Two novel bocaparvovirus species identified in wild Himalayan marmots

Bocaparvovirus (BOV) is a genetically diverse group of DNA viruses and a possible cause of respiratory, enteric, and neurological diseases in humans and animals. Here, two highly divergent BOVs (tentatively named as Himalayan marmot BOV, HMBOV1 and HMBOV2) were identified in the livers and feces of wild Himalayan marmots in China, by viral metagenomic analysis. Five of 300 liver samples from Himalayan marmots were positive for HMBOV1 and five of 99 fecal samples from these animals for HMBOV2. Their nearly complete genome sequences are 4,672 and 4,887 nucleotides long, respectively, with a standard genomic organization and containing protein-coding motifs typical for BOVs. Based on their NS1, NP1, and VP1, HMBOV1 and HMBOV2 are most closely related to porcine BOV SX/1-2 (approximately 77.0%/50.0%, 50.0%/53.0%, and 79.0%/54.0% amino acid identity, respectively). Phylogenetic analysis of these three proteins showed that HMBOV1 and HMBOV2 formed two distinctly independent branches in BOVs. According to these results, HMBOV1 and HMBOV2 are two different novel species in the Bocaparvovirus genus. Their identification expands our knowledge of the genetic diversity and evolution of BOVs. Further studies are needed to investigate their potential pathogenicity and their impact on Himalayan marmots and humans.     

Pre-zygotic embryological characters ofPlatycrater arguta, a rare and endangered species endemic to East Asia

Platycrater arguta Sieb. et Zucc. is a rare and endangered species endemic to East Asia. It produces two floral morphs viz. bisexual and male flowers. For bisexual flowers, simultaneous cytokinesis in the microsporocyte meiosis leads to a tetrahedral tetrad. The mature pollen grain is shed at 2-cell stage. The young anther wall is composed of epidermis, endothecium that develops fibrous thickenings at maturity, 1–2 middle layers and tapetum. The tapetum with uninucleate to binucleate cells, disintegrates in situ (glandular tapetum), yet in a small percentage of the anthers (about 37.6%), the tapetum does not disintegrate, causing complete male sterility. The ovules are anatropous, unitegmic, tenuinucellar and the formation of the embryo sac follows the monosporic, Polygonum type. Antipodal cells are lacking in the mature embryo sacs. Before fertilization, two polar nuclei fuse into a secondary nucleus. The formation of microsporangial wall, microsporogenesis and male gametogenesis in male flowers are analogous to those in the bisexual. Prezygotic embryological characters ofP. arguta were reported for the first time, revealing that its endangerment is correlated with the abortion of pollen of a part but not to the female development that is normal.     

Synthesis of 6-substituted 1-phenylbenzazepines and their dopamine D(1) receptor activities

A series of racemic 6-aryl substituted 1-phenylbenzazepines 7a-e, and 17a,b were prepared. All these compounds showed binding potencies compatible to or much higher than that of the prototypic (+/-)-SKF-38393 ((+/-)-1) and (+/-)-SKF-83959 (3) for the D(1) receptor. Among analogs of (+/-)-SKF-38393, compounds 7b, 7c and 7e possess 10-, 2- and 7-fold enhancement in binding for the D(1) receptor, respectively. Lower but compatible potency to that of (+/-)-1 was observed for compounds 7a and 7d. The optimal 6-substituents (m-tolyl, and 2'-naphthyl) were applied to the skeleton of (+/-)-SKF-83959 (3). The resulting compounds 17a,b displayed high affinity at the D(1) receptor, only slightly lower than that of 3. These two compounds also showed good binding at the D(2) receptor.     

N-Propylnoraporphin-11-O-yl carboxylic esters as potent dopamine D(2) and serotonin 5-HT(1A) receptor dual ligands

A small series of N-propylnoraporphin-11-O-yl carboxylic esters with variant ester lengths were synthesized and their binding potencies at dopamine receptors (D(1), D(2)) and serotonin receptors (5-HT(1A), 5-HT(2A)) were evaluated. Monoesters 3a-f showed binding potency of 100 nM or less for the D(2) receptor, and potency of 10-30 nM for the 5-HT(1A) receptor. Butyryl ester 3d was found to be the best compound possessing the highest potency for both receptors, with K(i) values of 55 and 12 nM for D(2) and 5-HT(1A) receptors, respectively. There is no correlation between the binding potency and the length of the monoesters, but the diesters 9 and 10 were inactive for the D(2) receptor. The dual binding profile of these monoesters for the D(2) and 5-HT(1A) receptors may be useful for the treatment of neuropsychiatric disorders.     

Synthesis and pharmacological investigation of novel 2-aminothiazole-privileged aporphines

A series of apomorphine ((-)-1, APO)-derived analogues ((+/-)-3, (-)-4-(-)-6) were designed and synthesized by hybridizing APO with a privileged 2-aminothiazole functionality which was lent from the orally available anti-parkinsonian drug, pramipexole (2). Among these hybridized compounds, catecholic aporphine (-)-6 shows good affinity at the D(2) receptor with K(i) of 328nM, slightly less potent (3-fold), but more selective against the D(1) receptor than that of the parent compound, APO. Although possessing reduced affinity at the D(2) receptor, aporphines 15 and 18 show significant potency at both the D(1) and 5-HT(1A) receptors. The former compound is equipotent at both receptors (K(i): 116 and 151nM, respectively), while the latter is 8-fold more potent at the D(1) (K(i): 78nM) than at the 5-HT(1A) receptors (K(i): 640nM). These results indicate that the catechol fragment is critical for the D(2) receptor binding of the anti-parkinsonian drug, APO ((-)-1), but not necessary for binding at the D(1) and 5-HT(1A) receptors.     

基于贝叶斯网络的DNA序列剪接位点预测

采用基于贝叶斯网络的建模方法,预测真核生物DNA序列中的剪接位点．分别建立了供体位点和受体位点模型,并根据两种位点的生物学特性,对模型的拓扑结构和上下游节点的选择进行了优化．通过贝叶斯网络的最大似然学习算法求出模型参数后,利用10分组交互验证方法对测试数据进行剪接位点预测。结果显示,受体位点的平均预测准确率为92．5％,伪受体位点的平均预测准确率为94．0％,供体位点的平均预测准确率为92．3％,伪供体位点的平均预测准确率为93．5％,整体效果要好于基于使用独立和条件概率矩阵、以及隐Markov模型的预测方法．表明利用贝叶斯网络对剪接位点建模是预测剪接位点的一种有效手段．     

胡萝卜光自养型愈伤组织的诱导培养及其光合特性

经诱导得到胡萝卜光自养型愈伤组织。以叶绿素为单位计算,获得的光自养型愈伤组织的光合活力达到甚至超过了整体植株叶片水平。同时测定愈伤组织光自养过程中光合特性的变化,结果表明其叶绿素含量逐渐上升、暗呼吸速率和Chl a/Chl b比值逐渐下降。并且用电子显微镜观察到愈伤组织中叶绿体结构逐渐发育的过程。     

Multivalent binding oligomers inhibit HIV Tat–TAR interaction critical for viral replication

We describe the development of a new type of scaffold to target RNA structures. Multivalent binding oligomers (MBOs) are molecules in which multiple sidechains extend from a polyamine backbone such that favorable RNA binding occurs. We have used this strategy to develop MBO-based inhibitors to prevent the association of a protein–RNA complex, Tat–TAR, that is essential for HIV replication. In vitro binding assays combined with model cell-based assays demonstrate that the optimal MBOs inhibit Tat–TAR binding at low micromolar concentrations. Antiviral studies are also consistent with the in vitro and cell-based assays. MBOs provide a framework for the development of future RNA-targeting molecules.     

Purification and characterization of a small cationic protein from the tobacco hornworm Manduca sexta

The prophenoloxidase (proPO) activation system is an important defense mechanism in arthropods, and activation of proPO to active phenoloxidase (PO) involves a serine proteinase cascade. Here, we report the purification and characterization of a small cationic protein CP8 from the tobacco hornworm, Manduca sexta, which can stimulate proPO activation. BLAST search showed that Manduca CP8 is similar to a fungal proteinase inhibitor-1 (AmFPI-1), an inducible serine proteinase inhibitor-1 (ISPI-1), and other small cationic proteins with unknown functions. However, we showed that Manduca CP8 did not inhibit proteinase activity, but stimulated proPO activation in plasma. When small amount (0.1 μg) of purified native CP8 or BSA was added to cell-free plasma samples and incubated for 20 min, low PO activity was observed in both groups. But significantly higher PO activity was observed in the CP8-group than in the BSA-group when more proteins (0.5 μg) were added and incubated for 20 min. However, when the plasma samples were incubated with proteins for 30 min, high PO activity was observed in both the CP8 and BSA groups regardless of the amount of proteins added. Moreover, when PO in the plasma was pre-activated with Micrococcus luteus, addition of CP8 did not have an effect on PO activity, and CP8/bacteria mixture did not stimulate PO activity to a higher level than did BSA/bacteria. These results suggest that CP8 helps activate proPO more rapidly at the initial stage. CP8 mRNA was specifically expressed in fat body and its mRNA level decreased when larvae were injected with saline or bacteria. However, CP8 protein concentration in hemolymph did not change significantly in larvae injected with saline or microorganisms.     

栽植生姜对不同种植模式下紫色土微生物生物量及水解酶活性的影响

研究了岷江下游紫色丘陵区玉米+红薯间作、大豆单作、生姜连作、水稻-紫云英轮作等4个典型种植模式下栽植生姜后土壤微生物生物量碳、氮、磷含量和水解酶活性的变化特征.结果表明： 栽植生姜显著降低了4个种植模式下土壤微生物生物量碳、氮和磷含量,但各种植模式之间存在较大差异.其中,玉米+红薯间作和水稻-紫云英轮作模式下土壤微生物生物量碳、氮的下降幅度明显低于大豆单作与生姜连作模式,但土壤微生物生物量磷下降幅度明显较高.栽植生姜显著降低了土壤酸性磷酸酶活性,其下降幅度以玉米+红薯间作模式最大,水稻-紫云英轮作模式最小;土壤转化酶活性在生姜连作模式下显著降低;土壤脲酶活性在大豆单作、生姜连作和水稻-紫云英轮作模式下均显著降低.相对于其他模式,栽植生姜使玉米+红薯间作模式下的土壤维持了较高的转化酶和脲酶活性.