Kinetic and structural properties of disulfide engineered phospholipase A2: insight into the role of disulfide bonding patterns

The family of secreted 14 kDa phospholipase A(2) (PLA2) enzymes have a common motif for the catalytic site but differ in their disulfide architecture. The functional significance of such structural changes has been analyzed by comparing the kinetic and spectroscopic properties of a series of disulfide mutants engineered into the sequence of pig pancreatic IB PLA2 to resemble the mammalian paralogues of the PLA2 family [Janssen et al. (1999) Eur. J. Biochem. 261, 197-207, 1999]. We report a detailed comparison of the functional parameters of pig iso-PLA2, as well as several of the human homologues, with these disulfide engineered mutants of pig IB PLA2. The crystal structure of the ligand free and the active site inhibitor-MJ33 bound forms of PLA2 engineered to have the disulfide bonding pattern of group-X (eng-X) are also reported and compared with the structure of group-IB and human group-X PLA2. The engineered mutants show noticeable functional differences that are rationalized in terms of spectroscopic properties and the differences detected in the crystal structure of eng-X. A major difference between the eng-mutants is in the calcium binding to the enzyme in the aqueous phase, which also influences the binding of the active site directed ligands. We suggest that the disulfide architecture of the PLA2 paralogues has a marginal influence on interface binding. In this comparison, the modest differences observed in the interfacial kinetics are attributed to the changes in the side chain residues. This in turn influences the coupling of the catalytic cycle to the calcium binding and the interfacial binding event.     

Thyroid dysfunction in adult long-term survivors after hemapoeitic stem-cell transplantation (HSCT).

Thyroid function was evaluated in 72 adult survivors (41 females and 31 males) at 16 to 56 years of age, 1.5 years mean time (range 0.2 - 9.8) after hemapoeitic stem cell transplantation (HSCT) with no known prior history of thyroid dysfunction. Thyroid stimulating hormone (TSH) and free thyroxin levels (FT4) were determined before and after stimulation with thyrotropin releasing hormone (TRH). Conditioning regimens for HSCT did not include TBI. Overt hypothyroidism (basal TSH > 8 microIU/ml, FT4 < 0.8 ng/dl) was observed in 6% of male patients and 5% of female patients; subclinical hypothyroidism (basal TSH 4 - 8 microIU/ml, low normal FT4 0.8 - 1.9 ng/dl) was observed in 13% of males and 5% of females. A significant number of euthyroid patients (40% males and 54% females) with normal basal TSH and FT4 levels overresponded to TRH stimulation; the finding being statistically significant (p < 0.005). A heavy TSH response after TRH stimulation indicates compensated subclinical dysfunction of the thyroid gland. Chemotherapy-only conditioning regimens may have an adverse effect on thyroid gland function not always detected by determination of basal TSH and FT4 levels. This finding warrants long-term evaluation of thyroid function in HSCT patients.     

中国白条天牛属比较形态学研究(鞘翅目,天牛科,沟胫天牛亚科,白条天牛族)

比较研究了白条天牛属10种的外部形态特征.结果表明:颊、触角、前胸背板斑纹、鞘翅斑纹、鞘翅基部瘤突、中胸后侧片、前足第1跗节等外部形态特征在白条天牛属中具有种间分类学意义.上颚基部、腹部第7节腹板形状、触角长度可用于雌雄鉴别.通过比较云斑白条天牛Batocera lineolata Chevrolat,1852与多斑白条天牛Batocera horsfieldi(Hope,1839)的外部形态与雄性外生殖器特征,结果表明上述两种均为有效种.     

Chloroplast phylogeographic patterns of Calligonum sect. Pterococcus (Polygonaceae) in arid Northwest China

To understand the impacts of past climatic change and geological events on the evolutionary history of Calligonum sect. Pterococcus, including C. aphyllum, C. rubicundum and C. leucocladum, a total of 128 individuals from 14 populations, mainly from arid Northwest China, were sampled. Two cpDNA intergenic spacer regions (rpl32‐trnL and ycf6‐psbM) were sequenced and 11 haplotypes were identified. Levels of genetic differentiation between populations was low in C. rubicundum (F_ST = 0.54317, p < 0.001) and C. aphyllum (F_ST = 0.55795), while much higher in C. leucocladum (F_ST = 0.95800, p < 0.001), possibly as an effct of differences in geographic distributions and habitats. Analysis of molecular variance (AMOVA) revealed that most of the total genetic variations occurred among species (72.97%). Among eleven identified haplotypes, only H1 and H2 were shared between C. aphyllum and C. rubicundum, while nine were private for one of the three species. The eleven identified haplotypes were divided into two major clades, but they did not yield three species‐specific lineages. Calligonum sect. Pterococcus therefore not appeared reciprocally monophyletic, more likely due to incomplete lineage sorting than hybridization. Mismatch distribution analysis suggested that only C. aphyllum has experienced recent demographic expansion. Divergence time among the 11 haplotypes was estimated at between 2.84 Ma and 0.06 Ma. Within the two clades, haplotype divergence began in early Pleistocene and mainly occurred during the middle to late Pleistocene and was most likely triggered by Quaternary climatic oscillations and increasing aridity of the region.     

不同水分胁迫对刺槐生理的影响

本文以二年生刺槐为供试材料,探讨水分胁迫对其生理的影响,为园林绿地中刺槐的灌溉提供理论指导.结果表明,水淹和干旱胁迫明显影响刺槐的生长.水淹11 d后,qP值、ETR值、Fv'/Fm'值和净光合速率的明显下降,qN值上升,19 d后植株死亡.中度干旱胁迫(土壤含水量在8%～15%之间)对植株生长有一定影响,表现为净光合速率的下降和qN值上升等;干旱胁迫(土壤含水量低于10%)严重抑制植株的生长,表现为qN值上升,而qP、ETR、Fv'/Fm'和净光合速率下降,处理19 d后干旱组植株死亡.轻度干旱(土壤含水量在15%～25%之间)适宜植物生长,表现为3次测定的qP、ETR、Fv'/Fm'、Fv/Fm和净光合速率都较高且稳定.土壤含水量日变化在15%～25%以内受轻度干旱胁迫是园林中刺槐的最佳灌溉方式,既不影响景观效果,同时也能节约灌溉用水.     

Patterns of nucleotide diversity in wild and cultivated rice

There are few reports of the patterns of polymorphism in the non-coding regions of plant genomes. In this study, we explored nucleotide diversity and linkage disequilibrium (LD) in 47 non-coding regions on chromosome 4 of wild and cultivated rice. The cultivated rice retained about 70% of the diversity of wild rice, which was verified by coalescent simulations with one population bottleneck for 198 combinations of duration and population sizes. Multi-locus likelihood analysis showed that the severity of the bottleneck ranged from 2.25 to 3.33, with an average value of 2.70; i.e., the diversity found in the cultivated rice could be explained by a founding population of 2,700 individuals if the initial domestication event occurred over a period of 1,000 years. LD decreased more rapidly in wild rice than in cultivated rice within 10 kb, and the LD observed in cultivated rice was increased at 100–140 kb by comparison with wild rice. The patterns of LD indicated the possibility of a haplotype block in cultivated rice but not in wild rice.     

P73蛋白在血管瘤组织中的表达及临床意义

目的探讨P73蛋白在血管瘤组织中的表达及在血管瘤发生、发展中的作用.方法采用免疫组织化学S-P法分别检测(各20例)增生期血管瘤、退化期血管瘤、正常皮肤组织中P73蛋白的表达,并用HPIAS-2000多媒体彩色病理图像分析系统定量分析、比较P73蛋白在增生期血管瘤、退化期血管瘤和正常皮肤组织中的表达.结果在增生期血管瘤、退化期血管瘤和正常皮肤,P73蛋白免疫组化阳性反应颗粒的平均光密度分别为:6.408±2.151,1.073±0.516,0.953±0.120;阳性面积率分别为:0.184±0.015,0.098±0.014,0.087±0.012.增生期血管瘤与退化期血管瘤、正常皮肤组分别相比,P73蛋白阳性表达的平均光密度和阳性面积率有显著性差异(P<0.001 ) ,消退期血管瘤与正常皮肤组之间,P73蛋白阳性表达的无显著性差异(P>0.05).结论 P73蛋白在血管瘤发生发展和毛细血管内皮细胞的异常过度增生中起着重要的作用.