Purification and identification of a protein kinase activity modulated by ionizing radiation

ATBF1 was first discovered as a suppressor of AFP expression in hepatocytes. It is present in brain, adult liver, lung, and gastro-intestinal tract. Recently, it has been reported that ATBF1 regulates myoblastic differentiation and interacts with v-Myb in regulation of its transactivation. Using the yeast two-hybrid system, we searched for protein-protein interactions to uncover new functions for ATBF1. We present here experimental evidence that ATBF1 is a new regulatory factor for STAT3-mediated signal transduction through its interaction with PIAS3. PIAS3 was thus identified as an ATBF1-binding protein. In co-transfection experiments, the full-length ATBF1 was found to form complexes with PIAS3 in Hep G2 cells. In the luciferase assay, ATBF1 was found to have no influence on STAT3 signaling induced by IL-6 stimulation, but it did synergistically enhance PIAS3 inhibition of activated STAT3. In conclusion, ATBF1 can suppress the IL-6-mediated cellular response by acting together with PIAS3.     

Quantitative evaluation of each catalytic subsite of cathepsin B for inhibitory activity based on inhibitory activity-binding mode relationship of epoxysuccinyl inhibitors by X-ray crystal structure analyses of complexes

To quantitatively estimate the inhibitory effect of each substrate-binding subsite of cathepsin B (CB), a series of epoxysuccinyl derivatives with different functional groups bound to both carbon atoms of the epoxy ring were synthesized, and the relationship between their inhibitory activities and binding modes at CB subsites was evaluated by the X-ray crystal structure analyses of eight complexes. With the common reaction in which the epoxy ring of inhibitor was opened to form a covalent bond with the SgammaH group of the active center Cys29, the observed binding modes of the substituents of inhibitors at the binding subsites of CB enabled the quantitative assessment of the inhibitory effect of each subsite. Although the single blockage of S1' or S2' subsite exerts only the inhibitory effect of IC50 = approximately 24 microM (k2 = approximately 1250 M(-1) s(-1)) or approximately 15 microM (k2 = approximately 1800 M(-1) s(-1)), respectively, the synchronous block of both subsites leads to IC50 = approximately 23 nM (k2 = 153,000 - 185,000 M(-1) s(-1)), under the condition that (i) the inhibitor possesses a P1' hydrophobic residue such as Ile and a P2' hydrophobic residue such as Ala, Ile or Pro, and (ii) the C-terminal carboxyl group of a P2' residue is able to form paired hydrogen bonds with the imidazole NH of His110 and the imidazole N of His111 of CB. The inhibitor of a Pn' > or = 3' substituent was not potentiated by collision with the occluding loop. On the other hand, it was suggested that the inhibitory effects of Sn subsites are independent of those of Sn' subsites, and the simultaneous blockage of the funnel-like arrangement of S2 and S3 subsites leads to the inhibition of IC50 = approximately 40 nM (k2 = approximately 66,600 M(-1) s(-1)) regardless of the lack of Pn' substituents. Here we present a systematic X-ray structure-based evaluation of structure-inhibitory activity relationship of each binding subsite of CB, and the results provide the structural basis for designing a more potent CB-specific inhibitor.     

Novel instrumented probe for measuring 3D pressure distribution along the vaginal canal

We developed an intravaginal instrumented probe (covered with a 10 × 10 matrix of capacitive sensors) for assessing the three-dimensional (3D) spatiotemporal pressure profile of the vaginal canal. The pressure profile was compared to the pelvic floor (PF) digital assessment, and the reliability of the instrument and repeatability of the protocol was tested. We also tested its ability to characterize and differentiate two tasks: PF maximum contraction and Valsalva maneuver (maximum intra-abdominal effort with downward movement of the PF). Peak pressures were calculated for the total matrix, for three major sub-regions, and for 5 planes and 10 rings throughout the vaginal canal. Intraclass correlation coefficients indicated excellent inter- and intra-rater reliability and intra-trial repeatability for the total and medial areas, with moderate reliability for the cranial and caudal areas. There was a moderate correlation between peak pressure and PF digital palpation [Spearman’s coefficient r = 0.55 (p < 0.001)]. Spatiotemporal profiles were completely different between tasks (2-way ANOVAs for repeated measures) with notably higher pressures (above 30 kPa) for the maximum contraction task compared to Valsalva (below 15 kPa). At maximum contraction, higher pressures occurred in the mid-antero-posterior zone, with earlier peak pressure onsets and more variable along the vaginal depth (from rings 3 to 10-caudal). During Valsalva, the highest pressures were observed in rings 4–6, with peak pressure onsets more synchronized between rings. With this protocol and novel instrument, we obtained a high-resolution and highly reliable innovative 3D pressure distribution map of the PF capable of distinguishing vaginal sub-regions, planes, rings and tasks.     

Single‐molecule force spectroscopy applied to heparin‐induced thrombocytopenia

Heparin‐induced thrombocytopenia (HIT), occurring up to approximately 1% to 5% of patients receiving the antithrombotic drug heparins, has a complex pathogenesis involving multiple partners ranging from small molecules to cells/platelets. Recently, insights into the mechanism of HIT have been achieved by using single‐molecule force spectroscopy (SMFS), a methodology that allows direct measurements of interactions among HIT partners. Here, the potential of SMFS in unraveling the mechanism of the initial steps in the pathogenesis of HIT at single‐molecule resolution is highlighted. The new findings ranging from the molecular binding strengths and kinetics to the determination of the boundary between risk and non‐risk heparin drugs or platelet‐surface and platelet‐platelet interactions will be reviewed. These novel results together have contributed to elucidate the mechanisms underlying HIT and demonstrate how SMFS can be applied to develop safer drugs with a reduced risk profile.     

Cross-resistance studies with V79 Chinese hamster cells adapted to the mutagenic or clastogenic effect of N-methyl-N′-nitro-N-nitrosoguanidine

When V79 cells are exposed to a single low dose of MNNG or MNU they acquire resistance to the mutagenic or to the clastogenic effect of the agents. Here the effect of MNNG pretreatment on mutagenesis (6-thioguanine resistance) and aberration formation in cells challenged with various mutagens/clastogens is reported. MNNG-adapted cells were resistant to the mutagenic effects of MNU and, to a lower extent, of EMS. No mutagenic adaptation was observed when MNNG-pretreated cells were challenged with MMS, ENU, MMC or UV.

Cells pretreated with a dose of MNNG which makes them resistant to the clastogenic effect of this compound were also resistant to the clastogenic activity of other methylating agents (MNU, MMS), but not so with respect to ethylating agents (EMS, ENU). Cycloheximide abolished the aberration-reducing effect of pretreatment. However, when given before the challenge dose of MNNG, MNU or MMS, it drastically enhanced the aberration frequency in both pretreated and non-pretreated cells. No significant enhancement of aberration frequency by cycloheximide was found for ethylating agents.

The results indicate that clastogenic adaptation is due to inducible cellular functions. It is concluded that mutagenic and clastogenic adaptation are probably caused by different adaptive repair pathways.     

Sunflower germination and growth behavior under various gamma radiation absorbed doses

Gamma radiation, various absorbed doses (0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5 krad) effects were evaluated on sunflower (Helianthus annus. L.) germination and growth characteristics. Sunflower healthy seeds were exposed to gamma radiation source Co⁶⁰ at nuclear institute for food and agriculture and exposed seeds were grown under controlled laboratory conditions. In comparison to control, gamma radiation absorbed doses affected the measured response positively i.e., radical length, plumule length, number of roots, seedling fresh weight, seedling dry weight, germination percentage, time of germination and diameter of hypocotyl of sunflower enhanced up to 83.15%, 70.32%, 73.03%, 4.80%, 3.26%, 72.0%, ? 18.88% and 12.58%, respectively. The time of germination, fresh weight and percent moisture contents enhanced insignificantly, however, the response was higher than control. All gamma radiation absorbed doses showed a stimulatory effect on sunflower germination and seedling growth characteristics. The low gamma radiation absorbed doses were found to be more effective versus higher doses for enhancing the germination and growth characteristics of sunflower. In view of positive effect of gamma radiation of sunflower germination and growth characteristics, it is concluded that this techniques could possibly be used for the enhancement of germination, growth and ultimately yield in sunflower in areas where germination is low due to unfavorable conditions.     

Radiation Hormesis and Cancer

Despite the long history of radiation hormesis and the public health concerns with low-level exposures to ionizing radiation, there has been surprisingly little formal evaluation of whether hormetic effects are displayed with respect to radiation exposure and cancer incidence (i.e., reduced cancer risk at low radiation doses compared to controls, enhanced cancer risk at higher doses) until relatively recently. This paper reviews data relevant to the question of radiation hormesis and cancer with particular emphasis on experimental studies in animal models exposed to low levels of ionizing radiation. Data exist that provide evidence both consistent with and/or supportive of radiation hormesis. Other biomedical research provides potentially important mechanistic insight: low dose exposures have the capacity to activate immune function to prevent the occurrence of tumor development and metastasis; low doses of radiation have been shown to reduce mutagenic responses and induce endogenous antioxidant responses. These findings are consistent with epidemiological data suggesting an inverse relationship between background radiation and cancer incidence and with occupational epidemiological investigations in which low-dose exposure groups display markedly lower standardized mortality rates than the referent or control group.     

地表UV-B辐射增强对土壤-大豆系统N2O排放的影响

通过大田试验和室外盆栽试验,采用人工增加紫外辐射的方法模拟UV-B辐射增强,用静态箱-气相色谱法测定N_2O排放通量,研究地表UV-B辐射增强对土壤-大豆系统N_2O排放的影响.结果表明:在相同的气象条件和田间管理措施下,UV-B辐射增强没有改变土壤-大豆系统N_2O排放通量的季节性变化规律.但从植株结荚到成熟,UV-B辐射增强降低了土壤-大豆系统N_2O排放通量,进而降低了N_2O的累积排放量.收割实验发现,在分枝开花期,UV-B辐射增强对土壤N_2O排放影响明显,降低了土壤N_2O排放通量;从结荚至鼓粒期,UV-B辐射增强主要通过降低植株地上部分N_2O排放通量来降低土壤-大豆系统的N_2O排放.UV-B辐射增强显著降低了植株的生物量,并影响到植株的氮代谢和土壤NH_4~+-N与微生物氮.UV-B辐射增强可能会导致农田生态系统N_2O排放量降低.