Assessing the effectiveness of a community intervention for monkeypox prevention in the Congo basin

Background

In areas where health resources are limited, community participation in the recognition and reporting of disease hazards is critical for the identification of outbreaks. This is particularly true for zoonotic diseases such as monkeypox that principally affect people living in remote areas with few health services. Here we report the findings of an evaluation measuring the effectiveness of a film-based community outreach program designed to improve the understanding of monkeypox symptoms, transmission and prevention, by residents of the Republic of the Congo (ROC) who are at risk for disease acquisition.

Methodology/Principal Findings

During 90 days, monkeypox outreach was conducted for ∼23,860 people in northern ROC. Two hundred seventy-one attendees (selected via a structured sample) were interviewed before and after participating in a small-group outreach session. The proportion of interviewees demonstrating monkeypox-specific knowledge before and after was compared. Significant gains were measured in areas of disease recognition, transmission, and mitigation of risk. The ability to recognize at least one disease symptom and a willingness to take a family member with monkeypox to the hospital increased from 49 and 45% to 95 and 87%, respectively (p<0.001, both). Willingness to deter behaviors associated with zoonotic risk, such as eating the carcass of a primate found dead in the forest, remained fundamentally unchanged however, suggesting additional messaging may be needed.

Conclusions/Significance

These results suggest that our current program of film-based educational activities is effective in improving disease-specific knowledge and may encourage individuals to seek out the advice of health workers when monkeypox is suspected.     

Nitrogen fertilizer and landscape position impacts on CO2 and CH4 fluxes from a landscape seeded to switchgrass

This study was conducted to evaluate the impacts of N fertilizer and landscape position on carbon dioxide (CO₂) and methane (CH₄) fluxes from a US Northern Great Plains landscape seeded to switchgrass (Panicum virgatum L.). The experimental design included three N levels (low, 0 kg N ha⁻¹; medium, 56 kg N ha⁻¹; and high, 112 kg N ha⁻¹) replicated four times. The experiment was repeated at shoulder and footslope positions. Soil CO₂ and CH₄ fluxes were monitored once every 2 weeks from May 2010 to October 2012. The CO₂ fluxes were 40% higher at the footslope than the shoulder landscape position, and CH₄ fluxes were similar in both landscape positions. Soil CO₂ and CH₄ fluxes averaged over the sampling dates were not impacted by N rates. Seasonal variations showed highest CO₂ release and CH₄ uptake in summer and fall, likely due to warmer and moist soil conditions. Higher CH₄ release was observed in winter possibly due to increased anaerobic conditions. However, year to year (2010–2012) variations in soil CO₂ and CH₄ fluxes were more pronounced than the variations due to the impact of landscape positions and N rates. Drought conditions reported in 2012, with higher annual temperature and lower soil moisture than long-term average, resulted in higher summer and fall CO₂ fluxes (between 1.3 and 3 times) than in 2011 and 2010. These conditions also promoted a net CH₄ uptake in 2012 in comparison to 2010 when there was net CH₄ release. Results from this study conclude that landscape positions, air temperature, and soil moisture content strongly influenced soil CO₂ fluxes, whereas soil moisture impacted the direction of CH₄ fluxes (uptake or release). However, a comprehensive life cycle analysis would be appropriate to evaluate environmental impacts associated with switchgrass production under local environmental conditions.     

Downstream bioprocessing of human pluripotent stem cell‐derived therapeutics

With the advancement in lineage‐specific differentiation from human pluripotent stem cells (hPSCs), downstream cell separation has now become a critical step to produce hPSC‐derived products. Since differentiation procedures usually result in a heterogeneous cell population, cell separation needs to be performed either to enrich the desired cell population or remove the undesired cell population. This article summarizes recent advances in separation processes for hPSC‐derived cells, including the standard separation technologies, such as magnetic‐activated cell sorting, as well as the novel separation strategies, such as those based on adhesion strength and metabolic flux. Specifically, the downstream bioprocessing flow and the identification of surface markers for various cell lineages are discussed. While challenges remain for large‐scale downstream bioprocessing of hPSC‐derived cells, the rational quality‐by‐design approach should be implemented to enhance the understanding of the relationship between process and the product and to ensure the safety of the produced cells.     

Fluid shear stress stimulates breast cancer cells to display invasive and chemoresistant phenotypes while upregulating PLAU in a 3D bioreactor

Breast cancer cells experience a range of shear stresses in the tumor microenvironment (TME). However most current in vitro three-dimensional (3D) models fail to systematically probe the effects of this biophysical stimuli on cancer cell metastasis, proliferation, and chemoresistance. To investigate the roles of shear stress within the mammary and lung pleural effusion TME, a bioreactor capable of applying shear stress to cells within a 3D extracellular matrix was designed and characterized. Breast cancer cells were encapsulated within an interpenetrating network hydrogel and subjected to shear stress of 5.4 dynes cm⁻² for 72 hr. Finite element modeling assessed shear stress profiles within the bioreactor. Cells exposed to shear stress had significantly higher cellular area and significantly lower circularity, indicating a motile phenotype. Stimulated cells were more proliferative than static controls and showed higher rates of chemoresistance to the anti-neoplastic drug paclitaxel. Fluid shear stress-induced significant upregulation of the PLAU gene and elevated urokinase activity was confirmed through zymography and activity assay. Overall, these results indicate that pulsatile shear stress promotes breast cancer cell proliferation, invasive potential, chemoresistance, and PLAU signaling.     

Diminished AMPK signaling response to fasting in thioredoxin-interacting protein knockout mice

Thioredoxin-interacting protein (Txnip) knockout (TKO) mice exhibit impaired response to fasting. Herein, we showed that activation of adenine monophosphate-activated protein kinase and cellular AMP levels were diminished in the heart and soleus muscle but not in gastrocnemius muscle of fasting TKO mice. Similarly, glycogen content in fasted TKO mice was increased in oxidative muscles but was not different in glycolytic muscles. These data suggest Txnip deficiency has a higher impact on oxidative muscle than glycolytic muscles and provide new insights into the metabolic role of Txnip.     

A genetic map of tetraploid <Emphasis Type="Italic">Paspalum notatum</Emphasis> Flügge (bahiagrass) based on single-dose molecular markers

Paspalum notatum Flügge is a warm-season forage grass with mainly diploid (2n = 20) and autotetraploid (2n = 40) representatives. Diploid races reproduce sexually and require crosspollination due to a self-incompatible mating system, while autotetraploids reproduce by aposporous apomixis. The objectives of this work were to develop a genetic linkage map of Paspalum notatum Flügge at the tetraploid level, identify the linkage/s group/s associated with apomixis and carry out a general characterization of its mode of inheritance. A pseudo test-cross F₁ family of 113 individuals segregating for the mode of reproduction was obtained by crossing a synthetic completely sexual tetraploid plant (Q4188) as female parent with a natural aposporous individual (Q4117) as pollen donor. Map construction was based on single-dose markers (SDAFs) segregating from both parents. Two linkage maps (female and male) were constructed. Within each map, homologous groups were assembled by detecting repulsion-phase linked SDAFs. Putative Q4188 and Q4117 homolog groups were identified by mapping shared single dose markers (BSDF). The Q4188 map consisted of 263 markers distributed on 26 co-segregation groups over a total genetic distance of 1.590.6 cM, while the Q4117 map contained 216 loci dispersed on 39 co-segregation groups along 2.265.7 cM, giving an estimated genome coverage of 88% and 83%, respectively. Seven and 12 putative homologous chromosomes were detected within Q4188 and Q4117 maps, respectively. Afterward, ten female and male homologous chromosomes were identified by mapping BSDFs. In the Q4117 map, a single linkage group was associated with apospory. It was characterized by restriction in recombination and preferential chromosome pairing. A BPSD marker mapping within this group allowed the detection of the female homolog and the putative four male groups of the set carrying apospory.     

Conditional deletion of Shp2 tyrosine phosphatase in thymocytes suppresses both pre-TCR and TCR signals

It is well known that T cell differentiation and maturation in the thymus is tightly controlled at multiple checkpoints. However, the molecular mechanism for the control of this developmental program is not fully understood. A number of protein tyrosine kinases, such as Zap-70, Lck, and Fyn, have been shown to promote signals required for thymocyte development, whereas a tyrosine phosphatase Src homology domain-containing tyrosine phosphatase (Shp)1 has a negative effect in pre-TCR and TCR signaling. We show in this study that Shp2, a close relative of Shp1, plays a positive role in T cell development and functions. Lck-Cre-mediated deletion of Shp2 in the thymus resulted in a significant block in thymocyte differentiation/proliferation instructed by the pre-TCR at the beta selection step, and reduced expansion of CD4(+) T cells. Furthermore, mature Shp2(-/-) T cells showed decreased TCR signaling in vitro. Mechanistically, Shp2 acts to promote TCR signaling through the ERK pathway, with impaired activation of ERK kinase observed in Shp2(-/-) T cells. Thus, our results provide physiological evidence that Shp2 is a common signal transducer for pre-TCR and TCR in promoting T cell maturation and proliferation.     

Ammonium Transformation in a Nitrogen-Rich Tidal Freshwater Marsh

The fate and transport of watershed-derived ammonium in a tidal freshwater marsh fringing the nutrient rich Scheldt River, Belgium, was quantified in a whole ecosystem ¹⁵N labeling experiment. In late summer (September) we added ¹⁵N-NH₄⁺ to the flood water entering a 3477 m² tidal freshwater marsh area, and traced the ammonium processing and retention in four subsequent tide cycles. In this paper we present the results for the water-phase components of the marsh system and compare them to a similar experiment conducted in spring/early summer (May). Changes in concentration and isotopic enrichment of NO₃⁻ + NO₂⁻, N₂O, N₂, NH₄⁺ and suspended particulate nitrogen (SPN) were measured in concert with a mass balance study. All analyzed N-pools were labeled, and 49% of the added ¹⁵NH₄⁺ was retained or transformed. The most important pool for ¹⁵N was nitrate, accounting for 17% of ¹⁵N-transformation. N₂, N₂O and SPN accounted for 2.4, 0.02 and 1.4%, respectively. The temporal and spatial patterns of ¹⁵N transformation in the water phase component of the system were remarkably similar to those observed in May, indicating good reproducibility of the whole ecosystem labeling approach, but the absolute ammonium transformation rate was 3 times higher in May. While the marsh surface area was crucial for nitrification in May this was less pronounced in September. Denitrification, on the other hand, appeared more important in September compared to May.