The BAF complex inhibitor pyrimethamine reverses HIV-1 latency in people with HIV-1 on antiretroviral therapy.

Reactivation of the latent HIV-1 reservoir is a first step toward triggering reservoir decay. Here, we investigated the impact of the BAF complex inhibitor pyrimethamine on the reservoir of people living with HIV-1 (PLWH). Twenty-eight PLWH on suppressive antiretroviral therapy were randomized (1:1:1:1 ratio) to receive pyrimethamine, valproic acid, both, or no intervention for 14 days. The primary end point was change in cell-associated unspliced (CA US) HIV-1 RNA at days 0 and 14. We observed a rapid, modest, and significant increase in (CA US) HIV-1 RNA in response to pyrimethamine exposure, which persisted throughout treatment and follow-up. Valproic acid treatment alone did not increase (CA US) HIV-1 RNA or augment the effect of pyrimethamine. Pyrimethamine treatment did not result in a reduction in the size of the inducible reservoir. These data demonstrate that the licensed drug pyrimethamine can be repurposed as a BAF complex inhibitor to reverse HIV-1 latency in vivo in PLWH, substantiating its potential advancement in clinical studies.

HIV-1 resistance against dolutegravir fluctuates rapidly alongside erratic treatment adherence: a case report.

OBJECTIVES: We report a case of incomplete HIV-1 suppression on a dolutegravir, lamivudine, and abacavir single-tablet regimen with the emergence of the H51Y and G118R integrase resistance mutations. METHODS: Integrase sequencing was performed retrospectively by Sanger and next-generation sequencing. Rates of emergence and decline of resistance mutations were calculated using next-generation sequencing data. Dolutegravir plasma concentrations were measured by ultra-performance liquid chromatography-tandem mass spectrometry. The effects of H51Y and G118R on infectivity, fitness, and susceptibility to dolutegravir were quantified using cell-based assays. RESULTS: During periods of non-adherence to treatment, mutations were retrospectively documented only by next-generation sequencing. Misdiagnosis by Sanger sequencing was caused by the rapid decline of mutant strains within the retroviral population. This observation was also true for a M184V lamivudine-resistant reverse transcriptase mutation found in association with integrase mutations on single HIV genomes. Resistance rebound upon treatment re-initiation was swift (>8000 copies per day). Next-generation sequencing indicated cumulative adherence to treatment. Compared to WT HIV-1, relative infectivity was 73%, 38%, and 43%; relative fitness was 100%, 35%, and 10% for H51Y, G118R, and H51Y+G118R viruses, respectively. H51Y did not change the susceptibility to dolutegravir, but G188R and H51Y+G118R conferred 7- and 28-fold resistance, respectively. CONCLUSION: This case illustrates how poorly-fit drug-resistant viruses wax and wane alongside erratic treatment adherence and are easily misdiagnosed by Sanger sequencing. We recommend next-generation sequencing to improve the clinical management of incomplete virological suppression with dolutegravir.

Added value of 18F-FDG-PET/CT and cardiac CTA in suspected transcatheter aortic valve endocarditis.

BACKGROUNDS: Transcatheter-implanted aortic valve infective endocarditis (TAVI-IE) is difficult to diagnose when relying on the Duke Criteria. Our aim was to assess the additional diagnostic value of 18F-fluorodeoxyglucose (18F-FDG) positron emission/computed tomography (PET/CT) and cardiac computed tomography angiography (CTA) in suspected TAVI-IE. METHODS: A multicenter retrospective analysis was performed in all patients who underwent 18F-FDG-PET/CT and/or CTA with suspected TAVI-IE. Patients were first classified with Duke Criteria and after adding 18F-FDG-PET/CT and CTA, they were classified with European Society of Cardiology (ESC) criteria. The final diagnosis was determined by our Endocarditis Team based on ESC guideline recommendations. RESULTS: Thirty patients with suspected TAVI-IE were included. 18F-FDG-PET/CT was performed in all patients and Cardiac CTA in 14/30. Using the Modified Duke Criteria, patients were classified as 3% rejected (1/30), 73% possible (22/30), and 23% definite (7/30) TAVI-IE. Adding 18F-FDG-PET/CT and CTA supported the reclassification of 10 of the 22 possible cases as "definite TAVI-IE" (5/22) or "rejected TAVI-IE" (5/22). This changed the final diagnosis to 20% rejected (6/30), 40% possible (12/30), and 40% definite (12/30) TAVI-IE. CONCLUSIONS: Addition of 18F-FDG-PET/CT and/or CTA changed the final diagnosis in 33% of patients and proved to be a valuable diagnostic tool in patients with suspected TAVI-IE.

[Infections during glucocorticoid use]. / Infecties bij gebruik van glucocorticoïden.

Glucocorticoid treatment increases the risk of opportunistic infection. Infections that can arise during glucocorticoid use, and for which preventative measures can be taken, include reactivation of latent tuberculosis and hepatitis B, pneumococcal and Pneumocystis jiroveci pneumonia, influenza, herpes zoster and Strongyloides stercoralis hyperinfection syndrome. The risk of such infections depends upon the duration of glucocorticoid use and dosage, as well as comorbidity and comedication. It is important to enquire about vaccinations, travel, exposure and previous infections when taking a case history. Possible infectious complications should be considered in patients who are receiving high-dose glucocorticoids treatment amounting to more than 420 mg PED per 4 weeks. Preventative measures are not usually required in patients who receive a short high-dosed treatment (30 mg PED in 7 days) or prednisolone at a dosage of < 15 mg/day.

The impact of infections on delisting patients from the liver transplantation waiting list.

Approximately 20% of the patients listed for liver transplantation die before transplantation can be accomplished. Understanding risk factors for waiting list mortality may help to improve survival and organ allocation. Infections are very common in patients with cirrhosis and are associated with significant morbidity and mortality. This study analysed the frequency and characteristics of infections in patients awaiting liver transplantation, identified risk factors for withdrawal from the waiting list and evaluated the impact of infections on the clinical outcome. A retrospective analysis of consecutive patients listed for liver transplantation in Rotterdam, the Netherlands from 2007 to 2014 was conducted. Infections occurred in 144 of 327 studied patients (44%). In this cohort, 23.4% of the patients on the liver transplantation waiting list were delisted or died before transplantation. Patients with an infection were 5.2 times more likely to become delisted than noninfected patients. In the 30 days after the first infection, patients were 33.8 times more likely to become delisted compared to noninfected patients. High age, high MELD score, refractory ascites and inappropriate antibiotic therapy were independent predictors for delisting due to infection. Infections occur frequently in patients on the liver transplantation waiting list. Emphasis on appropriate and timely antimicrobial therapy is required.

Dolutegravir as maintenance monotherapy: first experiences in HIV-1 patients.

BACKGROUND: Dolutegravir is recommended as part of combination ART (cART) for HIV-1-infected patients. Toxicities, drug interactions and costs related to cART still warrant the search for improved treatment options. Dolutegravir's high resistance barrier might make it suitable as antiretroviral maintenance monotherapy. The feasibility of this strategy is currently unknown. METHODS: This is a prospective case series on five consecutive HIV-1-infected patients on cART without previous virological failure who switched to dolutegravir monotherapy. All were HIV-RNA suppressed <50 copies/mL and had contraindications to current and alternative combinations of antiretroviral drugs. HIV-RNA was measured at baseline, week 4, week 8, week 12 and every 6 weeks thereafter. Patients would be switched back to their original cART upon confirmed HIV-RNA >50 copies/mL. RESULTS: The five patients had been HIV-RNA suppressed <50 copies/mL for ≥1.5 years prior to the initiation of dolutegravir monotherapy. All were on NNRTI-containing regimens at the switch. HIV-RNA remained <50 copies/mL at all timepoints in four patients. One patient, with end-stage renal disease and on calcium supplements, had a pre-cART HIV-RNA of 625 000 copies/mL with a CD4 nadir of 120 cells/mm(3) and had HIV-RNA of 8150 copies/mL at week 30. The dolutegravir Ctrough was 0.18 mg/L. This patient did not have acquired resistance or evidence of adherence problems and HIV-RNA was resuppressed after switching to his former cART. CONCLUSIONS: This case series indicates that dolutegravir monotherapy might be a valuable maintenance option in selected HIV-infected patients who are well suppressed on cART, if confirmed by future randomized clinical trials.

Aortic homograft endocarditis caused by Campylobacter jejuni.

We report the first case of homograft endocarditis caused by Campylobacter jejuni, which was treated successfully with antibiotic therapy and valve replacement. To our knowledge, only two other cases of C. jejuni endocarditis, involving native valves, have been reported in the medical literature.

Modelling treatment effects in a clinical Bayesian network using Boolean threshold functions.

OBJECTIVE: Appropriate antimicrobial treatment of infections in critically ill patients should be started as soon as possible, as delay in treatment may reduce a patient's prognostic outlook considerably. Ventilator-associated pneumonia (VAP) occurs in patients in intensive care units who are mechanically ventilated and is almost always preceded by colonisation of the respiratory tract by the causative microorganisms. It is very difficult to clinically diagnose VAP and, therefore, some form of computer-based decision support might be helpful for the clinician. MATERIALS AND METHODS: As diagnosing and treating VAP involves reasoning with uncertainty, we have used a Bayesian network as the primary tool for building a decision-support system. The effects of usage of antibiotics on the colonisation of the respiratory tract by various pathogens and the subsequent antibiotic choices in case of VAP were modelled using the notion of causal independence. In particular, the conditional probability distribution of the random variable that represents the overall coverage of pathogens by antibiotics was modelled in terms of the conjunctive effect of the seven different pathogens, usually referred to as the noisy-AND model. In this paper, we investigate different coverage models, as well as generalisations of the noisy-AND, called noisy-threshold models, and test them on clinical data of intensive care unit (ICU) patients who are mechanically ventilated. RESULTS: Some of the constructed noisy-threshold models offered further improvement of the performance of the Bayesian network in covering present causative pathogens by advising appropriate antimicrobial treatment. CONCLUSIONS: By reconsidering the modelling of interactions between the random variables in a Bayesian network using the theory of causal independence, it is possible to refine its performance. This was clearly shown for our Bayesian network concerning VAP, indicating that only specific noisy-threshold models might be appropriate for the modelling of the interaction between pathogens and antimicrobial treatment with respect to susceptibility. The results obtained also provide evidence that the noisy-OR and noisy-AND might not always be the best functions to model interactions among random variables.

Predicting pathogens causing ventilator-associated pneumonia using a Bayesian network model.

BACKGROUND: We previously validated a Bayesian network (BN) model for diagnosing ventilator-associated pneumonia (VAP). Here, we report on the performance of the model to predict microbial causes of VAP and to select antibiotics. METHODS: Pathogens were grouped into seven categories based upon the antibiotic susceptibility and epidemiological characteristics. Colonization of the upper respiratory tract was modelled in the BN and depended--in additional steps--on (i) duration of admission and ventilation, (ii) previous culture results and (iii) previous antibiotic use. A database with 153 VAP episodes and their microbial causes was used as reference standard. Appropriateness of antibiotic prescription, with fixed choices for pathogens predicted, was determined. RESULTS: One hundred and seven VAP episodes were monobacterial and 46 were caused by two pathogens. Using duration of admission and ventilation only, areas under the receiver operating curve (AUC) ranged from 0.511 to 0.772 for different pathogen groups, and model predictions significantly improved when adding information on culture results, but not when adding information on antibiotic use. The best performing model (with all information) had AUC values ranging from 0.859 for Acinetobacter spp. to 0.929 for Streptococcus pneumoniae. With this model, 91 (85%) and 29 (63%) of all pathogen groups were correctly predicted for monobacterial and polymicrobial VAP, respectively. With fixed antibiotic choices linked to pathogen groups, 92% of all episodes would have been treated appropriately. CONCLUSIONS: The BN models' performance to predict pathogens causing VAP improved markedly with information on colonization, resulting in excellent pathogen prediction and antibiotic selection. Prospective external validation is needed.

Effects of systemic antibiotic therapy on bacterial persistence in the respiratory tract of mechanically ventilated patients.

OBJECTIVE: Bacterial respiratory tract colonization predisposes critically ill patients to intensive care unit (ICU)-acquired infections. It is unclear to what extent systemic antibiotics affect colonization persistence. Persistence of respiratory tract colonization, and the effects of systemic antibiotics hereon, were determined in a cohort of ICU patients. DESIGN: Clinical and microbiological data were collected from 715 admitted mechanically ventilated ICU patients with bacterial growth documented in respiratory tract samples. First day of colonization, persistence of colonization and antibiotic effects hereon were analyzed for six groups of pathogens: Pseudomonas aeruginosa, Acinetobacter species, Enterobacteriaceae, Staphylococcus aureus, Streptococcus pneumoniae and Haemophilus influenzae. Systemic antibiotics were grouped into 'effective' and 'ineffective' antibiotics, based on in-vitro susceptibility data for the relevant bacteria. The effects of antibiotics were quantified as relative risk (RR) of bacterial persistence in the absence of effective antibiotics. MEASUREMENTS AND RESULTS: Persistence of colonization differed significantly between pathogens, ranging from 4 days (median) for H. influenzae and Strep. pneumoniae to 8 days for P. aeruginosa. Systemic antibiotics were administered on 7,102 (61%) of patient days. Antibiotic use was associated with non-persistence for all pathogens, except Acinetobacter species and P. aeruginosa. RR for non-persistence (as compared to ineffective or no antibiotics) ranged from 3.1 (95% CI 1.4-6.6) for H. influenzae to 0.5 (0.3-1.0) for Acinetobacter species. CONCLUSIONS: In mechanically ventilated patients, persistence dynamics of bacterial respiratory tract colonization, and the effects of (in-vitro) effective antibiotics hereon, are pathogen-specific.

A Bayesian decision-support system for diagnosing ventilator-associated pneumonia.

OBJECTIVE: To determine the diagnostic performance of a Bayesian Decision-Support System (BDSS) for ventilator-associated pneumonia (VAP). DESIGN: A previously developed BDSS, automatically obtaining patient data from patient information systems, provides likelihood predictions of VAP. In a prospectively studied cohort of 872 ICU patients, VAP was diagnosed by two infectious-disease specialists using a decision tree (reference diagnosis). After internal validation daily BDSS predictions were compared with the reference diagnosis. For data analysis two approaches were pursued: using BDSS predictions (a) for all 9422 patient days, and (b) only for the 238 days with presumed respiratory tract infections (RTI) according to the responsible physicians. MEASUREMENTS AND RESULTS: 157 (66%) of 238 days with presumed RTI fulfilled criteria for VAP. In approach (a), median daily BDSS likelihood predictions for days with and without VAP were 77% [Interquartile range (IQR) = 56-91%] and 14% [IQR 5-42%, p < 0.001, Mann-Whitney U-test (MWU)], respectively. In receiver operating characteristics (ROC) analysis, optimal BDSS cut-off point for VAP was 46%, and with this cut-off point positive predictive value (PPV) and negative predictive value (NPV) were 6.1 and 99.6%, respectively [AUC = 0.857 (95% CI 0.827-0.888)]. In approach (b), optimal cut-off for VAP was 78%, and with this cut-off point PPV and NPV were 86 and 66%, respectively [AUC = 0.846 (95% CI 0.794-0.899)]. CONCLUSIONS: As compared with the reference diagnosis, the BDSS had good test characteristics for diagnosing VAP, and might become a useful tool for assisting ICU physicians, both for routinely daily assessment and in patients clinically suspected of having VAP. Empirical validation of its performance is now warranted.

Clinical pulmonary infection score for ventilator-associated pneumonia: accuracy and inter-observer variability.

OBJECTIVE: Although quantitative microbiological cultures of samples obtained by bronchoscopy are considered the most specific tool for diagnosing ventilator-associated pneumonia, this labor-intensive invasive technique is not widely used. The Clinical Pulmonary Infection Score (CPIS), a diagnostic algorithm that relies on easily available clinical, radiographic, and microbiological criteria, could be an attractive alternative for diagnosing ventilator-associated pneumonia. Initially, the CPIS scoring system was validated upon 40 quantitative cultures of bronchoalveolar lavage fluid from 28 patients, and only few other studies have evaluated this scoring system since then. Therefore, little is known about the accuracy of this score. DESIGN: We compared the scores of a slightly adjusted CPIS with results from quantitative cultures of bronchoalveolar lavage fluid in 99 consecutive patients with suspicion of ventilator-associated pneumonia, using growth of > or =10(4) cfu/ml in bronchoalveolar lavage fluid as a cut-off for diagnosing ventilator-associated pneumonia. In addition, the CPIS were calculated for 52 patients by two different intensivists to determine the inter-observer variability. RESULTS: Ventilator-associated pneumonia was diagnosed in 69 (69.6%) patients. When using a CPIS >5 as diagnostic cutoff, the sensitivity of the score was 83% and its specificity was 17%. The area under the Receiver Operating Characteristic curve was 0.55. The level of agreement for prospectively measured Clinical Pulmonary Infection Score (< or =6 and >6) was poor (kappa =0.16). CONCLUSIONS: When compared to quantitative cultures of bronchoalveolar lavage fluid, the CPIS has a low sensitivity and specificity for diagnosing ventilator-associated pneumonia with considerable inter-observer variability.