Objective: In this study we performed single-cell analysis of the intracellular cytokine expression in peripheral CD4+ and CD8 + lymphocytes from patients with Hashimoto's thyroiditis (HT) to investigate the type-1 response separately for the two lymphocyte sub-populations. Design and methods: Twenty-nine patients affected by HT and 20 healthy subjects, matched for sex and age, were enrolled. After the analysis of the lymphocyte sub-populations, the intracellular content of interferon-g (IFN-g) in phorbolmyristate acetate (PMA)-stimulated CD4 + and CD8 + lymphocytes was assayed. Moreover, the CD4 + lymphocytes were also evaluated for the intracellular expression of IL-4. Results: No significant differences in CD3 + , CD4 + and CD8 + lymphocytes were found between HT patients and control subjects. However, the HT patients showed higher numbers of CD4 + IFN-g + , CD4+ IL-4 + and CD8 + IFN-g + ðt-test; P # 0:001Þ cells than the control subjects. Analysing the intracellular expression of IFN-g and IL-4 in relation to thyroid function, we found that the euthyroid patients (18 cases) showed more expression of IL-4 in CD4 + lymphocytes than the control subjects, without any significant modification of IFN-g expression in CD4+ and CD8 + lymphocytes. However, the hypothyroid patients (11 cases) showed an increase of IFN-g expression in both CD4 + and CD8 + lymphocytes with respect to the control subjects and the euthyroid patients. Moreover, the expression of IL-4 in CD4 + cells from hypothyroid patients was significantly lower than that seen in the euthyroid cases and comparable to that found in the control subjects. Conclusions: Our study has demonstrated that the peripheral CD4 + and CD8 + T lymphocytes from the HT patients show a type-1 activation strictly correlated to the occurrence of hypothyroidism. Further studies will be needed to clarify the exact role of peripheral lymphocytes in HT and whether they could provide a reliable marker of thyroid immune involvement.
The susceptibility of 253 Mycobacterium tuberculosis complex isolates to pyrazinamide (PZA) was assessed using the BACTECTM MGITTM 960 (M960) system. Resistant strains underwent paired repeat testing using 1) a critical concentration of 200 g/ml (PZA-200), and 2) a reduced inoculum of 0.25 ml. They were also examined using the BACTEC 460 (B460) reference method and investigated for pncA mutations. On M960, 37 isolates were resistant. In the PZA-200 assay, 20 of these were resistant and 17 susceptible, while 18 were resistant and 19 susceptible with reduced inoculum. The B460 assay and pncA sequencing confirmed results with reduced inoculum.
Background: Extrapulmonary tuberculosis (EPTB) remains difficult to diagnose because the clinical specimens to be examined are often paucibacillary and obtained with difficulty from inaccessible sites. An updated Xpert ® MTB/RIF Ultra (Ultra) test has been designed and licensed to improve sensitivity in the detection of Mycobacterium tuberculosis complex. The aim of the present study is to evaluate the performance of Ultra assay for the clinical diagnosis of EPTB in a low tuberculosis prevalence country. Methods: A retrospective analysis was performed at "A. O dei Colli" of Naples on consecutive extrapulmonary specimens for EPTB across a three-year period. All different types of extrapulmonary specimens were tested for EPTB by smear microscopy, culture and Ultra assay in accordance with relevant guidelines. Results: A total of 606 EPTB samples, 561 culture negative EPTB and 45 culture positive EPTB were included. Using culture as reference standard, the overall sensitivities and specificities of Ultra assay were 95.6% (95% CI 84.8 -99.5) and 97.5% (95% CI 95.8 -98.6) respectively. Sensitivity and specificity of Ultra for individual category of specimens were also performed. Conclusion: In a low-tuberculosis prevalence setting, Ultra assay confirms to have a good performance in the diagnosis of EPTB for all different extrapulmonary samples.
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