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SETTING: Patan Hospital, Lalitpur, Nepal.OBJECTIVES: To describe 1) the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) and its antibiotic sensitivity pattern; 2) the demographic and clinical characteristics associated with MRSA infections; and 3) the treatment outcomes of in-patients with MRSA infection among patients with S. aureus infection between January 2018 and December 2020.DESIGN: This was a cross-sectional study using electronic and paper-based hospital records of patients with S. aureus infection.RESULTS: Of the 1,804 patients with S. aureus infection, 1,027 patients (57%, 95% CI 55–59) had MRSA. The MRSA were susceptible to vancomycin (100%), linezolid (96%), doxycycline (96%), chloramphenicol (86%) and cotrimoxazole (70%), and resistant to erythromycin (68%), clindamycin (56%), gentamycin (58%), ciprofloxacin (92%) and ofloxacin (91%). The prevalence of MRSA was higher in 2019, among out-patients, and in respiratory samples, and lower in blood samples. Of the 142 in-patients with MRSA, 93% had a successful clinical outcome (cured/improved).CONCLUSION: More than 50% of patients with S. aureus infection had MRSA that were resistant to commonly available antibiotics. This calls for strengthening surveil-lance and good infection control practices in this hospital.

Introduction: Tajikistan is scaling up molecular diagnostic and digital technologies to strengthen its fight against drug-resistant TB (DR-TB). The study aimed to document national scale-up GeneXpert/GxAlert and Open MRS from 2012-2019 and compare time taken from TB diagnosis to treatment and quality of data recording before and after the introduction of GxAlert. Methodology: This was a longitudinal study that included a comparison of historical cohorts. Continuous variables were compared using Wilcoxon Rank-Sum test and categorical variables using the chi square test. Results: GeneXpert was introduced in 2011 and scaled up to 46 instruments in 43 (51%) diagnostic laboratories by May 2019. GxAlert was introduced in August 2018 and connected with all GeneXpert instruments by February 2019. Open MRS was introduced in 2014 and implemented in all 108 treatment centers by mid-2018. Time from diagnosis to treatment pre-GxAlert (range 0-749, median 3, days) was significantly longer than with GxAlert (range 0-273, median 3, days) (p 2 weeks was 16% (282/1740) pre-GxAlert and 11% (206/1902) with GxAlert (p < 0.001). Between 31%-34% of patients with DR-TB results in Open MRS did not have results available in GeneXpert/GxAlert systems. Where results were present in both systems, there were discrepancies in 8.2% of patients pre-GxAlert and 4.3% with GxAlert (p = 0.25). Conclusions: The scale-up of GeneXpert and digital technologies in Tajikistan was associated with a reduction in the proportion of patients with delays more than 2 weeks between diagnosis and treatment, but data quality recording improved only slightly.

Introduction: Poor human immunodeficiency virus (HIV) testing practices and underreporting of HIV-related data in TB information systems remain barriers to effective care for TB-HIV co-infected patients. HIV testing and recording practices in national TB program have not been formally evaluated in Armenia. This study aimed to assess the recording completeness of HIV testing and HIV status in the national TB program electronic database, and to determine trend in HIV testing and the association between HIV testing and treatment outcomes for all TB patients registered in Armenia (2015-2019). Methodology: A cohort study of TB patients using routine programmatic data from the national TB program of Armenia. Results: From 2015 to 2019, the electronic database was completed for HIV testing and HIV status by 48.1% and 97.5%, respectively. Of all registered TB patients 93.6% were tested for HIV. Of a total 4,674 patients, 1,085 (23.2%) had unsuccessful outcomes. Patients with HIV status “not tested” and “not recorded” compared to HIV “negatives” had 1.76 (95%CI 1.42-2.11) and 1.6 (95%CI 1.20-2.06) times higher risk of unsuccessful outcomes, respectively. Lost to follow-up was the most frequent unsuccessful outcome in HIV status “not tested” group. Conclusions: An analysis of nationwide data revealed incompleteness of the national TB electronic database for HIV data. Patients with HIV status “not tested” and “not recorded” had higher risk of unsuccessful TB treatment outcomes. Upgrade of the electronic database with information on key indicators of TB-HIV services will facilitate improved monitoring and reporting.

Introduction: MDR/RR-TB is a growing problem in Kyrgyzstan. In 2005, the country introduced standard or individualized treatment for 20-24 months. Because of poor treatment outcomes, in 2017 a short treatment with strict eligibility criteria was introduced. The aim of this study was to compare characteristics and treatment outcomes of MDR/RR-TB patients receiving short (9-12 months) treatment in 2017 with those receiving standard or individualized (20-24 months) treatment in 2016/2017. Methodology: A comparative cohort study using routine programmatic data. Characteristics, sputum culture conversion and treatment outcomes were compared between those on short treatment with those on standard/individualized treatment using the chi-square test, crude and adjusted risk ratios (RR and aRR). Results: The study included 274, 82 and 132 patients on standard, individualized and short treatment, respectively. There were more females, fewer migrants/homeless and unemployed and more new TB patients on short treatment compared with the other two groups. A favorable outcome (cure and treatment completed) was significantly higher in short treatment patients (83%) compared with those on standard (50%) or individualized (59%) treatment (p < 0.001). There was higher 1-month sputum culture conversion with short treatment (35%) compared with the other two groups (19% and 24%, p < 0.05). Short treatment (aRR 1.6, 1.4-1.8), female gender (aRR 1.2, 1.1-1.4), not being homeless (aRR 12.9, 4.5-17.3) and having new TB (aRR 1.3, 1.0-1.5) were independently associated with a favorable outcome. Conclusions: The treatment success was higher in selected MDR-TB patients given short treatment in Kyrgyzstan: this regimen should be scaled-up to all MDR-TB patients.

Alzheimer's disease (AD) is a progressive neurodegenerative disease with no cure. Huge efforts have been made to develop anti‐AD drugs in the past decades. However, all drug development programs for disease‐modifying therapies have failed. Possible reasons for the high failure rate include incomplete understanding of complex pathophysiology of AD, especially sporadic AD (sAD), and species difference between humans and animal models used in preclinical studies. In this study, sAD is modeled using human induced pluripotent stem cell (hiPSC)‐derived 3D brain organoids. Because the blood–brain barrier (BBB) leakage is a well‐known risk factor for AD, brain organoids are exposed to human serum to mimic the serum exposure consequence of BBB breakdown in AD patient brains. The serum‐exposed brain organoids are able to recapitulate AD‐like pathologies, including increased amyloid beta (Aβ) aggregates and phosphorylated microtubule‐associated tau protein (p‐Tau) level, synaptic loss, and impaired neural network. Serum exposure increases Aβ and p‐Tau levels through inducing beta‐secretase 1 (BACE) and glycogen synthase kinase‐3 alpha / beta (GSK3α/β) levels, respectively. In addition, single‐cell transcriptomic analysis of brain organoids reveals that serum exposure reduced synaptic function in both neurons and astrocytes and induced immune response in astrocytes. The human brain organoid‐based sAD model established in this study can provide a powerful platform for both mechanistic study and therapeutic development in the future., Brain organoids are exposed to human serum to mimic the serum exposure consequence of the blood–brain barrier (BBB) breakdown in Alzheimer's disease (AD) patient brains, which recapitulates several AD‐like pathologies, including increased amyloid beta (Aβ) and phosphorylated microtubule‐associated tau protein (p‐Tau) level, synaptic loss, and impaired neural network. This human brain organoid‐based sporadic AD (sAD) model provides a powerful platform for both mechanistic study and therapeutic development in the future.

Introduction: Ukraine has gaps in Tuberculosis (TB) service coverage, especially in key populations (KPs). We compared effectiveness of three different strategies for active TB detection among KPs and their linkage to TB treatment during three time periods. Methodology: The KPs included people who inject drugs (PWID), sex workers (SW), men who have sex with men (MSM) and groups at-risk of TB (ex-prisoners, Roma and homeless). The active case finding included decentralized symptom screening and specimen collection (2014, strategy-1), decentralized screening with patient referred for specimen collection (2015-2017, strategy-2) and strategy-2 plus GeneXpert (2018, strategy-3). Results: In total 680,760 KPs were screened, of whom 68% were PWID. TB case detection per 100,000 populations was 1,191 in strategy-1, 302 in strategy-2, and 235 in strategy-3. The number needed to screen (NNS) to identify one case was respectively 84, 332, and 425. TB detection was highest among homeless (range: 1,839-2,297 per 100,000 population). The lowest detection was among the MSM and SW. Between 2014 and 2018, 82-94% of all diagnosed TB patients in KPs started TB treatment. Conclusions: The active case finding in KPs increased detection of TB cases in Ukraine, and the majority of diagnosed KPs initiated TB treatment. Centralization of diagnosis reduced the effectiveness of TB screening. Each region in Ukraine should assess the composition and the needs of KPs which will allow for adoption of specific strategies to detect TB among KPs with high TB prevalence.

Introduction: People who inject drugs (PWID) are one of the key populations most vulnerable to HIV infection, with 28 times higher prevalence compared to the rest of the population. PWID are known to have many physical, psychological and lifestyle challenges that can influence access to care. Depression is common among PWID living with HIV. It has major effect on health-related quality of life (HRQoL) and is influencing adherence to antiretroviral therapy. This study was conducted to explore how anxiety and depression affect HRQoL among HIV-positive PWID in Ukraine. It will provide knowledge for the further policy development. Methodology: A descriptive cross-sectional study using data from interviewer- administrated questionnaires was performed. The questionnaire was based on the Hospital Anxiety and Depression Scale. The questionnaire on HRQoL was based on the SF-36. Results: Among the 90 HIV positive PWID 74% (67) and 61% (55) had anxiety and depression scores higher than 7 respectively, indicating that most patients had mental health problems. Average scores for general health (40), role limitations due to physical (44) and emotional health (34), vitality (41) and mental health (45) had mean scores less than 50 along with total physical (43) and mental health scores (35). Having an HIV positive partner or partner with unknown HIV status increases anxiety in HIV positive PWID. Conclusion: There are increased depressive and anxiety symptoms and poorer QoL among HIV-positive PWID in Ukraine. Strategies focusing on psychosocial support addressing QoL as part of HIV care could improve health outcomes for these comorbid and debilitating conditions.

Introduction: Adherence to tuberculosis (TB) treatment as an important determinant for the successful cure of patients can be increased by focusing on patient satisfaction. The objective of this study was to evaluate patient satisfaction with TB services, different aspects of patient satisfaction, and demographic, health and treatment characteristics associated with satisfaction. Methodology: Overall 505 randomly selected TB patients that received treatment during 2014-2015 in Armenia underwent a cross-sectional telephone survey. Patient satisfaction items were selected from the Patient Satisfaction Questionnaire (PSQ-18). The Consultation and Relational Empathy (CARE) and Patient Enablement Instrument (PEI) were also used. Treatment adherence was assessed using the Morisky Adherence Scale. Results: The respondents comprised 386 (76.4%) men and 119 (23.6%) women with a mean age of 45.5 ± 0.7 years. Nearly 99% (n = 500) of them were treatment-adherent. However, 45 (8.9%) mentioned the side effects as a reason for not following the treatment, revealing the non-adherence level of approximately 9%. About 93% of the patients were generally satisfied with the TB services, about 46% were satisfied with consultation and relational empathy and about 95% were satisfied with patient enablement. Being unsatisfied with TB services was associated with treatment non-adherence, inpatient treatment, drug-resistant TB, higher education, being unmarried, having a family income of below 50,000AMD (~120 USD) on average, being unsatisfied with consultation and empathy and place of residence. Conclusions: This study reports that TB patients are highly satisfied with TB care in Armenia. However, addressing specific characteristics associated with satisfaction may improve the TB program.

Compliance with treatment guidelines is essential to achieve successful outcomes in tuberculosis patients. Thus, we assessed if multidrug-resistant tuberculosis treatment practices from 2012–2018 in Uzbekistan were compliant with national guidelines in terms of regimens prescribed, weight-based drug dosages used, and documentation of treatment changes (such as prolongation of intensive phase, change of drugs, and their reasons) in the treatment card and Consilium form. A total of 1481 patients were included. Of them, only 25% received standardized regimens as per guidelines and the remaining received individualized regimens. There was an increasing trend in using standardized regimens from 2% in 2012 to 44% in 2018. Compliance to recommended weight-based drug dosages was observed in 85% of the patients during the intensive phase and 84% in the continuation phase—ranged 71–91% over the years. Prolongation of the intensive phase was done in 42% of patients. The treatment was changed in 44% of patients during the intensive phase and 34% of patients during the continuation phase. The documentation of treatment changes was suboptimal (42–75%) during the initial years (2012–2014), however, it improved significantly during later years (86–100%). Future research should explore reasons for non-compliance so that the quality of patient care can be improved.

Antimicrobial resistance (AMR) is the acquired ability of pathogens to withstand antimicrobial treatment. To bridge the gap in knowledge for implementing effective and targeted interventions in relation to the AMR in Armenia, we designed this study to explore the performance of AMR diagnostics and the profile of AMR in the Nork Infection Clinical Hospital (NICH) for the period of 2016–2019, particularly to (i) determine the proportions of antimicrobial resistance among all samples tested at the hospital laboratory, (ii) determine the proportion of resistance against specific antimicrobials, and (iii) identify factors associated with AMR. A cross-sectional study was conducted with a secondary data analysis that included all the patients tested for AMR in the laboratory of the NICH for the period of 2016–2019. For this period, only 107 (0.3%) patients out of 36,528 had their AMR test results available and of them, 87 (81%) had resistance at least to one tested antimicrobial. This study has provided some valuable information on the AMR situation in Armenia. The results call for immediate actions to control the access to and the use of antimicrobials, strengthen AMR surveillance, and improve laboratory capacity for the proper and fast identification of drug resistance through a comprehensive system.

Tuberculosis treatment is challenging, especially among people with drug-resistant forms of tuberculosis. The introduction of fully oral modified short treatment regimen has a great potential to shorten duration of treatment, improve safety and ultimately increase treatment success rate. In 2019 Georgia has piloted the modified fully oral shorter treatment regimen in a routine programmatic condition. Our study aimed to evaluate effectiveness and safety of the modified shorter treatment regimen in Georgia among the first 25 consecutively enrolled patients with rifampicin-resistant tuberculosis with proven sensitivity to fluoroquinolone and without prior exposure to second-line tuberculosis drugs. Regimen consisted of 9-month daily administration of bedaquilline, linezolid, levofloxacin, clofazimine and cycloserine. Study patients were enrolled between March-August 2019. We used a national electronic surveillance system, medical records and active TB drug-safety monitoring and management database to extract study related data. The mean age of the study participants was 48 years, 68% were male, 8% were HIV positive, 16% had diabetes and 12% tested positive for hepatitis C infection. The median time to culture conversion among 16 patients who were culture positive at treatment initiation was 1.0 (95% CI: 1.0-2.0) month. Of those, by the end of treatment 15 patients converted to negative. Out of the 25 patients in the study cohort 22 (88%) had successful treatment outcome, one patient (4%) died and two (8%) were lost to follow up. The regimen was largely well tolerated. Three patients (12%) experienced serious adverse events, of which in two cases were possibly related to TB drugs in the regimen. Seven patients developed adverse events of interest in eight instances, including musculoskeletal (twice), psychiatric, gastrointestinal disorders, hepatotoxicity, peripheral neuropathy, cardiotoxicity and myelosuppression (once each). In four patients (16%) the duration of the treatment was extended beyond nine months due to insufficient radiological improvements. Our findings demonstrate that good treatment outcomes are achievable in people with fluoroquinolone-sensitive tuberculosis within routine programmatic conditions using fully oral modified short treatment regimen. The extensive use of fully oral modified shorter treatment regimen in Georgia and other high priority countries in the World Health Organization European Region is warranted.

Considering the complexity of second-line anti-tuberculosis (TB) treatment regimens, the management of drug-resistant TB (DR-TB) in Georgia remains a major challenge. Since the introduction of new and repurposed anti-TB medications, the implementation of active TB Drug Safety Monitoring (aDSM) was a critical program component to help establish safety and manage all treatment related Serious Adverse Events (SAEs). In our study, we aimed to describe the occurrence, characteristics and timing of SAE among patients with Rifampicin Resistant and Multi-Drug Resistant TB (RR/MDR-TB) receiving new and/or repurposed anti-TB medications (bedaquiline, delamanid, linezolid, clofazimine, imipenem) during the period of 2016-2018 in Georgia and identify predictors of SAE. The data were obtained from the medical charts, electronic database and standardized aDSM reports During 2016-2018 period in total 970 people with RR/MDR-TB were notified in Georgia and 388 of them received new and/or repurposed TB drugs as part of their treatment regimen and all were included into the study. The results showed a total of 73 SAEs registered among 49 (12.6%) patients receiving new and/or repurposed drugs. The overall SAE incidence rate per 100 person-months was 1.16. The severity of the majority of the SAEs (46.6%) was grade III and 21.9% were grade IV. The most common SAE reported was hepatotoxicity, with an incidence of 0.26 per 100 person-month (n=16, 21.9%) followed by cardiotoxicity with an incidence of 0.16 per 100 person-month (n=10, 13.7%). Median time to SAE occurrence was 183 days (IQR 84 – 334) after treatment initiation. Resistance profile was the only predictor associated with occurrence of a SAEs. There was increased hazard of SAEs among patients with XDR-TB (adjusted HR=2.18, 95% CI: 1.12-4.23). Our findings on SAEs among patients treated with new or repurposed anti-TB drugs are echoing the findings available in the literature. They highlight the need for close monitoring of patients and underlines the importance of the aDSM during the whole treatment. Safety profile of the medications and combinations used are yet to be established and larger datasets comprised of patients receiving same treatment regimens need to be utilized.

BackgroundWHO’s directly observed therapy (DOT) strategy for tuberculosis (TB) treatment depends upon a well–organized healthcare system. This study sought to evaluate the effectiveness of self-administered drug intake supported by a family member versus in-clinic DOT.MethodsThis open–label, nationally-representative stratified cluster randomized controlled non–inferiority trial with two parallel equal arms involved drug–susceptible pulmonary TB patients in the continuation treatment phase. We randomly assigned outpatient–TB–centres (52 clusters) to intervention and control arms. The intervention included an educational/counseling session to enhance treatment adherence; weekly visits to outpatient–TB–centres to receive medication, and daily SMS medication reminders and phone calls to track adherence and record side effects. Controls followed clinical DOT at Outpatient–TB–centres. Both groups participated in baseline and 4–5 months follow–up surveys. The trial’s non–inferiority comparisons include: treatment success as the clinical (primary) outcome and medication adherence (self–reported), knowledge, depressive symptoms, stigma, quality of life, and social support as non–clinical (secondary) outcomes.ResultsPer–protocol analysis showed that the intervention (n = 187) and control (n = 198) arms achieved successful treatment outcome of 92.0 and 92.9%, respectively, indicating that the treatment success in the intervention group was non–inferior to DOT. Knowledge, depression, stigma, quality of life, and social support also showed non–inferiority, demonstrating substantial improvement over time for knowledge (change in the intervention = 1.05: 95%CL (0.49, 1.60); change in the control = 1.09: 95%CL (0.56, 1.64)), depression score (change in the intervention = − 3.56: 95%CL (− 4.99, − 2.13); change in the control = − 1.88: 95% CL (− 3.26, − 0.49)) and quality of life (change in the intervention = 5.01: 95%CL (− 0.64, 10.66); change in the control = 7.29: 95%CL (1.77, 12.81)). The intervention resulted in improved treatment adherence.ConclusionsThis socially empowering alternative strategy might be a preferable alternative to DOT available to patients in Armenia and in other countries. Further research evaluating cost effectiveness of the intervention and generalizability of the results is warranted.Trial registrationClinicaltrials.gov:NCT02082340, March 10, 2014.

Abnormal tau hyperphosphorylation and its aggregation into neurofibrillary tangles are a hallmark of tauopathies, neurodegenerative disorders that include Alzheimer’s disease (AD). Active and passive Tau-immunotherapy has been proposed as a therapeutic approach to AD with mixed results. One of the limitations of active immunotherapy may be associated with the mediocre immunogenicity of vaccines that are not inducing therapeutically potent titers of antibodies. The aim of this study was to test the efficacy of an anti-tau vaccine, AV-1980R/A composed of N terminal peptide of this molecule fused with an immunogenic MultiTEP platform and formulated in a strong adjuvant, Advax(CpG) in a Tg4510 mouse model of tauopathy. Experimental mice were immunized with AV-1980R/A and a control group of mice were injected with adjuvant only. Nontransgenic and tetracycline transactivator (tTA) transgenic littermates were included as baseline controls to contrast with the tau phenotype. Active immunization with AV-1980R/A induced very strong anti-tau humoral immune responses in both nontransgenic and transgenic mice with evidence of IgG in brains of AV-1980R/A vaccinated mice. These experimental animals displayed an improvement in short-term memory during a novel object recognition test. However, impairments in other behavioral tasks were not prevented by AV-1980R/A vaccinations. At the same time, high titers of anti-tau antibodies reduced hyperphosphorylated pSer396 tau but did not lower the level of other phosphorylated tau species in the brains of AV-1980R/A vaccinated mice. These data indicate that active immunotherapy with an N-terminal Tau epitope was only partially effective in improving cognition and reducing pathology in the stringent Tg4510 mouse model of tauopathy.

Background Alzheimer disease (AD) is characterized by the accumulation of beta-amyloid (Aβ) plaques and neurofibrillary tangles composed of hyperphosphorylated tau, which together lead to neurodegeneration and cognitive decline. Current therapeutic approaches have primarily aimed to reduce pathological aggregates of either Aβ or tau, yet phase 3 clinical trials of these approaches have thus far failed to delay disease progression in humans. Strong preclinical evidence indicates that these two abnormally aggregated proteins interact synergistically to drive downstream neurodegeneration. Therefore, combinatorial therapies that concurrently target both Aβ and tau might be needed for effective disease modification. Methods A combinatorial vaccination approach was designed to concurrently target both Aβ and tau pathologies. Tau22/5xFAD (T5x) bigenic mice that develop both pathological Aβ and tau aggregates were injected intramuscularly with a mixture of two MultiTEP epitope vaccines: AV-1959R and AV-1980R, targeting Aβ and tau, respectively, and formulated in AdvaxCpG, a potent polysaccharide adjuvant. Antibody responses of vaccinated animals were measured by ELISA, and neuropathological changes were determined in brain homogenates of vaccinated and control mice using ELISA and Meso Scale Discovery (MSD) multiplex assays. Results T5x mice immunized with a mixture of Aβ- and tau-targeting vaccines generated high Aβ- and tau-specific antibody titers that recognized senile plaques and neurofibrillary tangles/neuropil threads in human AD brain sections. Production of these antibodies in turn led to significant reductions in the levels of soluble and insoluble total tau, and hyperphosphorylated tau as well as insoluble Aβ42, within the brains of bigenic T5x mice. Conclusions AV-1959R and AV-1980R formulated with AdvaxCpG adjuvant are immunogenic and therapeutically potent vaccines that in combination can effectively reduce both of the hallmark pathologies of AD in bigenic mice. Taken together, these findings warrant further development of this vaccine technology for ultimate testing in human AD.

The treatment of childhood tuberculosis can be challenging due to the lack of pediatric drug formulations and monitoring of drug-toxicity in routine settings. There are no published studies from Uzbekistan on the adverse drug reactions (ADR) associated with anti-tuberculosis treatment in children. In this study, we aimed to investigate the ADR associated with anti-tuberculosis treatment in children. This was a cohort study using secondary program data of children treated at the city and regional tuberculosis clinics in Tashkent, Uzbekistan. Of the 302 patients evaluated, 135 (44.7%) reported ADR. New tuberculosis was registered in 277 (92%) patients and 262 (87%) had extrapulmonary tuberculosis. Factors associated with ADR included treatment at a regional hospital (adjusted odds ratio, aOR = 1.75, p = 0.026), female sex (aOR = 2.2, p = 0.004), and treatment with second-line drugs (aOR = 8.82, p &lt, 0.001). The most common ADRs were gastrointestinal disorders (28.5%) followed by hepatitis (8.9%) and dermatologic reactions (8.6%). Most of the ADRs were mild (55.6%) or moderate (43.7%), only one child had severe ADR. Patients with the identified risk factors should be closely monitored during the treatment. We also recommend expansion of ADR surveillance throughout the country for more representative data in the future.

Introduction: Tuberculosis and tobacco prove to be increasingly apparent world problems. Armenia is a developing country which is facing issues related to the high rates of tobacco consumption. Moreover, it is among the list of high multi-drug resistant (MDR) Tuberculosis TB burden countries. Treatment success rate in Armenia for sputum smear-positive cases never reached World Health Organization's (WHO) target of 85% in last 15 years. Data from different studies completed across the world suggests that there is an association between smoking and negative treatment outcomes. Methods: This retrospective study was designed to investigate aforementioned associations between TB treatment outcomes and smoking status of TB patients. Data for the study were derived from the national data available in the electronic database of the Armenian National TB Center. Results: Based on inclusion and exclusion criteria 992 TB patients registered in 2014 were enrolled in this study. All of them are were TB patients in which 387 were smokers and 605 were non-smokers. Notably, adjusted analysis showed that individuals who smoked during TB treatment had 1.61 higher odds of having unsuccessful TB treatment outcome. Additionally, consistent with the literature, statistically significant association was identified between TB treatment outcome and other well factors such as sputum smear status (OR=2.24, p

Previously, we reported that Alzheimer's disease (AD) epitope vaccines (EVs) composed of N-terminal β-amyloid (Aβ 42 ) B cell epitope fused with universal foreign T helper (Th) epitope(s) were immunogenic, potent, and safe in different amyloid precursor protein (APP) transgenic mice with early AD-like pathology. However, developing an effective therapeutic vaccine is much more challenging, especially when a self-antigen such as Aβ 42 is a target. Here, we directly compare the efficacy of anti-Aβ 42 antibodies in Tg2576 mice with low or high levels of AD-like pathology at the start of immunizations: 6–6.5 months for preventive vaccinations and 16–19 months for therapeutic vaccinations. EV in a preventive setting induced high levels of anti-Aβ antibodies, significantly reducing pathologic forms of Aβ in the brains of Tg2576 mice. When used therapeutically for immunesenescent Tg2576 mice, EV induced low levels of antibodies not sufficient for clearing of AD-like pathology. Separately, we demonstrated that EV was also not effective in 11–11.5-month-old Tg2576 mice with moderate AD-like pathology. However, we augmented the titers of anti-Aβ antibodies in transgenic (Tg) mice of the same age possessing the pre-existing memory Th cells and detected a significant decrease in diffuse and core plaques in cortical regions compared to control animals along with improved novel object recognition performance.

Pathological tau correlates well with cognitive impairments in Alzheimer’s disease (AD) patients and therefore represents a promising target for immunotherapy. Targeting an appropriate B cell epitope in pathological tau could in theory produce an effective reduction of pathology without disrupting the function of normal native tau. Recent data demonstrate that the N-terminal region of tau (aa 2-18), termed the “phosphatase activation domain (PAD)”, is hidden within native Tau in a ‘paperclip’-like conformation. Conversely, PAD is exposed in pathological tau and plays an essential role in the inhibition of fast axonal transport and tau polymerization. Thus, we hypothesized that anti-tau2-18 antibodies may safely and specifically reduce pathological tau and prevent further aggregation, which in turn would neutralize tau toxicity. Therefore, we evaluated the immunogenicity and therapeutic efficacy of our MultiTEP platform-based vaccine targeting tau2-18 formulated with AdvaxCpG adjuvant (AV-1980R/A) in PS19 tau transgenic mice. The AV-1980R/A induced extremely high antibody responses and the resulting sera recognized neurofibrillary tangles and plaque-associated dystrophic neurites in AD brain sections. In addition, under non-denaturing conditions AV-1980R/A sera preferentially recognized AD-associated tau. Importantly, vaccination also prevented age-related motor and cognitive deficits in PS19 mice and significantly reduced insoluble total and phosphorylated tau species. Taken together, these findings suggest that predominantly targeting misfolded tau with AV-1980R/A could represent an effective strategy for AD immunotherapy.

Introduction: Tuberculosis (TB) continues to be a global public health problem. People with weakened immune systems are more vulnerable to TB. It is one of the top 10 causes of death worldwide and is a leading cause of death for people living with HIV (PLWH). The aim of the current study was to perform programmatic data analysis of TB cases treated with the first-line drugs, registered in Armenia for the period of January 2017 – August 2018, and to identify gaps in TB care system in Armenia. Methodology: A retrospective cohort study using programmatic data from National TB Program. Results: Overall treatment success rate for the period of study was 79%. HIV had impact only on “died” outcome with odds ratio (OR) of 20.9. More than a third (34%) of all HIV-positive patients died during TB treatment and 45% of patients who had non-Armenian citizenship were lost to follow-up during the treatment (OR = 3.3). Treatment duration for the 8% of all cases (mainly with brain or bone localization) was > 9 months and lasted up to 500 days. Conclusions: Better collaboration and partial integration of TB and HIV services in Armenia is required. The access to care for non-Armenian citizens needs to be improved. The national TB treatment guideline needs to be updated based on scientific evidence. This study demonstrates that continuous analysis of the available data and tailoring of the system is required to address the needs of key populations and achieve universal care coverage.

Introduction: In efforts to reach the 2020 Tuberculosis targets, the WHO advocates for an outpatient, people-centered model of TB care. To this end, the TB care system in Armenia underwent structural and financing reforms in 2014. Financing mechanism for inpatient TB facilities was changed from a fee per bed/day to a mixed type of financing that includes fixed and variable costs eliminating incentives for unnecessary and extended hospitalizations. Unfortunately, outpatient facilities continue to be financed through per-capita mechanism, resulting in high number of referrals and draining resources. This study aimed to assess the implementation of these reforms within the Armenian TB care system. Methodology: This was a retrospective cross-sectional study using nationwide programme data and survey data collected from healthcare facilities. Results: In 2017 a total of 901 TB patients were registered in outpatient facilities. Only 7.6% of total TB cases were diagnosed in outpatient facilities and 30.9% of the presumptive TB cases were referred to inpatient facilities. The number of hospitalizations was reduced by 76% from 2013 to 2017. The average duration of stay reduced as well from 55+ days to 37 days. However, the proportion of smear negative TB patients remains high among all hospitalized patients (63.8%). Conclusions: Overall, the reform has been successful, however unnecessary hospitalizations persist. Our results indicate there a need to go upstream for a structural and financial reform of the outpatient sector to complete Armenia’s TB healthcare reform and improve both patient outcomes and efficient use of system resources.

Antimicrobial Resistance (AMR) and Healthcare Associated Infections (HAIs) are major global public health challenges in our time. This study provides a broader and updated overview of AMR trends in surgical wards of Mulago National Referral Hospital (MNRH) between 2014 and 2018. Laboratory data on the antimicrobial susceptibility profiles of bacterial isolates from 428 patient samples were available. The most common samples were as follows: tracheal aspirates (36.5%), pus swabs (28.0%), and blood (20.6%). Klebsiella (21.7%), Acinetobacter (17.5%), and Staphylococcus species (12.4%) were the most common isolates. The resistance patterns for different antimicrobials were: penicillins (40–100%), cephalosporins (30–100%), β-lactamase inhibitor combinations (70–100%), carbapenems (10–100%), polymyxin E (0–7%), aminoglycosides (50–100%), sulphonamides (80–100%), fluoroquinolones (40–70%), macrolides (40–100%), lincosamides (10–45%), phenicols (40–70%), nitrofurans (0–25%), and glycopeptide (0–20%). This study demonstrated a sustained increase in resistance among the most commonly used antibiotics in Uganda over the five-year study period. It implies ongoing hospital-based monitoring and surveillance of AMR patterns are needed to inform antibiotic prescribing, and to contribute to national and global AMR profiles. It also suggests continued emphasis on infection prevention and control practices (IPC), including antibiotic stewardship. Ultimately, laboratory capacity for timely bacteriological culture and sensitivity testing will provide a rational choice of antibiotics for HAI.

The DNA vaccine, AV-1959D, targeting N-terminal epitope of Aβ peptide, has been proven immunogenic in mice, rabbits, and non-human primates, while its therapeutic efficacy has been shown in mouse models of Alzheimer's disease (AD). Here we report for the first time on IND-enabling biodistribution and safety/toxicology studies of cGMP-grade AV-1959D vaccine in the Tg2576 mouse model of AD. We also tested acute neuropathology safety profiles of AV-1959D in another AD disease model, Tg-SwDI mice with established vascular and parenchymal Aβ pathology in a pre-clinical translational study. Biodistribution studies two days after the injection demonstrated high copy numbers of AV-1959D plasmid after single immunization of Tg2576 mice at the injection sites but not in the tissues of distant organs. Plasmids persisted at the injection sites of some mice 60 days after vaccination. In Tg2576 mice with established amyloid pathology, we did not observe short- or long-term toxicities after multiple immunizations with three doses of AV-1959D. Assessment of the repeated dose acute safety of AV-1959D in cerebral amyloid angiopathy (CAA) prone Tg-SwDI mice did not reveal any immunotherapy-induced vasogenic edema detected by magnetic resonance imaging (MRI) or increased microhemorrhages. Multiple immunizations of Tg-SwDI mice with AV-1959D did not induce T and B cell infiltration, glial activation, vascular deposition of Aβ, or neuronal degeneration (necrosis and apoptosis) greater than that in the control group determined by immunohistochemistry of brain tissues. Taken together, the safety data from two different mouse models of AD substantiate a favorable safety profile of the cGMP grade AV-1959D vaccine supporting its progression to first-in-human clinical trials.

Introduction: In former Soviet Union countries, tuberculosis (TB) financing largely relies on a hospital-centered model. The World Health Organization favors transformation to ambulatory treatment since it is cheaper and patient-centered. We explored policy and decision maker’s perspectives on: a) enabling factors for transformation in Armenia and b) challenges and ways forward in doing so in Ukraine and Tajikistan. Methodology: Qualitative study of key informants from government, donors and the national TB program. Results: 52 informants with a mean service record of 20 years were involved. Key enablers in Armenia included collaborative partnership and political will, carefully selecting an adapted financing scheme that avoided financial penalization of hospitals and health workers, and use of operational research. The operational challenges in Ukraine and Tajikistan hovered around the lack of technical capacity and guidance on “how to implement” alternative financing. Shortcomings in strategic planning, uncertainty/fear that existing hospital funding would be cut and reluctance to change were highlighted. Suggested ways forwards to change the current paradigm included country-level technical assistance, capacity building, regional exchanges and operational research. Conclusions: the perspectives of “those who decide” on transforming TB financing have been highlighted. Taking these perspectives on-board is vital for achieving the end-TB goals.

Novel dual vaccine, WSN-Aβ 1–10 , based on the recombinant influenza virus, expressing immunodominant B-cell epitope of β-amyloid, simultaneously induced therapeutically potent anti-Aβ and anti-influenza antibodies. In this study we showed that boosting of WSN-WT primed mice with WSN-Aβ 1–10 enhances anti-viral, but fails to induce anti-Aβ antibody responses. This inhibition is associated with expression of Aβ 1–10 within the context of an inactivated influenza virus vaccine. These results demonstrate that the use of an inactivated influenza virus as a carrier for AD vaccine may not be applicable due to the possible inhibition of anti-Aβ antibody response in individuals previously vaccinated or infected with influenza.

Co-ordination is crucial to find the best way to finance TB services to ensure quality care and cost-effectiveness http://ow.ly/AkdQL

Based on cost-effectiveness considerations and on the risk of nosocomial transmission of Mycobacterium tuberculosis strains, the World Health Organization (WHO) and other international institutions recommend to minimize unnecessary hospitalization of tuberculosis (TB) cases. As in other Former Soviet Union (FSU) countries, the organisations of TB services in Armenia are heavily based on vertical and specialized services. In Armenia, a country of 3.2 million inhabitants, TB is one of the major public health problems with overall notification and estimated mortality rates of 41 and 8.8 cases per 100,000 population in 2011, respectively. The prevalence of multidrug-resistant (MDR-TB) was 9.4% and 43.2% among new and previously treated cases, respectively, with 11.9% of them estimated to be extensively drug-resistant (XDR-TB). Specialized TB services include 9 in-patient TB departments, 73 out-patient TB units (called TB cabinets) based in general health-care facilities, and 31 laboratories providing sputum smear microscopy, while culture and drug susceptibility testing is performed in the National TB Reference Laboratory. Following WHO recommendations, the Armenian National TB Programme (NTP) expanded the basic package of services provided in out-patient settings, allowing free access/free of charge TB diagnosis and treatment to the population, to achieve the global TB targets in the WHO European Region under the National Strategic Plan 2007-2015 and Consolidated Action Plan to Prevent and Combat M/XDR-TB in the WHO European Region 2011-2015.

Although β-amyloid (Aβ) may be the primary driver of Alzheimer’s disease (AD) pathology, accumulation of pathological tau correlates with dementia in AD patients. Thus, the prevention/inhibition of AD may require vaccine/s targeting Aβ and tau simultaneously or sequentially. Since high antibody titers are required for AD vaccine efficacy, we have decided to generate vaccines, targeting Aβ (AV-1959R), Tau (AV-1980R) or Aβ/tau (AV-1953R) B cell epitopes, based on immunogenic MultiTEP platform and evaluate the immunogenicity of these vaccines formulated with AdvaxCpG, delta inulin, Alhydrogel®, Montanide-ISA51, Montanide-ISA720, MPLA-SM pharmaceutical grade adjuvants. Formulation of AV-1959R in AdvaxCpG induced the highest cellular and humoral immune responses in mice. The dual-epitope vaccine, AV-1953R, or the combination of AV-1959R and AV-1980R vaccines formulated with AdvaxCpG induced robust antibody responses against various forms of both, Aβ and tau pathological molecules. While anti-Aβ antibody titers after AV-1953R immunization were similar to that in mice vaccinated with AV-1959R or AV-1959R/AV-1980R combination, anti-tau titers were significantly lower after AV-1953R injection when compared to the AV-1980R or AV-1959R/AV-1980R. In silico 3D-modeling provided insight into the differences in immunogenicity of these vaccine constructs. In sum, AV-1959R and AV-1980R formulated with AdvaxCpG adjuvant were identified as promising immunogenic vaccines for ongoing pre-clinical assessment and future human clinical trials.

Background Previously we demonstrated that the resection of primary 4T1 tumors only slightly prolongs mouse survival, but importantly, creates a “window of opportunity” with attenuated suppressor cell and increased activated T cell populations. This suggests that additional activation of the immune system by immunostimulatory agents during this period may enhance anti-tumor immunity and potentially eradicate micro-metastatic disease in this stringent model. We hypothesized that the immunostimulator Immunomax®, which is comprised of a plant-derived polysaccharide, is non-toxic in humans and stimulates immune defense during the infectious diseases treatment, may have also anti-tumor activity and be beneficial in the adjuvant setting when endogenous anti-tumor responses are present and during the “window of opportunity” in post-resection metastatic breast cancer model. Here we provide the initial report that Immunomax® demonstrates the capacity to eliminate micro-metastatic disease in the post-resection, 4T1 mouse model of breast cancer. Methods The efficacy of Immunomax® was evaluated by analyzing survival rate and the number of spontaneous clonogenic tumor cells in the lung homogenates of mice. The frequencies of activated NK, CD4+ and CD8+ cells as well as myeloid-derived suppressor cells and Treg cells were evaluated using flow cytometry. Highly purified mouse and human dendritic and NK cells were sorted and the effect of Immunomax® on activation status of these cells was assessed by flow cytometry. The property of Immunomax® as TLR-4 agonist was determined by NF-κB/SEAP reporter gene assay, WB, RT-PCR. Results Immunomax® injections significantly prolonged overall survival and cured 31% of mice. This immunostimulator activates DCs via the TLR-4, which in turn stimulates tumoricidal NK cells and in vitro, completely inhibits growth of 4T1 cells. Incubation of PBMC from healthy donors with Immunomax® activates NK cells via activation of plasmacytoid DC leading significantly higher efficacy in killing of human NK-target cells K562 compared with non-treated cells. Conclusion This is the first demonstration that Immunomax® is a TLR-4 agonist and the first report of a documented role for this pharmaceutical grade immunostimulator in augmenting anti-tumor activity, suggesting that incorporation of Immunomax® into developing breast cancer therapeutic strategies may be beneficial and with less potential toxicity than checkpoint inhibitors. Electronic supplementary material The online version of this article (doi:10.1186/s12967-014-0322-y) contains supplementary material, which is available to authorized users.

We assessed the performance of decentralised tuberculosis (TB) out-patient centres in tuberculosis (TB) case notification and treatment success in Armenia. An average threshold case notification of ⩾37/100 000 was seen in centres that had higher numbers of presumptive TB patients, where more TB was diagnosed by in-patient facilities and where TB contacts were examined. The number of doctors and/or TB specialists at centres did not influence case notification. Onsite smear microscopy was significantly associated with a treatment success rate of ⩾85% for new TB patients. Addressing specific characteristics of TB centres associated with lower case notification and treatment success and optimising their location may improve performance.

It is believed that primary tumor resection modulates host-tumor immune interaction, but this has not been characterized in a stringent breast cancer tumor model. This report, using the 4T1 murine mammary tumor model, characterizes for the first time the dynamic longitudinal changes in immunosuppressive and effector components of the immune system after resection of an established orthotopic primary tumor with a defined natural history of developing lung metastases. More specifically, we analyzed changes of absolute numbers and frequencies of MDSC, regulatory T cells (Treg), as well as activated CD4 and CD8 positive T cells in spleens and, in some studies, lungs of 4T1 tumor-bearing mice and mice after primary tumor resection. Importantly, using mathematical analyses we established that primary resection of an orthotopic tumor had created a “window of opportunity” with decreased tumor-associated immune suppression that existed for approximately 10 days. Although tumor resection did slightly prolong survival, it did not affect the ultimate development of metastatic disease since animals with resected tumors or intact primary tumors eventually died by day 47 and 43, respectively. This window of opportunity likely occurs in humans providing a rationale and parameters for integration and testing of immunotherapeutic strategies in this critical “window of opportunity” to combat the development of metastatic disease.

Background The development of a safe and effective AD vaccine requires a delicate balance between providing an adequate anti-Aβ antibody response sufficient to provide therapeutic benefit, while eliminating an adverse T cell-mediated proinflammatory autoimmune response. To achieve this goal we have designed a prototype chemokine-based DNA epitope vaccine expressing a fusion protein that consists of 3 copies of the self-B cell epitope of Aβ42 (Aβ1–11) , a non-self T helper cell epitope (PADRE), and macrophage-derived chemokine (MDC/CCL22) as a molecular adjuvant to promote a strong anti-inflammatory Th2 phenotype. Methods and Findings We generated pMDC-3Aβ1–11-PADRE construct and immunized 3xTg-AD mouse model starting at age of 3–4 months old. We demonstrated that prophylactic immunizations with the DNA epitope vaccine generated a robust Th2 immune response that induced high titers of anti-Aβ antibody, which in turn inhibited accumulation of Aβ pathology in the brains of older mice. Importantly, vaccination reduced glial activation and prevented the development of behavioral deficits in aged animals without increasing the incidence of microhemorrhages. Conclusions Data from this transitional pre-clinical study suggest that our DNA epitope vaccine could be used as a safe and effective strategy for AD therapy. Future safety and immunology studies in large animals with the goal to achieve effective humoral immunity without adverse effects should help to translate this study to human clinical trials.

e13156 Background: Immunotherapy using the Venezuelan equine encephalitis replicon (VRP) vector system, with intrinsic tropism for dendritic cells, has shown exceptional promise. Previously, we rep...

It is widely accepted that the genetic instability of a tumor results in both intra- and inter-tumoral genetic heterogeneity, which leads to heterogeneous tumor-associated antigen (TAA) expression and contributes to the outgrowth of antigen-loss variants in response to immunologic selective pressure. Using a rat neu encoding, alphavirus based virus-like replicon particle (VRP) with restricted in vivo tropism for dendritic cells in a fully tolerant Fisher 344 rat mammary tumor (13762 MAT B III) model, we previously demonstrated the ability to cure a modest proportion, 20-40%, of rats with pre-existing mammary tumors and observed the outgrowth of antigen loss variant tumors in a proportion of animals that ultimately succumbed to their tumor. Thus, we hypothesized that strategies targeting multiple tumor antigens would lead to improved treatment efficacy. Expression of the cancer testes antigen, Brother of Regulator of Imprinted Sites (BORIS) was confirmed in the 13762 MAT B III tumor. Rats with pre-existing tumors were treated with VRP encoding either the TAA rat neu or the BORIS target antigen, alone or in combination. We found that targeting either rat neu or BORIS alone was effective in treating a proportion of animals. Interestingly, administration of both VRP agents at a single anatomic site drastically decreased the treatment efficacy, by approximately 50%, while separating their administration into two separate anatomic sites enhanced the treatment efficacy over that provided by each agent alone, to >56% cure rate. Herein, we will present data addressing potential mechanisms for these treatment observations. To our knowledge, this is the first report of multivalent anti-tumor immunotherapy utilizing solely the VRP vector system. These data represent an exceptionally promising platform upon which to build more efficacious immunotherapeutic strategies including those targeting multiple defined TAAs. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 4766.

Although broad clinical benefits of anti-tumor immunotherapy have not been demonstrated, the current data suggest such strategies could be beneficial if capable of inducing strong anti-cancer cellular immune responses. We report testing for the first time, the therapeutic efficacy of a dendritic cell (DC)-based immunotherapy strategy targeting a novel cancer testis (CT) antigen, Brother of Regulator of Imprinted Sites (BORIS), which is an epigenetically acting tumor promoting transcription factor expressed in various human and mouse cancer cells. In addition to therapeutic potency, we evaluated the cellular immune responses by assessing BORIS-specific CD4 T cell proliferation, T cell cytokine production both by intracellular cytokine staining and ELISpot analysis, and CTL responses. We evaluated the impact of this therapy on the development of metastatic disease, by analyzing the number of spontaneous clonogenic metastases after a 10-day culture of lung homogenates in presence of 6-thioguanine. The presence of CD4, CD8, Treg and myeloid derived suppressor cells (MDSCs) in the tumor and in the spleen was analyzed by flow cytometry. DC loaded with zink-finger deleted (mutated) BORIS vaccine (DC/mBORIS) was evaluated in the non-immunogenic, very aggressive and highly metastatic mouse model of 4T1 mammary carcinoma. Vaccination of mice after 4T1 tumor implantation inhibited tumor growth (18.75% mice remained tumor-free) and dramatically lowered number of spontaneous clonogenic metastases (50% mice remained metastases-free). Higher numbers of immune-effector CD4 and CD8 T cells infiltrated the tumors of vaccinated mice compared to control animals. Therapeutic vaccination with DC/mBORIS significantly decreased the numbers of MDSC infiltrating the tumor, but not splenic MDSC, likely associated with the effect of the immunotherapy on tumor burden rather than a direct effect on MDSCs. Currently, we are testing the suppressive activity of splenocytes from tumor-bearing mice as well as MDSC isolated from these splenocytes. These data suggest that mBORIS is an attractive target for immunotherapy having substantial efficacy in this highly aggressive tumor model when incorporated into a DC-based immunotherapy, and which may be enhanced even further in combination with agents to attenuate tumor-associated immune suppression. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr LB-319.