Encapsulation of Lipophilic Bioactive Molecules by Microchannel Emulsification

Currently, much effort is being invested in novel formulations of bioactive molecules, such as emulsions, for pharmaceutical, food, and cosmetic applications. Therefore, methods to produce emulsions with controlled-size droplets of uniform size distribution have been developed. On this concern, a microfluidic device called the microchannel (MC) was used in this work for emulsification. This is a novel method for producing monodispersed emulsion droplets with very narrow droplet size distribution and low energy input, due to the spontaneous droplet generation basically driven by the interfacial tension, unlike other conventional emulsification processes. This technology provides the formulation of oil-in-water (O/W) emulsions containing lipophilic active molecules with increased bioavailability, which may be readily absorbed by the human body. MC emulsification enables the preparation of highly monodispersed O/W emulsions, which may be applied as enhancer on active molecules delivery systems, as well as in foodstuff. In this study, formulations of O/W emulsions loaded with bioactive molecules, such as β-carotene and γ-oryzanol, were prepared by the MC emulsification process. Refined soybean oil containing the dissolved lipophilic molecule and either sugar ester or gelatin solution (1 wt.%) were used as the dispersed and continuous phases, respectively. The emulsification process conducted using the asymmetric straight-through MC plate enabled the production of monodispersed O/W emulsions, resulting in β-carotene-loaded O/W emulsions with average droplet size (d _av) of 27.6 μm and coefficient of variation (CV) of 2.3% and γ-oryzanol-loaded droplets with d _av of 28.8 μm and CV of 3.8%. The highly monodisperse β-carotene-loaded droplets were physically stable throughout the storage period observed, resulting in droplets with d _av 28.2 μm and CV of 2.9% after 4 months storage in darkness at 5 °C. Single micrometer-sized monodisperse emulsions loaded with β-carotene were successfully formulated using the grooved MC emulsification, resulting in droplets with d _av of 9.1 μm and CV of 6.2%. This work was funded by The Ministry of Agriculture, Forestry and Fisheries of Japan, through the Food Nanotechnology Project, and the Japan Society for the Promotion of Science.     

Encapsulation of drug reservoirs in fibers by emulsion electrospinning: morphology characterization and preliminary release assessment

In this paper, we prepared composite fibers via electrospinning from either W/O or O/W emulsion. SEM images demonstrate the beads-in-string structures in these fibers and proved this technique to be an effective method for microencapsulation. As a practical application, Ca-alginate microspheres, which serve as reservoirs for hydrophilic drugs, were prepared in a reverse emulsion and then incorporated into poly (l-lactic acid) (PLLA) fibers by electrospinning. With the bovine serum albumin (BSA) loaded into the microspheres, the beads-in-string structure thus entrapped hydrophilic proteins in hydrophobic polymeric matrix. In the in vitro release test, BSA, which was released from composite fibers, achieved prolonged release profiles and lower burst release rates than those from naked Ca-alginate microspheres. In comparison with other well-established techniques to prepare microcapsules, such as solvent evaporation and spray-drying techniques, emulsion electrospinning features partly competing, partly complementary characteristics. Extension to other emulsion systems will be able to fabricate new types of functional structures.     

Characteristics of Precipitation-formed Polyethylene Glycol Microgels Are Controlled by Molecular Weight of Reactants

This work describes the formation of poly(ethylene glycol) (PEG) microgels via a photopolymerized precipitation reaction. Precipitation reactions offer several advantages over traditional microsphere fabrication techniques. Contrary to emulsion, suspension, and dispersion techniques, microgels formed by precipitation are of uniform shape and size, i.e. low polydispersity index, without the use of organic solvents or stabilizers. The mild conditions of the precipitation reaction, customizable properties of the microgels, and low viscosity for injections make them applicable for in vivo purposes. Unlike other fabrication techniques, microgel characteristics can be modified by changing the starting polymer molecular weight. Increasing the starting PEG molecular weight increased microgel diameter and swelling ratio. Further modifications are suggested such as encapsulating molecules during microgel crosslinking. Simple adaptations to the PEG microgel building blocks are explored for future applications of microgels as drug delivery vehicles and tissue engineering scaffolds.     

Oxygen isotope fractionation between human phosphate and water revisited

The oxygen isotope composition of human phosphatic tissues (δ¹⁸O_P) has great potential for reconstructing climate and population migration, but this technique has not been applied to early human evolution. To facilitate this application we analyzed δ¹⁸O_P values of modern human teeth collected at 12 sites located at latitudes ranging from 4°N to 70°N together with the corresponding oxygen composition of tap waters (δ¹⁸O_W) from these areas. In addition, the δ¹⁸O of some raw and boiled foods were determined and simple mass balance calculations were performed to investigate the impact of solid food consumption on the oxygen isotope composition of the total ingested water (drinking water + solid food water). The results, along with those from three, smaller published data sets, can be considered as random estimates of a unique δ¹⁸O_W/δ¹⁸O_P linear relationship: δ¹⁸O_W = 1.54(±0.09) × δ¹⁸O_P−33.72(±1.51) (R² = 0.87: p [H₀:R² = 0] = 2 × 10⁻¹⁹). The δ¹⁸O of cooked food is higher than that of the drinking water. As a consequence, in a modern diet the δ¹⁸O of ingested water is +1.05 to 1.2‰ higher than that of drinking water in the area. In meat-dominated and cereal-free diets, which may have been the diets of some of our early ancestors, the shift is a little higher and the application of the regression equation would slightly overestimate δ¹⁸O_W in these cases.     

A new methodological approach to the establishment of sustainable agricultural ecology in drought vulnerable areas of eastern India

Rice (Oryza sativa L.), the staple food for much of the world, requires gallons of water during its growth phase from tillage to harvest through conventional farming and emits significant greenhouse gases. Hence, modified cultivation techniques are felt to be needed to increase rice productivity, minimize water and fertilizer consumption effectively, and mitigate greenhouse gas emissions to a certain extent. The present experiment was based on the Purulia district, identified as a drought-prone zone through the RS-GIS technique. Two methods, the conventional method (CVN) and the system of rice intensification (SRI), were studied with four varieties of rice, viz. var. MTU 1010, var. IET 4786, var. IET 17430 and var. IET 9947 on water resource vulnerability, associated greenhouse gas emissions (CH₄, N₂O) and global warming potential (GWP). The results indicated that SRI cultivation is the smart method compared to CVN in the drought-prone Purulia District due to its lower water requirements and greenhouse gas (GHG) emissions. There was higher CH₄ flux among the CVN plots than in the SRI plots (p < 0.05). The N₂O flux in the rice field plots of CVN (Rabi-1.54 ± 0.04 kg ha⁻¹, Kharif 1.51 ± 0.10 kg ha⁻¹) was lower than the SRI (Rabi-2.08 ± 0.07, Kharif- 1.83 ± 0.02 kg ha⁻¹) during Rabi season. The CH₄ flux for CVN (Rabi-94.32 ± 2.71 kg ha⁻¹; Kharif 110.33 ± 3.90 kg ha⁻¹) was higher than the SRI (Rabi-68.16 ± 2.37, Kharif- 61.72 ± 2.41 kg ha⁻¹) during Rabi season. The CO₂-eq emission (pGWP) and yield-scaled CO₂-eq emission (YpGWP) from the SRI were significantly lower when compared with CVN (p < 0.05). The result shows that the SRI technique is an effective agricultural method that farmers should be encouraged to follow in the drought-prone region to ensure less water consumption and a sustainable future for generations.     

Selective synthesis of triangular cluster oxido-sulfidocomplexes of Mo and W: High yield preparations of [Mo3O2S2(H2O)9], [W3O2S2(H2O)9], [W2MoO2S2(H2O)9] and their derivatization

Reaction of [Mo₂O₂(μ-S)₂(H₂O)₆]²⁺ with Mo(CO)₆ or metallic Mo under hydrothermal conditions (140 °C, 4 M HCl) gives oxido-sulfido cluster aqua complex [Mo₃(μ₃-S)(μ-O)₂(μ-S)(H₂O)₉]⁴⁺ (1). Similarly, [W₃(μ₃-S)(μ-O)₂(μ-S)(H₂O)₉]⁴⁺ (2) is obtained from [W₂O₂(μ-S)₂(H₂O)₆]²⁺ and W(CO)₆. While reaction of [Mo₂O₂(μ-S)₂(H₂O)₆]²⁺ with W(CO)₆ mainly proceeds as simple reduction to give 1, [W₂O₂(μ-S)₂(H₂O)₆]²⁺ with Mo(CO)₆ produces new mixed-metal cluster [W₂Mo(μ₃-S)(μ-O)₂(μ-S)(H₂O)₉]⁴⁺ (3) as main product. From solutions of 1 in HCl supramolecular adduct with cucurbit[6]uril (CB[6]) {[Mo₃O₂S₂(H₂O)₆Cl₃]₂CB[6]}Cl₂⋅18H₂O (4) was isolated and structurally characterized. The aqua complexes were converted into acetylacetonates [M₃O₂S₂(acac)₃(py)₃]PF₆ (M₃ = Mo₃, W₃, W₂Mo; 5a-c), which were characterized by X-ray single crystal analysis, electrospray ionization mass spectrometry and ¹H NMR spectroscopy. Crystal structure of (H₅O₂)(Me₄N)₄[W₃(μ₃-S)(μ₂-S)(μ₂-O)₂(NCS)₉] (6), obtained from 2, is also reported.     

Nitrous Oxide Emission from Grazed Grassland Under Different Management Systems

Nitrous oxide (N₂O) emissions from grazed grasslands are estimated to be approximately 28% of global anthropogenic N₂O emissions. Estimating the N₂O flux from grassland soils is difficult because of its episodic nature. This study aimed to quantify the N₂O emissions, the annual N₂O flux and the emission factor (EF), and also to investigate the influence of environmental and soil variables controlling N₂O emissions from grazed grassland. Nitrous oxide emissions were measured using static chambers at eight different grasslands in the South of Ireland from September 2007 to August 2009. The instantaneous N₂O flux values ranged from -186 to 885.6 μg N₂O-N m⁻² h⁻¹ and the annual sum ranged from 2 ± 3.51 to 12.55 ± 2.83 kg N₂O-N ha⁻¹ y⁻¹ for managed sites. The emission factor ranged from 1.3 to 3.4%. The overall EF of 1.81% is about 69% higher than the Intergovernmental Panel on Climate Change (IPCC) default EF value of 1.25% which is currently used by the Irish Environmental Protection Agency (EPA) to estimate N₂O emission in Ireland. At an N applied of approximately 300 kg ha⁻¹ y⁻¹, the N₂O emissions are approximately 5.0 kg N₂O-N ha⁻¹ y⁻¹, whereas the N₂O emissions double to approximately 10 kg N ha⁻¹ for an N applied of 400 kg N ha⁻¹ y⁻¹. The sites with higher fluxes were associated with intensive N-input and frequent cattle grazing. The N₂O flux at 17°C was five times greater than that at 5°C. Similarly, the N₂O emissions increased with increasing water filled pore space (WFPS) with maximum N₂O emissions occurring at 60–80% WFPS. We conclude that N application below 300 kg ha⁻¹ y⁻¹ and restricted grazing on seasonally wet soils will reduce N₂O emissions.     

Biodegradable and biocompatible polyampholyte microgels derived from chitosan, carboxymethyl cellulose and modified methyl cellulose

Two biocompatible and biodegradable polyampholyte microgels, namely chitosan-carboxymethyl cellulose (CS-CMC) and chitosan-modified methyl cellulose (CS-ModMC) were synthesized by an inverse microemulsion technique. The CS-CMC microgel system was pH-responsive while the CS-ModMC system possessed both pH and thermo-responsive properties. For CS-CMC system, the number of -OCH₂COOH and -NH₂ groups was determined to be 1.5 and 1.1 meq/g of microgel, respectively. In the pH range of 4-9, the zeta potential values varied from +10 to −40 mV, while the hydrodynamic radius varied from 160 nm in the swollen state (acidic and basic pH) to 110 nm in the “collapse” state (neutral pH). Furthermore, TEM micrographs confirmed the swelling/deswelling behaviour of CS-CMC microgel particles at acidic, neutral and basic conditions. For CS-ModMC system, the number of -OCH₂COOH and -NH₂ groups was determined to be 0.8 and 0.6 meq/g microgel, respectively. In the pH range of 4-9, the surface charge on the microgels varied from +25 to −60 mV and the hydrodynamic radii were 190 nm at low pH, 80 nm at neutral pH, to 120 nm at a high pH. In vitro drug release studies confirmed that CS-CMC microgels could encapsulate and release a model drug, thus they could potentially be used as biocompatible and biodegradable drug carriers.     

High-intensity Ultrasound as a Tool to Form Water in Oleogels Emulsions Structured by Lipids Oleogelators

This study aims to evaluate the structuration power and physical properties of oleogel emulsions formed by monoglycerides (MG), hardfat (HF), and lecithin (LE), with and without high-intensity ultrasound (HIU). Emulsion gels with and without HIU were prepared in mono-, bi- and ternary gelator blends (10% of the lipid phase) and with different water (W) proportions (W?=?1, 15, or 30%). All samples were analyzed according to their gel/emulsion formation, microstructure, melting behavior, rheology, texture, polymorphism, and oil binding capacity. In mono structured emulsions only MG was able to form structured emulsions that did not flow. Three synergic combinations of oleogels were found: MG:HF, HF:LE, and MG:HF:LE; nevertheless, the only synergic combination found in all water proportions was MG:HF. Sonicated MG:HF emulsions formed more organized and stable water droplets, and with 30 W even achieved similar physical properties as MG:HF oleogel (0 W). A soft structured emulsion was also found for sonicated LE and HF:LE with 30 W. Multicomponent emulsion gels formed by MG, HF, and LE are a good alternative for food applications because they can form different synergic combinations with good physical properties such as hardness (>?1 N), elastic modulus (>?1?×?10⁵ Pa), and oil binding capacity (>?99%), and these properties can be even improved by HIU.

     

Rapid detection of bacteriophages in starter culture using water-in-oil-in-water emulsion microdroplets

Bacteriophage contamination of starter culture and raw material poses a major problem in the fermentation industry. In this study, a rapid detection of lytic phage contamination in starter culture using water-in-oil-in-water (W/O/W) emulsion microdroplets was described. A model bacteria with varying concentrations of lytic phages were encapsulated in W/O/W emulsion microdroplets using a simple needle-in-tube setup. The detection of lytic phage contamination was accomplished in 1 h using the propidium iodide labeling of the phage-infected bacteria inside the W/O/W emulsion microdroplets. Using this approach, a detection limit of 10² PFU/mL of phages was achieved quantitatively, while 10⁴ PFU/mL of phages could be detected qualitatively based on visual comparison of the fluorescence images. Given the simplicity and sensitivity of this approach, it is anticipated that this method can be adapted to any strains of bacteria and lytic phages that are commonly used for fermentation, and has potential for a rapid detection of lytic phage contamination in the fermentation industry.     

Obesity does not increase external mechanical work per kilogram body mass during walking

Walking is the most common type of physical activity prescribed for the treatment of obesity. The net metabolic rate during level walking (W/kg) is ~10% greater in obese vs. normal weight adults. External mechanical work (W_ext) is one of the primary determinants of the metabolic cost of walking, but the effects of obesity on W_ext have not been clearly established. The purpose of this study was to compare W_ext between obese and normal weight adults across a range of walking speeds. We hypothesized that W_ext (J/step) would be greater in obese adults but W_ext normalized to body mass would be similar in obese and normal weight adults. We collected right leg three-dimensional ground reaction forces (GRF) while twenty adults (10 obese, BMI=35.6 kg/m² and 10 normal weight, BMI=22.1 kg/m²) walked on a level, dual-belt force measuring treadmill at six speeds (0.50–1.75 m/s). We used the individual limb method (ILM) to calculate external work done on the center of mass. Absolute W_ext (J/step) was greater in obese vs. normal weight adults at each walking speed, but relative W_ext (J/step/kg) was similar between the groups. Step frequencies were not different. These results suggest that W_ext is not responsible for the greater metabolic cost of walking (W/kg) in moderately obese adults.     

Thiotungstates containing the syn-[W2(O/S)2(μ-S)2] (E = O, S) and cuboidal [W2Cu2(μ3-S)4] cores

Reaction of [W^VIS₄]²⁻ with ethane-1,2-dithiol edtH₂ in the presence of the sulfide scavenger Cd²⁺ yielded the dinuclear tungstate syn-[{(edt)W^V(O/S)}₂(μ-S)₂]²⁻ (1), with the terminal S/O disordered over the two tungsten sites in the ratio 0.8:02. In the presence of thiocyanate, phosphine and CuI, the anionic cuboidal clusters of composition [{(SCN)₃W^V}₂{Cu^I(PPh₃)}₂(μ₃-S)₄]²⁻ (2) and (3, diphos = 1,2-bis(o-diphenylphosphinophenyl)ethane), and possibly via an intermediate [{(SCN)₃W^VS}₂(μ-S)₂]⁴⁻. The crystal and molecular structures of [Et₄N]₂1, [Et₄N]₂2 · H₂O and [Et₄N]₂3 · H₂O have been determined.     

Short-chain dehydrogenase/reductase,PsDeHase, from opium poppy: putative involvement in papaverine biosynthesis

Embryogenic synseeds were prepared in Albizia lebbeck by encapsulating cotyledon stage somatic embryos derived from in vitro maintained embryogenic cultures in different types of Ca-alginate beads. The germination rate of somatic embryos was affected significantly by the bead type, matrix composition and germination substrate. A matrix made of 3% Na₂-alginate complexed with 100 mM CaCl₂·2H₂O for a hardening period of 20 min provided uniform encapsulation of somatic embryo. Among different types of synseeds, type IIA, wherein somatic embryos encapsulated in a single layer of Ca-alginate matrix composed of MS medium supplemented with 2 g L^?1 activated charcoal and 1.0 µM gibberellic acid (GA₃) as reconstituted endosperm, was found to be the most efficient type having maximum germination rates (88.6?±?0.51%). Incorporation of GA₃ in the alginate beads stimulated greater germination of somatic embryos as against GA₃ supplementation in the germination substrate. Further, viability studies on short term cold (4 °C) storage of different types of embryogenic synseeds revealed that double layered synseeds (DLS) were found comparatively more robust to withstand longer storage durations than single layered synseeds as evident by greater germination rates of the former after 4–8 weeks of refrigerated storage. Also, the elevated levels of antioxidative enzymes (superoxide dismutase, catalase, ascorbate peroxidase and glutathione reductase) and leaf proline content in the plantlets derived from DLS reveals the possible role of alginate coatings in conferring alleviation to low temperature stress generated during different storage durations. Similar Inter simple sequence repeat profiles of embryogenic synseeds derived plantlets and mother tree nullifies the possible occurrence of somaclones, thereby establishing the efficacy of synseed technology for clonal propagation of A. lebbeck germplasm.

     

The tungsten(IV) aqua ion; reduction studies and detection of aquachloro complexes of [W3(μ3-O)(μ2-O)3]4+ by oxygen-17 and tungsten-183 NMR spectroscopy

Oxygen-17 and tungsten-183 NMR studies on solutions of orange tungsten(IV), prepared in 2 M p-toluenesulphonic acid (Hpts) solution via acid catalyzed hydrolysis of potassium hexachlorotungstate(IV) in 2 M Hpts followed by treatment with a single DOWEX 50W X2 cation-exchange column, confirm the formation of the species; [W₃(μ₃-O)(μ₂-O)₃(OH₂)₈Cl]³⁺ as major product in addition to [W₃(μ₃-O)(μ₂-O)₃(OH₂)₉]⁴⁺ under these conditions in a manner not possible by spectrophotometric means. Further characterization of the green W(III,III,IV) mixed-valence reduction product has also been carried out with in addition an estimation of the formal reduction potential for the W(IV)₃/W(III,III,IV) redox couple.     

Immobilization of Lactobacillus rhamnosus in polyvinyl alcohol/calcium alginate matrix for production of lactic acid

Immobilization of Lactobacillus rhamnosus ATCC7469 in poly(vinyl alcohol)/calcium alginate (PVA/Ca-alginate) matrix using “freezing–thawing” technique for application in lactic acid (LA) fermentation was studied in this paper. PVA/Ca-alginate beads were made from sterile and non-sterile PVA and sodium alginate solutions. According to mechanical properties, the PVA/Ca-alginate beads expressed a strong elastic character. Obtained PVA/Ca-alginate beads were further applied in batch and repeated batch LA fermentations. Regarding cell viability, L. rhamnosus cells survived well rather sharp immobilization procedure and significant cell proliferation was observed in further fermentation studies achieving high cell viability (up to 10.7 log CFU g⁻¹) in sterile beads. In batch LA fermentation, the immobilized biocatalyst was superior to free cell fermentation system (by 37.1%), while the highest LA yield and volumetric productivity of 97.6% and 0.8 g L⁻¹ h⁻¹, respectively, were attained in repeated batch fermentation. During seven consecutive batch fermentations, the biocatalyst showed high mechanical and operational stability reaching an overall productivity of 0.78 g L⁻¹ h⁻¹. This study suggested that the “freezing–thawing” technique can be successfully used for immobilization of L. rhamnosus in PVA/Ca-alginate matrix without loss of either viability or LA fermentation capability.

     

Encapsulation of <Emphasis Type="Italic">Pannonibacter phragmitetus</Emphasis> LSSE-09 in alginate–carboxymethyl cellulose capsules for reduction of hexavalent chromium under alkaline conditions

Cr(VI) was efficiently reduced to Cr(III) by Pannonibacter phragmitetus LSSE-09 encapsulated in liquid-core alginate–carboxymethyl cellulose capsules under alkaline conditions. Taking into account the physical properties of the capsules, the activity of encapsulated cells, and total Cr(III) concentration in the supernatant, optimal conditions (0.5% w/v sodium alginate; 2% w/v sodium carboxymethyl cellulose; 0.1 M CaCl₂; 30-min gelation time) for LSSE-09 encapsulation were determined. At optimal conditions, a relatively high reduction rate of 4.20 mg g_{(dry weight)}⁻¹  min⁻¹ was obtained. Total Cr(III) concentration in the supernatant was significantly decreased after reduction, because 63.7% of the formed soluble organo-Cr(III) compounds compared with those of free cells were captured by the relatively smaller porous structure of alginate capsules. The optimal pH value (9.0) for Cr(VI) reduction was not changed after encapsulation. In addition, encapsulated LSSE-09 showed no appreciable loss in activity after eight repeated cycles at 37°C, and 85.7% of its initial activity remained after 35-day storage at 4°C. The results suggest that encapsulated LSSE-09 in alginate–carboxymethyl cellulose capsules has potential biotechnological applications for the detoxification of Cr(VI)-contaminated wastewater.     

Nitrogen Transformations in Flowpaths Leading from Soils to Streams in Amazon Forest and Pasture

The modification of large areas of tropical forest to agricultural uses has consequences for the movement of inorganic nitrogen (N) from land to water. Various biogeochemical pathways in soils and riparian zones can influence the movement and retention of N within watersheds and affect the quantity exported in streams. We used the concentrations of NO₃ ⁻ and NH₄ ⁺ in different hydrological flowpaths leading from upland soils to streams to investigate inorganic N transformations in adjacent watersheds containing tropical forest and established cattle pasture in the southwestern Brazilian Amazon Basin. High NO₃ ⁻ concentrations in forest soil solution relative to groundwater indicated a large removal of N mostly as NO₃ ⁻ in flowpaths leading from soil to groundwater. Forest groundwater NO₃ ⁻ concentrations were lower than in other Amazon sites where riparian zones have been implicated as important N sinks. Based on water budgets for these watersheds, we estimated that 7.3–10.3 kg N ha⁻¹ y⁻¹ was removed from flowpaths between 20 and 100 cm, and 7.1–10.2 kg N ha⁻¹ y⁻¹ was removed below 100 cm and the top of the groundwater. N removal from vertical flowpaths in forest exceeded previously measured N₂O emissions of 3.0 kg N ha⁻¹ y⁻¹ and estimated emissions of NO of 1.4 kg N ha⁻¹ y⁻¹. Potential fates for this large amount of nitrate removal in forest soils include plant uptake, denitrification, and abiotic N retention. Conversion to pasture shifted the system from dominance by processes producing and consuming NO₃ ⁻ to one dominated by NH₄ ⁺, presumably the product of lower rates of net N mineralization and net nitrification in pasture compared with forest. In pasture, no hydrological flowpaths contained substantial amounts of NO₃ ⁻ and estimated N removal from soil vertical flowpaths was 0.2 kg N ha⁻¹ y⁻¹ below the depth of 100 cm. This contrasts with the extent to which agricultural sources dominate N inputs to groundwater and stream water in many temperate regions. This could change, however, if pasture agriculture in the tropics shifts toward intensive crop cultivation.     

The Importance of Termites to the CH<Subscript>4</Subscript> Balance of a Tropical Savanna Woodland of Northern Australia

Termites produce methane (CH₄) as a by-product of microbial metabolism of food in their hindguts, and are one of the most uncertain components of the regional and global CH₄ exchange estimates. This study was conducted at Howard Springs near Darwin, and presents the first estimate of CH₄ emissions from termites based on replicated in situ seasonal flux measurements in Australian savannas. Using measured fluxes of CH₄ between termite mounds and the atmosphere, and between soil and the atmosphere across seasons we determined net CH₄ flux within a tropical savanna woodland of northern Australia. By accounting for both mound-building and subterranean termite colony types, and estimating the contribution from tree-dwelling colonies it was calculated that termites were a CH₄ source of +0.24 kg CH₄-C ha⁻¹ y⁻¹ and soils were a CH₄ sink of −1.14 kg CH₄-C ha⁻¹ y⁻¹. Termites offset 21% of CH₄ consumed by soil resulting in net sink strength of −0.90 kg CH₄-C ha⁻¹ y⁻¹ for these savannas. For Microcerotermes nervosus (Hill), the most abundant mound-building termite species at this site, mound basal area explained 48% of the variation in mound CH₄ flux. CH₄ emissions from termites offset 0.1% of the net biome productivity (NBP) and CH₄ consumption by soil adds 0.5% to the NBP of these tropical savannas at Howard Springs.     

Hyaluronic acid-based microgels and microgel networks for vocal fold regeneration

Vocal fold scarring disrupts the viscoelastic properties of the lamina propria that are critical for normal phonation. There is a clinical need for the development of advanced biomaterials that approximate the mechanical properties of the lamina propria for in vivo vocal fold regeneration. We have developed hyaluronic acid (HA)-based microgels and cross-linked microgel networks with tunable degradation and mechanical properties. HA microgels were prepared by cross-linking HA derivatives carrying hydrazide (HAADH) and aldehyde (HAALD) functionalities within the inverse emulsion droplets. Alternatively, poly(ethylene glycol) dialdehyde (PEGDiALD) was employed in place of HAALD. Microgels based on HAADH/HAALD are more resistant to enzymatic degradation than those generated from HAADH/PEGDiALD. In vitro cytotoxicity studies using vocal fold fibroblasts indicate that microgels synthesized from HAADH/HAALD are essentially nontoxic, whereas microgels derived from HAADH/PEGDiALD exhibit certain adverse effects on the cultured cells at high concentration (> or =2 mg/mL). These microgels exhibit residual functional groups that can be used as reactive handles for covalent conjugation of therapeutic molecules. The presence of residual functional groups also allows for subsequent cross-linking of the microgels with other reactive polymers, giving rise to doubly cross-linked networks (DXNs) with tunable viscoelasticity. Mechanical measurements using a torsional wave apparatus indicate that HA-based DXNs exhibit elastic moduli that are similar to those of vocal fold lamina propria at frequencies close to the range of human phonation. These HA-based microgel systems are promising candidates for the treatment of vocal fold scarring, not just as biocompatible filler materials, but as smart entities that can repair focal defects, smooth the vocal fold margin, and potentially soften and dissolve scar tissue.     

Winter and summer nitrous oxide and nitrogen oxides fluxes from a seasonally snow-covered subalpine meadow at Niwot Ridge,Colorado

The soil emission rates (fluxes) of nitrous oxide (N₂O) and nitrogen oxides (NO + NO₂ = NO _x ) through a seasonal snowpack were determined by a flux gradient method from near-continuous 2-year measurements using an automated system for sampling interstitial air at various heights within the snowpack from a subalpine site at Niwot Ridge, Colorado. The winter seasonal-averaged N₂O fluxes of 0.047–0.069 nmol m⁻² s⁻¹ were ~15 times higher than observed NO _x fluxes of 0.0030–0.0067 nmol m⁻² s⁻¹. During spring N₂O emissions first peaked and then dropped sharply as the soil water content increased from the release of snowpack meltwater, while other gases, including NO _x and CO₂ did not show this behavior. To compare and contrast the winter fluxes with snow-free conditions, N₂O fluxes were also measured at the same site in the summers of 2006 and 2007 using a closed soil chamber method. Summer N₂O fluxes followed a decreasing trend during the dry-out period after snowmelt, interrupted by higher values related to precipitation events. These peaks were up to 2–3 times higher than the background summer levels. The integrated N₂O-N loss over the summer period was calculated to be 1.1–2.4 kg N ha⁻¹, compared to ~0.24–0.34 kg N ha⁻¹ for the winter season. These wintertime N₂O fluxes from subniveal soil are generally higher than the few previously published data. These results are of the same order of magnitude as data from more productive ecosystems such as fertilized grasslands and high-N-cycling forests, most likely because of a combination of the relatively well-developed soils and the fact that subnivean biogeochemical processes are promoted by the deep, insulating snowpack. Hence, microbially mediated oxidized nitrogen emissions occurring during the winter can be a significant part of the N-cycle in seasonally snow-covered subalpine ecosystems.