Effect of Salinity on Photosynthesis and Biochemical Characteristics in Mulberry Genotypes

Mulberry genotypes were subjected to salinity (0–12 mS cm^–1) in pot culture experiment. Chlorophyll and total carotenoid contents were reduced considerably by salinity. At low salinity, photosynthetic CO₂ uptake increased over the control, but it decreased at higher salinity. Contents of soluble proteins, free amino acids, soluble sugars, sucrose, starch, and phenols increased at salinity of 1–2 mS cm^–1 and decreased at higher salinity (8–12 mS cm^–1). Glycine betaine accumulated more than proline, the maximum accumulation of both was at salinity of 2–4 mS cm^–1. Among the genotypes studied, BC2-59 followed by S-30 showed better salinity tolerance than M-5.     

High-Pressure Inactivation of Hepatitis A Virus within Oysters

Previous results demonstrated that hepatitis A virus (HAV) could be inactivated by high hydrostatic pressure (HHP) (D. H. Kingsley, D. Hoover, E. Papafragkou, and G. P. Richards, J. Food Prot. 65:1605-1609, 2002); however, direct evaluation of HAV inactivation within contaminated oysters was not performed. In this study, we report confirmation that HAV within contaminated shellfish is inactivated by HHP. Shellfish were initially contaminated with HAV by using a flowthrough system. PFU reductions of >1, >2, and >3 log₁₀ were observed for 1-min treatments at 350, 375, and 400 megapascals, respectively, within a temperature range of 8.7 to 10.3°C. Bioconcentration of nearly 6 log₁₀ PFU of HAV per oyster was achieved under simulated natural conditions. These results suggest that HHP treatment of raw shellfish will be a viable strategy for the reduction of infectious HAV.     

Effect of bacterial vaginosis, Lactobacillus and Premarin estrogen replacement therapy on vaginal gene expression changes

The aim of the study was to investigate gene expression profiles of post-menopausal women receiving Premarin estrogen replacement therapy (ERT), compared to controls, and to examine any correlations between the bacterial vaginosis (BV) status of the subjects. Based upon an expected finding of a 50-60% difference between gene expression of host antimicrobials with alpha=0.05 (2-sided), beta=0.20 the calculation of 7 subjects per group, led to a sample size of 10 subjects receiving Premarin estrogen replacement therapy and 10 healthy, age-matched controls. Vaginal samples were collected at a single timepoint and processed for RNA recovery and Affymetrix array analysis, as well as Nugent scoring and denaturing gradient gel electrophoresis to identify bacteria. Lactobacillus iners was the most commonly detected species in the normal flora and this was confirmed with L. iners-specific PCR method. Vaginal swabs from 6 Premarin and 8 control vaginal samples provided a non-invasive means to analyze human gene expression. There was no significant up-regulation of cancer-associated gene expression in subject receiving Premarin ERT, but some evidence that the potentially protective innate immunity was reduced in patients with BV. Of those with a normal flora, there was a 2-fold down-regulation of carcinoma associated forkhead box A1 gene expression. BV was associated with 7-fold down-regulation of host antimicrobial colony stimulating factor, -9.83-fold for IL-1alpha, -8.33 for IL-1beta and -3.63 for IL-6. This is the first study to use gene arrays to correlate changes in host expression response to estrogen replacement therapy and BV.     

Dominant negative <Emphasis Type="Italic">Bmp5</Emphasis>mutation reveals key role of BMPs in skeletal response to mechanical stimulation

Background  

Over a hundred years ago, Wolff originally observed that bone growth and remodeling are exquisitely sensitive to mechanical forces acting on the skeleton. Clinical studies have noted that the size and the strength of bone increase with weight bearing and muscular activity and decrease with bed rest and disuse. Although the processes of mechanotransduction and functional response of bone to mechanical strain have been extensively studied, the molecular signaling mechanisms that mediate the response of bone cells to mechanical stimulation remain unclear.     

2-Arylbenzoxazoles as novel cholesteryl ester transfer protein inhibitors: optimization via array synthesis

2-Arylbenzoxazole 5 was identified as a hit from a fluorescence-based high-throughput screen for CETP inhibitors. The synthesis and SAR investigation employing array synthesis of the A- and B-rings are described.     

Genetic loss of Faah compromises male fertility in mice

Marijuana is the most commonly used illicit drug. Although there is some indication that reproductive functions in males are impaired in chronic marijuana users, the genetic evidence and underlying causes remain largely unknown. Herein we show that genetic loss of Faah, which encodes fatty acid amide hydrolase (FAAH), results in elevated levels of anandamide, an endocannabinoid, in the male reproductive system, leading to compromised fertilizing capacity of sperm. This defect is rescued by superimposing deletion of cannabinoid receptor 1 (Cnr1). Retention of Faah(-/-) sperm on the egg zona pellucida provides evidence that the capacity of sperm to penetrate the zona barrier is hampered by elevated anandamide levels. Collectively, the results show that aberrant endocannabinoid signaling via CNR1 impairs normal sperm function. Besides unveiling a new regulatory mechanism of sperm function, this study has clinical significance in male fertility.     

Terrestrial orchid conservation in the age of extinction

Background

Conservation through reserves alone is now considered unlikely to achieve protection of plant species necessary to mitigate direct losses of habitat and the pervasive impact of global climate change. Assisted translocation/migration represent new challenges in the face of climate change; species, particularly orchids, will need artificial assistance to migrate from hostile environments, across ecological barriers (alienated lands such as farmlands and built infrastructure) to new climatically buffered sites. The technology and science to underpin assisted migration concepts are in their infancy for plants in general, and orchids, with their high degree of rarity, represent a particularly challenging group for which these principles need to be developed. It is likely that orchids, more than any other plant family, will be in the front-line of species to suffer large-scale extinction events as a result of climate change.

Scope

The South West Australian Floristic Region (SWAFR) is the only global biodiversity hotspot in Australia and represents an ideal test-bed for development of orchid conservation principles. Orchids comprise 6 % of all threatened vascular plants in the SWAFR, with 76 out of the 407 species known for the region having a high level of conservation risk. The situation in the SWAFR is a portent of the global crisis in terrestrial orchid conservation, and it is a region where innovative conservation solutions will be required if the impending wave of extinction is to be averted. Major threatening processes are varied, and include land clearance, salinity, burning, weed encroachment, disease and pests. This is compounded by highly specialized pollinators (locally endemic native invertebrates) and, in the most threatened groups such as hammer orchids (Drakaea) and spider orchids (Caladenia), high levels of mycorrhizal specialization. Management and development of effective conservation strategies for SWAFR orchids require a wide range of integrated scientific approaches to mitigate impacts that directly influence ecological traits critical for survival.

Conclusions

In response to threats to orchid species, integrated conservation approaches have been adopted (including ex situ and translocation principles) in the SWAFR with the result that a significant, multidisciplinary approach is under development to facilitate conservation of some of the most threatened taxa and build expertise to carry out assisted migration to new sites. Here the past two decades of orchid conservation research in the SWAFR and the role of research-based approaches for managing effective orchid conservation in a global biodiversity hotspot are reviewed.Key words: Orchids, pollination, mycorrhiza, integrated conservation, terrestrial, threats, ex situ conservation, in situ conservation     

Arabidopsis Separase Functions beyond the Removal of Sister Chromatid Cohesion during Meiosis

Separase is a capase family protease that is required for the release of sister chromatid cohesion during meiosis and mitosis. Proteolytic cleavage of the α-kleisin subunit of the cohesin complex at the metaphase-to-anaphase transition is essential for the proper segregation of chromosomes. In addition to its highly conserved role in cleaving the α-kleisin subunit, separase appears to have acquired additional diverse activities in some organisms, including involvement in mitotic and meiotic anaphase spindle assembly and elongation, interphase spindle pole body positioning, and epithelial cell reorganization. Results from the characterization of Arabidopsis (Arabidopsis thaliana) separase (ESP) demonstrated that meiotic expression of ESP RNA interference blocked the proper removal of cohesin from chromosomes and resulted in the presence of a mixture of fragmented chromosomes and intact bivalents. The presence of large numbers of intact bivalents raised the possibility that separase may also have multiple roles in Arabidopsis. In this report, we show that meiotic expression of ESP RNA interference blocks the removal of cohesin during both meiosis I and II, results in alterations in nonhomologous centromere association, disrupts the radial microtubule system after telophase II, and affects the proper establishment of nuclear cytoplasmic domains, resulting in the formation of multinucleate microspores.The proper segregation of chromosomes during mitosis and meiosis is dependent on the systematic formation and subsequent removal of sister chromatid cohesion, which is required for homologous chromosome pairing, recombination, and repair (for review, see Onn et al., 2008; Peters et al., 2008). It is also required for the pairwise alignment of chromosomes on the metaphase I spindle and for the generation of tension across centromeres, thereby ensuring their bipolar attachment. In mitosis, cohesion is maintained by the cohesin complex, which consists of four evolutionally conserved proteins: Sister Chromatid Cohesion1 (SCC1), SCC3, Structural Maintenance of Chromosome1 (SMC1), and SMC3 (for review, see Nasmyth and Haering, 2005). During meiosis, SCC1 is largely replaced by its meiotic homolog REC8.The establishment of sister chromatid cohesion in yeast involves a multistep process (Milutinovich et al., 2007) that begins during telophase of the previous cell cycle when cohesin subunits associate with the chromatin, ultimately becoming enriched at discrete loci termed cohesin-associated regions (Blat and Kleckner, 1999; Laloraya et al., 2000). Cohesion is established during S-phase in a process that requires the Chromosome Transmission Fidelity protein (Ctf7), which is also known as Eco1 (Skibbens et al., 1999; Toth et al., 1999) and involves the replication fork (Kenna and Skibbens, 2003; Lengronne et al., 2006). In budding yeast (Saccharomyces cerevisiae) and fission yeast (Schizosaccharomyces pombe), cohesin complexes remain on the chromosomes until mitotic anaphase (Uhlmann et al., 1999, 2000; Tomonaga et al., 2000). In contrast, in vertebrates, most cohesin complexes are released from the chromosomes during prophase in a separase-independent process (Waizenegger et al., 2000; Losada et al., 2002). The small fraction of cohesin that remains primarily in centromeric regions is released to start anaphase (Sumara et al., 2000). The release of chromosome cohesion at the metaphase-to-anaphase transition is triggered by the Cys protease, separase (ESP1), which specifically cleaves the α-kleisin subunit (Ciosk et al., 1998; Uhlmann et al., 1999, 2000; Buonomo et al., 2000; Hauf et al., 2001). Prior to the metaphase-to-anaphase transition, securin inhibits the protease activity of separase. At the onset of anaphase, securin is degraded by the anaphase-promoting complex/cyclosome freeing separase, which cleaves SCC1, facilitating the release of cohesion and chromosome separation (Cohen-Fix et al., 1996; Ciosk et al., 1998).Studies on the distribution of cohesin proteins during meiosis in a number of organisms, including yeast, Caenorhabditis elegans, mammals, and Arabidopsis (Arabidopsis thaliana), have shown that similar to the situation during mitosis in animal cells, a significant amount of cohesin is either removed from or redistributed on prophase chromosomes in a separase-independent process (Pasierbek et al., 2001; Cai et al., 2003; Eijpe et al., 2003; Lee et al., 2003; Yu and Koshland, 2005). The final resolution of chiasmata, formed as the result of homologous chromosome recombination, and the separation of homologous chromosomes depends on separase cleavage of the meiotic α-kleisin subunit, REC8, along chromosome arms at anaphase I (Buonomo et al., 2000; Kitajima et al., 2003). Centromeric cohesion is protected by the conserved SGO family of proteins until anaphase II when separase cleavage of REC8 facilitates the separation of sister chromatids (Rabitsch et al., 2003; Katis et al., 2004; McGuinness et al., 2005).In addition to its highly conserved role in cleaving the α-kleisin subunit, separase appears to have acquired additional diverse activities in different organisms (Queralt and Uhlmann, 2005). For example, separase plays a role in DNA repair by promoting the redistribution of cohesin complexes to sites of DNA damage during mitotic interphase in budding and fission yeast (Nagao et al., 2004; Strom et al., 2004). Separase is also important for mitotic anaphase spindle assembly and elongation (Jensen et al., 2001; Papi et al., 2005; Baskerville et al., 2008), interphase spindle pole body positioning (Nakamura et al., 2002), and spindle formation during meiosis in yeast (Buonomo et al., 2003). It is also important for the proper positioning of the centrosomes during the first asymmetric mitotic division, eggshell development in C. elegans (Siomos et al., 2001; Rappleye et al., 2002), and for epithelial cell reorganization and dynamics in Drosophila melanogaster (Pandey et al., 2005). In zebra fish, a separase mutation causes genome instability and increased susceptibility to epithelial cancer (Shepard et al., 2007).Results from the characterization of Arabidopsis separase suggested that the protein also has multiple roles in plants (Liu and Makaroff, 2006). Seeds homozygous for a T-DNA insert in Arabidopsis ESP exhibited embryo arrest at the globular stage with the endosperm exhibiting a weak titan-like phenotype. Furthermore, expression of ESP RNA interference (RNAi) from the meiosis-specific DMC1 promoter disrupted the proper removal of the SYN1 cohesin protein from chromosomes during meiosis and resulted in the presence of a mixture of fragmented chromosomes and intact bivalents. The presence of large numbers of intact bivalents led the authors to suggest that in addition to its requirement for the removal of cohesin, ESP may also be required for either the proper attachment of the kinetochores to the spindle or spindle function. These findings, along with the observations that separase appears to have multiple roles in other organisms, led us to conduct a detailed characterization of meiosis in ESP RNAi plants.In this report, we show that meiotic expression of ESP RNAi blocks the release of sister chromatid cohesion during both meiosis I and II, results in nonhomologous centromere association, disrupts the radial microtubule system (RMS) after telophase II, and affects the proper establishment of nuclear cytoplasmic domains. Unlike the large majority of plant meiotic mutants that have been characterized to date, reduction of ESP levels during meiosis leads to the formation of multinucleate microspores.