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1.
The lyophilization of proteins in microplates, to assess and optimise formulations rapidly, has been applied for the first time to a therapeutic protein and, in particular, one that requires a cell-based biological assay, in order to demonstrate the broader usefulness of the approach. Factorial design of experiment methods were combined with lyophilization in microplates to identify optimum formulations that stabilised granulocyte colony-stimulating factor during freeze drying. An initial screen rapidly identified key excipients and potential interactions, which was then followed by a central composite face designed optimisation experiment. Human serum albumin and Tween 20 had significant effects on maintaining protein stability. As previously, the optimum formulation was then freeze-dried in stoppered vials to verify that the microscale data is relevant to pilot scales. However, to validate the approach further, the selected formulation was also assessed for solid-state shelf-life through the use of accelerated stability studies. This approach allows for a high-throughput assessment of excipient options early on in product development, while also reducing costs in terms of time and quantity of materials required.  相似文献   
2.
Striking uniformity exists in humans' preference to conceal sexual activity from conspecifics' view. Yet, little is known about the selective pressures acting upon its evolution. To investigate this question, we studied the cooperatively breeding Arabian babbler (Turdoides squamiceps), which has been suggested being the only other species where dominant individuals conceal sex regularly. We examined whether birds indeed conceal sex and tested different hypotheses postulating that sex concealment functions to avoid predators, signal dominance status, or to avoid social interference. The results showed that the birds concealed sex in all observed cases of copulation, did not prefer to copulate under shelters and concealed mating solicitations from adult conspecifics. In addition, subordinates did not attack dominants who courted the respective female. Hence, none of the tested hypotheses explains these results satisfactorily. We postulate that dominant Arabian babblers conceal sex to maintain cooperation with those helpers they prevent from mating. Empirical desiderata for testing this ‘Cooperation-Maintenance’ hypothesis are discussed.  相似文献   
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We tested the hypothesis that diversity and productivity of herbaceous plant communities in disturbed soil are related to the physical and biological heterogeneity of the landscape Our study was earned out on vegetation responses in porcupine diggings on a rocky slope in the central Negev desert in Israel We measured aboveground bio-mass and plant density per species in 150 porcupine diggings (15 cm deep and 15 to 20 cm wide) and in equally sized adjacent control samples in the undisturbed soil matrix We calculated mean annual biomass production, plant density and species richness for 10 sample areas along the slope In addition, we divided the plants into groups according to propagule size and dispersal mode We denoted two types of landscape heterogeneity, which we called physical and biological patchiness Physical patchiness was measured as the ratio of bare rock to soil surface Biological patchiness was the area of the soil covered by shrubs with associated soil mound and under-story relative to the total soil surface We also measured disturbance density, as the long term (17 yr) average density of newly made porcupine diggings We found that 1) the physical patchiness explained 30% of the variation of biological patchiness along the slope, while 2) the patterns of disturbance intensity and biological patchiness were similar (R-=0 386) 3) Biomass, density and species richness were significantly higher in diggings than m the soil matrix 4) Plant density in the matrix, but not m the diggings, was significantly correlated with physical patchiness, 5) species richness in diggings was significantly correlated with biological patchiness, but 6) biomass production in diggings and matrix was not affected by either physical or biological patchiness of the landscape 7) Disturbance density did not affect vegetation responses in diggings and matrix 8) A shift in the plant communities in the matrix towards plants with smaller seeds was associated with increasing physical patchiness, while m diggings there was an opposite shift 9) The proportion of wind dispersers was higher in diggings than outside, while the proportion of runoff dispersers was lower, 10) the densities of runoff dispersers in diggings and matrix were positively correlated with physical and biological patchiness 11) Physical and biological patchiness formed the two major gradients of species composition, explaining 30 and 25% respectively We conclude that the network of physical and biological patchiness and soil disturbance are important in the redistribution of resources and seeds, which control plant biomass, density, species richness and diversity The bare rock surface is the main source for runoff flow with associated soil, organic matter and nutrients The understory vegetation of shrubs provides seeds for creating and maintaining diversity The soil matrix absorbs runoff flow, and disturbances absorb runoff and trap seeds Thus, differences in landscape heterogeneity and their effects on resource and seed movement interact in controlling plant community productivity and diversity in the landscape  相似文献   
5.
Retaining biopharmaceutical proteins in a stable form is critical to their safety and efficacy, and is a major factor for optimizing the final product. Freeze‐dried formulations offer one route for improved stability. Currently the optimization of formulations for freeze‐drying is an empirical process that requires many time‐consuming experiments and also uses large quantities of product material. Here we describe a generic framework for the rapid identification and optimization of formulation excipients to prevent loss of protein activity during a lyophilization process. Using factorial design of experiment (DOE) methods combined with lyophilization in microplates a range of optimum formulations were rapidly identified that stabilized lactose dehydrogenase (derived from Lactobacillus leichmanii) during freeze‐drying. The procedure outlined herein involves two rounds of factorially designed experiments—an initial screen to identify key excipients and potential interactions followed by a central composite face designed optimization experiment. Polyethylene glycol (PEG) and lactose were shown to have significant effects on maintaining protein stability at the screening stage and optimization resulted in an accurate model that was used to plot a window of operation. The variation of freezing temperatures and rates of sublimation that occur across a microplate during freeze‐drying have been characterized also. The optimum formulation was then freeze‐dried in stoppered vials to verify that the microscale data was relevant to the effects observed at larger pilot scales. This work provides a generic approach to biopharmaceutical formulation screening where possible excipients can be screened for single and interactive effects thereby increasing throughput while reducing costs in terms of time and materials. Biotechnol. Bioeng. 2009; 104: 957–964. © 2009 Wiley Periodicals, Inc.  相似文献   
6.
BACKGROUND AND AIMS: Repair of damage to DNA of seed embryos sustained during long periods of quiescence under dry desert conditions is important for subsequent germination. The possibility that repair of embryo DNA can be facilitated by small amounts of water derived from dew temporarily captured at night by pectinaceous surface pellicles was tested. These pellicles are secreted during early seed development and form mucilage when hydrated. METHODS: Seeds of Artemisia sphaerocephala and Artemisia ordosica were collected from a sandy desert. Their embryos were damaged by gamma radiation to induce a standard level of DNA damage. The treated seeds were then exposed to nocturnal dew deposition on the surface of soil in the Negev desert highlands. The pellicles were removed from some seeds and left intact on others to test the ability of mucilage to support repair of the damaged DNA when night-time humidity and temperature favoured dew formation. Repair was assessed from fragmentation patterns of extracted DNA on agarose gels. KEY RESULTS: For A. sphaerocephala, which has thick seed pellicles, DNA repair occurred in seeds with intact pellicles after 50 min of cumulative night dew formation, but not in seeds from which the pellicles had been removed. For A. ordosica, which has thin seed pellicles, DNA repair took at least 510 min of cumulative night dewing to achieve partial recovery of DNA integrity. The mucilage has the ability to rehydrate after daytime dehydration. CONCLUSIONS: The ability of seeds to develop a mucilaginous layer when wetted by night-time dew, and to repair their DNA under these conditions, appear to be mechanisms that help maintain seed viability under harsh desert conditions.  相似文献   
7.
Adipose tissue injection as a free graft for the correction of soft-tissue defects is a widespread procedure in plastic surgery. The main problem in achieving long-term soft-tissue augmentation is partial absorption of the injected fat and hence the need for overcorrection and re-injection. The purpose of this study was to improve the viability of the injected fat by the use of interleukin-8. The rationale for the use of interleukin-8 was its abilities to accelerate angiogenesis and attract inflammatory cells and fibroblasts, providing the injected adipocytes more feeding vessels and a well-established graft bed to enhance their viability. Human adipose tissue, obtained by suction-assisted lipectomy, was re-injected into the subcutis in the scalp of nude mice. Interleukin-8 (0.25 ng) was injected subcutaneously to the scalp as a preparation of the recipient site 24 hours before the fat injection and was added to the fat graft itself (25 ng per 1 cc of injected fat). In the control group, pure fat without interleukin-8 was injected and no interleukin-8 was added for the preparation of the recipient site. One cubic centimeter of fat was injected in each animal in both the study and control groups. There were 10 animals in each group. The animals were euthanized 15 weeks after the procedure. Graft weight and volume were measured and histologic evaluation was performed. In addition, triglyceride content and adipose cell sizes were measured as parameters for fat cells viability. Histologic analysis demonstrated significantly less cyst formation in the group treated with interleukin-8. No significant differences were found between the groups with regard to graft weight and volume or the other histologic parameters investigated. No significant differences were demonstrated in adipose cell sizes and their triglyceride content. In conclusion, less cyst formation, indicating improved quality of the injected fat, can be obtained by the addition of interleukin-8. Further studies of various dosages of interleukin-8 and their long-term effect are required before these encouraging results could be applied clinically.  相似文献   
8.
Schismus arabicus, a desert annual grass, is one of the most common pasture annuals in the deserts of Israel and Asia. S. arabicus exhibits a unique set of adaptations and survival strategies, which enable it to germinate, develop and produce seeds even in years with annual rainfall of less than 100 mm. The current study examined whether an annual rhythm exists in the survival ability of S. arabicus seedlings exposed to desiccation. Our results indicate that survival of S. arabicus seedlings after six different periods of 7 to 42 days of desiccation depended on the month of germination of the caryopses (seeds). Seed germination was 80–100% in all experiments, regardless the month of germination; however, seedlings that germinated in different months varied in their root and shoot elongation rates. None of about 2,500 seedlings that germinated in July (in each of the 4 years) survived the desiccation treatment. The percentages of surviving seedlings in each month of June from 2002 to 2005 were less than 40%. In contrast, over 80% of the seedlings that germinated in each of the months of December and January survived after the desiccation periods of 7–42 days. Seedlings that survived were transferred to 5 L soil pots in which the seedlings developed into mature plants, completed their life cycle and produced seeds that germinated well. The current study demonstrated a novel phenomenon indicating that seedling survival in plants may depend on an annual periodicity according to the date of germination.  相似文献   
9.
Huang  Zhenying  Dong  Ming  Gutterman  Yitzchak 《Plant and Soil》2004,259(1-2):231-241
Psammochloa villosa (Trin.) Bor. (Poaceae), is distributed primarily in moving sand dunes of the Ordos Plateau, China. Freshly harvested caryopses (seeds) are in non-deep physiological dormancy (non-deep PD). Germination is slow and low and only over a narrow temperature range. A treatment of four weeks cold stratification at 3 to 5 °C in darkness was required to break non-deep PD, allowing germination to reach high percentages at higher temperatures and without light requirement. Rate and percentages of germination were increased by scarifying the caryopsis coat and by artificial removal of different proportions of the endosperm. However, seedling dry weight and increases in root and shoot lengths, were significantly influenced by the proportion of the endosperm that remained on the caryopses. Higher percentages of seedling emergence were obtained from the shallowly buried caryopses, ranging from 0.5–2.0 cm, and the depth of the caryopses in the sand affected the time of germination. The deeper the caryopses were buried, the more that remained ungerminated and in enforced dormancy. The caryopses germinated when the upper sand layer was removed and buried caryopses were at a suitable sand depth for germination, or when the sand was aerated. In natural habitats, germinated seedlings may be wholly exposed to the air by sand erosion and thus be exposed to drought stress. However, young seedlings have the ability to tolerate desiccation and to recover after rehydration. Root length at the `point of no return' is 4 mm.  相似文献   
10.
Zaady  Eli  Gutterman  Yitzchak  Boeken  Bertrand 《Plant and Soil》1997,190(2):247-252
We studied the effect of intact, crushed or sterile cyanobacterial soil crust from the Negev Desert highlands of Israel as substrates for the germination of seeds of three annual plant species from local populations that produce mucilaginous seeds t Plantago coronopus, Reboudia pinnata and t Carrichtera annua. Mucilaginous seeds of these species were wetted on local intact cyanobacterial soil crust which inhibited their germination in comparison with their germination on filter paper. However, the percentage of germination of each species differed on sterile and crushed soil crust after 72 h of wetting. Germination of t Plantago coronopus seeds was inhibited mechanically and biologically on intact soil crust, since it was significantly higher on both crushed and sterile soil crust. Germination of t Reboudia pinnata seeds was slightly mechanically and biologically inhibited on intact soil crust, but germination on live crushed crust was higher than on sterile crushed crust. By contrast, germination of t Carrichtera annua seeds was not mechanically inhibited by live soil crust but there was significant biological acceleration of germination on intact soil crust relative to crushed soil crust. Each of the three species exhibited different requirements for germination. Germination of t P. coronopus is promoted by disturbances and removing the crust components (including its biological and structural effects). t R. pinnata and t C. annua germination is less affected by crust disturbance but is generally inhibited after removing the live components of the cyanobacterial crust.These species differences suggest different functions of the mucilage of the seed coat for the three species.  相似文献   
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