Prediction of recurrence-free survival using a protein expression-based risk classifier for head and neck cancer.

Loco-regional recurrence in 50% of oral squamous cell carcinoma (OSCC) patients poses major challenge for oncologists. Lack of biomarkers that can predict disease aggressiveness and recurrence risk makes the scenario more dismal. On the basis of our earlier global proteomic analyses we identified five differentially expressed proteins in OSCC. This study aimed to develop protein biomarkers-based prognostic risk prediction model for OSCC. Sub-cellular expression of five proteins, S100A7, heterogeneous nuclear ribonucleoproteinK (hnRNPK), prothymosin α (PTMA), 14-3-3ζ and 14-3-3σ was analyzed by immunohistochemistry in test set (282 Indian OSCCs and 209 normal tissues), correlated with clinic-pathological parameters and clinical outcome over 12 years to develop a risk model for prediction of recurrence-free survival. This risk classifier was externally validated in 135 Canadian OSCC and 96 normal tissues. Biomarker signature score based on PTMA, S100A7 and hnRNPK was associated with recurrence free survival of OSCC patients (hazard ratio=1.11; 95% confidence interval 1.08, 1.13, P<0.001, optimism-corrected c-statistic=0.69) independent of clinical parameters. Biomarker signature score stratified OSCC patients into high- and low-risk groups with significant difference for disease recurrence. The high-risk group had median survival 14 months, and 3-year survival rate of 30%, whereas low-risk group survival probability did not reach 50%, and had 3-year survival rate of 71%. As a powerful predictor of 3-year recurrence-free survival in OSCC patients, the newly developed biomarkers panel risk classifier will facilitate patient counseling for personalized treatment.

Erratum.

[This corrects the article on p. 166 in vol. 4, PMID: 25215193.].

Assessment of the genotype MTBDRplus assay for rifampin and isoniazid resistance detection on sputum samples in Cote d'Ivoire.

We conducted an evaluation study on the GenoType MTBDRplus assay's ability to detect mutations conferring resistance to rifampin and isoniazid directly from sputum taken from 120 smear positive pulmonary patients from tuberculosis (TB) centers in Cote d'Ivoire. The sputum was decontaminated by N-acetyl-l-cysteine (NALC) and comparatively analyzed with the MTBDRplus assay version 2.0 and the mycobacterial growth indicator tube (MGIT) 960 automated drug susceptibility testing (MGIT-DST). The Gene-Xpert Mycobacterium tuberculosis (MTB)/rifampicin (RIF) assay was performed for 21 sputa with absence of hybridization for at least one rpoB wild-type probes. Four and seven, respectively, discordant and concordant results were also analyzed. The mutations in the rpoB gene were 21 (17.5%), 20 (16.7%), 7 (5.8%), and 10 (8.3%), respectively, for D516V, H526Y, H526D, and S531L. S315T mutation in katG gene associated or not with mutation in promoter of inhA was detected in 76 (63.3%) of the sputum. Compared to MGIT-DST, the sensitivity and specificity of the MTBDRplus for rifampin resistance detection were 100% (75-100%) and 73.2% (61.3-84%), respectively. For isoniazid resistance detection, the sensitivity and specificity were, respectively, 95% (90-|99) and 95.1% (88.5-100%). Interpretation of 16 sputa without hybridization of rpoB wild-type probe 8 compared to those obtained with MGIT-DST and GeneXpert MTB/RIF was discordant and concordant, respectively, for 11 and 5.

Use of rapid molecular test for multidrug-resistant tuberculosis detection among relapse cases in Cote d'Ivoire.

UNLABELLED: Tuberculosis is explicitly recognized as a major global public health problem. In Côte d'Ivoire, relapse cases represent 66.5% of patients eligible for retreatment according to the National Tuberculosis Control Program. This study objective was to detect multidrug-resistance tuberculosis among relapse cases. Patients were recruited in tuberculosis centers in routine. A standardized questioning was administrated. Two sputum samples were collected and transported at Institut Pasteur. Sputum samples were decontaminated by NALC method. The DNA extraction was realized with 500µl of decontaminated sputum sample with smear-positive. MTBDRplus assay version 2.0 was performed according to the manufacturer's instruction. An internal quality control program with positive and negative controls was implemented for interpretation of results. In total 146 relapse cases with smear positive were studied. Out of selected patients, 130 had received the 2RHZE/4RH regimen and 16, the 2RHZES/1RHZE/5HRE. In group of relapse cases previously treated with 2RHZE/4RH regimen, 40 (31.3%, IC95%: [0.23; 0.39]) had punctual mutations at codon 526 in rpoB gene. Although, in patients under treated with 2RHZES/1RHZE/5HRE, a mutation in rpoB gene was identified in 12 of 16 sputum samples. Thirteen mutations conferring a resistance to Isoniazid were observed of which 9 in katG gene and 4 in katG and promoter region of inhA gene. The comparison (Chi-square with Yates correction) of resistance rates to Rifampin estimated showed a statistically significant difference. CONCLUSION: Use of a rapid method to detect drug-resistance in recurrent TB cases has permitted to identify patients eligible for first-line drugs or not.

Manganese amendment and Pleurotus ostreatus treatment to convert tomato pomace for inclusion in poultry feed.

If freshly dried tomato pomace could be fed to poultry, its naturally occurring alpha-tocopherol would retard lipid oxidation during further processing, long-term frozen storage, and heating of poultry meat; however, the high fiber content in this agricultural by-product adversely affects its use. Experiments were conducted to investigate the chemical composition and in vitro true digestibility of amended (without and with 487 micromol of manganese/g) tomato pomace substrate after treatment with the white-rot fungus Pleurotus ostreatus. In treated pomace without manganese, protein content was improved by 3.1%, cellulose and hemicellulose decreased over time, but lignin degradation was not detected. In addition, treated pomace without manganese showed a significant reduction of in vitro true digestibility. Manganese in pomace inhibited fungal growth and did not enhance lignin degradation. Under the conditions of the experiment, P. ostreatus improved the nutritional composition of tomato pomace; however, it did not reduce lignin. It is possible that manganese amendment at the level used affected gaseous conditions (O(2) consumption and CO(2) evolution rates), important factors that must be considered when attempting to enhance accelerated lignin degradation by P. ostreatus.

Inhibition of cytokine production and cytokine-stimulated T-cell activation by FK506 (tacrolimus)1.

Insofar as it exerted its immunosuppressive effect by inhibiting cytokine expression, we assessed the effect of FK506 (Tacrolimus) on cytokine-stimulated T-cell activation. Human T cells, treated with FK506, or controls were stimulated with the mitogens PHA + PMA, Con A, and the "CD3-bypass" stimulation regimen, PMA + ionomycin. T-cell proliferation was quantitated by measuring the uptake of tritiated thymidine, and mRNA expression was assessed by RT-PCR. FK506, in a concentration-dependent fashion, inhibited T-cell proliferation and steady-state mRNA expression of IL-2 and IL-7; half-maximal suppression was obtained at 10(-7) to 5 x 10(-8) M. We tested whether FK506 antiproliferative effect could be overcome with exogenously reconstituted rIL-2 and/or rIL-7. Neither rIL-2 nor rlL-7, individually in conjunction with suboptimal concentrations of PHA or Con A, or in combination without any costimulus, was capable of abrogating FK506 antiproliferative effect, indicating that FK506 also acted by inhibiting cytokine-stimulated T-cell activation. To confirm this, T cells were treated with FK506 and stimulated by rIL-2 and rIL-7, individually in conjunction with suboptimal concentration of PHA and Con A. In addition, T cells were stimulated with rIL-2 and rIL-7 without any costimuli. FK506 inhibited T-cell activation stimulated by rIL-2 and by rIL-7, individually and in combination. This confirms that, in exerting its antiproliferative effect, FK506 acts at two levels, by inhibiting cytokine availability and by suppressing cytokine effect on target cells, and explains the beneficial effect of FK506 in attenuating ongoing immune responses.

Vaginal calculi in a young woman.

A case of a large vaginal stone in a 26-year-old woman is presented. Vaginal stones are relatively rare. Various causes can lead to vaginal stone formation. In our case, the calculi was due to vaginal outlet obstruction in childhood. The diagnosis is usually easy to be done. In our patient, X-ray films using frontal and lateral views of the urinary system permitted to establish the right diagnosis. Treatment can vary, depending on the cause of the stone formation and the consistency of the calculi. In this case, the treatment was surgical with simple evolution. It permitted a normal social and sexual life to this young patient.

Opposing effects of rapamycin and cyclosporin A on activation-induced Ca(2+) release.

Insofar as Ca(2+) plays a major role in T cell activation, we investigated the effect of the immunosuppressants cyclosporin A and rapamycin on T cell proliferation and on the activation-induced increase in [Ca(2+)](i). Both cyclosporin A and rapamycin inhibited mitogen (concanavalin A and phytohemagglutinin) and ionomycin+phorbol myristate acetate (PMA)-driven T cell proliferation (Ca(2+)-dependent). However, only rapamycin suppressed T cell proliferation stimulated by anti-CD28 antibody (Ab)+PMA, and recombinant interleukin-6-stimulated proliferation of the interleukin-6 dependent B9 cells (Ca(2+)-independent). These differences were associated with a different effect of both drugs on Ca(2+) release, as cyclosporin A attenuated while rapamycin augmented the mitogen-induced elevation in [Ca(2+)](i). Collectively, this supports the notion that Ca(2+) is required in early stages of T cell activation, and that cyclosporin A blocked only Ca(2+)-dependent while rapamycin blocked both Ca(2+)-dependent and -independent events of T cell activation.

Pretreatment with glucocorticoids enhances T-cell effector function: possible implication for immune rebound accompanying glucocorticoid withdrawal.

Glucocorticoids (GCs) exert their immunosuppressive/antiproliferative effects largely through inhibition of cytokine expression, and paradoxically upregulate the expression of (proinflammatory) cytokine receptors on select nonlymphoid cells. Clinically, withdrawal of GCs was frequently associated with worsening of the outcome of heightened immunity disorders, thereby implicating enhanced cytokine and cytokine receptor expression as a possible consequence of acute/short-term GCs withdrawal. In view of the significance of this complication of GC therapy, we addressed the effect of GC withdrawal on cytokine receptor expression and subsequent T-cell effector function, using the proliferation of human T cells as biological readout. To mimic GC withdrawal, T cells were treated with GCs or controls, stimulated, and incubated for 16-20 h at 37 degrees C, washed, and reactivated for a further 4-48 h. Surface marker expression was assessed by FACS analysis, and proliferation was determined by measuring the cellular uptake of tritiated thymidine. Dexamethasone (DEX) and prednisolone (PRED), in a concentration-dependent manner, inhibited T-cell proliferation induced by anti-CD28 Ab + PMA. However, pretreatment of T cells activated with mitogens, cross-linking antibodies, or PMA + ionomycin ("CD3-bypass" stimulation regimen), but not resting T cells, with DEX or PRED resulted in a marked increase in IL-IR, IL-2R alpha, and IL-6R expression, which was accompanied by a significant enhancement in T-cell proliferation. This effect of GCs was neither stimulus specific nor did it result from alteration in cell viability, and was paralleled by augmentation in cytokine (rIL-2) effects on DEX-pretreated and preactivated T cells. Taken together, our results underline the dual effects of GCs in regulating T-cell activation and cytokine expression. In essence, GCs directly inhibited T-cell proliferation by suppressing cytokine production, and, by enhancing cytokine receptor expression, pretreatment with GCs augmented T-cell proliferation.