A Phase 1/2 study of the PD-L1 inhibitor, BGB-A333, alone and in combination with the PD-1 inhibitor, tislelizumab, in patients with advanced solid tumours.

BACKGROUND: Many patients do not respond or eventually relapse on treatment with programmed cell death protein-1 (PD-1)/programmed death-ligand 1 (PD-L1) checkpoint inhibitors due to secondary or acquired resistance; therefore, there is a need to investigate novel PD-1/PD-L1 inhibitors. METHODS: This open-label, non-randomised study investigated the safety and anti-tumour activity of BGB-A333, a PD-L1 inhibitor, alone and in combination with tislelizumab in patients with advanced solid tumours with progression during/after standard therapy. The primary objectives were to determine the recommended Phase 2 dose (RP2D), safety and tolerability for BGB-A333 alone and in combination with tislelizumab (Phase 1a/1b) and to determine the overall response rate (ORR) with BGB-A333 plus tislelizumab (Phase 2). RESULTS: Overall, 39 patients across Phase 1a (N = 15), 1b (N = 12) and 2 (N = 12) were enroled. In Phase 1a, an RP2D of 1350 mg was determined. In Phase 1a and 1b/2, serious treatment-emergent adverse events (TEAEs) were reported in five and eight patients, respectively. Two patients experienced TEAEs that led to death. In Phase 2, the ORR was 41.7% (n = 5/12; 95% confidence interval: 15.17%, 72.33%). CONCLUSIONS: TEAEs reported with BGB-A333 were consistent with other PD-L1 inhibitors. Encouraging preliminary anti-tumour activity was observed with BGB-A333 in combination with tislelizumab. CLINICAL TRIAL REGISTRATION: NCT03379259.

Protein over-expression in Escherichia coli triggers adaptation analogous to antimicrobial resistance.

BACKGROUND: The E. coli pET system is the most widely used protein over-expression system worldwide. It relies on the assumption that all cells produce target protein and it is generally believed that integral membrane protein (IMP) over-expression is more toxic than their soluble counterparts. RESULTS: Using GFP-tagged proteins, high level over-expression of either soluble or IMP targets results in > 99.9% cell loss with survival rate of only < 0.03%. Selective pressure generates three phenotypes: large green, large white and small colony variants. As a result, in overnight cultures, ~ 50% of the overall cell mass produces no protein. Genome sequencing of the phenotypes revealed genomic mutations that causes either the loss of T7 RNAP activity or its transcriptional downregulation. The over-expression process is bactericidal and is observed for both soluble and membrane proteins. CONCLUSIONS: We demonstrate that it is the act of high-level over-expression of exogenous proteins in E. coli that sets in motion a chain of events leading to > 99.9% cell death. These results redefine our understanding of protein over-production and link it to the adaptive survival response seen in the development of antimicrobial resistance.

Efflux drug transporters at the forefront of antimicrobial resistance.

Bacterial antibiotic resistance is rapidly becoming a major world health consideration. To combat antibiotics, microorganisms employ their pre-existing defence mechanisms that existed long before man's discovery of antibiotics. Bacteria utilise levels of protection that range from gene upregulation, mutations, adaptive resistance, and production of resistant phenotypes (persisters) to communal behaviour, as in swarming and the ultimate defence of a biofilm. A major part of all of these responses involves the use of antibiotic efflux transporters. At the single cell level, it is becoming apparent that the use of efflux pumps is the first line of defence against an antibiotic, as these pumps decrease the intracellular level of antibiotic while the cell activates the various other levels of protection. This frontline of defence involves a coordinated network of efflux transporters. In the future, inhibition of this efflux transporter network, as a target for novel antibiotic therapy, will require the isolation and then biochemical/biophysical characterisation of each pump against all known and new antibiotics. This depth of knowledge is required so that we can fully understand and tackle the mechanisms of developing antimicrobial resistance.

Hemizygous loss of function mutations in CLCN5 causing end-stage kidney disease without Dent disease phenotype.

Dent disease type 1 is suspected in the presence of a complete phenotype of low molecular weight (LMW) proteinuria, hypercalciuria and at least one of the following: nephrocalcinosis, nephrolithiasis, haematuria, hypophosphatemia or chronic kidney disease (CKD). We present two brothers who presented with CKD alone. In the absence of typical clinical features, further assessment of LMW proteinuria and hypercalciuria was not undertaken. Whole-genome sequencing revealed hemizygous loss of function mutations in chloride voltage-gated channel 5 (CLCN5) consistent with Dent disease. Dent disease should, therefore, be considered in patients with an incomplete phenotype, including unexplained CKD alone.

Aspirations and compromises: changes in homestead space relations of the extreme poor after disaster.

BACKGROUND: Construction of houses in homesteads and their settings occur in the context of traditional perceptions and practices in the rural culture of Bangladesh. Functional spaces inside and around the house are produced according to need over time. Inhabitants construct their houses with locally available resources and knowledge. After devastating disasters houses are delivered as products by the development agencies to quickly cater to the needs of the sufferers. The extreme poor are the receivers and inhabitants of these new houses, which can cause significant changes in the physical and environmental characteristics of the neighborhood. In this regard the building and dwelling values of the inhabitants in relation with these houses may be changed or lost. But these values are otherwise inherent characters of the rural houses in the habitations that are shaped by the aspirations of the dwellers. METHODS AND FINDINGS: This paper investigates how relief houses serve the needs of the extreme poor after disasters and how these houses gradually blend with the surrounding environment matching with the aspirations of the inhabitants. The methodology followed was observation of the backgrounds of the pre and post disaster situations, focus group discussions, drawings sessions and interviews with the inhabitants, craftsmen and locals, use of secondary sources, and visits to the houses during and after construction to understand the techniques and space value. CONCLUSIONS: The present practice of distribution of relief houses without involvement of the owners either in the information sharing or building processes and without understanding owners' perceptions about dwellings, may compromise the compatibility and hence the sustainability of relief houses. Hence, houses may only be used as temporary or transitional shelters to sustain life in the disaster phase, and will not be used as "houses" long term.

Plasmapheresis in nephrology: an update.

PURPOSE OF REVIEW: In the past, recommendations for the use of plasmapheresis were based on findings reported from pilot studies or anecdotes. New results from several randomized controlled trials have changed the indications for the use of plasma exchange. RECENT FINDINGS: A large randomized controlled study of patients with antineutrophil cytoplasmic antibody associated vasculitis showed benefit of plasmapheresis in those with severe renal disease. Patients receiving plasmapheresis compared with methylprednisolone as adjuvant therapy were more likely to be alive and dialysis independent. Plasmapheresis, following publication of a recent randomized controlled trial, should no longer be used for patients with myeloma and acute renal failure. Standard therapy with five to seven plasma exchanges was compared with standard therapy alone. There was no difference in those patients reaching the composite endpoints between the two treatments. New indications include desensitization protocols, using plasmapheresis and intravenous immunoglobulin, which have allowed transplantation across immunological barriers. Highly sensitized patents and ABO incompatible patients compared with their potential donors are now being transplanted with excellent results. Studies still need to be done to assess the best desensitization protocol. SUMMARY: The use of plasmapheresis requires further validation by randomized clinical trials. Recent published trials should alter practice but further studies are required.

Crystal structure of the potassium channel KirBac1.1 in the closed state.

The KirBac1.1 channel belongs to the inward-rectifier family of potassium channels. Here we report the structure of the entire prokaryotic Kir channel assembly, in the closed state, refined to a resolution of 3.65 angstroms. We identify the main activation gate and structural elements involved in gating. On the basis of structural evidence presented here, we suggest that gating involves coupling between the intracellular and membrane domains. This further suggests that initiation of gating by membrane or intracellular signals represents different entry points to a common mechanistic pathway.