclonal antibodies, primed CD45RO−T clones for high CCL3/macrophage‐inflammatory protein‐1α (MIP‐1α) and CCL4/MIP‐1β levels. In CD4+and CD8+clones from two patients deficient for IL‐12Rβ1 (IL‐12Rβ1−/−), production of CCL3/MIP‐1α and CCL4/MIP‐1β was defective. CD4+clones from two patients deficient for interferon‐γ (IFN‐γ) R1 (IFN‐γR1−/−) produced somewhat decreased CCL4/MIP‐1β levels. IL‐12 failed to prime CD4+or CD8+healthy clones for high CCL5/regulated on activation, normal T expressed and secreted (RANTES) production, although its secretion was impaired in CD4+clones from IL‐12Rβ1−/−and IFN‐γR1−/−patients. CCR5 surface expression was up‐regulated in resting peripheral blood mononuclear cells and CD4+clones from both kinds of patients, rendering them more susceptible to CCR5‐dependent (R5) HIV‐1 infection. Neutralization of IFN‐γ increased CCR5 expression and decreased CC‐chemokine secretion by CD4+clones from healthy and IL‐12Rβ1−/−individuals, suggesting an IFN‐γ‐dependent control of CCR5 expression. These data provide the first documented analysis of chemokine secretion and chemokine receptor expression on T cells from IL‐12 and IFN‐γ receptor‐deficient patients and dissect the role of IL‐12 and IFN‐γ on inducing inflammatory chemokine secretion and down‐regulating CCR5 expression in human T cells.

Phytohemagglutinin (PHA)-derived T lymphoblasts or T cell clones from patients genetically deficient in IL-12Rβ1 (IL-12Rβ1^–/–) or IFN-γR1 (IFN-γR1^–/–) produced two- to threefold reduced IFN-γ levels compared to the corresponding cells from healthy individuals after anti-CD3 and PMA stimulation. Moderate IFN-γ production was observed in PHA-derived T lymphoblasts or T cell clones derived from healthy subjects in the presence of anti-IFN-γR1 or anti-IL-12 mAb, whereas it was negligible in the presence of both mAb. However, when anti-IFN-γR1 and/or anti-IL-12 mAb were added during restimulation, the cells produced normal levels of IFN-γ, indicating that both IFN-γ and IL-12 had an effect on the priming phase. Moderate production of IFN-γ was partially enhanced only in IFN-γR1^–/– T cell clones generated in the presence of IL-12, but was almost completely abolished when IL-12Rβ1^–/– and IFN-γR1^–/– T cell clones were generated in the presence of anti-IFN-γR1 or anti-IL-12 mAb, respectively. IL-4 production was enhanced in T cell clones from IL-12Rβ1^–/–,but not from IFN-γR1^–/– patients, whereas IL-10 and IL-2 production did not differ significantly in polyclonal T cells or clones from healthy and deficient individuals. These results indicate that IL-12Rβ1- and IFN-γR1-dependent signals co-ordinately regulate IFN-γ, but not IL-2 and IL-10 production, whereas only IL-12 negatively controls IL-4 production by in vitro-generated T cell clones. Thus, although IL-12 and IFN-γ signals are each sufficient for moderate production of IFN-γ by human T cells, both are needed for optimal IFN-γ production, and in the absence of both IFN-γ production is completely abrogated.

As T cell response to tumor-associated antigens may be impaired by the acidic microenvironment typical of solid tumors, we assessed the effect of extracellular pH (pHe) on the activation and proliferation of human T lymphocytes and generation of the cytotoxic response. T lymphocytes stimulated with anti-CD3 mAb or PHA at low pHe were unable to secrete IL-2 and IFN-γand their ability to progress through the cell cycle was impaired. T lymphocytes also displayed up-regulation of IFN-γR2 chain and CTLA-4 expression, rendering them sensitive to negative regulatory signals. Agonistic mAb against CD28, but not against CD2, completely restored cytokine production and cell cycle progression, but down-regulated IFN-γR2 and CTLA-4 expression. The anti-CD28mAb rescued the CTL response of allogeneic anti-tumor cultures generated at low pHe. Following anti-CD28 mAb treatment, T cells synthesized cyclooxygenase-2 (Cox-2) protein, which is involved in the early phases of T cell activation. This rescue of T cell activation was independent of the inducible 6-phosphofructo-2-kinase (iPFK-2) pathway, which stimulates proliferation in hypoxic and acidic conditions. The restoration of proliferative and cytotoxic T cell responses by CD28-triggering provides insight into the mechanisms by which B7 enhances the T cell anti-tumor response in vivo.

The sensitivity of Mycobacterium tuberculosis, the pathogen that causes tuberculosis (TB), to antibiotic prodrugs is dependent on the efficacy of the activation process that transforms the prodrugs into their active antibacterial moieties. Various oxidases of M. tuberculosishave the potential to activate the prodrug ethionamide. Here, we used medicinal chemistry coupled with a phenotypic assay to select the N-acylated 4-phenylpiperidine compound series. The lead compound, SMARt751, interacted with the transcriptional regulator VirS of M. tuberculosis, which regulates the mymAoperon encoding a monooxygenase that activates ethionamide. SMARt751 boosted the efficacy of ethionamide in vitro and in mouse models of acute and chronic TB. SMARt751 also restored full efficacy of ethionamide in mice infected with M. tuberculosisstrains carrying mutations in the ethAgene, which cause ethionamide resistance in the clinic. SMARt751 was shown to be safe in tests conducted in vitro and in vivo. A model extrapolating animal pharmacokinetic and pharmacodynamic parameters to humans predicted that as little as 25 mg of SMARt751 daily would allow a fourfold reduction in the dose of ethionamide administered while retaining the same efficacy and reducing side effects.

Renal phosphate excretion was measured in 23 cases of Cushing's syndrome using the phosphate excretion index (PEI) described by Nordin & Fraser(1960). The majority of the patients had an increased PEI associated with a tendency towards hypophosphoraemia. Regression of the syndrome due to intrasellar implantation of 90Y or to removal of an adrenal neoplasm led to prompt correction of both the PEI and the phosphoraemia. In patients with adrenal insufficiency the PEI values fell below the lower limit of the normal.

A study of calcium and phosphorus metabolism has been carried out on 13 acromegalic patients, in various stages of the disease. This study was repeated in nine patients following implantation of the pituitary gland with 90Y and in another two patients after deep X-ray therapy and suction removal of a pituitary adenoma respectively.Increased urinary calcium and phosphorus excretion was found in all the patients in whom the disease was in an active phase of evolution. The calcium tolerance test revealed a marked decrease of calcium retention in certain subjects, while in others, calcium retention was found to be increased. Such changes were not found in patients in whom the disease was in a quiescent phase. The blood calcium, phosphorus and alkaline phosphatase were found to be either normal or slightly increased.The implantation of the pituitary gland with 90Y and deep X-ray therapy induced a marked decrease of hypercalciuria, both spontaneous and induced, and of hyperphosphaturia, together with a definite improvement, of the clinical picture and glucose metabolism. It is concluded that the changes in calcium and phosphorus metabolism described above depend either directly or indirectly on a pituitary factor. They may therefore prove a reliable index for assessing pituitary growth hormone activity.

This first-time-in-human (FTIH) study evaluated the safety, tolerability, pharmacokinetics, and food effect of single and repeat oral doses of GSK3036656, a leucyl-tRNA synthetase inhibitor. In part A, GSK3036656 single doses of 5 mg (fed and fasted), 15 mg, and 25 mg and placebo were administered. In part B, repeat doses of 5 and 15 mg and placebo were administered for 14 days once daily.

In this study the air gamma dose rate map of Piemonte, a region in the North-West of Italy, was produced from gamma spectrometry soil data. Soil samples collected in 110 different sites of Piemonte were analysed with Hyperpure Germanium (HPGe) detectors (30% relative efficiency), which allow the evaluation of the activity concentrations of natural radionuclides and 137Cs. Then, using the available mathematical models, the gamma absorbed dose rate in air due to radionuclides was calculated. The contribution of the cosmic radiation to the total absorbed dose rate, which depend on the site altitude was also evaluated and added to the soil contribution. Finally, the map of the whole region was obtained by fitting the dose rate values of the different sites with kriging algorithms.

Four different techniques for the assessment of environmental gamma dose are discussed and compared: high pressure ionisation chamber (Reuter-Stokes type), TLD dosemeters (GR-200), in situ gamma ray spectrometry and dose evaluation from the activity concentration of radionuclides in the soil. Soil samples gathered from a field near Turin (North-West Italy) have been analysed with the p-type HPGe, and their radionuclide concentrations have been used to evaluate the dose at 1 m above the soil surface. At the same location the dose rate was evaluated with the in situ gamma ray spectrometry (with a portable n-type HPGe) and with the ionisation chamber and the TLD dosemeters. The contribution of cosmic rays was added to the dose rate calculated from the soil samples and the in situ gamma spectrometry. The results obtained with these four techniques agree well within 20%.