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1.

Background

The isolation of lymphocytes – and removal of platelets (PLTs) and red blood cells (RBCs) – from an initial blood sample prior to culture is a key enabling step for effective manufacture of cellular therapies. Unfortunately, currently available methods suffer from various drawbacks, including low cell recovery, need for complex equipment, potential loss of sterility and/or high materials/labor cost.

Methods

A newly developed system for selectively concentrating leukocytes within precisely designed, but readily fabricated, microchannels was compared with conventional density gradient centrifugation with respect to: (i) ability to recover lymphocytes while removing PLTs/RBCs and (ii) growth rate and overall cell yield once expanded in culture.

Results

In the optimal embodiment of the new microfluidic approach, recoveries of CD3+, CD19+ and CD56+ cells (85%, 89% and 97%, respectively) were significantly higher than for paired samples processed via gradient-based separation (51%, 53% and 40%). Although the removal of residual PLTs and RBCs was lower using the new approach, its enriched T-cell fraction nevertheless grew at a significantly higher rate than the gradient-isolated cells, with approximately twice the cumulative cell yield observed after 7 days of culture.

Discussion

The standardization of each step of cellular therapy manufacturing would enable an accelerated translation of research breakthroughs into widely available clinical treatments. The high-throughput approach described in this study – requiring no ancillary pumping mechanism nor expensive disposables to operate – may be a viable candidate to standardize and streamline the initial isolation of lymphocytes for culture while also potentially shortening the time required for their expansion into a therapeutic dose.  相似文献   
2.
1. Although leaf morphology and chemistry show profound changes as trees age, the consequences of such changes to herbivory have received little attention, particularly late in the ontogeny of canopy trees. 2. Using a mobile aerial lift for canopy access, patterns of leaf damage were evaluated in canopy‐dominant mature sugar maple (Acer saccharum Marsh) trees ranging from ~20 to 70 cm in diameter, corresponding to an age range of ~40–180 years. 3. Herbivore damage patterns varied in relation to both vertical canopy position (among upper‐, mid‐, and lower‐canopy positions) and with tree size. Damage types attributable to herbivores active on leaf surfaces, including leaf skeletonizers and leaf cutters (both principally Lepidoptera), and leaf stippling inducers (Hemiptera) showed decreases with tree size, and with increasing height in the canopy. In contrast, leaf damage from the most abundant gall‐forming arthropod in the system, the eriophyid mite Vasates aceriscrumena, increased markedly with tree size. 4. The results indicate that herbivory patterns vary with both canopy stratum and with tree size in sugar maple, and that the relative strength of vertical stratification and tree ontogeny effects are similar in magnitude. The predominant patterns are of a decrease in herbivory with increasing height in the canopy and with tree size, but certain galling arthropods exhibit the reverse trends.  相似文献   
3.
ABSTRACT Little information on foraging habitats of sympatric species of skunks in Texas, USA, is available. We compared 11 western spotted skunks (Spilogale gracilis) and 10 striped skunks (Mephitis mephitis) using radiotelemetry data to assess habitat use during foraging at broad levels of selection in a fragmented habitat. Western spotted skunks used areas with more large mesquite (Prosopis glandulosa) trees than did striped skunks and randomly selected points. Striped skunk habitat use was not different from randomly chosen locations. Contrary to previous research, both species appear to avoid agricultural habitat. A habitat management plan may be difficult to implement for striped skunks in Texas because they did not favor any available habitat. Conservation of western spotted skunks in west-central Texas should focus on areas with older mesquite trees, areas that are now often brush controlled for management of livestock.  相似文献   
4.
目的研究亚甲基蓝/光化学法辅助治疗慢性牙周炎对牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)检出率的变化。方法经伦理委员会同意后,本研究拟筛选出45例慢性牙周炎患者(每位患者经过详细牙周检查及X线检查后确认为牙周炎,有4个以上≥5 mm的牙周袋并分布在2个以上口腔区域),随机分3组:其中A组在SRP之后接受1次PERIOWAVE光化学治疗,B组在SRP后接受1次PERIOWAVE治疗以及在6周后再一次接受光化学治疗,而SRP组仅接受SRP治疗,各组患者取治疗前后相同4个位点龈下菌斑,聚合酶链式反应(Poly-merase chain reaction,PCR)检测Pg,观察Pg菌的检出率,采用卡方检验,并计算χ2值。结果 SRP组、A组、B组3组各组治疗后均较治疗前检出率降低(P<0.001),A组与SRP组治疗后比较差异无统计学意义[A组:40%(24/60),SRP组:45%(27/60),χ2=1.4815,P>0.05],B组与SRP治疗后比较检出率降低[SRP组:45%(27/60),B组:20%(12/60)χ2=8.547,P<0.01];B组与A组治疗后比较检出率降低[A组:40%(24/60),B组:20%(12/60),χ2=5.7143,P<0.05]。结论亚甲基蓝/光化学法辅助治疗慢性牙周炎后牙龈卟啉单胞菌的检出率较治疗前降低;结合临床试验结果,尚可认为2次激光疗效较1次激光好。  相似文献   
5.
6.
1. We studied the effects of different temperature regimes on leaf litter processing in three forested Appalachian headwater streams of different pH (mean pH = 4.2, 6.5, 7.5).
2. We compared leaf breakdown rates, microbial biomass and macroinvertebrate shredder density and biomass between two 12-week processing periods (October–January and November–February) in each stream. Leaf processing rates were calculated both as k (day–1) and k d (degree day–1).
3. There were no significant differences in processing rates ( k day–1) between the two study periods for any leaf species in any stream. The average difference in temperature between the two study periods was 175 degree days. Shredder density was significantly higher during the earlier study period on 40% of the sample dates, but shredder biomass was not significantly different between the two study periods. ATP concentration was significantly higher during the early study period for 60% of the sample dates.
4. More significant differences in these variables (shredder density and biomass, ATP concentration) were seen among the three study streams than between the two study periods. This indicates that in this study other factors, particularly stream pH, contributed more to processing rate variation than did differences in thermal regime.  相似文献   
7.
Microsatellites are continuing to increase in popularity for genetic studies. A major drawback, however, is they often need to be isolated de novo. The traditional method (shotgun cloning) generally results in thousands of recombinant clones, which are then screened by Southern hybridization. Results are typically around 2% positive for mammals, but less than 0.4% for birds. Shotgun cloning is therefore generally not an effective method for isolating microsatellites from avian genomes. Several alternative methods have been developed, but the decision as to which method to use is often arbitrary. Here we present results for two microsatellite isolation techniques from four avian species.  相似文献   
8.
Many spiders use silk to construct webs that must function for days at a time, whereas many other species renew their webs daily. The mechanical properties of spider silk can change after spinning under environmental stress, which could influence web function. We hypothesize that spiders spinning longer‐lasting webs produce silks composed of proteins that are more resistant to environmental stresses. The major ampullate (MA) silks of orb web spiders are principally composed of a combination of two proteins (spidroins) called MaSp1 and MaSp2. We expected spider MA silks dominated by MaSp1 to have the greatest resistance to post‐spin property change because they have high concentrations of stable crystalline β‐sheets. Some orb web spiders that spin three‐dimensional orb webs, such as Cyrtophora, have MA silks that are predominantly composed of MaSp1. Hence, we expected that the construction of three‐dimensional orb webs might also coincide with MA silk resistance to post‐spin property change. Alternatively, the degree of post‐spin mechanical property changes in different spider silks may be explained by factors within the spider's ecosystem, such as exposure to solar radiation. We exposed the MA silks of ten spider species from five genera (Nephila, Cyclosa, Leucauge, Cyrtophora, and Argiope) to ecologically high temperatures and low humidity for 4 weeks, and compared the mechanical properties of these silks with unexposed silks. Using species pairs enabled us to assess the influence of web dimensionality and MaSp composition both with and without phylogenetic influences being accounted for. We found neither the MaSp composition nor the three‐dimensionality of the orb web to be associated with the degree of post‐spin mechanical property changes in MA silk. The MA silks in Leucauge spp. are dominated by MaSp2, which we found to have the least resistance to post‐spin property change. The MA silk in Argiope spp. is also dominated by MaSp2, but has high resistance to post‐spin property change. The ancestry of Argiope is unresolved, but it is largely a tropical genus inhabiting hot, open regions that present similar stressors to silk as those of our experiment. Ecological factors thus appear to influence the vulnerability of orb web spider MA silks to post‐spin property change. © 2012 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 106 , 580–588.  相似文献   
9.
1. Declining abundances of forage fish and the introduction and establishment of non‐indigenous species have the potential to substantially alter resource and habitat exploitation by top predators in large lakes. 2. We measured stable isotopes of carbon (δ13C) and nitrogen (δ15N) in field‐collected and archived samples of Lake Ontario lake trout (Salvelinus namaycush) and five species of prey fish and compared current trophic relationships of this top predator with historical samples. 3. Relationships between δ15N and lake trout age were temporally consistent throughout Lake Ontario and confirmed the role of lake trout as a top predator in this food web. However, δ13C values for age classes of lake trout collected in 2008 ranged from 1.0 to 3.9‰ higher than those reported for the population sampled in 1992. 4. Isotope mixing models predicted that these changes in resource assimilation were owing to the replacement of rainbow smelt (Osmerus mordax) by round goby (Neogobius melanostomus) in lake trout diet and increased reliance on carbon resources derived from nearshore production. This contrasts with the historical situation in Lake Ontario where δ13C values of the lake trout population were dominated by a reliance on offshore carbon production. 5. These results indicate a reduced capacity of the Lake Ontario offshore food web to support the energetic requirements of lake trout and that this top predator has become increasingly reliant on prey resources that are derived from nearshore carbon pathways.  相似文献   
10.
An effective visual signal elicits a strong receiver response. The visual receptors of most insects are sensitive to ultraviolet (UV), blue and green light. The decorations of certain orb web spiders may be described as a type of visual signal, a sensory trap, as they exploit visual biases in insects. We filtered UV and blue light from the decorations of Argiope keyserlingi , under field conditions, using plastic sheets to test if the UV and blue light reflected affects the type of prey caught. We found that houseflies, blowflies, stingless bees, honeybees and vespid wasps were caught more frequently in webs with decorations than webs without, while ichneumonid wasps were caught less frequently. Blowflies, stingless bees, honeybees and vespid wasps were caught more often in unfiltered decorated webs. These insects also have receptor sensitivities in the blue and UV. We showed that exploiting visual sensory biases plays an integral role in attracting insects to orb web decorations. Whether UV light, blue light, or both, are the most important cue, however, requires further study.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 94 , 221–229.  相似文献   
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