Molecular cloning and characterization of a granulin-containing cysteine protease SPCP3 from sweet potato (Ipomoea batatas) senescent leaves

Granulins are a family of evolutionarily ancient proteins that are involved in regulating cell growth and division in animals. In this report a full-length cDNA, SPCP3, was isolated from senescent leaves of sweet potato (Ipomoea batatas). SPCP3 contains 1389 nucleotides (462 amino acids) in its open reading frame, and exhibits high amino acid sequence homologies (ca. 64-73.6%) with several plant granulin-containing cysteine proteases, including potato, tomato, soybean, kidney bean, pea, maize, rice, cabbage, and Arabidopsis. Gene structural analysis shows that SPCP3 encodes a putative precursor protein. Via cleavage of the N-terminal propeptide, it generates a protein with 324 amino acids (from the 139th to the 462nd amino acid residues), which contains two main domains: the conserved catalytic domain with the putative catalytic residues (the 163rd Cys, 299th His and 319th Asn) and the C-terminal granulin domain (from the 375th to the 462nd amino acid residues). Semi-quantitative RT-PCR and protein gel blot hybridization showed that SPCP3 gene expression was enhanced significantly in natural senescent leaves and in dark- and ethephon-induced senescent leaves, but was almost undetectable in mature green leaves, veins, and roots. Phylogenic analysis showed that SPCP3 displayed close association with a group of plant granulin-containing cysteine proteases which have been implied to be involved in programmed cell death. In conclusion, sweet potato SPCP3 is a functional, senescence-associated gene. Its mRNA and protein levels were significantly enhanced in natural and induced senescing leaves. The physiological role and/or function of SPCP3 associated with programmed cell death during leaf senescence were also discussed.     

Effect of 2-mercaptoethanol and cysteine supplementation during in vitro maturation on the developmental competence of oocytes from hormone-stimulated lambs

The objective was to determine the effect of 2-mercaptoethanol and cysteine on in vitro developmental competence of oocytes from lambs (4-8-week old) stimulated with eCG and pFSH. Oocytes were matured in medium (TCM199) with no supplement (Control group) or with 100muM 2-mercaptoethanol and 600muM cysteine (GSH group). Oocytes from adult sheep were also included (Adult group). The addition of 2-mercaptoethanol and cysteine did not improve nuclear maturation or microtubule configuration 12, 15, 18, or 24h after placement in maturation medium. Sperm head decondensation and male pronucleus formation were evaluated at 6, 12, and 18h after commencement of IVF; sperm decondensation appeared earlier in the GSH group (6h after the start of IVF). There were differences (P<0.05) between the Control group and the GSH and Adult groups for: fertilization rate at both 12h (55.4, 77.0, and 80.6%, respectively) and 18h (67.9, 86.9, and 88.7%); parthenogenesis rate at both 12h (25.0, 10.8, and 5.6%) and 18h (28.3, 9.8, and 4.5%); and polyspermy rate at 18h (26.4, 4.9, and 5.7%). Blastocyst rate at 7d was higher in the GSH group than the Control group (23.9% vs. 14.9%, P<0.05), but both were lower (P<0.05) than the Adult group (38.3%). The addition of 2-mercaptoethanol and cysteine improved sperm decondensation and rates of fertilization and the blastocyst development to 7d, with no effect on blastocyst rate at 9d.     

A Preliminary Study about the Potential Effects of Heavy Metals on the Human Male Reproductive Parameters in HIV-Infected Population in China

Due to the inconsistent effects of human immunodeficiency virus (HIV) on the human male reproduction in previous studies and the impacts of environmental exposures, such as heavy metals, on male reproduction receiving little attention in HIV-infected population, the aim of present study was to investigate whether heavy metals have potential effects on reproductive parameters in HIV-infected men. The current study assessed the associations between semen quality or serum hormone and concentration of the three heavy metal toxicants (lead (Pb), cadmium (Cd), and zinc (Zn)) in seminal, urine, and serum, and 50 HIV-infected men were recruited in the present study. Concentrations of Pb, Cd, and Zn were measured in three fluids by graphite furnace atomic absorption spectrophotometer. Semen analyses were performed according to World Health Organization criteria. Serum samples were analyzed for follicle-stimulating hormone, luteinizing hormone, and testosterone. HIV RNA viral load was determined by HIV virus loads kit. Spearman’s rank correlations were used for correlation analyses. The results showed that the concentrations of Pb, Cd, and Zn were significantly correlated with semen quality and serum hormone. HIV-1 virus loads were significantly associated with increased seminal Pb. However, HIV-1 virus loads were not statistically associated with semen quality and serum hormone. Our findings suggested that environmental heavy metals had potential effects on reproductive parameters in HIV-infected men in China.     

The regulation of glycerol 3-phosphate oxidase of rat brownadipose tissue mitochondria by long-chain free fatty acids

Added free fatty acids inhibit oxidation of glycerol 3-phosphate, succinate and NADH in brown-adipose tissue mitochondria from 10-day-old rats. The most pronounced is the inhibitory effect of glycerol 3-phosphate cytochrome c reductase (GP-cyto. c reductase). Contrary to other reductases, GP-cyto. c reductase activity of freshly isolated mitochondria is already inhibited by the fraction of endogenous free fatty acids. Both added and endogenous free fatty acids inhibition of GP-cyto. c reductase is fully reversible by the removal of free fatty acids by bovine serum albumine treatment. The inhibition of GP-cyto. c reductase is of strictly non-competitive type. The most inhibitory are unsaturated long-chain free fatty acids-oleic and linoleic acid. Results are discussed with regards to the regulatory importance of free fatty acids in brown-adiposetissue during intensive non-shivering thermogenesis.     

Production of a value-added hydroxy fatty acid, 7,10-dihydroxy-8(<Emphasis Type="Italic">E</Emphasis>)-octadecenoic acid,from high oleic safflower Oil by <Emphasis Type="Italic">Pseudomonas aeruginosa</Emphasis> PR3

Hydroxy fatty acids (HFAs), originally obtained in small amounts from plant systems, are good examples of structurally modified lipids, and they render special properties such as higher viscosity and reactivity compared to normal fatty acids. Based on these properties, HFAs possess high industrial potential in a wide range of applications. Recently, various microbial strains were tested for the production of HFAs from different unsaturated fatty acids since HFA production is limited to plant systems. Among the microbial strains tested, Pseudomonas aeruginosa PR3 has been well studied for the production of 7,10-dihydroxy-8(E)-octadecenoic acid (DOD) from oleic acid. Previously, we reported that strain PR3 could utilize triolein instead of oleic acid as a substrate for the production of DOD (Appl. Microbiol. Biotechnol. 2007, 74: 301–306). In this study, we focused on utilization of vegetable oil as a substrate for DOD production by PR3. Consequently, strain PR3 efficiently utilized high oleic safflower oil as a substrate for DOD production. Optimal initial medium pH and incubation time were pH 8.0 and 72 h, respectively. Optimal carbon and nitrogen sources were fructose and glutamine, respectively. Results from this study demonstrate that normal vegetable oils could be used as efficient substrates for the production of value-added HFAs by microbial bioconversion.     

The Roles of p38 MAPK/MSK1 Signaling Pathway in the Neuroprotection of Hypoxic Postconditioning Against Transient Global Cerebral Ischemia in Adult Rats

Postconditioning has regenerated interest as a mechanical intervention against cerebral ischemia/reperfusion injury, but its molecular mechanisms remain unknown. We previously reported that hypoxic postconditioning (HPC) ameliorated neuronal death induced by transient global cerebral ischemia (tGCI) in hippocampal CA1 subregion of adult rats. This study tested the hypothesis that p38-mitogen-activated protein kinase (p38 MAPK)/mitogen- and stress-response kinase 1 (MSK1) signaling pathway plays a role in the HPC-induced neuroprotection. Male Wistar rats were subjected to 10 min ischemia induced by applying the four-vessel occlusion method. HPC with 120 min was applied at 24 h after reperfusion. Immunohistochemistry and Western blot were used to detect the expression of phosphorylation of p38 MAPK and MSK1, as well as cleaved caspase-3. We found that HPC induced a significant increase of phosphorylated p38 MAPK and MSK1 in neurons of hippocampal CA1 region and a significant decrease in glial cells after tGCI as well. Furthermore, HPC attenuated caspase-3 cleavation triggered by tGCI in CA1 region. Moreover, p38 MAPK inhibition by SB203580 significantly decreased the phosphorylation of MSK1, increased cleaved caspase-3 expression, and abolished the neuroprotection of HPC. These findings suggested that p38 MAPK/MSK1 signaling axis contributed to HPC-mediated neuroprotection against tGCI, at least in part, by regulating the activation of caspase-3.     

Effects of extreme temperature on seedling establishment of nonnative invasive plants

Climate change and plant invasion are two of the most important ecological issues facing the world today. Extreme events are likely to play an important role in plant invasion. For example, tolerance to temperature stress is critical for plant germination and survival of seedlings. Nonnative invasive species tend to differ from co-occurring native species in several traits. Increased mean temperatures are known to enhance the risk of plant invasions, but few experimental studies have linked plant invasion to both increasing mean temperature and extreme (low and high) temperatures. Ten plant species from Asteraceae (six nonnative invasive and four native species) were chosen and six temperatures (extremely low, average winter, average annual, average summer, high and extremely high) were used to test the effects of extreme temperatures on plant invasion in southern China. The results showed that nonnative invasive plant species (IS) germinated more readily and the seedlings grew better than those of native plant species (NS) at high temperatures, suggesting that global warming may facilitate invasion. Extreme temperatures decreased the seed germination rate and seedling growth of both IS and NS, although NS were more tolerant of extremely low temperatures (5/0 °C). IS, in turn, were more tolerant of extremely high temperatures (40/35 °C). Extreme high temperatures may increase the risk of plant invasion because IS seedlings are better able to become established, whereas low temperatures may hinder invasion. In addition, the species-specific differences in plant origin (IS and NS) and temperature tolerance were correlated with other climatic factors and should be considered in managing invasive species in a changing world.     

Encapsulated in silica: genome, proteome and physiology of the thermophilic bacterium Anoxybacillus flavithermus WK1

Background

Gram-positive bacteria of the genus Anoxybacillus have been found in diverse thermophilic habitats, such as geothermal hot springs and manure, and in processed foods such as gelatin and milk powder. Anoxybacillus flavithermus is a facultatively anaerobic bacterium found in super-saturated silica solutions and in opaline silica sinter. The ability of A. flavithermus to grow in super-saturated silica solutions makes it an ideal subject to study the processes of sinter formation, which might be similar to the biomineralization processes that occurred at the dawn of life.

Results

We report here the complete genome sequence of A. flavithermus strain WK1, isolated from the waste water drain at the Wairakei geothermal power station in New Zealand. It consists of a single chromosome of 2,846,746 base pairs and is predicted to encode 2,863 proteins. In silico genome analysis identified several enzymes that could be involved in silica adaptation and biofilm formation, and their predicted functions were experimentally validated in vitro. Proteomic analysis confirmed the regulation of biofilm-related proteins and crucial enzymes for the synthesis of long-chain polyamines as constituents of silica nanospheres.

Conclusions

Microbial fossils preserved in silica and silica sinters are excellent objects for studying ancient life, a new paleobiological frontier. An integrated analysis of the A. flavithermus genome and proteome provides the first glimpse of metabolic adaptation during silicification and sinter formation. Comparative genome analysis suggests an extensive gene loss in the Anoxybacillus/Geobacillus branch after its divergence from other bacilli.     

A NEW PALEOCENE BIRD FROM ANHUI, CHINA

redspecimenrepersentsanewgenusandspecies.Wanshuinaliigen.etsp.nov.SpecimenShaftofarighthumerus,andthedistalendofaIefttibiotarsusanda1efttarsometatarsus,Vlo529.DistributionQianshan,AnhuiProvince;Paleocene,DoumuFormation.DiagnosisSizesma1l-medium,humeruss1enderandlong,mediancrestventraleelongated,tibiotarsuscompressedanteroposteriorly,externalcondy1eroundinlateraIview,grooveforperoneusprofundusdeepandlong;internalcondylelongerthanex-ternalcondyle,adepressionwelldeve1opedin1owerborder0finter…