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The effects of nitrogen (N) deposition on forests largely depend on its fate after entering the ecosystem. While several studies have addressed the forest fate of N deposition using 15N tracers, the long‐term fate and redistribution of deposited N in tropical forests remains unknown. Here, we applied 15N tracers to examine the fates of deposited ammonium (NH4+) and nitrate (NO3‐) separately over 3 years in a primary and a secondary tropical montane forest in southern China. Three months after 15N tracer addition, over 60% of 15N was retained in the forests studied. Total ecosystem retention did not change over the study period, but between 3 months and 3 years following deposition 15N recovery in plants increased from 10% to 19% and 13% to 22% in the primary and secondary forests, respectively, while 15N recovery in the organic soil declined from 16% to 2% and 9% to 2%. Mineral soil retained 50% and 35% of 15N in the primary and secondary forests, with retention being stable over time. The total ecosystem retention of the two N forms did not differ significantly, but plants retained more 15NO3‐ than 15NH4+ and the organic soil more 15NH4+ than NO3‐. Mineral soil did not differ in 15NH4+ and 15NO3‐ retention. Compared to temperate forests, proportionally more 15N was distributed to mineral soil and plants in these tropical forests. Overall, our results suggest that atmospherically deposited NH4+ and NO3‐ is rapidly lost in the short term (months) but thereafter securely retained within the ecosystem, with retained N becoming redistributed to plants and mineral soil from the organic soil. This long‐term N retention may benefit tropical montane forest growth and enhance ecosystem carbon sequestration. [ABSTRACT FROM AUTHOR]

Nitrogen (N) deposition is projected to substantially increase in the tropics over the coming decades, which is expected to lead to enhanced N saturation and gaseous N emissions from tropical forests (via NO, N 2 O, and N 2). However, it is unclear how N deposition in tropical forests influences both the magnitude of gaseous loss of nitrogen and its partitioning into the N 2 and N 2 O loss mechanisms. Here, for the first time, we employed the acetylene inhibition technique and the 15N-nitrate labeling method to quantify N 2 and N 2 O emission rates for long-term experimentally N-enriched treatments in primary and secondary tropical montane forest. We found that during laboratory incubation under aerobic conditions long-term increased N addition of up to 100 kg N ha−1 yr−1 at Jianfengling forest, China, did not cause a significant increase in either N 2 O or N 2 emissions, or N 2 O/N 2. However, under anaerobic conditions, N 2 O emissions decreased and N 2 emissions increased with increasing N addition in the secondary forest. These changes may be attributed to substantially greater N 2 O reduction to N 2 during denitrification, further supported by the decreased N 2 O/N 2 ratio with increasing N addition. No such effects were observed in the primary forest. In both forests, N addition decreased the contribution of denitrification while increasing the contribution of co-denitrification and heterotrophic nitrification to N 2 O production. Denitrification was the predominant pathway to N 2 production (98–100%) and its contribution was unaffected by N addition. Despite the changes in the contributions of denitrification to N 2 O gas emissions, we detected no change in the abundance of genes associated with denitrification. While the mechanisms for these different responses are not yet clear, our results indicate that the effects of N deposition on gaseous N loss were ecosystem-specific in tropical forests and that the microbial processes responsible for the production of N gases are sensitive to N inputs. Graphical abstract Image 1 Highlights • The effects of N addition on N gas losses were studied for two tropical forests. • The effects on N gas losses was significant in one forest while not in the other. • N addition increased N 2 O reduction to N 2 by denitrification in secondary forest. • Microbial processes contributions to N 2 O productions are sensitive to N additions. • The abundances of denitrification genes did not change after long term N additions. [ABSTRACT FROM AUTHOR]

A number of product development partnerships are actively developing new vaccines to combat infectious diseases in developing countries. To be effective, the products under development should not only be safe, efficacious, and affordable, but they should also have additional desirable technical attributes, including enhanced stability, efficient packaging, and improved ease of delivery. New technologies are now available to achieve these attributes; however, many of the technologies have yet to be adopted by the vaccine industry. This commentary discusses the opportunities and challenges associated with advancing such attributes, especially vaccine thermostability and dose sparing strategies, and the critical issues that must be addressed to bridge the gap between technology development and product development. © 2012 Wiley Periodicals, Inc. and the American Pharmacists Association J Pharm Sci 102:29-33, 2013 [ABSTRACT FROM AUTHOR]

Abstract: Epidermal powder immunization (EPI) can efficiently deliver powdered protein vaccines to the epidermis. A phase I clinical trial was conducted to evaluate powdered trivalent influenza vaccine delivered using the PowderJect ND5.2 delivery system. Subjects received either Fluvirin® IM injection (15μg of each influenza strain), a single EPI vaccination (15μg of each influenza strain) or two adjacent EPI (total of 30μg of each influenza strain). Systemic reactogenicity was similar between control and EPI vaccines. Site reactions following EPI were primarily mild and self-limiting. Seroconversions, titer increases and geometric mean titers to all strains were equivalent or higher in EPI-immunized groups than in controls. Powdered influenza vaccine delivered by EPI is safe and elicits humoral immune responses in humans. [Copyright &y& Elsevier]

Epidermal powder immunization (EPI) of mice with an influenza vaccine elicited consistently a higher hemagglutination inhibition (HI) antibody titers than intramuscular (IM) injection using the same dose of vaccine. The epidermal Langerhans cells (LCs) at the site of EPI were found to play an important role in the immune responses. Indeed, depletion of LCs from the immunization site prior to EPI caused a significant reduction in the antibody response. Transfer of LCs isolated from the EPI sites to naive mice induced a robust antigen-specific antibody response. Cytokines produced by target site cells appear to be important for the augmented immune responses induced by EPI. LTR72, a genetically detoxified heat-labile toxin from Escherichia coli with a strong adjuvant effect in EPI, was found to bind the keratinocytes of the epidermis, but not the LCs, and caused the production of elevated TNF-α and IL-12 cytokines in emigrating epidermal cells. These results have important implications for the development of a more efficacious human influenza vaccine. [Copyright &y& Elsevier]

Antigen presentation is a critical step for the immune system to mount an antigen-specific immune response to invading pathogens. Delivering antigens directly to antigen presenting cells (APCs) is a promising way to improve the immunogenicity and effectiveness of vaccines. Epidermal powder immunization (EPI) delivers vaccine to the epidermis, the superficial layer of skin containing a large number of APCs, and has been shown to elicit augmented antibody responses and cellular immunity to the split influenza vaccine in small animals, nonhuman primates, and humans. EPI is safe and effective and holds promise for future human immunization with influenza and other vaccines. [Copyright &y& Elsevier]

Epidermal powder immunization (EPI) with an influenza vaccine and an adjuvant such as QS-21, LTR72, or cholera toxin elicited augmented serum and mucosal antibody responses in mice. Rhesus macaques, which have an immune system and skin structure similar to humans, were used to further evaluate the immunogenicity of the influenza vaccine following EPI. EPI of rhesus macaques with an influenza vaccine and QS-21 adjuvant elicited significantly higher serum hemagglutination inhibition (HI) titers than antigen alone administered by EPI or by intramuscular (IM) injection using a needle and syringe. In the absence of QS-21, EPI and IM injection elicited comparable HI titers in the monkeys. This study suggests that EPI is a promising technique for administering human vaccine and that QS-21 augments the immunogenicity of co-administered influenza vaccine. [Copyright &y& Elsevier]

The non-toxic B subunit of cholera toxin (CTB) and E. coli heat-labile toxin mutant proteins with reduced toxicity (LTR72) or no toxicity (LTK63) were used as adjuvants for epidermal powder immunization (EPI) with an influenza vaccine. When administered by EPI, CTB, LTR72 and LTK63 significantly augmented antibody responses to the influenza vaccine and protection against a lethal challenge in a mouse model. The antigen dose could be reduced by 125-fold. These adjuvants were well-tolerated both locally and systemically following EPI. These results suggest that EPI with influenza vaccine and a non-toxic bacterial enterotoxin hold promise for human vaccination. [Copyright &y& Elsevier]

Describes epidermal powder immunization (EI) which effectively delivers powdered vaccines to the epidermis by using a helium-powered, needle free PowderJect system. Comparison between EI and needle injection; Challenge experiments; Immunogenicity of influenza vaccine.

Abstract: All vaccines are susceptible to damage by elevated temperatures and many are also damaged by freezing. The distribution, storage, and use of vaccines therefore present challenges that could be reduced by enhanced thermostability, with resulting improvements in vaccine effectiveness. Formulation and processing technologies exist that can improve the stability of vaccines at temperature extremes, however, customization is required for individual vaccines and results are variable. Considerations affecting decisions about stabilization approaches include development cost, manufacturing cost, and the ease of use of the final product. Public sector agencies can incentivize vaccine developers to prioritize stabilization efforts through advocacy and by implementing policies that increase demand for thermostable vaccines. [Copyright &y& Elsevier]

Denitrification is a major process contributing to the removal of nitrogen (N) from ecosystems, but its rate is difficult to quantify. The natural abundance of isotopes can be used to identify the occurrence of denitrification and has recently been used to quantify denitrification rates at the ecosystem level. However, the technique requires an understanding of the isotopic enrichment factor associated with denitrification, which few studies have investigated in forest soils. Here, soils collected from two tropical and two temperate forests in China were incubated under anaerobic or aerobic laboratory conditions for two weeks to determine the N and oxygen (O) isotope enrichment factors during denitrification. We found that at room temperature (20 °C), NO 3 − was reduced at a rate of 0.17 to 0.35 μg N g −1 h −1 , accompanied by the isotope fractionation of N ( 15 ε ) and O ( 18 ε ) of 31‰ to 65‰ (48.3 ± 2.0‰ on average) and 11‰ to 39‰ (18.9 ± 1.7‰ on average), respectively. The N isotope effects were, unexpectedly, much higher than reported in the literature for heterotrophic denitrification (typically ranging from 5‰ to 30‰) and in other environmental settings (e.g., groundwater, marine sediments and agricultural soils). In addition, the ratios of Δδ 18 O:Δδ 15 N ranged from 0.28 to 0.60 (0.38 ± 0.02 on average), which were lower than the canonical ratios of 0.5 to 1 for denitrification reported in other terrestrial and freshwater systems. We suggest that the isotope effects of denitrification for soils may vary greatly among regions and soil types and that gaseous N losses may have been overestimated for terrestrial ecosystems in previous studies in which lower fractionation factors were applied. [ABSTRACT FROM AUTHOR]

The quantification of large-scale leaf-age-dependent leaf area index has been lacking in tropical and subtropical evergreen broadleaved forests (TEFs), despite the recognized importance of leaf age in influencing leaf photosynthetic capacity in this biome. Here, we simplified the canopy leaves of TEFs into three age cohorts (i.e., young, mature, and old, with different photosynthesis capacities; i.e., Vc,max) and proposed a novel neighbor-based approach to develop the first gridded dataset of a monthly leaf-age-dependent leaf area index (LAI) product (referred to as Lad-LAI) at 0.25 ∘ spatial resolution over the continental scale during 2001–2018 from satellite observations of sun-induced chlorophyll fluorescence (SIF) that was reconstructed from MODIS and TROPOMI (the TROPOspheric Monitoring Instrument). The new Lad-LAI products show good performance in capturing the seasonality of three LAI cohorts, i.e., young (LAI young ; the Pearson correlation coefficient of R=0.36), mature (LAI mature ; R=0.77), and old (LAI old ; R=0.59) leaves at eight camera-based observation sites (four in South America, three in subtropical Asia, and one in the Democratic Republic of the Congo (DRC)) and can also represent their interannual dynamics, validated only at the Barro Colorado site, with R being equal to 0.54, 0.64, and 0.49 for LAI young , LAI mature , and LAI old , respectively. Additionally, the abrupt drops in LAI old are mostly consistent with the seasonal litterfall peaks at 53 in situ measurements across the whole tropical region (R=0.82). The LAI seasonality of young and mature leaves also agrees well with the seasonal dynamics of the enhanced vegetation index (EVI; R=0.61), which is a proxy for photosynthetically effective leaves. Spatially, the gridded Lad-LAI data capture a dry-season green-up of canopy leaves across the wet Amazonian areas, where mean annual precipitation exceeds 2000 mm yr -1 , consistent with previous satellite-based analyses. The spatial patterns clustered from the three LAI cohorts also coincide with those clustered from climatic variables over the whole TEF region. Herein, we provide the average seasonality of three LAI cohorts as the main dataset and their time series as a supplementary dataset. These Lad-LAI products are available at 10.6084/m9.figshare.21700955.v4 (Yang et al., 2022). [ABSTRACT FROM AUTHOR]

Abstract: New vaccine platform and delivery technologies that can have significant positive impacts on the effectiveness, acceptability, and safety of immunizations in developing countries are increasingly available. Although donor support for vaccine technology development is strong, the uptake of proven technologies by the vaccine industry and demand for them by purchasers continues to lag. This article explains the challenges and opportunities associated with accelerating the availability of innovative and beneficial vaccine technologies to meet critical needs in resource-poor settings over the next decade. Progress will require increased dialog between the public and private sectors around vaccine product attributes; establishment of specifications for vaccines that mirror programmatic needs; stronger encouragement of vaccine developers to consider novel technologies early in the product development process; broader facilitation of research and access to technologies through the formation of centers of excellence; the basing of vaccine purchase decisions on immunization systems costs rather than price per dose alone; possible subsidization of early technology adoption costs for vaccine producers that take on the risks of new technologies of importance to the public sector; and the provision of data to purchasers, better enabling them to make informed decisions that take into account the value of specific product attributes. [Copyright &y& Elsevier]

Aluminum-containing adjuvants have been used to enhance the immune response against killed, inactivated, and subunit antigens for more than seven decades. Nevertheless, we are only beginning to gain important insight as to what may be some very fundamental parameters for optimizing their use. For example, there is evidence that the conventional approach of maximizing antigen binding (amount and/or strength) may not result in an optimal immune response. Adsorption of antigen onto the adjuvant has recently been suggested to decrease the thermal stability of some antigens; however, whether adsorption-induced alterations to the structure and/or stability of the antigen have consequences for the elicited immune response is unclear. Finally, the thermal stability of vaccines with aluminum-containing adjuvants is not robust. Optimizing the stability of these vaccines requires an understanding of the freeze sensitivity of the adjuvant, freeze and heat sensitivity of the antigen in the presence of the adjuvant, and perhaps most important, how (or whether) various approaches to formulation can be used to address these instabilities. This review attempts to summarize recent findings regarding issues that may dictate the success of vaccines with aluminum-containing adjuvants. © 2010 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 100:388-401, 2011 [ABSTRACT FROM AUTHOR]

Biochar, produced as a by-product of pyrolysis/gasification of waste biomass, shows great potential to reduce the environment impact, address the climate change issue, and establish a circular economy model. Despite the promising outlook, the research on the benefits of biochar remains highly debated. This has been attributed to the heterogeneity of biochar itself, with its inherent physical, chemical and biological properties highly influenced by production variables such as feedstock types and treating conditions. Hence, to enable meaningful comparison of results, establishment of an agreed international standard to govern the production of biochar for specific uses is necessary. In this study, we analyzed four key uses of biochar: 1) in agriculture and horticulture, 2) as construction material, 3) as activated carbon, and 4) in anaerobic digestion. Then the guidelines for the properties of biochar, especially for the concentrations of toxic heavy metals, for its environmental friendly application were proposed in the context of Singapore. The international status of the biochar industry code of practice, feedback from Singapore local industry and government agencies, as well as future perspectives for the biochar industry were explained. Unlabelled Image • Waste to biochar is a sustainable partway to circular economy. • Four key uses of the Singapore biochar industry are analyzed and reviewed. • The code of practice for biochar application in Singapore is proposed. • Future perspective of the research, innovation and development for biochar industry is discussed. [ABSTRACT FROM AUTHOR]

A Cr–Mn–Ni–N series high strength metastable austenitic stainless steel (14Cr10Mn) was used to investigate the effect of cold rolling reduction on mechanical properties at various low testing temperatures. Microstructures of tested samples were observed by transmission electron microscope (TEM) and electron back scatter diffraction (EBSD) at the gauge section. The experimental results show that the size of the rib-like α′- martensite formed during tensile testing is sensitive to temperature but insensitive to cold rolling reduction. The results also show that tensile strength is more sensitive to temperature than yield strength, while the effect of cold rolling is completely opposite. The cold rolling inhibits the martensite transformation kinetics of the tensile process after promoting it at a certain temperature condition, and the promotion effect reaches a maximum value approximately 4% when the cold rolling reduction is 10%. However, it also suppresses the α′- martensite content finally formed during tensile testing due to the high stress-strain state when the cold rolling reduction exceeds 10%, while a larger cold reduction promotes the production of α′- martensite. In addition, lowering temperature can effectively promote the formation of ε-martensite at the end of the Stage I of work hardening, and becomes a transitional phase of subsequent transformation to α′- martensite, which directly leads to the enhancement of strength in the stage II of work hardening. [ABSTRACT FROM AUTHOR]

Abstract According to manufacturers, inactivated poliovirus vaccines (IPVs) are freeze sensitive and require storage between 2°C and 8°C, whereas oral poliovirus vaccine requires storage at −20 °C. Introducing IPV into ongoing immunization services might result in accidental exposure to freezing temperatures and potential loss of vaccine potency. To better understand the effect of freezing IPVs, samples of single-dose vaccine vials from Statens Serum Institut (VeroPol) and multi-dose vaccine vials from Sanofi Pasteur (IPOL) were exposed to freezing temperatures mimicking what a vaccine vial might encounter in the field. D-antigen content was measured to determine the in vitro potency by ELISA. Immunogenicity testing was conducted for a subset of exposed IPVs using the rat model. Freezing VeroPol had no detectable effect on in vitro potency (D-antigen content) in all exposures tested. Freezing of the IPOL vaccine for 7 days at −20 °C showed statistically significant decreases in D-antigen content by ELISA in poliovirus type 1 (p < 0.0001) and type 3 (p = 0.048). Reduction of poliovirus type 2 potency also approached significance (p = 0.062). The observed loss in D-antigen content did not affect immunogenicity in the rat model. Further work is required to determine the significance of the loss observed and the implications for vaccine handling policies and practices. [ABSTRACT FROM AUTHOR]

In order to understand the mechanism of substitution for nickel with manganese in metastable austenitic stainless steels 14Cr10Mn and 16Cr6Mn, the transformation behavior of ε- and α′- martensite and the corresponding mechanical properties were investigated in this study. The microstructures and tensile and hardness properties of the cold-rolled steels under different rolling reductions at ambient temperature were measured using X-ray diffraction (XRD), scanning electron microscope (SEM) together with an electron back scatter diffraction (EBSD) and transmission electron microscope (TEM). The results show that α′- martensite is not easy to be transformed from austenite, and the martensite transformation γ → ε → α′ in 14Cr10Mn and 16Cr6Mn metastable stainless steels is found during cold rolling. The results also reveal that ε- martensite acts as an hardness reinforcing phase, and high total martensite volume fraction and ratio of ε- and α′- martensite content are helpful to achieve higher tensile strength with sigmoidal shape stress-strain curves. In addition, a model considering cold rolling reduction was developed to predict the contents of ε- and α′- martensite in the steels. [ABSTRACT FROM AUTHOR]

Double mutant heat-labile toxin (dmLT) is a promising adjuvant for oral vaccine administration. The aims of our study were to develop sensitive methods to detect low concentrations of dmLT and to use the assays in preformulation studies to determine whether dmLT remains stable under conditions encountered by an oral vaccine. We developed a sandwich ELISA specific for intact dmLT and a sensitive SDS-PAGE densitometry method, and tested stability of dmLT in glass and plastic containers, in saliva, at the pH of stomach fluid, and in high-osmolarity buffers. The developed ELISA has a quantification range of 62.5 to 0.9 ng/mL and lower limit of detection of 0.3 ng/mL; the limit of quantification of the SDS-PAGE is 10 μg/mL. This work demonstrates the application of dmLT assays in preformulation studies to development of an oral vaccine containing dmLT. Assays reported here will facilitate the understanding and use of dmLT as an adjuvant. [ABSTRACT FROM AUTHOR]

An influenza pandemic remains a major public health concern. A key strategy to prevent a pandemic is to stockpile and pre-position stable influenza vaccine to allow rapid deployment in response to an outbreak. However, most influenza vaccines today are formulated as liquids that are stable only within a temperature range of 2°C to 8°C and require use of a cold chain, making vaccine transportation, distribution, and storage complicated and expensive, particularly for developing countries. To support the National Strategy for Pandemic Influenza preparedness in the United States and internationally, we developed two lead dry formulations of stable H1N1 influenza subunit vaccines using freeze-drying technology. The stable formulations contain an excipient combination of a disaccharide, such as sucrose or trehalose, and glycine, in addition to a surfactant and phosphate buffer. The freeze-dried vaccines were shown to be safe and remained immunogenic in an in vivo study in mice. Moreover, the lead formulations demonstrated no significant loss of activity after 40 months at storage temperatures of 25°C and 37°C. This stability can be particularly attractive as it could eliminate the need to use a cold chain for vaccine deployment and facilitate integration of vaccine distribution with general drug distribution where appropriate. These freeze-dried thermostable influenza subunit vaccines could also reduce the frequency of vaccine stockpile turnover, offering a cost-effective option for pandemic preparedness. [ABSTRACT FROM AUTHOR]

Vaccination is the most effective means of preventing influenza. However, the cost of producing annual seasonal influenza vaccines puts them out of reach for most developing countries. While live attenuated influenza vaccines are among the most efficacious and can be manufactured at low cost, they may require lyophilization to be stable enough for developing-country use, which adds a significant cost burden. The development of a liquid live attenuated seasonal influenza vaccine that is stable for around a year—the duration of an annual influenza season—would significantly improve not only the production output but also the use and accessibility of influenza vaccines in low-resource settings. In this study, potential stabilizing excipients were screened and optimized using the least stable influenza vaccine strain presently known, H1N1 (A/California/07/2009), as a model. The stability-conferring properties of the lead formulations were also tested with a Type B strain of influenza virus (B/Brisbane/60/2008). Stability was also evaluated with higher titers of influenza virus and exposure to agitation and freeze–thaw stresses to further confirm the stability of the lead formulations. Through this process, we identified a liquid formulation consisting of sucrose phosphate glutamate buffer with 1% arginine and 0.5% recombinant human serum albumin that provided storage stability of one year at 2–8 °C for the influenza A and B strains tested. [ABSTRACT FROM AUTHOR]

Rotavirus infection, which can be prevented by vaccination, is responsible for a high burden of acute gastroenteritis disease in children, especially in low-income countries. An appropriate formulation, packaging, and delivery device for oral rotavirus vaccine has the potential to reduce the manufacturing cost of the vaccine and the logistical impact associated with introduction of a new vaccine, simplify the vaccination procedure, and ensure that the vaccine is safely and accurately delivered to children. Single-dose prefilled presentations can be easy to use; however, they are typically more expensive, can be a bottleneck during production, and occupy a greater volume per dose vis-à-vis supply chain storage and medical waste disposal, which is a challenge in low-resource settings. Multi-dose presentations used thus far have other issues, including increased wastage of vaccine and the need for separate delivery devices. In this study, the goals were to evaluate both the technical feasibility of using preservatives to develop a liquid multi-dose formulation and the primary packaging alternatives for orally delivered, liquid rotavirus vaccines. The feasibility evaluation included evaluation of commonly used preservatives for compatibility with rotavirus vaccines and stability testing of rotavirus vaccine in various primary containers, including Lameplast's plastic tubes, BD's oral dispenser version of Uniject™ (Uniject DP), rommelag's blow-fill-seal containers, and MEDInstill's multi-dose vial and pouch. These presentations were compared to a standard glass vial. The results showed that none of the preservatives tested were compatible with a live attenuated rotavirus vaccine because they had a detrimental effect on the viability of the virus. In the presence of preservatives, vaccine virus titers declined to undetectable levels within 1 month. The vaccine formulation without preservatives maintained a stability profile over 12 months in all primary containers that was similar to its profile in standard glass vials. This study demonstrates that there are multiple options for the primary container for rotavirus vaccines intended for oral delivery. Selection of an optimal primary container should take into consideration additional factors, including stability as well as cold chain volume, usability, cost, and manufacturing feasibility. [ABSTRACT FROM AUTHOR]

Purpose: Stable vaccines with long shelf lives and reduced dependency on the cold chain are ideal for stockpiling and rapid deployment during public emergencies, including pandemics. Spray drying is a low-cost process that has potential to produce vaccines stable at a wide range of temperatures. Our aim was to develop a stable formulation of a recombinant H1N1 influenza hemagglutinin vaccine candidate and take it to pilot-scale spray-drying production. Methods: Eight formulations containing different excipients were produced and assayed for antigen stability, powder characteristics, and immunogenicity after storage at a range of temperatures, resulting in the identification of four promising candidates. A pilot-scale spray-drying process was then developed for further testing of one formulation. Results: The pilot-scale process was used to reproducibly manufacture three batches of the selected formulation with yields >90%. All batches had stable physical properties and in vitro potency for 6 months at temperatures from −20°C to +50°C. Formulations stored for 3 months elicited immunogenic responses in mice equivalent to a frozen lot of bulk vaccine used as a stability control. Conclusions: This study demonstrates the feasibility of stabilizing subunit vaccines using a spray-drying process and the suitability of the process for manufacturing a candidate product. [ABSTRACT FROM AUTHOR]

Tropical rainforest soil is an important source of atmospheric nitrous oxide (N2O). However, there is still considerable uncertainty about the spatial and temporal variability of N2O fluxes. To understand these fluxes, we quantified the annual N2O emissions from three tropical mountain rainforests (primary mountain rainforest, PMR; secondary mountain rainforest, SMR; and Podocarpus imbricatus plantation, PIP) in the Jianfengling National Natural Reserve on Hainan Island, China. The average of N2O emissions in this area was 2.52 ± 0.33 kg N–N2O ha−1 yr−1 (3.52 kg N–N2O ha−1 yr−1 in the wet season and 1.62 kg N–N2O ha−1 yr−1 in the dry season) during our study period, with highly seasonal variations. The mean N2O emission rates were significantly higher during the wet season (68% of the total average) than the dry season (32% of the total average) (P < 0.05). PIP had the highest N2O emission rate at 3.49 ± 0.61 kg N–N2O ha−1 yr−1 (4.74 kg N–N2O ha−1 yr−1 in the wet season and 2.32 kg N–N2O ha−1 yr−1 in the dry season), followed by SMR at 3.03 ± 0.64 kg N–N2O ha−1 yr−1 (4.16 kg N–N2O ha−1 yr−1 in the wet season and 1.97 kg N–N2O ha−1 yr−1 in the dry season), and then PMR at 1.53 ± 0.49 kg N–N2O ha−1 yr−1 (2.21 kg N–N2O ha−1 yr−1 in the wet season and 0.94 kg N–N2O ha−1 yr−1 in the dry season). We observed a significant Gaussian relationship between the N2O fluxes and soil temperature for SMR and PIP but no significant relationship in PMR. There was a significant exponential relationship between the N2O fluxes and water filled pore space (WFPS) in SMR and PIP but not in PMR. [ABSTRACT FROM AUTHOR]

[Display omitted] • A municipal solid waste to energy/resources system was proposed. • The performances of the gasification and dehumidification system were analyzed. • The performance decline of the desiccant coated heat exchanger was evaluated. • Mechanical tests of char-mortar mixtures were conducted. Energy consumption and waste management are two significant challenges for human societies. To realize zero waste management, a pilot-scale demonstration of the conversion of municipal solid waste to energy/resource, which is comprised of a gasification system and a dehumidification air-conditioning to produce heat, cooling, and construction-used char, is investigated and analyzed. The waste heat (60 °C hot water) from a gasification system is applied to drive a dehumidification system for cooling, while the char from the gasification is added to the construction material. The effect of cycle time and air flow rate on the performance of the dehumidification system is also analyzed. The waste to energy efficiency of 57.2%, the maximum average moisture removal of 11.6 g/kg DA , and thermal coefficient of performance of 0.76 are obtained under the climate conditions of 34 °C, 67.5% relative humidity in Singapore. Through comparison between the brand new and used desiccant coated heat exchanger, it is noted that there is a 29.37% decrease of average moisture removal corresponding with a 17.1% loss of thermal coefficient of performance. A 14.6% compressive strength increment after seven days of curing age and 10.3% of compressive strength increment after 28 days are investigated by studying the mechanical behavior of a mortar containing char. [ABSTRACT FROM AUTHOR]

Highlights: [•] FDTs could provide safe and efficient delivery of oral vaccines. [•] Created a formulation containing live attenuated enterotoxigenic E. coli vaccine. [•] Freeze-drying produced a robust, compact FDT with high bacterial viability. [•] FDTs obtained by freeze drying directly in blister sheets can allow efficiency in manufacturing. [•] Vaccines in FDTs will especially be useful in treatment of traveler's diarrhea for travelers to developing countries. [ABSTRACT FROM AUTHOR]

Abstract: HSV-2-gD2 DNA vaccine was precisely delivered to immunologically sensitive regions of the skin epithelia using dry-coated microprojection arrays. These arrays delivered a vaccine payload to the epidermis and the upper dermis of mouse skin. Immunomicroscopy results showed that, in 43±5% of microprojection delivery sites, the DNA vaccine was delivered to contact with professional antigen presenting cells in the epidermal layer. Associated with this efficient delivery of the vaccine into the vicinity of the professional antigen presenting cells, we achieved superior antibody responses and statistically equal protection rate against an HSV-2 virus challenge, when compared with the mice immunized with intramuscular injection using needle and syringe, but with less than 1/10th of the delivered antigen. [ABSTRACT FROM AUTHOR]

Abstract: There is an unmet medical need for a prophylactic vaccine against herpes simplex virus (HSV). DNA vaccines and cutaneous vaccination have been tried for many applications, but few reports combine this vaccine composition and administration route. We compared DNA administration using the Nanopatch™, a solid microprojection device coated with vaccine comprised of thousands of short (110μm) densly-packed projections (70μm spacing), to standard intramuscular DNA vaccination in a mouse model of vaginal HSV-2 infection. A dose-response relationship was established for immunogenicity and survival in both vaccination routes. Appropriate doses administered by Nanopatch™ were highly immunogenic and enabled mouse survival. Vaginal HSV-2 DNA copy number day 1 post challenge correlated with survival, indicating that vaccine-elicited acquired immune responses can act quickly and locally. Solid, short, densely-packed arrays of microprojections applied to the skin are thus a promising route of administration for DNA vaccines. [ABSTRACT FROM AUTHOR]

Abstract: A combination of unique stabilizers and mild spray drying process conditions was employed to produce heat-stable measles vaccine powder. Live attenuated measles vaccine from Serum Institute of India was formulated with pharmaceutically approved stabilizers, including sugars, proteins, amino acids, polymers, surfactants, and plasticizers, as well as charged ions. In addition, the effects of buffer salt and pH on the storage stability of measles virus were examined. The potency of the dried vaccine stored at several temperatures was quantified by TCID50 assay on Vero cells. As a comparison to other process methods, lead formulations were also subjected to freeze drying and foam drying. The optimized measles vaccine formulation tested at 37°C was stable for approximately 8 weeks (i.e. time for 1logTCID50 loss). The measles titer decreased in a bi-phasic manner, with initial rapid loss within the first week but relative stability thereafter. Key stabilizers identified during the formulation screening processes were l-arginine, human serum albumin, and a combination of divalent cations. Spray drying was identified as the optimal processing method for the preparation of dried vaccine, as it generally resulted in negligible process loss and comparable, if not better storage stability, with respect to the other processes. Processing methods and formulation components were developed that produced a measles vaccine stable for up to 8 weeks at 37°C, which surpassed the WHO requirement for heat stability of 1 week at that temperature. [Copyright &y& Elsevier]

Abstract: Cold chain requirements for vaccine storage and distribution are both economic and logistical burdens for immunization programs, especially those in lower-resource settings. Inadvertent exposure of vaccines to both heat and freezing temperatures within such cold chains are frequently occurring problems in both developing and industrialized countries. Here we report on a new hepatitis B vaccine formulation that is stable against repeated freezing at −20°C and is also stable for 12 months at 37°C. The thermostable vaccine contains all the components of the original vaccine plus 7.5% (v/v) propylene glycol, 40mM phosphate, and 40mM histidine with a final pH of 5.2. The propylene glycol is responsible for the freeze stability while the other components are essential for the heat stability. This formulation was found to be well tolerated in rabbits without any significant local or systemic side effects. The improved stability of this hepatitis B vaccine could be a key factor in ensuring vaccine effectiveness, extending immunization coverage, simplifying immunization logistics, and reducing the costs associated with the cold chain. [Copyright &y& Elsevier]

Abstract: Vaccines containing aluminum salt adjuvants are prone to inactivation following exposure to freeze–thaw stress. Many are also prone to inactivation by heat. Thus, for maximum potency, these vaccines must be maintained at temperatures between 2°C and 8°C which requires the use of the cold chain. Nevertheless, the cold chain is not infallible. Vaccines are subject to freezing during both transport and storage, and frozen vaccines are discarded (under the best circumstances) or inadvertently administered despite potentially reduced potency. Here we describe an approach to minimize our reliance on the proper implementation of the cold chain to protect vaccines from freeze–thaw inactivation. By including PEG 300, propylene glycol, or glycerol in a hepatitis B vaccine, particle agglomeration, changes in the fluorescence emission spectrum – indicative of antigen tertiary structural changes – and losses of in vitro and in vivo indicators of potency were prevented following multiple exposures to −20°C. The effect of propylene glycol was examined in more detail and revealed that even at concentrations too low to prevent freezing at −10°C, −20°C, and −80°C, damage to the vaccine could be prevented. A pilot study using two commercially available diphtheria, tetanus toxoid, and acellular pertussis (DTaP) vaccines suggested that the same stabilizers might protect these vaccines from freeze–thaw agglomeration as well. It remains to be determined if preventing agglomeration of DTaP vaccines preserves their antigenic activity following freeze–thaw events. [Copyright &y& Elsevier]

Abstract: The purpose of this study was to evaluate whether a single peptide containing a major T cell epitope might induce peripheral tolerance in a complex allergen model. C57BL/6 mice were sensitized by intraperitoneal injection of house dust mite extract (HDM), and exposed to antigen via trachea instillation. Der p 1 peptide was administered by i.v. before or after sensitization. Lung lavage fluids were analyzed for cellular infiltration. Respiratory exposure of sensitized mice to antigen results in airway inflammation and eosinophilia. Intravenous administration of a single peptide protected sensitized mice from these changes. Further, the emergence of antigen-specific CD25+CD4+ and IL-10 secreting cell populations in DO11.10 mice was demonstrated after peptide administration. Thus, intravenous delivery of a single peptide epitope is capable of inducing peripheral tolerance and protection in a complex allergy model, possibly through regulatory T cells and bystander suppression. [Copyright &y& Elsevier]

To evaluate the protective effect of epidermal powder immunization (EPI) against equine herpesvirus-1 (EHV-1) infection, we prepared a powder vaccine in which formalin-inactivated virions were embedded in water-soluble, sugar-based particles. A PowderJect device was used to immunize mice with the powder vaccine via their abdominal skin. We found that twice-immunized mice were protected against challenge with the wild-type virus. This protective effect was equivalent to or better than that observed in mice immunized with other types of vaccines, including a gene gun-mediated DNA vaccine containing the glycoprotein D (gD) gene or conventional inactivated virus vaccines introduced via intramuscular or intranasal injections. These findings indicate that the powder vaccine is a promising approach for the immunological control of EHV-1 infection, either alone or as a part of prime-boost vaccination strategies. [Copyright &y& Elsevier]

Summary Langerhans cells in the epidermis of skin are potent antigen-presenting cells that trigger the immune system to respond to invading microorganisms. We have previously shown that epidermal powder immunization with a powdered inactivated influenza virus vaccine, by targeting the Langerhans cell-rich epidermis, was more efficacious than deeper tissue injection using a needle and syringe. We now report enhanced humoral and cellular immune responses to recombinant hepatitis B surface antigen following epidermal powder immunization. We observed that epidermal powder immunization with unadjuvanted hepatitis B surface antigen elicited an antibody titre equivalent to that induced by the alum-adjuvanted vaccine delivered by intramuscular injection, suggesting that epidermal powder immunization can overcome the need for adjuvantation. We demonstrated that synthetic CpG oligonucleotides (CpG DNA) could be coformulated with hepatitis B surface antigen and delivered by epidermal powder immunization to further augment the antibody response and modulate T helper cell activities. Epidermal powder immunization of hepatitis B surface antigen formulated with CpG DNA formulations resulted in 1.5-2.0 logs higher IgG antibody titres than alum-adjuvanted commercial vaccines administered by intramuscular injection. Formulation of hepatitis B surface antigen with CpG DNA elicited an augmented IgG2a antibody response and increased frequency of IFN-γ secreting cells. In addition, CpG DNA was found to activate epidermal Langerhans cells and stimulate the production of TNF-α and IL-12 cytokines by epidermal cells, explaining its strong adjuvant activity following epidermal powder immunization. These results show that epidermal powder immunization is a safe and effective method to deliver hepatitis B surface antigen and the addition of new adjuvants, such as CpG DNA, may further enhance the efficacy of this vaccine. [ABSTRACT FROM AUTHOR]

Parenterally administered rotavirus vaccines may overcome the low efficacy observed in resource-poor regions that use live oral formulations. We have reported work on a trivalent nonreplicating rotavirus vaccine (NRRV) for parenteral administration consisting of the recombinant tetanus toxoid P2 CD4 epitope fused to a truncated VP8* fragment (P2-VP8*) for the P[4], P[6], and P[8] serotypes of rotavirus adjuvanted with aluminum. An essential part of developing this vaccine candidate was devising quantification methods for each antigen in the trivalent NRRV in the presence of aluminum adjuvant. This report describes the development of quantitative inhibition enzyme-linked immunosorbent assays (ELISAs) for in vitro antigenicity determination of the adjuvanted trivalent NRRV using serotype-specific monoclonal antibodies (mAbs) against each of the P2-VP8* antigens. Adjuvanted trivalent vaccine samples are titrated and incubated with a constant concentration of specific mAbs against each NRRV P2-VP8* antigen variant. Unbound mAbs are measured by ELISA to indirectly quantify the amount of each antigen present in the trivalent vaccine. Sensitive, specific, and reproducible inhibition ELISAs were developed and qualified for each antigen and used for final product quantification and release testing without desorption of the vaccine antigen. [ABSTRACT FROM AUTHOR]