Federica Cugnata

First independent evaluation of QuantiFERON-TB Plus performance

Tuberculosis elimination requires an effective strategy to diagnose and treat people infected with Mycobacterium tuberculosis who would otherwise be at high risk of developing and transmitting active disease [1, 2]. The diagnostic tools for latent tuberculosis infection (LTBI) are the tuberculin skin test (TST) and the T-cell interferon-γ release assays (IGRAs). Two IGRAs are commercially available, QuantiFERON-TB Gold In-Tube (QFT-GIT) (Qiagen, Hilden, Germany) and T-SPOT.TB (Oxford Immunotec, Abingdon, UK). Compared to the TST, IGRAs offer operational advantages and higher specificity in the bacille Calmette–Guérin (BCG)-vaccinated population [3], and they are at least as sensitive for LTBI [4]. However, IGRAs have limitations: reduced sensitivity in children and immunocompromised subjects, including HIV-infected individuals [3, 4]; failure to discriminate between active tuberculosis and LTBI; and poor correlation with the risk of progression to active disease [3]. QuantiFERON-TB Plus improves sensitivity for active TB and maintains high specificity among unvaccinated controls http://ow.ly/XjYPK

First evaluation of QuantiFERON-TB Gold Plus performance in contact screening

Identifying latently infected individuals is crucial for the elimination of tuberculosis (TB). We evaluated for the first time the performance of a new type of interferon-γ release assay, QuantiFERON-TB Plus (QFT-Plus), which includes an additional antigen tube (TB2), stimulating both CD4+ and CD8+ T-cells in contacts of TB patients. Contacts were screened for latent TB infection by tuberculin skin test, QFT-Plus and QuantiFERON-TB Gold in Tube (QFT-GIT). In 119 TB contacts, the overall agreement between QFT-Plus and QFT-GIT was high, with a Cohens κ of 0.8. Discordant results were found in 12 subjects with negative QFT-GIT and positive QFT-Plus results. In analyses of markers of TB exposure and test results, the average time spent with the index case was the strongest risk factor for positivity in each of these tests. The difference in interferon-γ production between the two antigen tubes (TB2−TB1) was used as an estimate of CD8+ stimulation provided by the TB2. TB2−TB1 values >0.6 IU·mL−1 were significantly associated with proximity to the index case and European origin. QFT-Plus has a stronger association with surrogate measures of TB exposure than QFT-GIT in adults screened for latent TB infection. Interferon-γ response in the new antigen tube used an indirect estimate of specific CD8+ response correlates with increased Mycobacterium tuberculosis exposure, suggesting a possible role in identifying individuals with recent infection. QuantiFERON-TB Plus improved the diagnostic accuracy for latent TB infection in the setting of contact screening http://ow.ly/2Az0300SDg3

Validation study of the Italian version of the Insomnia Severity Index (ISI).

To test the factorial structure of the Italian version of the Insomnia Severity Index (ISI) using a confirmatory approach and to assess its psychometric properties. ISI questionnaire was completed by 272 patients (average age 41.28, range 18–73) with insomnia diagnosis performed by a sleep medicine physician and retrospectively enrolled in the study. All patients underwent Cognitive Behavioral Treatment for Insomnia (CBT-I) and completed sleep diaries before starting the treatment. Data from sleep diaries were analyzed for assessing concurrent validity of the ISI. Confirmatory factor analysis (CFA) for ordinal Likert-type items was applied to compare four competing models proposed in the literature. 244 patients, out of the 272, completed the ISI at the end of CBT-I. A comparison of ISI score before and after treatment was performed. The CFA analysis confirmed the presence of three main factors conceptualized as severity and impact of the disease along with sleep satisfaction. Significant correlations of the first three items of the questionnaire, investigating three different subtypes of insomnia, and the subjective measures from the sleep diaries were found, thus supporting the concurrent validity of the test. Sleep efficiency (SE) had a significant inverse correlation with the severity and satisfaction factors and with ISI’s total score. After CBT-I treatment, a significant reduction of ISI’s scores was observed, thus confirming the effectiveness of the CBT-I treatment. The internal reliability coefficient was 0.75. The ISI questionnaire maintains good psychometric properties in the Italian version, thus confirming that this instrument is reliable for detecting insomnia severity and identifying patients’ symptoms.

Abnormal neutrophil signature in the blood and pancreas of presymptomatic and symptomatic type 1 diabetes

BACKGROUND Neutrophils and their inflammatory mediators are key pathogenic components in multiple autoimmune diseases, while their role in human type 1 diabetes (T1D), a disease that progresses sequentially through identifiable stages prior to the clinical onset, is not well understood. We previously reported that the number of circulating neutrophils is reduced in patients with T1D and in presymptomatic at-risk subjects. The aim of the present work was to identify possible changes in circulating and pancreas-residing neutrophils throughout the disease course to better elucidate neutrophil involvement in human T1D. METHODS Data collected from 389 subjects at risk of developing T1D, and enrolled in 4 distinct studies performed by TrialNet, were analyzed with comprehensive statistical approaches to determine whether the number of circulating neutrophils correlates with pancreas function. To obtain a broad analysis of pancreas-infiltrating neutrophils throughout all disease stages, pancreas sections collected worldwide from 4 different cohorts (i.e., nPOD, DiViD, Siena, and Exeter) were analyzed by immunohistochemistry and immunofluorescence. Finally, circulating neutrophils were purified from unrelated nondiabetic subjects and donors at various T1D stages and their transcriptomic signature was determined by RNA sequencing. RESULTS Here, we show that the decline in β cell function is greatest in individuals with the lowest peripheral neutrophil numbers. Neutrophils infiltrate the pancreas prior to the onset of symptoms and they continue to do so as the disease progresses. Of interest, a fraction of these pancreas-infiltrating neutrophils also extrudes neutrophil extracellular traps (NETs), suggesting a tissue-specific pathogenic role. Whole-transcriptome analysis of purified blood neutrophils revealed a unique molecular signature that is distinguished by an overabundance of IFN-associated genes; despite being healthy, said signature is already present in T1D-autoantibody-negative at-risk subjects. CONCLUSIONS These results reveal an unexpected abnormality in neutrophil disposition both in the circulation and in the pancreas of presymptomatic and symptomatic T1D subjects, implying that targeting neutrophils might represent a previously unrecognized therapeutic modality. FUNDING Juvenile Diabetes Research Foundation (JDRF), NIH, Diabetes UK.

Validation of a 14-drug microtitre plate that includes delamanid and bedaquiline for susceptibility testing of M. tuberculosis

UKMYC5 is a 96-well microtitre plate designed by the Comprehensive Resistance Prediction for Tuberculosis: an International Consortium (CRyPTIC) to enable the measurement of minimum inhibitory concentrations (MICs) of 14 different anti-TB compounds for >30,000 clinical tuberculosis isolates. Unlike the MYCOTB plate, on which UKMYC5 is based, the plate included two new (bedaquiline and delamanid) and two repurposed (clofazimine and linezolid) compounds. UKMYC5 plates were tested by seven laboratories on four continents using a panel of 19 external quality assessment (EQA) strains, including H37Rv. To assess the optimal combination of reading method and incubation time, MICs were measured from each plate by two readers using three methods (mirrored-box, microscope and Vizion™ Digital viewing system) after 7, 10, 14 and 21 days incubation. In addition, all EQA strains were whole-genome sequenced and phenotypically characterized by 7H10/7H11 agar proportion method (APM) and MGIT960. We conclude that the UKMYC5 plate is optimally read using the VizionTM system after 14 days incubation, achieving an inter-reader agreement of 97.9% and intra- and inter-laboratory reproducibilities of 95.6% and 93.1%, respectively. The mirrored-box had similar reproducibilities. Strains classified as resistant by APM, MGIT960 or the presence of mutations known to confer resistance consistently record elevated MICs compared with those strains classified as susceptible. Finally, the UKMYC5 plate records intermediate MICs for one strain which the APM measured MICs close the applied critical concentration, providing early evidence that the UKMYC5 plate can quantitatively measure the magnitude of resistance to anti-TB compounds due to specific genetic variation.Background Universal access to drug susceptibility testing is key to ending TB. UKMYC5 is a 96-well microtitre plate designed by the Comprehensive Resistance Prediction for Tuberculosis: an International Consortium (CRyPTIC) which has potential to determine, at low cost, the MICs for 14 different anti-TB drugs, including several new and repurposed compounds. It is a dry-format plate and therefore easy to transport and store. Objectives Determine how long to incubate the plates before reading, and the optimal reading method. Establish the reproducibility of the UKMYC5 plate and compare it to established methods. Methods UKMYC5 plates were tested by seven laboratories on four continents using a panel of 19 external quality assessment (EQA) strains, including H37Rv. MICs were measured from each plate by two readers using three methods (mirrored-box, microscope and Vizion™ Digital viewing system) at four different timepoints. All EQA strains were whole-genome sequenced and phenotypically characterized by MGIT960, 7H10/7H11 agar and resazurin microtitre assay. Results The optimum duration to incubate a plate is 14 days. The within-and between-laboratory reproducibilities for the best performing methods (mirrored-box and Vizion) were both 95% and 92%, respectively. One site was identified as requiring re-training and one drug (para-aminosalicylic acid) produced inconsistent results. Conclusions MICs measured using the UKMYC5 microtitre plate (i) are reproducible, (ii) compare well with the results of several established methods and (iii) correlate with the presence or absence of genetic mutations that confer resistance. This study provides the evidence that this assay can be deployed by TB reference laboratories world-wide as a diagnostic and research tool.

Latent tuberculous infection among foreign-born individuals applying to shelters in the metropolitan area of Milan

SETTING Screening for latent tuberculous infection (LTBI) of groups at high risk of active tuberculosis (TB) is a key component of the End TB Strategy. OBJECTIVE To conduct a retrospective descriptive analysis of LTBI rates among foreign-born individuals applying to shelters in the metropolitan area of Milan, Italy. DESIGN All foreign-born individuals registering for accommodation centres in the city of Milan from November 2009 to April 2017 were screened for active TB and LTBI. Individuals aged <36 years with a tuberculin skin test (TST) induration of >10 mm were offered confirmatory testing with QuantiFERON®-TB Gold In-Tube (QFT-GIT). RESULTS Of the 2666 TST-positive migrants aged <36 years who underwent LTBI confirmation testing, 1322 (49.6%) tested negative, 1339 (50.2%) were positive and five (0.2%) had indeterminate results. In the multivariate analysis, TB incidence in the country of origin and age were significantly associated with QFT-GIT positivity. Although estimated TB incidence in Eritrea, Morocco and Romania was 100/100 000 person-years (py), the probability of being QFT-GIT-positive in individuals from these countries were not statistically significantly different from individuals from countries with TB incidence > 250/100 000 person-years. CONCLUSION Our data showed a high proportion of LTBI among individuals coming from intermediate TB burden countries.

Coping Mechanisms, Psychological Distress, and Quality of Life Prior to Cancer Genetic Counseling

Background: Breast Cancer susceptibility genes 1 and 2 are implicated in hereditary breast and ovarian cancer and women can test for the presence of these genes prior to developing cancer. The goal of this study is to examine psychological distress, quality of life, and active coping mechanisms in a sample of women during the pre-test stage of the genetic counseling process, considering that pre-test distress can be an indicator of post-test distress. We also wanted to identify if subgroups of women, defined based on their health status, were more vulnerable to developing distress during the genetic counseling process. Methods: This study included 181 female participants who accessed a Cancer Genetic Counseling Clinic. The participants were subdivided into three groups on the basis of the presence of a cancer diagnosis: Affected patients, Ex-patients, and Unaffected participants. Following a self-report questionnaire, a battery of tests was administered to examine psychological symptomatology, quality of life, and coping mechanisms. Results: The results confirm that the genetic counseling procedure is not a source of psychological distress. Certain participants were identified as being more vulnerable than others; in the pre-test phase, they reported on average higher levels of distress and lower quality of life. These participants were predominantly Ex-patients and Affected patients, who may be at risk of distress during the counseling process. Conclusions: These findings highlight that individuals who take part in the genetic counseling process are not all the same regarding pre-test psychological distress. Attention should be paid particularly to Ex-patients and Affected patients by the multidisciplinary treating team.

“Health in the Mirror”: An Unconventional Approach to Unmet Psychological Needs in Oncology

Background: The introduction of aesthetic care programs for cancer patients inside hospitals could help patients cope with the side effects of both disease and treatment. The specific objective of this study is to evaluate whether a complementary and supportive program, called “Health in the Mirror,” has a positive effect on participants by analyzing certain psychological variables. Methods: Eighty-eight female cancer patients were included in this analysis. The support program is composed of three group aesthetic interventions that address both physical and psychological aspects that accompany cancer and its treatment. Patients were asked to complete a battery of tests in order to measure the impact of the program on certain psychological variables including anxiety, depression, body image, self-esteem, and quality of life. Outcome variables were measured at three different time-points: prior to participation, on the last day of the program, and after a 3-month follow-up. Results: Participating in the psychosocial support program “Health in the Mirror” determines an improvement in the psychological variables measured. Results revealed a significant reduction in depressive symptoms, anxiety and body image issues, as well as an improvement in self-esteem levels; this suggests that participating in this program could facilitate better adjustment to disease and treatment. Discussion: This study legitimizes the importance of implementing supportive and complementary therapies together with conventional therapies; the therapeutic approach to cancer cannot be restricted solely to medical care, but it must consider the patient as a whole person with needs that are not only physical or medical, but also psychological, social, and existential.

Identification of Socio-demographic and Psychological Factors Affecting Women’s Propensity to Breastfeed: An Italian Cohort

Background: Exclusive breastfeeding until 6 months postpartum is a World Health Organization objective and benefits have been demonstrated for both mother and infant. It is important to clarify which factors influence breastfeeding intentions. Our objective was to assess and identify socio-demographic and psychological factors associated with breastfeeding intention in a sample of pregnant Italian women. Materials and Methods: This prospective study included 160 pregnant women. The following psychological constructs were measured using standardized questionnaires: anxiety, prenatal attachment, adult attachment, personality traits, and intention to breastfeed. Socio-demographic data were also collected using a self-report questionnaire. Assessment took place after the 20th gestational week. Results: Self-employment, age and feeding received as an infant were significantly related to breastfeeding intention. Regarding psychological factors, we also found that Neuroticism was negatively associated with mother’s breastfeeding intentions. Relationships between psychological constructs and breastfeeding attitude were examined and represented within a graphical modeling framework. Conclusion: It may be possible to identify women that are less inclined to breastfeed early on in pregnancy. This may aid healthcare staff to pay particular attention to women who show certain socio-demographic and psychological characteristics, so as to fulfill more focused programs.