The Union and Médecins Sans Frontières approach to operational research.

Operational research (OR) has become a hot topic at national meetings, international conferences and donor fora. The International Union Against Tuberculosis and Lung Disease (The Union) and Médecins Sans Frontières (MSF) Operational Centre Brussels strongly promote and implement OR with colleagues in low- and middle-income countries. Here we describe how the two organisations define OR, and explain the guiding principles and methodology that underpin the strategy for developing and expanding OR in those countries. We articulate The Union's and MSF's approach to supporting OR, highlighting the main synergies and differences. Then, using the Malawi National Tuberculosis Control Programme as an example, we show how OR can be embedded within tuberculosis control activities, leading to changes in policy and practice at the national level. We discuss the difficult, yet vitally important, issue of capacity building, and share our vision of a new paradigm of product-related training and performance-based OR fellowships as two ways of developing the necessary skills at country level to ensure research is actually performed. Finally, we highlight the need to consider and incorporate into practice the ethical components of OR. This is a key moment to be involved in OR. We are confident that in partnership with interested stakeholders, including the World Health Organization, we can stimulate the implementation of quality, relevant OR as an integral part of health service delivery that in turn will lead to better health for people, particularly for those living in the poorer parts of the world

Impact of molecular genetic methods on the initiation of chemotherapy in multiple drug resistant tuberculosis patients in Arkhangelsk Region

In the Arkhangelsk Region, the prevalence of multiple drug-resistant tuberculosis is one of the highest in the world. In 2016, the portion of multiple drug resistant tuberculosis made 33.1% among new cases and 59.5% among relapses. Using new molecular genetic diagnostic techniques allows reducing the time for diagnostics of tuberculosis and drug resistance and should result in the earlier start of adequate treatment. The goal of the study is to assess the impact of new diagnostic molecular genetic methods on the time period from the first referral for medical care till the start of MDR-TB treatment. It was assumed that the introduction of molecular genetic tests would lead to early initiation of treatment in MDR TB patients (the research project of the International Union Against Tuberculosis and Lung Diseases and Tuberculosis Control Program of Arkhangelsk Region on The PROVE-IT LPA; Policy Relevant Outcomes from Validating Evidence on Impact of Line Probe Assays). Subjects and Methods. The results of the diagnostic procedure using cultures were compared with the results of the procedure based on molecular genetic tests aimed to detect MDR-TB. 295 MDR TB patients were enrolled into the study, of them, 163 had culture and 132 had molecular genetic tests. Results. The use of molecular genetic tests in smear-positive patients (AFB+) resulted in the reduction of the time period before initiation of MDRTB treatment by 50 and 66 days (median) versus culture by BacTAlert and absolute concentration on Lowenstein-Jensen medium respectively (p <0.001). Patients with a negative smear (AFB-), in whom MDR TB was detected by molecular genetic methods started treatment by 78 days earlier (median) versus patients who had culture (Lowenstein-Jensen, p < 0.001). Despite the significant reduction in the time period, even using molecular genetic methods, it took 24 days for cases with AFB+ and 62 days for cases with AFB- to be notified and start treatment of MDR TB

A novel approach in the treatment of neuroendocrine gastrointestinal tumors: Additive antiproliferative effects of interferon-γ and meta-iodobenzylguanidine

BACKGROUND: Therapeutic options to effectively inhibit growth and spread of neuroendocrine gastrointestinal tumors are still limited. As both meta-iodobenzylguanidine (MIBG) and interferon-γ (IFNγ) cause antineoplastic effects in neuroendocrine gastrointestinal tumor cells, we investigated the antiproliferative effects of the combination of IFNγ and non-radiolabeled MIBG in neuroendocrine gut STC-1 and pancreatic carcinoid BON tumor cells. METHODS AND RESULTS: IFNγ receptors were expressed in both models. IFNγ dose- and time-dependently inhibited the growth of both STC-1 and of BON tumor cells with IC(50)-values of 95 ± 15 U/ml and 135 ± 10 U/ml, respectively. Above 10 U/ml IFNγ induced apoptosis-specific caspase-3 activity in a time-dependent manner in either cell line and caused a dose-dependent arrest in the S-phase of the cell cycle. Furthermore, IFNγ induced cytotoxic effects in NE tumor cells. The NE tumor-targeted drug MIBG is selectively taken up via norepinephrine transporters, thereby specifically inhibiting growth in NE tumor cells. Intriguingly, IFNγ treatment induced an upregulation of norepinephrine transporter expression in neuroendocrine tumors cells, as determined by semi-quantitative RT-PCR. Co-application of sub-IC(50 )concentrations of IFNγ and MIBG led to additive growth inhibitory effects, which were mainly due to increased cytotoxicity and S-phase arrest of the cell cycle. CONCLUSION: Our data show that IFNγ exerts antiproliferative effects on neuroendocrine gastrointestinal tumor cells by inducing cell cycle arrest, apoptosis and cytotoxicity. The combination of IFNγ with the NE tumor-targeted agent MIBG leads to effective growth control at reduced doses of either drug. Thus, the administration of IFNγ alone and more so, in combination with MIBG, is a promising novel approach in the treatment of neuroendocrine gastrointestinal tumors

Specificity of a whole blood IGRA in German nursing students

<p>Abstract</p> <p>Background</p> <p>Interferon-gamma release assays (IGRA) are used for tuberculosis (TB) screening in healthcare workers (HCWs). However, data on specificity of IGRA in serial testing of HCWs is sparse. Therefore the specificity and the negative predictive value of the IGRA - QuantiFERON-TB Gold In-Tube (QFT) - in German nursing students was investigated.</p> <p>Methods</p> <p>194 nursing students at the start of their professional career were tested with the QFT. 14 nursing students were excluded from the specificity analysis, due to exposure to mycobacterium tuberculosis. Two of these subjects were QFT- positive. None of them developed disease during the year of follow-up. A study group of 180 students, all with very low risk of prior TB infection, remained in the specificity analysis. Subjects were monitored for at least two years with respect to the development of active TB disease. IGRA was performed at the start of the training and after one year.</p> <p>Results</p> <p>The mean age of the study group (n = 180) was 23 years (range 18-53) with 70.9% female and 99.4% German born. The specificity of QFT was 98.9% (178/180; 95% CI 0.96-0.99); lowering the cut-off from 0.35 IU/ml to 0.1 IU/ml would have decreased specificity only slightly to 97.8% (176/180; 95% CI 0.94-0.99). Of the 154 nursing students available for re-testing, one student who initially scored positive reverted to negative, and one student initially negative converted to positive. None of the monitored group with initially negative QFT results developed TB disease, indicating a high negative predictive value of the IGRA in this population.</p> <p>Conclusions</p> <p>Following our data, QFT can serve as an effective tool in pre-employment TB screenings for HCWs. As its negative results were stable over time, specificity of the QFT in serial testing of HCWs is high. As the risk of acquiring TB infection in the German healthcare system appears to be low, our data supports the recommendation of performing TB screening only in those HCWs with known contact to TB patients or infectious materials.</p

Is IP-10 a Better Biomarker for Active and Latent Tuberculosis in Children than IFNγ?

Background: The blood based interferon-gamma release assays (IGRA) for the diagnosis of tuberculosis do not discriminate between active TB disease and latent TB infection (LTBI). The search for distinguishing biomarkers therefore continues, as the accurate diagnosis of tuberculosis is particularly challenging in children. IFN-c-inducible protein 10 (IP-10/CXCL10) has recently been evaluated as a marker for active TB in adults with promising results. Aim: To investigate this new biomarker for active TB and LTBI in paediatrics. Method: We measured IP-10 levels using ELISA in supernatants of whole blood samples stimulated with TB-specificantigens and negative control antigen. Results: IP-10 is produced in high levels following mycobacterial antigen stimulation in active TB (n = 17) and LTBI (n = 16) compared to controls (n = 16) and to IFN-c. The baseline levels of IP-10 are increased in active TB and in LTBI, but there is no significant difference of stimulated levels of IP-10 between active TB and LTBI. Conclusions: IP-10 is a biomarker for tuberculosis in children. However like IFNc, IP-10 also does not distinguish between active TB and LTBI

ВЛИЯНИЕ ВНЕДРЕНИЯ МОЛЕКУЛЯРНО-ГЕНЕТИЧЕСКИХ МЕТОДОВ НА СРОКИ НАЧАЛА ХИМИОТЕРАПИИ БОЛЬНЫХТУБЕРКУЛЕЗОМ С МЛУ МБТ В АРХАНГЕЛЬСКОЙ ОБЛАСТИ

In the Arkhangelsk Region, the prevalence of multiple drug-resistant tuberculosis is one of the highest in the world. In 2016, the portion of multiple drug resistant tuberculosis made 33.1% among new cases and 59.5% among relapses. Using new molecular genetic diagnostic techniques allows reducing the time for diagnostics of tuberculosis and drug resistance and should result in the earlier start of adequate treatment.The goal of the study is to assess the impact of new diagnostic molecular genetic methods on the time period from the first referral for medical care till the start of MDR-TB treatment. It was assumed that the introduction of molecular genetic tests would lead to early initiation of treatment in MDR TB patients (the research project of the International Union Against Tuberculosis and Lung Diseases and Tuberculosis Control Program of Arkhangelsk Region on The PROVE-IT LPA; Policy Relevant Outcomes from Validating Evidence on Impact of Line Probe Assays).Subjects and Methods. The results of the diagnostic procedure using cultures were compared with the results of the procedure based on molecular genetic tests aimed to detect MDR-TB. 295 MDR TB patients were enrolled into the study, of them, 163 had culture and 132 had molecular genetic tests.Results. The use of molecular genetic tests in smear-positive patients (AFB+) resulted in the reduction of the time period before initiation of MDRTB treatment by 50 and 66 days (median) versus culture by BacTAlert and absolute concentration on Lowenstein-Jensen medium respectively (p &lt;0.001).Patients with a negative smear (AFB-), in whom MDR TB was detected by molecular genetic methods started treatment by 78 days earlier (median) versus patients who had culture (Lowenstein-Jensen, p &lt; 0.001). Despite the significant reduction in the time period, even using molecular genetic methods, it took 24 days for cases with AFB+ and 62 days for cases with AFB- to be notified and start treatment of MDR TB.Распространенность туберкулеза (ТБ) с множественной лекарственной устойчивостью возбудителя (МЛУ) в Архангельской области – одна из самых высоких в мире. В 2016 г. доля МЛУ-ТБ составила 33,1% среди новых случаев и 59,5% среди рецидивов ТБ. Использование новых молекулярно-генетических диагностических методов (МГМ) позволяет сократить время диагностики ТБ и лекарственной устойчивости возбудителя заболевания и должно привести к раннему началу адекватного лечения.Цель исследования: оценка влияния новых МГМ диагностики на изменение времени от первого обращения за медицинской помощью до начала лечения МЛУ-ТБ. Предполагали, что внедрение МГМ приведет к раннему началу лечения больных с МЛУ-ТБ (научный проект Международного союза борьбы с туберкулезом и болезнями легких и туберкулезной программы Архангельской области «The PROVE-IT LPA; Policy Relevant Outcomes from Validating Evidence on Impact of Line Probe Assays – значимые результаты подтверждения данных о влиянии метода гибридизации линейными зондами)».Материалы и методы. Результаты использования диагностического алгоритма с применением культуральных методов были сопоставлены с результатами алгоритма, основанного на МГМ выявления МЛУ-ТБ. В исследование включено 295 больных с МЛУ-ТБ, из которых у 163 мокрота исследована культуральными методами, а у 132 – с помощью МГМ.Результаты. В группе больных с положительным результатом микроскопии мокроты (КУМ+) применение МГМ привело к сокращению срока до начала лечения МЛУ-ТБ на 50 и 66 дней (медиана) по сравнению с культуральными методами BacTAlert и методом абсолютных концентраций на среде Левенштейна – Йенсена (ЛЙ) соответственно (p &lt; 0,001).Больные с отрицательным результатом микроскопии мокроты (КУМ-), у которых МЛУ-ТБ был выявлен с помощью МГМ, начинали лечение на 78 дней раньше (медиана) по сравнению с больными, у которых применялись культуральные методы (ЛЙ, p &lt; 0,001). Несмотря на значительное сокращение сроков, даже с использованием МГМ потребовалось 24 дня для случаев КУМ+ и 62 дня для случаев с КУМ- для регистрации и начала лечения МЛУ-ТБ

Xpert MTB/RIF Ultra assay for tuberculosis disease and rifampicin resistance in children

Background Every year, an estimated one million children and young adolescents become ill with tuberculosis, and around 226,000 of those children die. Xpert MTB/RIF Ultra (Xpert Ultra) is a molecular World Health Organization (WHO)‐recommended rapid diagnostic test that simultaneously detects Mycobacterium tuberculosis complex and rifampicin resistance. We previously published a Cochrane Review 'Xpert MTB/RIF and Xpert MTB/RIF Ultra assays for tuberculosis disease and rifampicin resistance in children'. The current review updates evidence on the diagnostic accuracy of Xpert Ultra in children presumed to have tuberculosis disease. Parts of this review update informed the 2022 WHO updated guidance on management of tuberculosis in children and adolescents. Objectives To assess the diagnostic accuracy of Xpert Ultra for detecting: pulmonary tuberculosis, tuberculous meningitis, lymph node tuberculosis, and rifampicin resistance, in children with presumed tuberculosis. Secondary objectives To investigate potential sources of heterogeneity in accuracy estimates. For detection of tuberculosis, we considered age, comorbidity (HIV, severe pneumonia, and severe malnutrition), and specimen type as potential sources. To summarize the frequency of Xpert Ultra trace results. Search methods We searched the Cochrane Infectious Diseases Group Specialized Register, MEDLINE, Embase, three other databases, and three trial registers without language restrictions to 9 March 2021. Selection criteria Cross‐sectional and cohort studies and randomized trials that evaluated Xpert Ultra in HIV‐positive and HIV‐negative children under 15 years of age. We included ongoing studies that helped us address the review objectives. We included studies evaluating sputum, gastric, stool, or nasopharyngeal specimens (pulmonary tuberculosis), cerebrospinal fluid (tuberculous meningitis), and fine needle aspirate or surgical biopsy tissue (lymph node tuberculosis). For detecting tuberculosis, reference standards were microbiological (culture) or composite reference standard; for stool, we also included Xpert Ultra performed on a routine respiratory specimen. For detecting rifampicin resistance, reference standards were drug susceptibility testing or MTBDRplus. Data collection and analysis Two review authors independently extracted data and, using QUADAS‐2, assessed methodological quality judging risk of bias separately for each target condition and reference standard. For each target condition, we used the bivariate model to estimate summary sensitivity and specificity with 95% confidence intervals (CIs). We stratified all analyses by type of reference standard. We summarized the frequency of Xpert Ultra trace results; trace represents detection of a very low quantity of Mycobacterium tuberculosis DNA. We assessed certainty of evidence using GRADE. Main results We identified 14 studies (11 new studies since the previous review). For detection of pulmonary tuberculosis, 335 data sets (25,937 participants) were available for analysis. We did not identify any studies that evaluated Xpert Ultra accuracy for tuberculous meningitis or lymph node tuberculosis. Three studies evaluated Xpert Ultra for detection of rifampicin resistance. Ten studies (71%) took place in countries with a high tuberculosis burden based on WHO classification. Overall, risk of bias was low. Detection of pulmonary tuberculosis Sputum, 5 studies Xpert Ultra summary sensitivity verified by culture was 75.3% (95% CI 64.3 to 83.8; 127 participants; high‐certainty evidence), and specificity was 97.1% (95% CI 94.7 to 98.5; 1054 participants; high‐certainty evidence). Gastric aspirate, 7 studies Xpert Ultra summary sensitivity verified by culture was 70.4% (95% CI 53.9 to 82.9; 120 participants; moderate‐certainty evidence), and specificity was 94.1% (95% CI 84.8 to 97.8; 870 participants; moderate‐certainty evidence). Stool, 6 studies Xpert Ultra summary sensitivity verified by culture was 56.1% (95% CI 39.1 to 71.7; 200 participants; moderate‐certainty evidence), and specificity was 98.0% (95% CI 93.3 to 99.4; 1232 participants; high certainty‐evidence). Nasopharyngeal aspirate, 4 studies Xpert Ultra summary sensitivity verified by culture was 43.7% (95% CI 26.7 to 62.2; 46 participants; very low‐certainty evidence), and specificity was 97.5% (95% CI 93.6 to 99.0; 489 participants; high‐certainty evidence). Xpert Ultra sensitivity was lower against a composite than a culture reference standard for all specimen types other than nasopharyngeal aspirate, while specificity was similar against both reference standards. Interpretation of results In theory, for a population of 1000 children: • where 100 have pulmonary tuberculosis in sputum (by culture): ‐ 101 would be Xpert Ultra‐positive, and of these, 26 (26%) would not have pulmonary tuberculosis (false positive); and ‐ 899 would be Xpert Ultra‐negative, and of these, 25 (3%) would have tuberculosis (false negative). • where 100 have pulmonary tuberculosis in gastric aspirate (by culture): ‐ 123 would be Xpert Ultra‐positive, and of these, 53 (43%) would not have pulmonary tuberculosis (false positive); and ‐ 877 would be Xpert Ultra‐negative, and of these, 30 (3%) would have tuberculosis (false negative). • where 100 have pulmonary tuberculosis in stool (by culture): ‐ 74 would be Xpert Ultra‐positive, and of these, 18 (24%) would not have pulmonary tuberculosis (false positive); and ‐ 926 would be Xpert Ultra‐negative, and of these, 44 (5%) would have tuberculosis (false negative). • where 100 have pulmonary tuberculosis in nasopharyngeal aspirate (by culture): ‐ 66 would be Xpert Ultra‐positive, and of these, 22 (33%) would not have pulmonary tuberculosis (false positive); and ‐ 934 would be Xpert Ultra‐negative, and of these, 56 (6%) would have tuberculosis (false negative). Detection of rifampicin resistance Xpert Ultra sensitivity was 100% (3 studies, 3 participants; very low‐certainty evidence), and specificity range was 97% to 100% (3 studies, 128 participants; low‐certainty evidence). Trace results Xpert Ultra trace results, regarded as positive in children by WHO standards, were common. Xpert Ultra specificity remained high in children, despite the frequency of trace results. Authors' conclusions We found Xpert Ultra sensitivity to vary by specimen type, with sputum having the highest sensitivity, followed by gastric aspirate and stool. Nasopharyngeal aspirate had the lowest sensitivity. Xpert Ultra specificity was high against both microbiological and composite reference standards. However, the evidence base is still limited, and findings may be imprecise and vary by study setting. Although we found Xpert Ultra accurate for detection of rifampicin resistance, results were based on a very small number of studies that included only three children with rifampicin resistance. Therefore, findings should be interpreted with caution. Our findings provide support for the use of Xpert Ultra as an initial rapid molecular diagnostic in children being evaluated for tuberculosis

Whole Blood Interferon-Gamma Responses to Mycobacterium tuberculosis Antigens in Young Household Contacts of Persons with Tuberculosis in Uganda

Due to immunologic immaturity, IFN-gamma-producing T cell responses may be decreased in young children compared to adults, thus we hypothesized that IFN-gamma responses to mycobacterial antigens in household contacts exposed to Mycobacterium tuberculosis (Mtb) would be impaired in young children relative to adults. The objective of this study was to compare whole blood IFN-gamma production in response to mycobacterial antigens between children and adults in Uganda.We studied household contacts of persons with culture-positive pulmonary tuberculosis (TB) enrolled in a cohort study conducted in Kampala, Uganda. Whole blood IFN-gamma production in response to Mtb culture-filtrate antigens was measured by ELISA and compared between infants (<2 years old, n = 80), young children (2 <5 years old, n = 216), older children (5 <15 years old, n = 443) and adults (> or =15 years old, n = 528). We evaluated the relationship between IFN-gamma responses and the tuberculin skin test (TST), and between IFN-gamma responses and epidemiologic factors that reflect exposure to Mtb, and the effect of prior BCG vaccination on IFN-gamma responses. Young household contacts demonstrated robust IFN-gamma responses comparable to those of adults that were associated with TST and known risk factors for infection. There was no effect of prior BCG immunization on the IFN-gamma response.Young children in a TB endemic setting can mount robust IFN-gamma responses generally comparable to those of adults, and as in adults, these responses correlated with the TST and known epidemiologic risk factors for Mtb infection