Control of glucocorticoid and progesterone receptor subcellular localization by the ligand-binding domain is mediated by distinct interactions with tetratricopeptide repeat proteins

The TPR proteins FKBP52, FKBP51, Cyp40, and PP5 are found in steroid receptor (SR) complexes, but their receptor-specific preferences and roles remain unresolved. We have undertaken a systematic approach to this problem by examining the contribution of all four TPRs to the localization properties of glucocorticoid (GR) and progesterone (PR) receptors. The GR of L929 cells was found in the cytoplasm in a complex containing PP5 and FKBP51, while the GR of WCL2 cells was nuclear and contained PP5 and FKBP52. Cyp40 did not interact with the GR in either cell line. To test whether FKBP interaction determined localization, we overexpressed Flag-tagged FKBP51 in WCL2 cells and Flag-FKBP52 in L929 cells. In WCL2 cells, the GR exhibited a shift to greater cytoplasmic localization that correlated with recruitment of Flag-FKBP51. In contrast, Flag-FKBP52 was not recruited to the GR of L929 cells, and no change in localization was observed, suggesting that both cell-type-specific mechanisms and TPR abundance contribute to the SR-TPR interaction. As a further test, GR-GFP and PR-GFP constructs were expressed in COS cells. The GR-GFP construct localized to the cytoplasm, while the PR-GFP construct was predominantly nuclear. Similar to L929 cells, the GR in COS interacted with PP5 and FKBP51, while PR interacted with FKBP52. Analysis of GR-PR chimeric constructs revealed that the ligand-binding domain of each receptor determines both TPR specificity and localization. Lastly, we analyzed GR and PR localization in cells completely lacking TPR. PR in FKBP52 KO cells showed a complete shift to the cytoplasm, while GR in FKBP51 KO and PP5 KO cells showed a moderate shift to the nucleus, indicating that both TPRs contribute to GR localization. Our results demonstrate that SRs have distinct preferences for TPR proteins, a property that resides in the LBD and which can now explain long-standing differences in receptor subcellular localization.     

High-throughput quantification of microbial birth and death dynamics using fluorescence microscopy

Background: Microbes live in dynamic environments where nutrient concentrations fluctuate. Quantifying fitness in terms of birth rate and death rate in a wide range of environments is critical for understanding microbial evolution and ecology. Methods: Here, using high-throughput time-lapse microscopy, we have quantified how Saccharomyces cerevisiae mutants incapable of synthesizing an essential metabolite (auxotrophs) grow or die in various concentrations of the required metabolite. We establish that cells normally expressing fluorescent proteins lose fluorescence upon death and that the total fluorescence in an imaging frame is proportional to the number of live cells even when cells form multiple layers. We validate our microscopy approach of measuring birth and death rates using flow cytometry, cell counting, and chemostat culturing. Results: For lysine-requiring cells, very low concentrations of lysine are not detectably consumed and do not support cell birth, but delay the onset of death phase and reduce the death rate compared to no lysine. In contrast, in low hypoxanthine, hypoxanthine-requiring cells can produce new cells, yet also die faster than in the absence of hypoxanthine. For both strains, birth rates under various metabolite concentrations are better described by the sigmoidal-shaped Moser model than the well-known Monod model, while death rates can vary with metabolite concentration and time. Conclusions: Our work reveals how time-lapse microscopy can be used to discover non-intuitive microbial birth and death dynamics and to quantify growth rates in many environments.     

广西凌云县罗楼地区早三叠世牙形石

罗楼地区下三叠统,经赵金科（１９５９）研究显示,最低层位为Ｖishnuites marginalis菊石带（Gyronitan阶）,张舜新（１９９０）研究广西凤山金下三叠统形石时,未发现最高的牙形石带,当前研究发现,罗楼地区下三叠统牙形石丰富,可识别出Ｈindeodus parvus,Isarciclla staeschei ,Neospathodus waageni,Neospathodus novaehel-landiae,Neospathodus homeri-Neospathodus triangularis,Chiosella timorensis6个牙形石带（组合带）,从而表明该地区不但存在下三叠统底部层位Ｏtoceratan阶,而且存在下三叠统最高的牙形石带,牙形石组合面貌和岩相显示,该牙形石动物群生活环境与田东县作登地区同属盆地相环境。     

Genomic and proteomic perspectives in cell culture engineering.

In the last few years, the number of biologics produced by mammalian cells have been steadily increasing. The advances in cell culture engineering science have contributed significantly to this increase. A common path of product and process development has emerged in the last decade and the host cell lines frequently used have converged to only a few. Selection of cell clones, their adaptation to a desired growth environment, and improving their productivity has been key to developing a new process. However, the fundamental understanding of changes during the selection and adaptation process is still lacking. Some cells may undergo irreversible alteration at the genome level, some may exhibit changes in their gene expression pattern, while others may incur neither genetic reconstruction nor gene expression changes, but only modulation of various fluxes by changing nutrient/metabolite concentrations and enzyme activities. It is likely that the selection of cell clones and their adaptation to various culture conditions may involve alterations not only in cellular machinery directly related to the selected marker or adapted behavior, but also those which may or may not be essential for selection or adaptation. The genomic and proteomic research tools enable one to globally survey the alterations at mRNA and protein levels and to unveil their regulation. Undoubtedly, a better understanding of these cellular processes at the molecular level will lead to a better strategy for 'designing' producing cells. Herein the genomic and proteomic tools are briefly reviewed and their impact on cell culture engineering is discussed.     

Expression of a synthetic E. coli heat-labile enterotoxin B sub-unit (LT-B) in maize

We have produced the B subunit of the enterotoxigenic Escherichia coli (ETEC) heat-labile enterotoxin (LT-B) in transgenic maize seed. LT-B is a model antigen that induces a strong immune response upon oral administration and enhances immune responses to conjugated and co-administered antigens. Using a synthetic LT-B gene with optimized codon sequence, we examined the role of promoters and the SEKDEL endoplasmic reticulum retention motif in LT-B accumulation in callus and in kernels. Two promoters, the constitutive CaMV 35S promoter and the maize 27 kDa gamma zein promoter, which directs endosperm-specific gene expression in maize kernels, regulated LT-B expression. Ganglioside-dependent ELISA analysis showed that using the constitutive promoter, maximum LT-B level detected in callus was 0.04% LT-B in total aqueous-extractable protein (TAEP) and 0.01% in R₁ kernels of transgenic plants. Using the gamma zein promoter, LT-B accumulation reached 0.07% in R₁ kernels. The SEKDEL resulted in increased LT-B levels when combined with the gamma zein promoter. We monitored LT-B levels under greenhouse and field conditions over three generations. Significant variability in gene expression was observed between transgenic events, and between plants within the same event. A maximum of 0.3% LT-B in TAEP was measured in R₃ seed of a transgenic line carrying CaMV 35S promoter/LT-B construct. In R₃ seed of a transgenic line carrying the gamma zein promoter/LT-B construct, up to 3.7% LT-B in TAEP could be detected. We concluded that maize seed can be used as a production system for functional antigens.     

Growth of hybridoma cells under different agitation conditions

Summary The effect of mechanical agitation on hybridoma cell growth was examined in laboratory scale vessels. At an agitation rate four times that required to keep the cells in suspension, both growth rate and growth extent were reduced. However, using the minimum agitation rate required to suspend cells, no adverse effect on cell growth was observed even with a turbine agitator.     

西藏锐顶天牛属一新种（鞘翅目：天牛科）

本记述锐顶天牛属一新种，即墨脱锐颚天牛Cleptometopus motuoensis sp．nov．。模式标本保存于西南农业大学植保系昆虫标本馆。     

Optimal Management of Two-Species Bioeconomics Systems

1IntroductionOptimalmanagementofsystemsofrenewableresourcesisanimportantbranchofMathemati-calEcology.Theoptimalmanagementofbiologicalspecies,asatypicalrepresentativeoftheopti-malmanagementofrenewableresourcesisbeingattachedmuchmoreinportancebyBioeconomistandbecameanindependentbranchinthe1980's.C.W.ClarkandR.W.Mcklvey,etc,didalotofresearchworkinthisfield.Themajorcharacteristicoftheoptimalmanagementofbioeconomicsystemistooptimizetheharvestrevenueonthebasisofecologicalbalance(e.g.,permanentex…     

A new natural allotriploid,Lycoris × hubeiensis hybr. nov. (Amaryllidaceae), identified by morphological,karyological and molecular data

A new natural hybrid, Lycoris × hubeiensis K. Liu, is confirmed and described based on morphological, karyological and molecular data. The new natural hybrid displays remarkable morphological differences relative to the putative parents L. radiata (2n = 22 = 22A) and L. aurea (2n = 14 = 8m + 6T), and has a karyotype of 3n = 29 = 4m + 22A + 3T. Two chloroplast intergenic regions of ndhF‐rpl32R and rpl32F‐trnL were sequenced with three haplotypes found in the three taxa. The putative hybrid and L. radiata shared the same haplotype (H1), while L. aurea had two other haplotypes (H2 and H3). Combined with the morphological, karyological and molecular data, we conclude that the hybrid is an allotriploid with L. radiata being the maternal parent offering an unreduced gamete and L. aurea being the male parent.     

几种化学药剂对悬铃木碳氮比值和花芽分化的影响

四种化学药剂处理的悬铃木,其冬季糖和氮含量均高于对照,春季则相反;并对新梢伸长有促进,对粗度有抑制效应;其中经ＧＡ３和乙硫氨酸处理的Ｃ／Ｎ比值增大,新梢成花量增加;乙烯利和２硫尿嘧啶处理的Ｃ／Ｎ比值减小,新梢成花量显著减少。从而为选用减灭悬铃木球果药剂提供理论依据