Negative Cell Cycle Regulation and DNA Damage-inducible Phosphorylation of the BRCT Protein 53BP1

In a screen designed to discover suppressors of mitotic catastrophe, we identified the Xenopus ortholog of 53BP1 (X53BP1), a BRCT protein previously identified in humans through its ability to bind the p53 tumor suppressor. X53BP1 transcripts are highly expressed in ovaries, and the protein interacts with Xp53 throughout the cell cycle in embryonic extracts. However, no interaction between X53BP1 and Xp53 can be detected in somatic cells, suggesting that the association between the two proteins may be developmentally regulated. X53BP1 is modified via phosphorylation in a DNA damage-dependent manner that correlates with the dispersal of X53BP1 into multiple foci throughout the nucleus in somatic cells. Thus, X53BP1 can be classified as a novel participant in the DNA damage response pathway. We demonstrate that X53BP1 and its human ortholog can serve as good substrates in vitro as well as in vivo for the ATM kinase. Collectively, our results reveal that 53BP1 plays an important role in the checkpoint response to DNA damage, possibly in collaboration with ATM

Coordinated changes in mRNA turnover, translation, and RNA processing bodies in bronchial epithelial cells following inflammatory stimulation.

Bronchial epithelial cells play a pivotal role in airway inflammation, but little is known about posttranscriptional regulation of mediator gene expression during the inflammatory response in these cells. Here, we show that activation of human bronchial epithelial BEAS-2B cells by proinflammatory cytokines interleukin-4 (IL-4) and tumor necrosis factor alpha (TNF-alpha) leads to an increase in the mRNA stability of the key chemokines monocyte chemotactic protein 1 and IL-8, an elevation of the global translation rate, an increase in the levels of several proteins critical for translation, and a reduction of microRNA-mediated translational repression. Moreover, using the BEAS-2B cell system and a mouse model, we found that RNA processing bodies (P bodies), cytoplasmic domains linked to storage and/or degradation of translationally silenced mRNAs, are significantly reduced in activated bronchial epithelial cells, suggesting a physiological role for P bodies in airway inflammation. Our study reveals an orchestrated change among posttranscriptional mechanisms, which help sustain high levels of inflammatory mediator production in bronchial epithelium during the pathogenesis of inflammatory airway diseases

Study on the Structure and Related Parameters of Underwater Ozone Generator

AbstractIn order to control eutrophication for water body better, underwater ozone generator has been developed by electro- hydraulic power impulse technology. By one that in a pair of electrodes was designed as hollow structure, the charge and discharge circuit of the device and the structure of the electric chamber were analyzed. And not only the optimal related electrical parameters, such as voltage, frequency of discharge and electric capacity, but also the best parameters of electrodes and the structure of electric chamber have been established. The result shows that the processes of product ozone and purify water are all complete in water by underwater ozone generator based on electro-hydraulic power impulse technology. The related parameters of the device which have been established have provided a basis for improving the efficiency of ozone generation & water treatment and applicability of the ozone generator

GRK5 intronic (CA)n polymorphisms associated with type 2 diabetes in Chinese Hainan Island.

A genome-wide association study had showed G-protein-coupled receptor kinase 5 (GRK5) rs10886471 was related to the risk of type 2 diabetes mellitus (T2DM) through upregulated GRK5 mRNA expression. Rs10886471 is located in the intron region of GRK5. However, the mechanism by which intronic SNP affects gene expression remains unclear, whether the effect on gene expression depends on the intronic short tandem repeat (STR) (CA)n splicing regulator or not. Here we investigated the STR (CA)n polymorphism in rs10886471 and further discussed its role in the T2DM risk of Chinese Hainan Island individuals. A total of 1164 subjects were recruited and classified into a normal fasting glucose (NFG) group, an impaired fasting glucose (IFG) group, an impaired glucose tolerance (IGT) group, and a T2DM group. STR (CA)n polymorphisms were detected through polymerase chain reaction and sequencing. Five intronic (CA)n alleles, (CA)15 to (CA)19, were identified in GRK5 rs10886471. Only the (CA)16 allele was significantly associated with increased prediabetes and T2DM risk [odds ratio (OR)>1, P<0.05]. Conversely, multiple alleles without any (CA)16 protected against prediabetes and T2DM (0<OR<1, P<0.05). In summary, rs10886471 acts as both an SNP and an STR. The rs10886471 intronic SNP causes GRK5 overexpression the subsequent risk of T2DM may be due to the rs10886471 intronic STR (CA)n splicing enhancer. Further studies should focus on verifying these finding using a large sample size and analyzing the splicing mechanism of intronic (CA)n in rs10886471

Structure-Guided Strategies of Targeted Therapies for Patients with <i>EGFR</i>-Mutant Non–Small Cell Lung Cancer

Oncogenic mutations within the EGFR kinase domain are well-established driver mutations in non–small cell lung cancer (NSCLC). Small-molecule tyrosine kinase inhibitors (TKIs) specifically targeting these mutations have improved treatment outcomes for patients with this subtype of NSCLC. The selectivity of these targeted agents is based on the location of the mutations within the exons of the EGFR gene, and grouping mutations based on structural similarities has proved a useful tool for conceptualizing the heterogeneity of TKI response. Structure-based analysis of EGFR mutations has influenced TKI development, and improved structural understanding will inform continued therapeutic development and further improve patient outcomes. In this review, we summarize recent progress on targeted therapy strategies for patients with EGFR-mutant NSCLC based on structure and function analysis

Structure-Guided Strategies of Targeted Therapies for Patients with EGFR-Mutant Non&ndash;Small Cell Lung Cancer

Oncogenic mutations within the EGFR kinase domain are well-established driver mutations in non&ndash;small cell lung cancer (NSCLC). Small-molecule tyrosine kinase inhibitors (TKIs) specifically targeting these mutations have improved treatment outcomes for patients with this subtype of NSCLC. The selectivity of these targeted agents is based on the location of the mutations within the exons of the EGFR gene, and grouping mutations based on structural similarities has proved a useful tool for conceptualizing the heterogeneity of TKI response. Structure-based analysis of EGFR mutations has influenced TKI development, and improved structural understanding will inform continued therapeutic development and further improve patient outcomes. In this review, we summarize recent progress on targeted therapy strategies for patients with EGFR-mutant NSCLC based on structure and function analysis

Frequency distribution of rs10886471 (CA)<i>_n</i> alleles in the four groups.

<p>Not all comparisons are shown. The allelic frequencies of (CA)<i>₁₆</i> in the IFG, IGT, and T2DM groups wasmuch higher than that in the NFG group (^△<b>X</b><b>²</b> = 12.300, <i>P</i> = 0.000; ^□<b>X</b><b>²</b> = 13.672, <i>P</i> = 0.000;^▪<b>X</b><b>²</b> = 14.476,<i>P</i> = 0.000, respectively). The allelic frequency of (CA)<i>₁₆</i> was significantly lower than that of (CA)<i>₁₇</i>, but higher than those of (CA)<i>₁₈</i> and (CA)<i>₁₉</i> among the four groups (^△△<b>X</b><b>²</b> = 16.190, <i>P</i> = 0.001; ^□□<b>X</b><b>²</b> = 10.221,<i>P</i> = 0.017; ^▪▪<b>X</b><b>²</b> = 8.265, <i>P</i> = 0.041, respectively).</p

Association of GRK5 intronic (CA)_n repeat polymorphisms with prediabetes and T2DM (n = 1164).

a<p>Reference allele; <b>^b</b>Lacking the (CA)<i>₁₆</i> allele and contains (CA)<i>₁₅</i>, (CA)<i>₁₇</i>,(CA)<i>₁₈</i>, and (CA)<i>₁₉</i>; OR, odds ratio; 95%CI, 95% confidence interval.</p

Intronic (CA)<i>n</i> polymorphism in <i>GRK5</i> rs10886471.

<p>(A) Amplified fragments in the <i>GRK5</i> rs10886471 studied region. The PCR products were electrophoresed in 2% agarose gel and were then photographed under UV light. Bands 1–12 were from the NFG group, bands 13–24 from were from the IFG group, bands 25–36 were from the IGT group, and bands 37–48 were from the T2DM group. The PCR marker was in lane M. (B) SNPSTR marker in <i>GRK5</i> rs10886471. PCR sequencing demonstrated a STR (CA)<i>_n</i> with one tightly linked SNP (C/T) in <i>GRK5</i> rs10886471. (C) Sequence analysis using the physical map of <i>GRK5</i> rs10886471. The SNPSTR marker is located in the intron of <i>GRK5</i> rs10886471.</p

The mechanism of intronic (CA)<i>_n</i> splicing regulator in rs10886471.

<p>The heterogeneous nuclear ribonucleoprotein L (hnRNP L) is specifically bound to diverse CA elements. ^{10, 13} It contains four RNA recognition motifs (RRMs) that bind to CA repeats. The crystal structures of hnRNP L RRMs at 2.0 and 1.8 Å has been elucidated. ²⁰ The intronic (CA)<i>_n</i> repeats in <i>GRK5</i> rs10886471 act as splicing enhancers or repressors and their yin–yang effect on T2DM depends on the CA repeat number. Intronic SNPs that affect gene expression may be mediated by LD with intronic STR (CA)<i>_n</i> regulators.</p