An evaluation of antigen-driven expansion and differentiation of hapten-specific B lymphocytes purified from aged mice

Hapten-specific, trinitrophenyl antigen-binding B cells (TNP-ABC) were purified from inbred strains of mice representative of short-, intermediate-, and long-lived animals. Such populations of B cells were stimulated by either thymus-independent or thymus-dependent antigens in vitro and evaluated for both proliferation and differentiation into antibody-secreting cells. In the thymus-dependent system, the three strains of mice were selected on the basis of all being able to interact appropriately with the same T helper cell line. The results indicate that TNP-ABC purified from aged animals of all three strains responded to both forms of antigenic stimulation similar to TNP-ABC selected from young, littermate control animals. These results are discussed in terms of concepts of intrinsic B cell defects during the aging process.     

Establishment of the biological control agent Hypocosmia pyrochroma for Dolichandra unguis-cati (Bignoniaceae) is limited by microclimate

Cat's claw creeper, Dolichandra unguis-cati (Bignoniaceae), a perennial woody vine native to tropical America, is a target for biological control in Australia and South Africa. The cat's claw creeper leaf-tying moth Hypocosmia pyrochroma (Lepidoptera: Pyralidae) from tropical South America was released as a biological control agent for cat's claw creeper in Australia from 2007 to 2010. A total of 2,277 adults, 837 pupae and 77,250 larvae were released at 40 sites in Queensland and New South Wales. Releases were made mostly in open fields (85%), and at limited sites (15%) in insect-proof cages erected over naturally occurring cat's claw creeper infestations in the field. Sampling was conducted annually in spring and autumn to monitor the establishment and dispersal of H. pyrochroma. Establishment of H. pyrochroma was first noticed in 2012 at three release sites and since then the number of established sites has increased to 80 in 2020. Establishment was evident on both ‘short-pod’ and ‘long-pod’ forms of cat's claw creeper and was more widespread in sites where releases were made within insect-proof field cages (50%) than in sites with open field releases (9%). The moth was active from late spring to late autumn with peak larval activity in late summer. To date, all field establishments have been in areas predicted by a CLIMEX model as climatically suitable but restricted mostly to riparian environment (93% of establishment), where the moth has continued to spread from 1.5 to 23 km from release sites. In contrast, there is the only limited establishment and spread in non-riparian corridors, highlighting the role of microclimate (riparian) as a limiting factor for establishment and spread. Future efforts will focus on redistribution of the agent to river/creek systems where the moth is currently not present.     

Reducing Stock-Outs of Life Saving Malaria Commodities Using Mobile Phone Text-Messaging: SMS for Life Study in Kenya

Background

Health facility stock-outs of life saving malaria medicines are common across Africa. Innovative ways of addressing this problem are urgently required. We evaluated whether SMS based reporting of stocks of artemether-lumefantrine (AL) and rapid diagnostic tests (RDT) can result in reduction of stock-outs at peripheral facilities in Kenya.

Methods/Findings

All 87 public health facilities in five Kenyan districts were included in a 26 week project. Weekly facility stock counts of four AL packs and RDTs were sent via structured incentivized SMS communication process from health workers’ personal mobile phones to a web-based system accessed by district managers. The mean health facility response rate was 97% with a mean formatting error rate of 3%. Accuracy of stock count reports was 79% while accuracy of stock-out reports was 93%. District managers accessed the system 1,037 times at an average of eight times per week. The system was accessed in 82% of the study weeks. Comparing weeks 1 and 26, stock-out of one or more AL packs declined by 38 percentage-points. Total AL stock-out declined by 5 percentage-points and was eliminated by the end of the project. Stock-out declines of individual AL packs ranged from 14 to 32 percentage-points while decline in RDT stock-outs was 24 percentage-points. District managers responded to 44% of AL and 73% of RDT stock-out signals by redistributing commodities between facilities. In comparison with national trends, stock-out declines in study areas were greater, sharper and more sustained.

Conclusions

Use of simple SMS technology ensured high reporting rates of reasonably accurate, real-time facility stock data that were used by district managers to undertake corrective actions to reduce stock-outs. Future work on stock monitoring via SMS should focus on assessing response rates without use of incentives and demonstrating effectiveness of such interventions on a larger scale.     

Opportunistic Predation of Wild Turkey Nests by Wild Pigs

Wild pigs (Sus scrofa; i.e., feral hogs, feral swine) are considered an invasive species in the United States. Where they occur, they damage agricultural crops and wildlife habitat. Wild pigs also depredate native wildlife, particularly ground-nesting bird species during nesting season. In areas inhabited by wild turkeys (Meleagris gallopavo), nest destruction caused by wild pigs may affect recruitment. There is debate whether wild pigs actively seek ground-nesting bird nests or depredate them opportunistically. To address this debate, in 2016 we examined the movements of wild pigs relative to artificial wild turkey nests (i.e., control [no artificial nests], moderate density [12.5–25 nests/km²], and high density [25–50 nests/km²]) throughout the nesting season (i.e., early, peak, and late) in south-central Texas, USA. We found no evidence that wild pigs learned to seek and depredate wild turkey nests relative to nest density or nesting periods. Despite wild pigs being important nest predators, depredation was not a functional response to a pulsed food resource and can only be associated with overlapping densities of wild pigs and nests. Protecting reproductive success of wild turkeys will require reducing wild pig densities in nesting habitat prior to nesting season. © 2019 The Wildlife Society.     

Characterization of the complex formed between a potent neutralizing ovine-derived polyclonal anti-TNFα Fab fragment and human TNFα

TNFα (tumour necrosis factor α) is an early mediator in the systemic inflammatory response to infection and is therefore a therapeutic target in sepsis. AZD9773 is an ovine-derived, polyclonal anti-TNFα Fab fragment derived from a pool of serum and currently being developed as a treatment for severe sepsis and septic shock. In the present study, we show that although AZD9773 has a modest affinity for TNFα in a binding assay, the K_i in a cell-based assay is approximately four orders of magnitude lower. We show using SEC (size exclusion chromatography) that the maximum size of the complex between AZD9773 and TNFα is consistent with approximately 12 Fabs binding to one TNFα trimer. A number of approaches were taken to map the epitopes recognized by AZD9773. These revealed that a number of different regions on TNFα are involved in binding to the polyclonal Fab. The data suggest that there are probably three epitopes per monomer that are responsible for most of the inhibition by AZD9773 and that all three can be occupied at the same time in the complex. We conclude that AZD9773 is clearly demonstrated to bind to multiple epitopes on TNFα and suggest that the polyclonal nature may account, at least in part, for the very high potency observed in cell-based assays.     

A nuclear lamina‐chromatin‐Ran GTPase axis modulates nuclear import and DNA damage signaling

The Ran GTPase regulates nuclear import and export by controlling the assembly state of transport complexes. This involves the direct action of RanGTP, which is generated in the nucleus by the chromatin‐associated nucleotide exchange factor, RCC1. Ran interactions with RCC1 contribute to formation of a nuclear:cytoplasmic (N:C) Ran protein gradient in interphase cells. In previous work, we showed that the Ran protein gradient is disrupted in fibroblasts from Hutchinson–Gilford progeria syndrome (HGPS) patients. The Ran gradient disruption in these cells is caused by nuclear membrane association of a mutant form of Lamin A, which induces a global reduction in heterochromatin marked with Histone H3K9me3 and Histone H3K27me3. Here, we have tested the hypothesis that heterochromatin controls the Ran gradient. Chemical inhibition and depletion of the histone methyltransferases (HMTs) G9a and GLP in normal human fibroblasts reduced heterochromatin levels and caused disruption of the Ran gradient, comparable to that observed previously in HGPS fibroblasts. HMT inhibition caused a defect in nuclear localization of TPR, a high molecular weight protein that, owing to its large size, displays a Ran‐dependent import defect in HGPS. We reasoned that pathways dependent on nuclear import of large proteins might be compromised in HGPS. We found that nuclear import of ATM requires the Ran gradient, and disruption of the Ran gradient in HGPS causes a defect in generating nuclear γ‐H2AX in response to ionizing radiation. Our data suggest a lamina–chromatin–Ran axis is important for nuclear transport regulation and contributes to the DNA damage response.