A number of new tuberculosis (TB) vaccines have been or are entering clinical trials, which include genetically modified mycobacteria, mycobacterial antigens delivered by viral vectors, or ...mycobacterial antigens in adjuvant. Some of these vaccines aim to replace the existing BCG vaccine but others will be given as a boosting vaccine following BCG vaccination given soon after birth. It is clear that the existing BCG vaccines provide incomplete and variable protection against pulmonary TB. This review will discuss what we have learnt over the last 20 years about how the BCG vaccine induces specific and non-specific immunity, what factors influence the immune responses induced by BCG, and progress toward identifying correlates of immunity against TB from BCG vaccination studies. There is still a lot to learn about the BCG vaccine and the insights gained can help the development of more protective vaccines.
GIAO NMR shift calculation has been applied to the challenging task of reliably assigning stereochemistry with quantifiable confidence when only one set of experimental data are available. We have ...compared several approaches for assigning a probability to each candidate structure and have tested the ability of these methods to distinguish up to 64 possible diastereoisomers of 117 different molecules, using NMR shifts obtained in rapid and computationally inexpensive single-point calculations on molecular mechanics geometries without time-consuming ab initio geometry optimization. We show that a probability analysis based on the errors in each 13C or 1H shift is significantly more successful at making correct assignments with high confidence than are probabilities based on the correlation coefficient and mean absolute error parameters. Our new probability measure, which we have termed DP4, complements the probabilities obtained from our previously developed CP3 parameter, which applies to the case of assigning a pair of diastereoisomers when one has both experimental data sets. We illustrate the application of DP4 to assigning the stereochemistry or structure of 21 natural products that were originally misassigned in the literature or that required extensive synthesis of diastereoisomers to establish their stereochemistry.
Since the emergence of the biopharmaceutical industry in the 1980's, Escherichia coli, has played an important role in the industrial production of recombinant proteins and plasmid DNA for ...therapeutic use. Currently, advanced biopharmaceutical products, including rationally designed recombinant proteins and viral-vector gene therapies, offer unprecedented promise for the long-term management, and even cure of disease. As such, E. coli remains an important production host for the biopharmaceutical industry. This review provides insight into the industrially relevant strain engineering approaches used to enhance both the quantity and quality of these therapeutic products.
The bromodomain (BrD) is a conserved protein modular domain found in many chromatin- and transcription-associated proteins that has the ability to recognize acetylated lysine residues. This activity ...allows bromodomains to play a vital role in many acetylation-mediated protein–protein interactions in the cell, ranging from substrate recruitment for histone acetyltransferases (HATs) to aiding in multiple-protein complex assembly for gene transcriptional activation or suppression in chromatin. In recent years, considerable efforts have been made to develop chemical inhibitors of these bromodomains in an effort to probe their cellular functions. Potent and selective inhibitors have been extensively developed for one group of the bromodomain family termed BET proteins that consist of tandem bromodomains followed by an extra terminal domain. Drug developers are actively designing inhibitors of other bromodomains and working to move the most successful inhibitors into the clinic.
The long-term efficacy and safety of mepolizumab for treatment of severe eosinophilic asthma are well established. Here, we examine the clinical impact of stopping mepolizumab after long-term use.
...COMET (NCT02555371) was a randomised, double-blind, placebo-controlled, parallel-group, multicentre study. Patients who had completed COLUMBA (NCT01691859) or COSMEX (NCT02135692) and received continuous mepolizumab treatment for ≥3 years were randomised 1:1 to stop (switch to placebo) or continue subcutaneous mepolizumab 100 mg every 4 weeks for 52 weeks. Primary end-point: time to first clinically significant exacerbation; secondary end-points: time to first exacerbation requiring hospitalisation/emergency department visit, time to decrease in asthma control (≥0.5-point increase in Asthma Control Questionnaire-5 score from COMET baseline) and blood eosinophil count ratio to COMET baseline. Safety was assessed.
Patients stopping (n=151)
continuing (n=144) mepolizumab had significantly shorter times to first clinically significant exacerbation (hazard ratio 1.61, 95% CI 1.17-2.22; p=0.004) and decrease in asthma control (hazard ratio 1.52, 95% CI 1.13-2.02; p=0.005), and higher blood eosinophil counts at week 52 (270
40 cells·µL
; ratio (stopping
continuing) 6.19, 95% CI 4.89-7.83; p<0.001). Differences in efficacy outcomes between groups were observed when assessed from week 12 (16 weeks after last mepolizumab dose). Exacerbations requiring hospitalisation/emergency department visit were rare. Adverse events in patients continuing mepolizumab were consistent with previous studies. For patients who stopped mepolizumab, the safety profile was consistent with other eosinophilic asthma populations.
Patients who stopped mepolizumab had an increase in exacerbations and reduced asthma control
those who continued.
GIAO NMR chemical shifts have been calculated for a set of 28 pairs of diastereoisomers in order to test the ability of NMR shift calculation to distinguish between diastereomeric structures. We ...compare the performance of several different parameters for quantifying the agreement between calculated and experimental shifts from the point of view of assigning structures and introduce a new parameter, CP3, based on comparing differences in calculated shift with differences in experimental shift, which is significantly more successful at making correct structure assignments with high confidence than are the currently used parameters of the mean absolute error and the correlation coefficient. Using our new parameter in conjunction with Bayes’ theorem, stereostructure assignments can be made with quantifiable confidence using shifts obtained in single point calculations on molecular mechanics geometries without computationally expensive ab initio geometry optimization.
Background In patients with severe eosinophilic asthma, local maturation rather than systemic recruitment of mature cells might contribute to persistent airway eosinophilia. Group 2 innate lymphoid ...cells (ILC2s) are a major source of type 2 cytokines (IL-5 and IL-13) and can facilitate eosinophilic inflammatory responses in mouse models of asthma in the absence of CD4+ lymphocytes. This study investigated the potential role of ILC2s in driving chronic airway eosinophilia in patients with severe asthma, despite regular high-dose oral corticosteroid therapy. Methods In a cross-sectional study we enumerated blood and sputum ILC2s (lin− CD45+ 127+ ST2+ ) and levels of intracellular IL-5 and IL-13 in patients with severe asthma (n = 25), patients with steroid-naive mild atopic asthma (n = 19), and nonatopic control subjects (n = 5). Results were compared with numbers of CD4+ lymphocytes, eosinophil lineage–committed progenitors (eosinophilopoietic progenitor cells EoPs), and mature eosinophils. Results Significantly greater numbers of total and type 2 cytokine–producing ILC2s were detected in blood and sputum of patients with severe asthma compared to mild asthmatics. In contrast, intracellular cytokine expression by CD4 cells and EoPs within the airways did not differ between the asthmatic groups. In patients with severe asthma, although sputum CD4+ cells were more abundant than ILC2s and EoPs, proportionally, ILC2s were the predominant source of type 2 cytokines. In addition, there were significantly greater numbers of sputum IL-5+ IL-13+ ILC2s in patients with severe asthma whose airway eosinophilia was greater than 3%, despite normal blood eosinophil numbers (<300/μL). Conclusions Our findings suggest that ILC2s can promote the persistence of airway eosinophilia in patients with severe asthma through uncontrolled localized production of the type 2 cytokines IL-5 and IL-13, despite high-dose oral corticosteroid therapy.
Interreligious engagement (IE) has been experienced and theorized mainly as the pursuit of a shared respectful awareness of the beliefs, practices, and social experiences of multiple religious ...communities. In rare instances, it has been possible to create architecture specifically to foster IE, as in the “tri-faith” Abrahamic campus in Omaha and the Berlin House of One. The theme is: Here we are, accepting that we share the world. Another form of IE that deserves to attract more interest is multireligious collaboration in civic work (addressing homelessness, urban blight, illiteracy, etc.). Some adherents of the intrinsically cosmopolitan “world” religions are actively cosmopolitan to the extent of seeking this engagement. The theme is: Let us share the work of the world, including sharing our religiously inflected processing of what the practical issues facing us are. There is a new initiative of this sort in my city, Jackson, Mississippi, named (from M. L. King) the “Beloved Community”. An architectural thought experiment may prove helpful in articulating the ideals for such an endeavor. What would be the physical desiderata for its headquarters? Let us imagine a new downtown building, The Meeting, dedicated to housing meetings where mixed religious groups learn about civic issues and coordinate efforts to address them. Full interreligious sharing of a space seems to require a neutral design lacking any definite religious inspiration. But there are nonsectarian ways to create an appreciably special, non-ordinary space, as in courtrooms and classrooms. Could a civic IE headquarters be special, expressive of practical optimism, and contain a sufficient religious allusion to qualify as a “next-to-sacred space” in which religious actors felt supported in the civic extension of their religious lives? I offer suggestions for discussion, including (1) a pavilion-style building suggestive of being set up for a special purpose—not soaringly grandiose but with a vertical feature such as a central roof lantern; (2) at least one major porch, with benches and tables; (3) an outside water fountain with public water supply (a historical allusion to the Islamic sabil); (4) inside, right-sized meeting rooms around the glass-walled periphery; (5) a big “living room” lounge in the center, usable for larger meetings, with access to a kitchen, and with a big project board for tracking work completed and work in hand next to a large map of the city; (6) a moderate descent of several steps into each meeting room so that there is a feeling of commitment in attending a meeting and a sense of challenge in going forth from one; (7) otherwise a main floor levelness and openness facilitating movement in and out, as in a train station; and (8) upstairs small offices for religious and other qualifying organizations. Answering the aesthetic and practical questions these suggestions raise takes us into imagining civic IE more concretely.
The decrease in sequencing cost and increased sophistication of assembly algorithms for short-read platforms has resulted in a sharp increase in the number of species with genome assemblies. However, ...these assemblies are highly fragmented, with many gaps, ambiguities, and errors, impeding downstream applications. We demonstrate current state of the art for de novo assembly using the domestic goat (Capra hircus) based on long reads for contig formation, short reads for consensus validation, and scaffolding by optical and chromatin interaction mapping. These combined technologies produced what is, to our knowledge, the most continuous de novo mammalian assembly to date, with chromosome-length scaffolds and only 649 gaps. Our assembly represents a ∼400-fold improvement in continuity due to properly assembled gaps, compared to the previously published C. hircus assembly, and better resolves repetitive structures longer than 1 kb, representing the largest repeat family and immune gene complex yet produced for an individual of a ruminant species.
Facilities involved in the manufacture of pharmaceutical products are an under-investigated source of pharmaceuticals to the environment. Between 2004 and 2009, 35 to 38 effluent samples were ...collected from each of three wastewater treatment plants (WWTPs) in New York and analyzed for seven pharmaceuticals including opioids and muscle relaxants. Two WWTPs (NY2 and NY3) receive substantial flows (>20% of plant flow) from pharmaceutical formulation facilities (PFF) and one (NY1) receives no PFF flow. Samples of effluents from 23 WWTPs across the United States were analyzed once for these pharmaceuticals as part of a national survey. Maximum pharmaceutical effluent concentrations for the national survey and NY1 effluent samples were generally <1 μg/L. Four pharmaceuticals (methadone, oxycodone, butalbital, and metaxalone) in samples of NY3 effluent had median concentrations ranging from 3.4 to >400 μg/L. Maximum concentrations of oxycodone (1700 μg/L) and metaxalone (3800 μg/L) in samples from NY3 effluent exceeded 1000 μg/L. Three pharmaceuticals (butalbital, carisoprodol, and oxycodone) in samples of NY2 effluent had median concentrations ranging from 2 to 11 μg/L. These findings suggest that current manufacturing practices at these PFFs can result in pharmaceuticals concentrations from 10 to 1000 times higher than those typically found in WWTP effluents.
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