Spatial Distribution of Mountain Pine Beetle Outbreaks in Relation to Climate and Stand Characteristics： A Dendroecological Analysis

Principal components analysis, followed by K-means cluster analysis, was used to detect variations in the timing and magnitude of Pinus contorta Dough ex Loud. growth releases attributed to mountain pine beetle outbreaks in 31 stands of central British Columbia. Four major growth release patterns were identified from 1970 to 2000. Variations in the timing of growth releases among clustered stands corresponded well to aerial survey data indicating the timing of beetle outbreaks in the study area. Redundancy analysis was used to determine how variations in the timing and magnitude of growth releases attributed to beetle outbreaks changed with variations in climate or stand conditions over the study area. The first RDA axis, which accounted for 39% of the variations in growth patterns among stands, was significantly （P〈0.05） correlated with gradients in the percentage of pine in stands killed by mountain pine beetle, summer aridity, variation in summer precipitation, distance from initial infestation site, average pine age, and maximum August temperatures. The second RDA axis explained 6% of the variations and was significantly correlated with gradients in the beetle climate suitability index, extreme cold month temperatures, and site index. Comparisons of growth release patterns with aerial survey data and redundancy analyses indicated that dendrochronological techniques are useful for identifying mountain pine beetle outbreaks in central British Columbia, particularly among stands that had a density high enough to produce a growth release signal. Provided future studies account for interannual weather fluctuations, identification of growth increases due to stand thinning caused by beetle outbreaks will be useful for reconstructing the history of beetle outbreaks over much longer time periods.     

Back to the future of oridonin： again, compound from medicinal herb shows potent antileukemia efficacies in vitro and in vivo

Traditional Chinese Medicine （TCM） has been widely and successfully used in treating illnesses ranging from inflammation to cancer, and compounds from medicinal herbs and minerals are playing more and more important roles in taming various kinds of diseases [1 ], exemplified by artemisinin and arsenic trioxide （ATO）. Artemisinin （or Qinghaosu in Chinese） is isolated from a plant called sweet wormwood （Artemisia annua; or Qinghao in Chi- nese） which has been used as an antipyretic remedy for more than 1500 years in China. Artemisinin has impressive parasiticidal properties in vitro and in vivo, and is now one of the most important class of antimalarial agents [2, 3]. ATO is a common, naturally occurring substance which had been used in China for a long time as a therapeutic agent for some severe diseases with the ancient philosophy of ＇treating an evil with a toxic＇ [4]. In 1990s, ATO was shown to be able to cause partial differentiation at low dose and apoptosis at high concentration of acute promyelocytic leukemia （APL） cells [5], and induce complete remission in 90% of patients with relapsed or refractory APL [6-8]. These paradigms suggest that TCM is the treasure house not only for the Chinese people, but also for the whole human beings. It is our responsibility to develop evidence-based therapeutic approaches from TCM for diseases with poor prognosis.     

Two-Dimensional Electrophoretic Analysis of Soluble Leaf Proteins of a Salt-sensitive （Triticum aestivum） and a Salt-tolerant （T. durum） Cultivar in Response to NaCI Stress

In this research, 3-day-old etiolated wheat seedlings of Triticum aestivum L. cv. Ceyhan-99 （salt-sensitive） and T. durum Desf. cv. Firat-93 （salt-tolerant） were grown in control and salt （150 mmol/L NaCl） treatments at a 15/25℃ temperature regime in the light for 12 days. Soluble proteins extracted from the first leaf tissues of two cultivars were analyzed by twodimensional （2-D） electrophoresis in order to detect NaCl-induced changes. The soluble leaf protein profiles of cultivars were observed to be similar. However, quantitative differences in 74 proteins were detected in the salt treatment group, compared to the control. Among the 74 protein spots, 14 were common for two cultivars. As a result of NaCl treatment, two low-molecular-weight （LMW） proteins （28.9 and 30.0 kDa） and one intermediate-molecular-weight （IMW） protein （44.3 kDa） in cv. Ceyhan-99 and six LMW proteins （18.6, 19.4, 25.7, 25.9, 26 and 27.6 kDa） in cv. Firat-93 were newly synthesized. The newly synthesized proteins were specific to each cultivar. In the Firat-93 cultivar, four proteins with LMW （24.8-27.9 kDa） were completely lost in NaCl treatment. Moreover, these four protein spots were not observed in both protein profiles of cv. Ceyhan-99. Most of these proteins were in acidic character （pl 〈6.0-6.9） and low molecular weight （〈31.6 kDa）. It is suggested that the newly synthesized or completely lost LMW proteins may be important for cultivars differing in sensitivity towards NaCl.     

Phylogenetic Analysis of Brine Shrimp （Artemia） in China Using DNA Barcodinge

DNA barcoding is a powerful approach for characterizing species of organisms, especially those with almost identical morphological features, thereby helping to to establish phylogenetic relationships and reveal evolutionary histories. In this study, we chose a 648-bp segment of the mitochondrial gene, cytochrome c oxidase subunit 1 （COI）, as a standard barcode region to establish phylogenetic relationships among brine shrimp （Artemia） species from major habitats around the world and further focused on the biodiversity of Artemia species in China, especially in the Tibetan Plateau. Samples from five major salt lakes of the Tibetan Plateau located at altitudes over 4,000 m showed clear differences from other Artemia populations in China. We also observed two consistent amino acid changes, 153A/V and 183L/F, in the COI gene between the high and low altitude species in China. Moreover, indels in the COI sequence were identified in cyst and adult samples unique to the Co Qen population from the Tibetan Plateau, demonstrating the need for additional investigations of the mitochondrial genome among Tibetan Artemia populations.     

Cloning and Sequence Analysis of Genome from the Inner Mongolia Strain of the Endogenous Betaretroviruses （enJSRV）

In order to amplify the complete genome of enJSRV from the strain of Inner Mongolia （enJSRV-NM）, we used enJSRV-specific and JSRV-specific DNA probes in dot blot hybridization. Seven pairs of primers were designed based on Genbank sequences. Seven fragments were obtained by PCR and were cloned into the PMD19-T vectors. The recombinant plasmids were sequenced and analyzed. The results showed that the genome was 7 942 bp in length and contained four overlapping open reading frames corresponding to the gag, pro, pol and env genes as well as an additional open reading frame （orf-x） that overlaps the 3＇ end of the pol gene. The nucleotide acid sequences of the enJSRV-NM loci were compared with the sequences of South Africa enJS56A1 strain （Accession No.AF153615） and USA JSRV21 strain （Accession No. AF105220）. The nucleotide acid identities were 99.2% and 92.3% respectively. Two zinc fingers were found in the NC region in the predicted amino acid sequence. However, the YXXM motif, which is a reliable molecular marker for the infectious exogenous virus, was not found in the TM region. It was found that the enJSRV-NM region was 90%-98% identical at the amino acid level to its exogenous infectious counterparts in most of the retroviral genome. This is the first nucleotide sequence of enJSRV reported in P.R China. The resource work has provided a wide range of information useful not only for expression genomics and annotation of genomic DNA sequence, but also for further research on the clinical diagnosis of OPA.     

Influence of physical and chemical factors during foraging and culture of the symbiont fungus in Atta sexdens rubropilosa （Hymenoptera： Formicidae）

The aim of this paper is to investigate the influence of physical and chemical factors on transport and use of substrate for Atta sexdens rubropilosa workers. Three types of rectangular fragments were used to study the physical influence factors： filter paper with paraffin, filter paper without paraffin and polyester film. To study the chemical factors, some fragments were impregnated with organic extract of orange albedo, others were soaked with soybean oil and for the remaining ones nothing was applied. The fonowing parameters were evaluated： （i） attractiveness of substrate for transport and number of loading workers per treatment; （ii） foraged material incorporation; （iii） rejection by numbers of fragments deposited in the garbage or beside the fungus garden. All the polyester film fragments carried out to the fungus garden were subsequently rejected. We verified that chemical factors of the substrate were more quickly detected by the workers, whereas physical factors were used as a criterion in the decision-making to reject or accept the substrate collected.     

Analysis of forestry impacts and biodiversity in two Pyrenean forests through a comparison of moth communities （Lepidoptera, Heterocera）

We have compared the biodiversity variations in moth communities between unmanaged forests and commercial forests in a mountainous environment （Pyrenees France）. The aim was to evaluate the impact of forestry activities on moth diversity. The data collected from the insects were analysed with a Bayesian specific similarity index （noted SSP index） and by statistical biodiversity indexes comparison. It was seen that diversity and richness were decreased in the plantation compared to the unmanaged forest. Interestingly, the composition of the communities of moths was shown to be not only related to the presence/absence of host plants, but also to be in relation with changes in the differences in forest architecture （i.e. the relative coverage by the different vegetation levels）. However, the moth community in the commercial forest still has a high capacity to converge with the moth community present in more natural forests. We report here a list of 177 species of moths, providing information on the distribution of some uncommon species, poorly studied as yet in the Pyrenees.     

Seasonal changes of resistance to fenvalerate and cypermethrin in the larval parasitoid Cotesia plutellae （Hymenoptera： Braconidae）

The seasonal changes of insecticide resistance and stability in hymenopteran Cotesia plutellae, collected from Jianxin, Fuzhou-City, and Shangjie, Minhou-County, Fujian, China, were assessed by using a dry residual film method. The resistance to two insecticides in the field populations of C. plutellae was not stable under insecticide-free conditions in the insectarium. Compared with susceptible F11 progeny of C. plutellae in the insectarium, the resistance ratios （RR） in F0 parents were 18.4 for fenvalerate and 11.4 for cypermethrin based on LC50 at 9 hours, and 32.8 for fenvalerate and 28.5 for cypermethrin based on LC50 at 24 hours when the parasitoids were left in contact with the insecticides for 1 hour and mortalities were recorded at 9 and 24 hours, respectively. However, the RR in a field population of C. plutellae were 9.2 for fenvalerate and 12.7 for cypermethrin, if the parasitoids were left in contact with the insecticides for 24 hours. The resistances to the two pyrethroids in other field populations collected from Jianxin and Shangjie from November 2000 and July 2004 were also determined. Significant seasonal variations of resistance to the two insecticides in the field populations of C. plutellae were found. The RR were 3.0-18.4 for fenvalerate and 4.8-20.6 for cypermethrin in Jianxin populations from November 2000 to April 2002 based on LC50 at 9 h, and 2.3-13.6 for fenvalerate and 3.6-16.0 for cypermethrin in Shangjie populations from May 2002 to July 2004 based on LC50 at 24 hours. The resistance levels were high in spring and autumn and decreased sharply in summer. In addition, significant recovery from the knocked-down caused by the insecticides was found in the F0 and field populations of C. plutellae which were resistant to fenvalerate and cypermethrin if the parasitoids were left in contact with the pyrethroids for 1 hour. However, no recovery was found in susceptible F11 progeny.     

Human Calprotectin： Effect of Calcium and Zinc on its Secondary and Tertiary Structures, and Role of pH in its Thermal Stability

Calprotectin, a heterodimeric complex belonging to the S 100 protein family, has been found predominantly in the cytosolic fraction of neutrophils. In the present study, human calprotectin was purified from neutrophils using two-step ion exchange chromatography. The purified protein was used for circular dichroism study and fluorescence analysis in the presence of calcium and zinc at physiological concentrations, as well as for assessment of its inhibitory activity on the K562 leukemia cell line. The thermal stability of the protein at pH 7.0 （physiological pH） and 8.0 （similar to intestinal pH） was also compared. The results of cell proliferation analysis revealed that human calprotectin initiated growth inhibition of the tumor cells in a dose- dependent manner. The intrinsic fluorescence emission spectra of human calprotectin （50 ktg/ml） in the presence of calcium and zinc ions show a reduction in fluorescence intensity, reflecting a conformational change within the protein with exposure of aromatic residues to the protein surface that is important for the biological function of calprotectin. The far ultraviolet-circular dichroism spectra of human calprotectin in the presence of calcium and zinc ions at physiological concentrations show a decrease in the m-helical content of the protein and an increase in [3- and other structures. Our results also show that increasing the pH level from 7.0 to 8.0 leads to a marked elevation in the thermal stability of human calprotectin, indicating a significant role for pH in the stability of calprotectin in the gut.     

Expression of PIN Genes in Rice （Oryza sativa L.）： Tissue Specificity and Regulation by Hormones

Twelve genes of the PIN family in rice were analyzed for gene and protein structures and an evolutionary relationship with reported AtPINs in Arabidopsis. Four members of PIN1 （designated as OsPINla-d）, one gene paired with AtPIN2 （OsPIN2）, three members of PIN5 （OsPIN5a-c）, one gene paired with AtPIN8 （OsPIN8）, and three monocot-specific PINs （OsPIN9, OsPINIOa, and b） were identified from the phylogenetic analysis. Tissue-specific expression patterns of nine PIN genes among them were investigated using RT-PCR and GUS reporter. The wide variations in the expression domain in different tissues of the PIN genes were observed. In general, PIN genes are up-regulated by exogenous auxin, while different responses of different PIN genes to other hormones were found.     

Analysis of plastid and nuclear DNA data in plant phylogenetics——evaluation and improvement

Correct combination of plastid(cp)and nuclear(nr)DNA data for plant phylogenetic reconstructions is not a new issue,but with an increasing number of nrDNA loci being used,it is of ever greater practical concern.For accurately reconstructing the phylogeny and evolutionary history of plant groups,correct treatment of phylogenetic incongruence is a vital step in the proper analysis of cpDNA and nrDNA data.We first evaluated the current status of analyzing cpDNA and nrDNA data by searching all articles published in the journal Systematic Botany between 2005 and 2011.Many studies combining cpDNA and nrDNA data did not rigorously assess the combinability of the data sets,or did not address in detail possible reasons for incongruence between the two data sets.By reviewing various methods,we outline a procedure to more accurately analyze and/or combine cpDNA and nrDNA data,which includes four steps:identifying significant incongruence,determining conflicting taxa,providing possible interpretations for incongruence,and reconstructing the phylogeny after treating incongruence.Particular attention is given to explanation of the cause of incongruence.We hope that our procedure will help raise awareness of the importance of rigorous analysis and help identify the cause of incongruence before combining cpDNA and nrDNA data.     

Analysis of Genetic Diversity and Relationships in a Tunisian Fig （Ficus carica） Germplasm Collection by Random Amplified Microsatellite Polymorphisms

The random amplified mirosatellite polymorphism method was performed in a set of Tunisian fig landraces using eighteen primer combinations. A total of sixty three random amplified microsatellite polymorphism （RAMPO） markers were scored and used either to assess the genetic diversity in these cultivars or to detect cases of mislabeling. Opportunely, data proved that the designed procedure constitutes an attractive and fast method with low costs and prevents radio exposure. As a result, we have identified the primer combinations that are the most efficient to detect genetic polymorphism in this crop. Therefore, the derived unweighted pair-group method with arithmetic averages （UPGMA） dendrogram illustrates the genetic divergence among the landraces studied and exhibits a typically continuous variation. Moreover, no evident correlation between the sexes of trees was observed. In addition, using these markers, discrimination between landraces has been achieved. Thus, random amplified mirosatellite polymor- phism is proved to be powerful for characterizing the local fig germplasm.     

Effect of planting date and nitrogen fertilization on photosynthesis and soluble carbohydrate contents of cotton in relation to silverleaf whitefly （Bemisia tabaci biotype “B”） populations

The impacts of planting date and nitrogen fertilization on cotton （Gossypium hirsutum L.） photosynthesis and soluble carbohydrate contents in relation to silverleaf whitefly, Bemisia tabaci （Gennadius） biotype “B”, populations were examined in field experiments. Cotton planted in late April and early June was treated with 0, 112, 168 and 224 kg/N hectare in soil using urea fertilizer. The mean photosynthetic rate of April-planted cotton was 4%-20% higher than that of June-planted cotton early in the season, but 10%- 18% lower than that of June-planted cotton late in the season. The photosynthetic rates for both planting dates were positively correlated with levels of added nitrogen. While levels of glucose for both planting dates were positively correlated with nitrogen levels, fructose and sucrose levels were not. The mean levels of fructose were up to 40% lower, while that of sucrose were up to 59% higher, in April-planted cotton than in June-planted cotton. Levels of photosynthetic rate or stomatal conductance were not correlated with adult whitefly densities for either planting date. Levels of glucose and fructose were positively correlated with whitefly densities only for June-planted cotton late in the season.     

Dysfunction of Wnt signaling and synaptic ：lisassembly in neurodegenerative diseases

The molecular mechanisms that regulate synapse formation have been well documented. However, little is known about the factors that modulate synaptic stability. Synapse loss is an early and invariant feature of neurodegenerative diseases including Alzheimer＇s lAD） and Parkinson＇s disease. Notably, in AD the extent of synapse loss correlates with the severity of the disease. Hence, understanding the molecular mechanisms that underlie synaptic maintenance is crucial to reveal potential targets that will allow the development of ther- apies to protect synapses. Writs play a central role in the formation and function of neuronal circuits. Moreover, Wnt signaling compo- nents are expressed in the adult brain suggesting their role in synaptic maintenance in the adult. Indeed, blockade of Wnts with the Wnt antagonist Dickkopf-1 （Dkkl） causes synapse disassembly in mature hippocampal cells. Dkkl is elevated in brain biopsies from AD patients and animal models. Consistent with these findings, Amyloid-β （Aβ） oUgomers induce the rapid expression of Dkkl. Importantly, Dkkl neutralizing antibodies protect synapses against Aβ toxicity, indicating that Dkkl is required for Aβ-mediated synapse loss. In this review, we discuss the role of Wnt signaling in synapse maintenance in the adult brain, particularly in relation to synaptic loss in neurodegenerative diseases.     

Differences in Incubation Length and Hatchling Morphology among Five Species of Oviparous Phrynocephalus Lizards （Agamidae） from China

We incubated eggs of five Phrynocephalus species （P. albolineatus, P. axillaries, P. grumgrzimailoi, P. helioscopus and P. przewalskii） at three constant temperatures （24℃, 28℃ and 32℃） to examine differences in incubation length and hatchling morphology among species and among temperature treatments. We combined data from this study with those reported previously for P. frontalis and P. versicolor to examine whether embryonic stage at laying is a causal factor for interspecific variation in incubation length, and whether the phylogenetic relationship inferred from hatchling morphology is consistent with the relationship based on mitochondrial DNA data. Mean values for incubation length differed among the five species studied herein and, in all these five species, incubation length decreased at a decreasing rate as temperature increased. In none of the five species did hatchling size （snout-vent length and body mass） and other morphological variables differ among the three temperature treatments. The seven oviparous Phrynocephalus lizards found in China differ from each other in hatchling morphology, and embryonic stage at laying is a causal factor of inter- and intra-specific variation in incubation length. The phylogenetic relationship inferred from hatchling morphology is not always consistent with the currently known relationship based on mitochondrial DNA data. Data from this study and those reported previously allow the conclusion that any Phrynocephalus species may have its unique position along the axis defined by hatchling morphology.     

Effects of Gibberellic Acid on Primary Terpenoids and △^9 -Tetrahydrocannabinol in Cannabis sativa at Flowering Stage

Plants synthesize an astonishing diversity of isoprenoids, some of which play essential roles in photosynthesis, respiration, and the regulation of growth and development. Two independent pathways for the biosynthesis of isoprenoid precursors coexist within the plant cell： the cytosolic mevalonic acid （MVA） pathway and the plastidial methylerythritol phosphate （MEP） pathway. However, little is known about the effects of plant hormones on the regulation of these pathways. In the present study we investigated the effect of gibberellic acid （GA3） on changes in the amounts of many produced terpenoids and the activity of the key enzymes, 1-deoxy-D-xylulose 5-phosphate synthase （DXS） and 3-hydroxy-3-methylglutaryl coenzyme A reductase （HMGR）, in these pathways. Our results showed GA3 caused a decrease in DXS activity in both sexes that it was accompanied by a decrease in chlorophylls, carotenoids and Ag-tetrahydrocannabinol （THC） contents and an increase in α-tocopherol content. The treated plants with GA3 showed an increase in HMGR activity. This increase in HMGR activity was followed by accumulation of stigmasterol and β-sitosterol in male and female plants and campestrol in male plants. The pattern of the changes in the amounts of sterols was exactly similar to the changes in the HMGR activity. These data suggest that GA3 can probably influence the MEP and MVA pathways oppositely, with stimulatory and inhibitory effects on the produced primary terpenoids in MVA and DXS pathways, respectively.     

Structure variations of TBA G-quadruplex induced by 2＋-O-methyl nucleotide in K＋ and Ca2＋ environments

Thrombin binding aptamer （TBA）, a 15-mer oligonucleo- tide of d（GGTTGGTGTGGTTGG） sequence, folds into a chair-type antiparallel G-quadruplex in the K＋ environ- ment, and each of two G-tetrads is characterized by a syn-anti-syn-anti glycosidic conformation arrangement. To explore its folding topology and structural stability, 2＇-0- methyl nucleotide （OMe） with the C31-endo sugar pucker conformation and anti glycosidic angle was used to selectively substitute for the guanine residues of G-tetrads of TBA, and these substituted TBAs were characterized using a circular dichroism spectrum, thermally differential spectrum, ultra- violet stability analysis, electrophoresis mobility shift assay, and thermodynamic analysis in K＋ and Ca2＋ environments. Results showed that single substitutions for syn-dG residues destabilized the G-quadruplex structure, while single substi- tutions for anti-dG residues could preserve the G-quadruplex in the K＋ environment. When one or two G-tetrads were modified with OMe, TBA became unstructured. In contrast, in Ca2＋ environment, the native TBA appeared to be un- structured. When two G-tetrads were substituted with OMe, TBA seemed to become a more stable parallel G-4 structure. Further thermodynamic data suggested that OMe-substitu- tions were an enthalpy-driven event. The results in this study enrich our understanding about the effects of nucleo- tide derivatives on the G-quadruplex structure stability in different ionic environments, which will help to design G-quadruplex for biological and medical applications.