Identification of Kaposi's Sarcoma-Associated Herpesvirus LANA Regions Important for Episome Segregation,Replication, and Persistence

Kaposi''s sarcoma-associated herpesvirus (KSHV) latency-associated nuclear antigen (LANA) is a 1,162-amino-acid protein that mediates the maintenance of episomal viral genomes in latently infected cells. The two central components of episome persistence are DNA replication with each cell division and the segregation of DNA to progeny nuclei. LANA self-associates to bind KSHV terminal-repeat (TR) DNA and to mediate its replication. LANA also simultaneously binds to TR DNA and mitotic chromosomes to mediate the segregation of episomes to daughter nuclei. The N-terminal region of LANA binds histones H2A and H2B to attach to mitotic chromosomes, while the C-terminal region binds TR DNA and also associates with chromosomes. Both the N- and C-terminal regions of LANA are essential for episome persistence. We recently showed that deletion of all internal LANA sequences results in highly deficient episome maintenance. Here we assess independent internal LANA regions for effects on episome persistence. We generated a panel of LANA mutants that included deletions in the large internal repeat region and in the unique internal sequence. All mutants contained the essential N- and C-terminal regions, and as expected, all maintained the ability to associate with mitotic chromosomes in a wild-type fashion and to bind TR DNA, as assessed by electrophoretic mobility shift assays (EMSA). Deletion of the internal regions did not reduce the half-life of LANA. Notably, deletions within either the repeat elements or the unique sequence resulted in deficiencies in DNA replication. However, only the unique internal sequence exerted effects on the ability of LANA to retain green fluorescent protein (GFP) expression from TR-containing episomes deficient in DNA replication, consistent with a role in episome segregation; this region did not independently associate with mitotic chromosomes. All mutants were deficient in episome persistence, and the deficiencies ranged from minor to severe. Mutants deficient in DNA replication that contained deletions within the unique internal sequence had the most-severe deficits. These data suggest that internal LANA regions exert critical roles in LANA-mediated DNA replication, segregation, and episome persistence, likely through interactions with key host cell factors.     

Later Passages of Neural Progenitor Cells from Neonatal Brain Are More Permissive for Human Cytomegalovirus Infection

Congenital human cytomegalovirus (HCMV) infection is the most frequent infectious cause of birth defects, primarily neurological disorders. Neural progenitor/stem cells (NPCs) are the major cell type in the subventricular zone and are susceptible to HCMV infection. In culture, the differentiation status of NPCs may change with passage, which in turn may alter susceptibility to virus infection. Previously, only early-passage (i.e., prior to passage 9) NPCs were studied and shown to be permissive to HCMV infection. In this study, NPC cultures derived at different gestational ages were evaluated after short (passages 3 to 6) and extended (passages 11 to 20) in vitro passages for biological and virological parameters (i.e., cell morphology, expression of NPC markers and HCMV receptors, viral entry efficiency, viral gene expression, virus-induced cytopathic effect, and release of infectious progeny). These parameters were not significantly influenced by the gestational age of the source tissues. However, extended-passage cultures showed evidence of initiation of differentiation, increased viral entry, and more efficient production of infectious progeny. These results confirm that NPCs are fully permissive for HCMV infection and that extended-passage NPCs initiate differentiation and are more permissive for HCMV infection. Later-passage NPCs being differentiated and more permissive for HCMV infection suggest that HCMV infection in fetal brain may cause more neural cell loss and give rise to severe neurological disabilities with advancing brain development.     

Human H7N9 Influenza A Viruses Replicate in Swine Respiratory Tissue Explants

Recently, novel H7N9 influenza viruses have caused an unprecedented outbreak in humans. Pigs are an important intermediate host for influenza; thus, we assessed the replication ability of three human H7N9 viruses (A/Anhui/1/2013, A/Shanghai/1/2013, A/Shanghai/2/2013) in swine tissue explants. All viruses tested replicated efficiently in explants from tracheas and bronchi, with limited replication in alveolar cells. Swine respiratory tissue explants can serve as an efficient model for screening replication potential of newly emerging H7N9 viruses.     

Molecular Diagnosis of an Ocular Toxocariasis Patient in Vietnam

An ocular Toxocara canis infection is reported for the first time in Vietnam. A 34-year-old man residing in a village of Son La Province, North Vietnam, visited the National Eye Hospital (NEH) in August 2011. He felt a bulge-sticking pain in his left eye and loss of vision occurred over 3 months before visiting the hospital. The eye examination in the hospital showed damage of the left eye, red eye, retinal fibrosis, retinal detachment, inflammation of the eye tissues, retinal granulomas, and a parasitic cyst inside. A larva of Toxocara was collected with the cyst by a medical doctor by surgery. Comparison of 264 nucleotides of internal transcribed spacer 2 (ITS2) of ribosomal DNA was done between our Vietnamese Toxocara canis and other Toxocara geographical isolates, including Chinese T. canis, Japanese T. canis, Sri Lankan T. canis, and Iranian T. canis. The nucleotide homology was 97-99%, when our T. canis was compared with geographical isolates. Identification of a T. canis infection in the eye by a molecular method was performed for the first time in Vietnam.     

Phytoplankton–zooplankton dynamics in periodic environments taking into account eutrophication

In this paper, we derive and analyze a mathematical model for the interactions between phytoplankton and zooplankton in a periodic environment, in which the growth rate and the intrinsic carrying-capacity of phytoplankton are changing with respect to time and nutrient concentration. A threshold value: “Predator’s average growth rate” is introduced and it is proved that the phytoplankton–zooplankton ecosystem is permanent (both populations survive cronically) and possesses a periodic solution if and only if the value is positive. We use TP (Total Phosphorus) concentration to mark the degree of eutrophication. Based on experimental data, we fit the growth rate function and the environmental carrying capacity function with temperature and nutrient concentration as independent variables. Using measured data of temperature on water bodies we fit a periodic temperature function of time, and this leads the growth rate and intrinsic carrying-capacity of phytoplankton to be periodic functions of time. Thus we establish a periodic system with TP concentration as parameter. The simulation results reveal a high diversity of population levels of the ecosystem that are mainly sensitive to TP concentration and the death-rate of zooplankton. It illustrates that the eruption of algal bloom is mainly resulted from the increasing of nutrient concentration while zooplankton only plays a role to alleviate the scale of algal bloom, which might be used to explain the mechanism of algal bloom occurrence in many natural waters. What is more, our results provide a better understanding of the traditional manipulation method.     

A comparison of the strength of biodiversity effects across multiple functions

In order to predict which ecosystem functions are most at risk from biodiversity loss, meta-analyses have generalised results from biodiversity experiments over different sites and ecosystem types. In contrast, comparing the strength of biodiversity effects across a large number of ecosystem processes measured in a single experiment permits more direct comparisons. Here, we present an analysis of 418 separate measures of 38 ecosystem processes. Overall, 45 % of processes were significantly affected by plant species richness, suggesting that, while diversity affects a large number of processes not all respond to biodiversity. We therefore compared the strength of plant diversity effects between different categories of ecosystem processes, grouping processes according to the year of measurement, their biogeochemical cycle, trophic level and compartment (above- or belowground) and according to whether they were measures of biodiversity or other ecosystem processes, biotic or abiotic and static or dynamic. Overall, and for several individual processes, we found that biodiversity effects became stronger over time. Measures of the carbon cycle were also affected more strongly by plant species richness than were the measures associated with the nitrogen cycle. Further, we found greater plant species richness effects on measures of biodiversity than on other processes. The differential effects of plant diversity on the various types of ecosystem processes indicate that future research and political effort should shift from a general debate about whether biodiversity loss impairs ecosystem functions to focussing on the specific functions of interest and ways to preserve them individually or in combination.     

Genomic regions associated with the nitrogen limitation response revealed in a global wheat core collection

Modern wheat (Triticum aestivum L.) varieties in Western Europe have mainly been bred, and selected in conditions where high levels of nitrogen-rich fertilizer are applied. However, high input crop management has greatly increased the risk of nitrates leaching into groundwater with negative impacts on the environment. To investigate wheat nitrogen tolerance characteristics that could be adapted to low input crop management, we supplied 196 accessions of a wheat core collection of old and modern cultivars with high or moderate amounts of nitrogen fertilizer in an experimental network consisting of three sites and 2 years. The main breeding traits were assessed including grain yield and grain protein content. The response to nitrogen level was estimated for grain yield and grain number per m² using both the difference and the ratio between performance at the two input levels and the slope of joint regression. A large variability was observed for all the traits studied and the response to nitrogen level. Whole genome association mapping was carried out using 899 molecular markers taking into account the five ancestral group structure of the collection. We identified 54 main regions involving almost all chromosomes that influence yield and its components, plant height, heading date and grain protein concentration. Twenty-three regions, including several genes, spread over 16 chromosomes were involved in the response to nitrogen level. These chromosomal regions may be good candidates to be used in breeding programs to improve the performance of wheat varieties at moderate nitrogen input levels.     

Pb and Cd Bioaccumulations in the Habitat and Preys of Red-Crowned Cranes (Grus japonensis) in Zhalong Wetland, Northeastern China

Pb and Cd concentrations in the habitat and preys of the red-crowned crane (i.e., reed rhizomes and three typical aquatic animal families (Perccottus glehni Dybowski, Carassius auratus Linnaeus, and Viviparidae)) were analyzed to examine the impact of these hazards on red-crowned cranes in northeastern China. Results indicated that Pb and Cd concentrations in the preys of the red-crowned cranes were elevated via food chain. Most of the detected Pb and Cd contents in the sediments were above the natural background level, ranging from 9.85 to 129.72 ppm and 1.23 to 10.63 ppm (dry weight), respectively. Cd geo-accumulation index at all sites were larger than 3, even reached 5.22, suggesting serious pollution in this region. Three common water animal families were detected to contain heavy metals, following the order of increasing concentrations: primary consumers (i.e., Viviparidae and Carassius auratus Linnaeus)?<?secondary consumers (i.e., Perccottus glehni Dybowski). Pb and Cd concentrations in the buffer zone are significantly higher than in the core area and being elevated in the food chain. The molten feathers of the red-crowned cranes showed the highest toxic metal concentrations of Pb (2.09 to 5.81 ppm) and Cd (1.42 to 3.06 ppm) compared with the feces produced by cranes and residual eggshell left by water fowls. Exceptionally high Pb and Cd concentrations in the cranes and their preys were thought to be associated with their habitat.     

Arctigenin Exhibits Relaxation Effect on Bronchus by Affecting Transmembrane Flow of Calcium

Arctigenin, a lignan extract from Arctium lappa (L.), exhibits anti-inflammation, antioxidation, vasodilator effects, etc. However, the effects of arctigenin on bronchus relaxation are not well investigated. This study aimed to investigate how arctigenin regulates bronchus tone and calcium ion (Ca²⁺) flow. Trachea strips of guinea pigs were prepared for testing the relaxation effect of arctigenin to acetylcholine, histamine, KCl, and CaCl₂, respectively. Furthermore, l-type calcium channel currents were detected by patch–clamp, and intracellular Ca²⁺ concentration was detected by confocal microscopy. The results showed that arctigenin exhibited relaxation effect on tracheae to different constrictors, and this was related to decreasing cytoplasmic Ca²⁺ concentration by inhibiting Ca²⁺ influx partly through l-type calcium channel as well as promoting Ca²⁺ efflux. In summary, this study provides new insight into the mechanisms by which arctigenin exhibits relaxation effect on bronchus and suggests its potential use for airway disease therapy.