Using a Mass Balance to Determine the Potency Loss during the Production of a Pharmaceutical Blend

The manufacture of a blend containing the active pharmaceutical ingredient (API) and inert excipients is a precursor for the production of most pharmaceutical capsules and tablets. However, if there is a net water gain or preferential loss of API during production, the potency of the final drug product may be less than the target value. We use a mass balance to predict the mean potency loss during the production of a blend via wet granulation and fluidized bed drying. The result is an explicit analytical equation for the change in blend potency a function of net water gain, solids losses (both regular and high-potency), and the fraction of excipients added extragranularly. This model predicts that each 1% gain in moisture content (as determined by a loss on drying test) will decrease the API concentration of the final blend at least 1% LC. The effect of pre-blend solid losses increases with their degree of superpotency. This work supports Quality by Design by providing a rational method to set the process design space to minimize blend potency losses. When an overage is necessary, the model can help justify it by providing a quantitative, first-principles understanding of the sources of potency loss. The analysis is applicable to other manufacturing processes where the primary sources of potency loss are net water gain and/or mass losses.     

Leaf senescence and decline of end-of-season gas exchange in five temperate deciduous tree species grown in elevated CO2 concentrations

We measured rates of leaf senescence and leaf level gas exchange during autumnal senescence for seedlings of five temperate forest tree species under current and elevated atmospheric CO₂ concentrations and low- and high-nutrient regimes. Relative indices of whole canopy carbon gain, water loss and water use efficiency through the senescent period were calculated based on a simple integrative model combining gas exchange per unit leaf area and standing canopy area per unit time. Seedlings grown under elevated [CO₂] generally had smaller canopies than their current [CO₂]-grown counterparts throughout most of the senescent period. This was a result of smaller pre-senescent canopies or accelerated rates of leaf drop. Leaf-level photosynthetic rates were higher under elevated [CO₂] for grey birch canopies and for low-nutrient red maple and high-nutrient ash canopies, but declined rapidly to values below those of their current [CO₂] counterparts by midway through the senescent period. CO₂ enrichment reduced photosynthetic rates for the remaining species throughout some or all of the senescent period. As a result of smaller canopy sizes and reduced photosynthetic rates, elevated [CO₂]-grown seedlings had lower indices of whole canopy end-of-season carbon gain with few exceptions. Leaf level transpiration rates were highly variable during autumnal senescence and neither [CO₂] nor nutrient regime had consistent effects on water loss per unit leaf area or integrated whole canopy water loss throughout the senescent period. Indices of whole canopy, end-of-season estimates of water use efficiency, however, were consistently lower under CO₂ enrichment, with few exceptions. These results suggest that whole canopy end-of-season gas exchange may be altered significantly in an elevated [CO₂] world, resulting in reduced carbon gain and water use efficiency for many temperate forest tree seedlings. Seedling growth and survivorship, and ultimately temperate forest regeneration, could be reduced in CO₂-enriched forests of the future.     

Impact of management practices on the tallgrass prairie

Summary The ELM ecosystem-level grassland model simulates the flow of water, heat, nitrogen, and phosphorus through the ecosystem and the biomass dynamics of plants, consumers, and the decomposers. This model was adapted to a tallgrass prairie site in northeastern Oklahoma, USA, the Osage Site of the U.S. International Biological Program Grassland Biome. Several range management manipulations were simulated by the model and the results compared to field data and literature information: (1) altering the grazing intensity, grazing system, and grazing time period; (2) adding nitrogen and phosphorus to the grassland; (3) adding water during the growing season; and (4) spring burning of the prairie.The model showed that cattle weight gain per head, above-ground and belowground plant production, transpiration water loss, standing dead biomass, and the net nitrogen balance decrease with increasing grazing intensity, while soil water content and bare soil water loss increase. A moderately stocked year-round cow-calf grazing system is more beneficial to the grassland than a more highly stocked seasonal steer grazing system because the former increases the aboveground and belowground primary production and the plant nutrient uptake rates. Range manipulations, such as fire, which stimulate uniform grazing of a pasture, increase primary production, cattle weight gains, and nutrient uptake of plants and animals. Model results indicated that adding fertilizer was the best strategy for increasing cattle weight gains per head, while adding water would produce the greatest increase in primary production. Simulation of yearly and triennial spring burns suggests that these treatments increase primary production, plant nutrient uptake, and cattle weight gain per head. Burning increases the nitrogen losses from the systems; however, these losses are greater with annual burns. The model results also suggest the spatial grazing pattern of cattle must be considered to correctly represent the impact of grazing on the prairie.The model is used to describe the behavior of the tallgrass prairie ecosystem, evaluate alternative management strategies, and identify future scientific research and management studies.     

Intron exclusion and the mystery of intron loss

Mechanisms for loss and gain of introns are elusive. Reported here is a new pattern of intron loss which features a random loss of a single intron in a multiple-intron gene with its neighboring introns remained, which process is defined as intron exclusion. Intron exclusion is reminiscent of removal of a limited stretch of non-homologous sequence in a homologous recombination (HR) triggered by a double strand break (DSB), and therefore lends further evidence for a theory of intron loss through HR between a cDNA and its genomic intron-containing locus. Thus, a model for intron loss is formulated.     

Mutational equilibrium model of genome size evolution

The paper describes a mutational equilibrium model of genome size evolution. This model is different from both adaptive and junk DNA models of genome size evolution in that it does not assume that genome size is maintained either by positive or stabilizing selection for the optimum genome size (as in adaptive theories) or by purifying selection against too much junk DNA (as in junk DNA theories). Instead the genome size is suggested to evolve until the loss of DNA through more frequent small deletions is equal to the rate of DNA gain through more frequent long insertions. The empirical basis for this theory is the finding of a strong correlation and of a clear power-function relationship between the rate of mutational DNA loss (per bp) through small deletions and genome size in animals. Genome size scales as a negative 1.3 power function of the deletion rate per nucleotide. Such a relationship is not predicted by either adaptive or junk DNA theories. However, if genome size is maintained at equilibrium by the balance of mutational forces, this empirilical relationship can be readily accommodated. Within this framework, this finding would imply that the rate of DNA gain through large insertions scales up a quarter-power function of genome size. On this view, as genome size grows, the rate of growth through large insertions is increasing as a quarter power function of genome size and the rate of DNA loss through small deletions increases linearly, until eventually, at the stable equilibrium genome size value, rates of growth and loss equal each other. The current data also suggest that the long-term variation is genome size in animals is brought about to a significant extent by changes in the intrinsic rates of DNA loss through small deletions. Both the origin of mutational biases and the adaptive consequences of such a mode of evolution of genome size are discussed.     

Photosynthetic induction and leaf carbon gain in the tropical understorey epiphyte,Aspasia principissa

Gas exchange of the understorey epiphyte Aspasia principissa was studied in fluctuating light conditions both in the laboratory and in the field, testing the hypothesis that vascular epiphytes differ from most terrestrial understorey plants in showing a higher priority for water conservation. Consequently, a slow response of stomatal conductance to sudden increases in incident photon flux density (PFD) was expected, as was a fast loss of induction after such a light fleck. Results were only partly consistent with these expectations. Full induction of photosynthesis was indeed very slow and was not reached before, respectively, 40 and 60 min of saturating PFD in the field and the laboratory. In contrast, kinetics of induction loss were comparable with those of most terrestrial species studied to date. The overall impact of light flecks on in situ carbon gain again fulfilled expectations, being rather limited: the observed carbon gain was only approx. 66% of the potential carbon gain estimated from a square-wave response model. It is concluded that in the drought-prone epiphytic habitat of a moist lowland forest, water conservation takes priority over carbon gain, which severely limits the use of light flecks for CO(2) fixation in vascular epiphytes.     

Optimization of leaf morphology in relation to leaf water status: A theory

The leaf economic traits such as leaf area, maximum carbon assimilation rate, and venation are all correlated and related to water availability. Furthermore, leaves are often broad and large in humid areas and narrower in arid/semiarid and hot and cold areas. We use optimization theory to explain these patterns. We have created a constrained optimization leaf model linking leaf shape to vein structure that is integrated into coupled transpiration and carbon assimilation processes. The model maximizes net leaf carbon gain (NPP_leaf) over the loss of xylem water potential. Modeled relations between leaf traits are consistent with empirically observed patterns. As the results of the leaf shape–venation relation, our model further predicts that a broadleaf has overall higher NPP_leaf compared to a narrowleaf. In addition, a broadleaf has a lower stomatal resistance compared to a narrowleaf under the same level of constraint. With the same leaf area, a broadleaf will have, on average, larger conduits and lower total leaf xylem resistance and thus be more efficient in water transportation but less resistant to cavitation. By linking venation structure to leaf shape and using water potential as the constraint, our model provides a physical explanation for the general pattern of the covariance of leaf traits through the safety–efficiency trade‐off of leaf hydraulic design.     

Solar furnaces or swamp coolers: costs and benefits of water use by solar-tracking flowers of the alpine snow buttercup, Ranunculus adoneus

Solar tracking or heliotropism simultaneously raises organ temperature and light interception. For leaves and flowers carbon gain is maximized at the expense of water loss. In this study I explore how costs and benefits associated with water use by solar-tracking flowers of the alpine snow buttercup, Ranunculus adoneus change with ambient temperature. First, I test whether heliotropism increases the water cost of reproduction in the snow buttercup under extant alpine conditions. I then explore whether water use for evaporative cooling in solar-tracking flowers reduces the risk of over-heating as temperatures increase. Solar tracking, by elevating floral temperature and irradiance causes a 29% increase in water uptake by flowers. Gas exchange measurements suggest that the extra water taken up by solar-tracking flowers is released through transpiration. Transpirational cooling in turn allows solar-tracking flowers to gain advantages of enhanced light interception and warmth while reducing the risk of over-heating. Transpiration reduces excess temperature in solar-tracking flowers, but at a water cost. Results show that even in cool alpine habitats, flower heliotropism has water costs to balance its reproductive advantages. Plants with solar-tracking flowers may tolerate hotter conditions if soil moisture is plentiful, but not under drought.     

Partial consumption of prey: the significance of prey water loss on estimates of biomass intake

Summary A number of studies on the feeding behaviour of sucking predators have estimated the weight of biomass the predator extracts from the prey by measuring the weight change occurring in the prey. This method does not consider that a proportion of the prey weight change is lost to the immediate environment. I examined the spider Diaea sp. feeding on the fruit fly Drosophila immigrans and found that the prey lost approximately 28% more weight than the predator gained. This difference was largely explained by water loss from the prey. My results suggest that water loss, which is not available to the predator, is an important part of prey weight loss. To avoid overestimating predator biomass gain it is necessary to measure the predator weight gain directly or take into account water loss as a component of prey weight change.     

Calculating food consumption in the laboratory: A formula to adjust for natural weight loss

We derive a formula for correcting the weight of food eaten by an animal for natural weight loss (or gain) of the food, such as through moisture loss and food respiration. The formula is derived using a differential equation to model the loss in fresh weight based on a constant consumption rate and assumptions about the form of the function for weight loss over time of the aliquot. The standard formula is shown to over-estimate the weight of eaten food compared to our theoretically derived formula. This over-estimation is negligible (< 0.2%) for moisture loss below about 10% but increases sharply to 8% when moisture loss is 50%. We illustrate the application of these two formulae using a laboratory study of consumption rate by larvae of a phytophagous insect: the Tasmanian eucalyptus leaf beetle, Chrysophtharta bimaculata (Olivier) (Coleoptera: Chrysomelidae).     

Hydrological properties of the epiphyte mass of a montane tropical rain forest,Colombia

The effect of a well-developed epiphytic vegetation on the interception of rainfall was investigated in an upper montane rain forest at an altitude of 3370 m in the Cordillera Central of the Colombian Andes. Total epiphyte mass was estimated at 12 tonnes dry weight per hectare, most of it being bryophytes and dead organic matter. The hydrological properties of the epiphyte mass were studied using detached branches. Water gain (during rainfall) and water loss (after experimental saturation) were determined gravimetrically in the field.Rain falling on epiphyte-covered branches was efficiently captured. Drainage of water (held temporarily in excess of the retention capacity) was very gradual. Water loss through evaporation was slow. The observations are in accordance with previously reported data for rainfall interception in the forest studied. The potentially large influence of the exceptionally high canopy storage capacity is restricted due to the clumped distribution of epiphytes and associated low turnover rates of water in the epiphyte mass.     

Hydric balance and locomotor performance of an anuran (Rhinella marina) invading the Australian arid zone

Invasive species often encounter environmental conditions well outside those found in their native geographic ranges, and thus provide ideal model systems with which to explore responses to novel abiotic challenges. Within Australia, the invasive cane toad Rhinella marina has colonized areas that are considerably more arid than those found within its native range. Has the colonization of these novel environments been accompanied by shifts in physiology and/or locomotor performance? We measured rates of evaporative water loss, water gain, and effects of desiccation on locomotor performance of cane toads from two invasion fronts: one mesic (the wet‐dry tropics) and one arid (the semi‐desert). The two populations diverged substantially. Contrary to intuition (but consistent with intra‐specific comparisons between other toad populations from mesic vs arid areas), rates of evaporative water loss were lower (not higher) in toads from the mesic population. However, arid‐zone toads gained water more rapidly through their ventral surfaces, and rates of water loss and gain were highly correlated within individual toads from the arid‐zone population. Rates of water exchange in laboratory‐acclimated toads from the semi‐arid zone did not differ from those of free‐ranging conspecifics from the same population, suggesting that divergences between mesic and semi‐arid toads reflect genetic changes that have occurred during the species’ Australian invasion. Mesic and semi‐arid toads showed similar locomotor performance (endurance, distance per hop) when fully hydrated, but locomotor performance declined much more rapidly with desiccation in the mesic toads. Thus, within the short (decades‐long) timespan of the cane toad's Australian invasion, there has been substantial population divergence in the ability to withstand desiccating conditions. If we are to accurately predict the distributions (and hence impacts) of invading organisms, we will need to include adaptation potential in risk assessment schemes.     

Knot formation reduces water exchange by adult males of the hairworm,Paragordius varius (Nematomorpha: Gordioidea)

To examine how aggregation by hairworms may enhance survival in freshwater, we determined water balance characteristics of Paragordius varius in groups of different sizes. P. varius is hyperosmotic resulting in high body water content and functions down to one-half of water stores. Absence of a critical transition temperature implies a watertight, low-porosity cuticle. Aggregated worms lose water slowly, as a physiological consequence of reduced motor activity. The water balance strategy shifts from a reliance on high dehydration tolerance for isolated individuals, affording high water loss rates, to suppressed activation energy when aggregated, wherein blocking water gain is important when water loss is slower. Low water loss rate derives from stillness and aggregating that facilitate mating or anti-predator defense, rather than as a behavior to regulate water loss. Presence of hairworms in streams is an indicator of high-quality water that is necessary to maintain water balance.     

A mathematical model for human brain cooling during cold-water near-drowning

A two-dimensionalmathematical model was developed to estimate the contributions ofdifferent mechanisms of brain cooling during cold-water near-drowning.Mechanisms include 1) conductive heat loss through tissue to the water at the head surface and in theupper airway and 2) circulatorycooling to aspirated water via the lung and via venous return from thescalp. The model accounts for changes in boundary conditions, bloodcirculation, respiratory ventilation of water, and head size. Resultsindicate that conductive heat loss through the skull surface or theupper airways is minimal, although a small child-sized head willconductively cool faster than a large adult-sized head. However,ventilation of cold water may provide substantial brain cooling throughcirculatory cooling. Although it seems that water breathing is requiredfor rapid "whole" brain cooling, it is possible that conductivecooling may provide some advantage by cooling the brain cortexperipherally and the brain stem centrally via the upper airway.

     

Canopy Carbon Gain and Water Use: Analysis of Old-growth Conifers in the Pacific Northwest

This report summarizes our current knowledge of leaf-level physiological processes that regulate carbon gain and water loss of the dominant tree species in an old-growth forest at the Wind River Canopy Crane Research Facility. Analysis includes measurements of photosynthesis, respiration, stomatal conductance, water potential, stable carbon isotope values, and biogenic hydrocarbon emissions from Douglas-fir (Pseudotsuga menziesii), western hemlock (Tsuga heterophylla), and western red cedar (Thuja plicata). Leaf-level information is used to scale fluxes up to the canopy to estimate gross primary production using a physiology-based process model. Both light-saturated and in situ photosynthesis exhibit pronounced vertical gradients through the canopy, but are consistently highest in Douglas-fir, intermediate in western hemlock, and lowest in western red cedar. Net photosynthesis and stomatal conductance are strongly dependent on vapor-pressure deficit in Douglas-fir, and decline through the course of a seasonal drought. Foliar respiration is similar for Douglas-fir and western hemlock, and lowest for western red cedar. Water-use efficiency varied with species and tree height, as indexed using stable carbon isotopes values for foliage. Leaf water potential is most negative for Douglas-fir and similar for western hemlock and western red cedar. Terpene fluxes from foliage equal approximately 1% of the net carbon loss from the forest. Modeled estimates based on physiological measurements show gross primary productivity (GPP) to be about 22 Mg C m^–2 y^–1. Physiological studies will be necessary to further refine estimates of stand-level carbon balance and to make long-term predictions of changes in carbon balance due to changes in forest structure, species composition, and climate.     

Group influence on water conservation in the giant Madagascar hissing-cockroach, Gromphadorhina portentosa (Dictyoptera: Blaberidae)

Abstract. . Under laboratory conditions the hissing-cockroach Gromphadorhina portentosa (Schaum) forms clusters, which appears to be an adaptive behaviour to help reduce water loss. Adults grouped together retain water nearly twice as effectively as isolated individuals. This 'group effect' complements the cockroach's large body size (small surface area to volume ratio) to lower the rate of water loss still further. Despite a modest 28% tolerance limit for weight loss during dehydration, adult females survive absolute drying conditions of 0 % relative humidity without food and free water for at least a month, showing their impressive capacity for water retention. Rates of water loss of immature adults correlate with size, and no transition temperature is detected in females. To replenish water stores, cockroaches drank liquid water; there is no evidence for water gain by water vapour absorption. The profound impact exerted by the 'group' for water conservation suggests that members of this species live huddled together in nature, particularly during the long tropical dry season in order to conserve water, and this adds to previous evidence for the existence of a probable social structure.     

Effects of water and nitrogen addition on species turnover in temperate grasslands in northern China

Global nitrogen (N) deposition and climate change have been identified as two of the most important causes of current plant diversity loss. However, temporal patterns of species turnover underlying diversity changes in response to changing precipitation regimes and atmospheric N deposition have received inadequate attention. We carried out a manipulation experiment in a steppe and an old-field in North China from 2005 to 2009, to test the hypothesis that water addition enhances plant species richness through increase in the rate of species gain and decrease in the rate of species loss, while N addition has opposite effects on species changes. Our results showed that water addition increased the rate of species gain in both the steppe and the old field but decreased the rates of species loss and turnover in the old field. In contrast, N addition increased the rates of species loss and turnover in the steppe but decreased the rate of species gain in the old field. The rate of species change was greater in the old field than in the steppe. Water interacted with N to affect species richness and species turnover, indicating that the impacts of N on semi-arid grasslands were largely mediated by water availability. The temporal stability of communities was negatively correlated with rates of species loss and turnover, suggesting that water addition might enhance, but N addition would reduce the compositional stability of grasslands. Experimental results support our initial hypothesis and demonstrate that water and N availabilities differed in the effects on rate of species change in the temperate grasslands, and these effects also depend on grassland types and/or land-use history. Species gain and loss together contribute to the dynamic change of species richness in semi-arid grasslands under future climate change.