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We have examined the quantum dynamics of a tightly bound planar (2-D) quantum dot subject to Landau quantization in a high magnetic field. The dot is taken to be set on a 2-D host sheet in a perpendicular magnetic field, and it is represented by a 2-D Dirac delta-function potential, which would support just a single subband state in the absence of the magnetic field. The integral equation for the Schro?dinger Green's function of this system is solved in closed form analytically, and we have studied the dispersion relation for the single-particle subband energy spectrum. We find that it has a plethora of single-particle subband modes induced by Landau quantization.

The human LGI1 gene is a leucine-rich, repeat-containing gene that was cloned from the t(10:19) breakpoint of the T98G glioblastoma cell line. The LGI1 gene maps to 10q24, a region of peak LOH in malignant gliomas, and is inactivated during the transition from low to high-grade brain tumors. Here we report detailed studies of the genomic structure of the LGI1 gene and its promoter. We have also cloned and characterized the mouse lgil gene, which is 97% homologous to the human gene at the amino acid level and 91% homologous at the nucleotide level. LGI1 contains 8 exons, where each of the four leucine-rich repeat units is contained in an individual 72-bp exon. The cysteine-rich regions flanking the LRR and the single trans-membrane domain do not occupy individual exons. Approximately 5-kb of the genomic region 5' to LGI1 was sequenced, but conventional CAAT and TATA motifs were not present within this sequence. A 597-bp fragment of this 5' sequence was cloned upstream of a promoterless luciferase gene and was shown to be sufficient to drive transcription. SSCP analysis of the coding region of LGI1 in 20 glioblastomas and five cell lines did not reveal any mutations. Because LGI1 expression is considerably downregulated in gliomas, we also investigated whether this was owing to changes in the methylation status of the promoter. Southern blot analysis and 5-azacytidine treatment did not show any appreciable difference in methylation status between normal brain and glioblastomas.

Although the induction of drug-metabolizing systems by phenobarbital has been recognized for about 40 years, the mechanism by which cytochrome P450 gene expression is increased is still not well understood. A 163-bp fragment at about -2.2 Kb in CYP2B2 has been shown to mediate phenobarbital induction in primary rat hepatocytes (Trottier, et al. [1995] Gene 158:263-268) and by an in situ transient transfection assay in rat liver (Park, Y., et al. [1996]. J. Biol. Chem. 271:23725-23728). Deletion mutations of this fragment indicated that the 88-bp stretch from -2258 to -2170 was the minimal sequence that could mediate phenobarbital induction in the in situ system if single copies of the deleted fragments fused to the CYP2C1 proximal promoter were assayed. If three copies of the fragments were present, 5' and 3' deletions defined a minimal 37-bp core fragment, which, although necessary for phenobarbital responsiveness, was not sufficient unless additional sequence was present at either end, suggesting that redundant elements were present in the two flanking regions. Site-specific mutagenesis of an NF-1 site within the 88-bp fragment and linker scanning mutagenesis across the fragment indicated that the NF-1 site and a region to the 5' side of the site contributed to the magnitude of the response, but neither the NF-1 mutations nor any of the linker scanning mutations eliminated the response to phenobarbital. Mutation in a region 3' of the NF-1 site resulted in elevated basal expression without substantial effects on phenobarbital-induced expression. Binding of NF-1 to the 37-bp core fragment was established by gel-shift competition studies and by supershifts of the protein-DNA complexes by antisera to NF-1. Additional protein-DNA complexes were detected in the regions flanking the NF-1 site. These studies indicate that the CYP2B2 phenobarbital-responsive enhancer contains multiple constitutive and phenobarbital-responsive elements. Binding of nuclear proteins from control or phenobarbital-treated animals in vitro to this region was very similar. The only difference detected was a complex that was substantially reduced by phenobarbital treatment and mapped to the 3' side of the NF-1 site.

The yeast Kluyveromyces marxianus is a potential microbial catalyst for fuel ethanol production from a wide range of biomass substrates. To improve its growth and ethanol yield at elevated temperature under microaerophilic conditions, K. marxianus NRRL Y-1109 was irradiated with UV-C using automated protocols on a robotic platform for picking and spreading irradiated cultures and for processing the resulting plates. The plates were incubated under anaerobic conditions on xylose or glucose for 5 mo at 46 °C. Two K. marxianus mutant strains (designated 7-1 and 8-1) survived and were isolated from the glucose plates. Both mutant strains, but not wild type, grew aerobically on glucose at 47 °C. All strains grew anaerobically at 46 °C on glucose, galactose, galacturonic acid, and pectin; however, only 7-1 grew anaerobically on xylose at 46 °C. Saccharomyces cerevisiae NRRL Y-2403 did not grow at 46 °C on any of these substrates. With glucose as a carbon source, ethanol yield after 3 d at 46 °C was higher for 8-1 than for wild type (0.51 and 0.43 g ethanol/g glucose, respectively). With galacturonic acid as a carbon source, the ethanol yield after 7 d at 46 °C was higher for 7-1 than for wild type (0.48 and 0.34 g ethanol/g galacturonic acid, respectively). These mutant strains have potential application in fuel ethanol production at elevated temperature from sugar constituents of starch, sucrose, pectin, and cellulosic biomass.

With the fast development of economy, the mining industry in China has become seriously concerned about the future supply of its professionals in mineral processing and allied disciplines. Educating mining professionals become the more and more important thing. This paper outlines the mining education background in the Kunming University of Science and Technology (KUST). This paper then summaries the current situation of educating the mining professionals in the university, which shows that the professionals shall be trained by multiple ways. At the same time, some new thoughts on educating the mining professionals in KUST are put forward.

This chapter details the recent advances made on bioconversion of lignocellulosic biomass to butanol, a superior biofuel that can be used in internal combustion engines or transportation industry. It should be noted that butanol producing cultures cannot tolerate or produce more than 20–30 g/L of acetone-butanol-ethanol (ABE) in batch reactors of which butanol is of the order of 13–18 g/L. This is due to toxicity of butanol to the culture. In order to overcome this challenge, two approaches have been applied: (1) developing more butanol tolerant strains using genetic engineering techniques and (2) employing process engineering approaches to simultaneously recover butanol from the fermentation broth thus not allowing butanol concentrations in the reactor to accumulate beyond culture’s tolerance. By the application of the first approach, a number of butanol producing strains have been developed; however, none of these accumulated greater than 1,200 mg/L (1.2 g/L) butanol, while using the second approach total ABE up to 461 g/L has been produced. Attempts to improve the newly developed strains are continuing. Lignocellulosic substrates have been used to produce butanol due to their abundant availability and economical prices usually in the range of $24–60/ton as opposed to corn prices which have been in the range of $153–218/ton during recent months. It should be noted that lignocellulosic substrates require separate hydrolysis prior to fermentation. In a more recent approach, hydrolysis and fermentation (and simultaneous recovery) have been integrated or combined to reduce the cost of butanol production from cellulosic substrates. Using such an approach, up to 192 g/L ABE was produced from 430 g/L cellulosic biomass/sugars. Additionally, this chapter provides details of process integration and simultaneous product recovery technologies for butanol production.

Whether energetic electrons (10s of keV) in the magnetosheath can be directly transported into the magnetosphere and further energized through radial diffusion is significant in understanding the physical mechanisms for producing the radiation belt electrons (>100s of keV) in the magnetosphere. In this study, we analyze more than two hundred magnetopause crossing events using the energetic electron and magnetic field measurements from Geotail and compare the flux and phase space density (PSD) of the energetic electrons on both sides of the magnetopause. It is found that for most of the events (>70%), the fluxes and PSDs of energetic electrons in the magnetosheath are less than those in the magnetosphere, suggesting that the energetic electrons in the magnetosheath cannot be a direct source sufficient for the energetic electrons inside the magnetosphere. In fact, our analysis suggests a possible leakage of the energetic electrons from inside to outside the magnetopause. By investigating the average energetic electron flux distribution in the magnetosheath, we find that the energetic electron fluxes are higher near the bow shock and the magnetopause than in between. The high energetic electron flux near the bow shock can be understood as due to energization of electrons when they go through the bow shock. The relatively low flux of the energetic electrons in between indicates that it is difficult for the energetic electrons to travel from the bow shock to the magnetopause and vice versa, possibly because the energetic electrons near the bow shock and the magnetopause are all on open magnetic field lines and these two relatively intense energetic electron populations in the magnetosheath rarely get mixed.

Five reactor systems (free cell batch, free cell continuous, entrapped cell immobilized, adsorbed cell packed bed, and cell recycle membrane reactors) were compared for ethanol production from xylose using Escherichia coli FBR5. In the free cell batch and free cell continuous reactors (continuous stirred tank reactor-CSTR) productivities of 0.84 gL−1 h−1 and 1.77 gL−1 h−1 were achieved, respectively. A cell recycle membrane reactor resulted in the highest productivity of 55.56 gL−1 h−1, which is an increase of 66-fold (e.g., 6614%) over the batch reactor. Calcium alginate gel CSTR resulted in a productivity of 2.04 gL−1 h−1 whereas adsorbed cell packed bed reactor resulted in a productivity of 4.39 gL−1 h−1. In the five reactor systems, ethanol concentrations ranged from 18.9 to 40.30 gL−1 with metabolic yields from 0.44 to 0.51. Published 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012

This study reports the production and characterization of a novel antibacterial polypeptide, designated laparaxin, which is secreted by Lactobacillus paracasei NRRL B-50314. Crude laparaxin has antibacterial activity against a wide variety of Gram-positive bacteria, including: lactic acid bacteria (Lactococcus lactis and Lactobacillus buchneri), food-borne pathogens (Listeria monocytogenes), gastrointestinal pathogens (Enterococcus faecalis), and opportunistic pathogens (Staphylococcus aureus methicillin-sensitive (MSSA) and methicillin resistant (MRSA) strains, a hetero-vancomycin-intermediate methicillin resistant strain (Hetero VISA also MRSA) MM66, and homogeneous vancomycin intermediate (Homo VISA). Using L. lactis as an indicator strain, the inhibitory activity of crude laparaxin was detected originally in early log phase, and the activity maximizes at the early stationary phase and remains stable after prolonged incubation. Laparaxin activity is stable after 30 min of incubation at 94°C. Higher concentrations of inhibitory activity are produced when glucose, fructose and sucrose are used as carbon-sources in growth media. Crude laparaxin has potential applications in food and feed industries, as well as in clinical and veterinary medicine.

Currently a total of 17 states in the US use bio-ethanol to replace methyl tert-butyl ether (MTBE) as a safer oxygenate for gasoline formulations (www.eia.gov ). Moreover, E15 (15% ethanol blend) was recently approved by the U.S. Environmental Protection Agency for use in select automobiles from 2001 and newer model years, and in all flexible fuel vehicles (FFVs). It has been projected that in the US by the year 2035, 63% of the bio-ethanol produced in the US will be used for E10 and E15, while 37% will be used in FFVs operating on E85 (85% ethanol blended with 15% gasoline), which should significantly reduce US consumption of petroleum- based transportation fuel (http://www.

The spatial distribution of >38 keV electron fluxes in the central plasma sheet (CPS) and the statistical relationship between the CPS electron fluxes and the upstream solar wind and interplanetary magnetic field (IMF) parameters are investigated quantitatively using measurements from the Geotail satellite (1998-2004) at geocentric radial distances of 9-30 RE in the night side. The measured electron fluxes increase with closer distance to the center of the neutral sheet, and exhibit clear dawn-dusk asymmetry, with the lowest fluxes at the dusk side and increasing toward the dawn side. The asymmetry persists along the Earth's magnetotail region (at least to Geotail's apogee of 30 RE during the period of interest). Both the individual case and the statistical analysis on the relationship between >38 keV CPS electron fluxes and solar wind and IMF properties show that larger (smaller) solar wind speed and southward (northward) IMF B(z) imposed on the magnetopause result in higher (lower) energetic electron fluxes in the CPS with a time delay of about 1 hour, while the influence of solar wind ion density on the energetic electrons fluxes is insignificant. Interestingly, the energetic electron fluxes at a given radial distance correlate better with IMF B(z) than with the solar wind speed. Based on these statistical analyses, an empirical model is developed for the first time to describe the 2-D distribution (along and across the Earth's magnetotail) of the energetic electron fluxes (>38 keV) in the CPS, as a function of the upstream solar wind and IMF parameters. The model reproduces the observed energetic electron fluxes well, with a correlation coefficient R equal to 0.86. Taking advantage of the time delay, full spatial distribution of energetic electron fluxes in the CPS can be predicted about 2 hours in advance using the real-time solar wind and IMF measurements at the L1 point: 1 hour for the solar wind to propagate to the magnetopause and a 1 hour delay for the best correlation. Such a prediction helps us to determine whether there are enough electrons in the CPS available to be transported inward to enhance the outer electron radiation belt.

Lactobacillus buchneri strain NRRL B-30929 was a contaminant obtained from a commercial ethanol fermentation. This facultative anaerobe is unique because of its rapid growth on xylose and simultaneous fermentation of xylose and glucose. The strain utilizes a broad range of carbohydrate substrates and possesses a high tolerance to ethanol and other stresses, making it an attractive candidate for bioconversion of biomass substrates to various bioproducts. The genome sequence of NRRL B-30929 will provide insight into the unique properties of this lactic acid bacterium.

Acremonium zeae, one of the most prevalent fungal colonists of preharvest corn, possesses a suite of hemicellulolytic activities including xylanase, xylosidase, and arabinofuranosidase. Two enzymes with arabinofuranosidase activity were purified from cell-free culture supernatants of A. zeae grown on oat spelt xylan. A 47 kDa enzyme (AF47) was optimally active at 37 °C and pH 6.0, and had a specific activity for 4-nitrophenyl-α-L-arabinofuranoside (4NPA) of 6.2 U/mg. A 30 kDa enzyme (AF30) was optimally active at 50 °C and pH 4.5, and had a specific activity for 4NPA of 12.4 U/mg. AF47 hydrolyzed 4-nitrophenyl-β-D-xylopyranoside, 4-nitrophenyl-β-D-glucopyranoside, and 4-nitrophenyl-β-D-cellobioside, as well as producing reducing sugars from corn fiber, wheat, and oat spelt arabinoxylan. AF30 had little detectable activity on the 4-nitrophenyl substrates, except for 4NPA, but activity on arabinoxylans from corn fiber, wheat, and oat spelt was at least 7-fold higher than AF47, with specific activities of 109, 358, and 153 U/mg, respectively. A combination of the two enzymes released 61 and 88% of the total arabinose from corn fiber and wheat arabinoxylans. The arabinofuranosidases produced by A. zeae may have industrial application for the enzymatic hydrolysis of recalcitrant lignocellulosic feedstocks such as corn fiber and wheat straw.

[1] A new three-dimensional asymmetric magnetopause model has been developed for corrected GSM coordinates and parameterized by the solar wind dynamic and magnetic pressures (Pd + Pm), the interplanetary magnetic field (IMF) Bz, and the dipole tilt angle. On the basis of the magnetopause crossings from Geotail, IMP 8, Interball, TC1, Time History of Events and Macroscale Interactions during Substorms (THEMIS), Wind, Cluster, Polar, Los Alamos National Laboratory (LANL), GOES, and Hawkeye, and the corresponding upstream solar wind parameters from ACE, Wind, or OMNI, this model is constructed by the Levenberg-Marquardt method for nonlinear multiparameter fitting step-by-step over the divided regions. The asymmetries of the magnetopause and the indentations near the cusps are appropriately described in this new model. In addition, the saturation effect of IMF Bz on the subsolar distance and the extrapolation for the distant tail magnetopause are also considered. On the basis of this model, the power law index for the subsolar distance versus Pd + Pm is a bit less than −1/6, the northward IMF Bz almost does not influence the magnetopause, and the dipole tilt angle is very important to the north–south asymmetry and the location of indentations. In comparison with the previous empirical magnetopause models based on our database, the new model improves prediction capability to describe the three-dimensional structure of the magnetopause. It is shown that this new model can be used to quantitatively study how Pd + Pm compresses the magnetopause, how the southward IMF Bz erodes the magnetopause, and how the dipole tilt angle influences the north–south asymmetry and the indentations.

The first step of the butanol pathway involves an acetyl-CoA acetyltransferase (ACoAAT), which controls the key branching point from acetyl-CoA to butanol. ACoAAT, also known as thiolase (EC 2.3.1.9), is encoded by the thl gene and catalyzes ligation of two acetyl-CoA into acetoacetyl-CoA. Bioinformatics analyses suggest there are no thl in the genomes of lactic acid bacteria (LAB), in this study we aimed to introduce the thl gene into selected LAB strains and analyze the fermentation products. The thl gene from Clostridium beijerinckii P260 was amplified by genomic PCR using gene-specific primers designed from the published genome sequences of C. beijerinckii NCIMB 8025. The 1.2 kb thl gene was cloned into the pETBlue vector and overexpressed in Escherichia coli Tuner (DE3) pLacI cells. Functional enzyme activity was detected spectrophotometrically by measuring the decrease in absorbance at 303 nm, which reflects the change in acetoacetyl-CoA concentrations. The thl gene was subsequently introduced into Lactococcus lactis and Lactobacillus buchneri strains, and GC analysis indicated about 28 mg/L and 66 mg/L of butanol was produced in the recombinant strains, respectively. This study reports the first step toward developing a butanolgenic LAB through the introduction of the butanol pathway into butanol-tolerant strains of LAB.

Aims: A Lactobacillus buchneri strain NRRL B-30929 can convert xylose and glucose into ethanol and chemicals. The aims of the study were to survey three strains (NRRL B-30929, NRRL 1837 and DSM 5987) for fermenting 17 single substrates and to exam NRRL B-30929 for fermenting mixed substrates from biomass hydrolysates. Methods and Results: Mixed acid fermentation was observed for all three L. buchneri strains using various carbohydrates; the only exception was uridine which yielded lactate, acetate and uracil. Only B-30929 is capable of utilizing cellobiose, a desired trait in a potential biocatalyst for biomass conversion. Flask fermentation indicated that the B-30929 strain can use all the sugars released from pretreated hydrolysates, and producing 1·98–2·35 g l−1 ethanol from corn stover hydrolysates and 2·92–3·01 g l−1 ethanol from wheat straw hydrolysates when supplemented with either 0·25× MRS plus 1% corn steep liquor or 0·5× MRS. Conclusions: The L. buchneri NRRL B-30929 can utilize mixed sugars in corn stover and wheat straw hydrolysates for ethanol and other chemical production. Significance and Impact of the Study: These results are valuable for future research in engineering L. buchneri NRRL B-30929 for fermentative production of ethanol and chemicals from biomass.

The emergence of antibiotic-resistant bacteria may limit the effectiveness of antibiotics to treat bacterial contamination in fuel ethanol plants, and therefore, new antibacterial intervention methods and tools to test their application are needed. Using shake-flask cultures of Saccharomyces cerevisiae grown on saccharified corn mash and strains of lactic acid bacteria isolated from a dry-grind ethanol facility, a simple model to simulate bacterial contamination and infection was developed. Challenging the model with 108 CFU/mL Lactobacillus fermentum decreased ethanol yield by 27% and increased residual glucose from 6.2 to 45.5 g/L. The magnitude of the effect was proportional to the initial bacterial load, with 105 CFU/mL L. fermentum still producing an 8% decrease in ethanol and a 3.2-fold increase in residual glucose. Infection was also dependent on the bacterial species used to challenge the fermentation, as neither L. delbrueckii ATCC 4797 nor L. amylovorus 0315-7B produced a significant decrease in ethanol when inoculated at a density of 108 CFU/mL. In the shake-flask model, treatment with 2 µg/mL virginiamycin mitigated the infection when challenged with a susceptible strain of L. fermentum (MIC for virginiamycin ≤2 ppm), but treatment was ineffective at treating infection by a resistant strain of L. fermentum (MIC = 16 ppm). The model may find application in developing new antibacterial agents and management practices for use in controlling contamination in the fuel ethanol industry. Biotechnol. Bioeng. 2009;103: 117–122. Published 2008 Wiley Periodicals, Inc.

A three-plasmid yeast expression system utilizing the portable small ubiquitin-like modifier (SUMO) vector set combined with the efficient endogenous yeast protease Ulp1 was developed for production of large amounts of soluble functional protein in Saccharomyces cerevisiae. Each vector has a different selectable marker (URA, TRP, or LEU), and the system provides high expression levels of three different proteins simultaneously. This system was integrated into the protocols on a fully automated plasmid-based robotic platform to screen engineered strains of S. cerevisiae for improved growth on xylose. First, a novel PCR assembly strategy was used to clone a xylose isomerase (XI) gene into the URA-selectable SUMO vector and the plasmid was placed into the S. cerevisiae INVSc1 strain to give the strain designated INVSc1-XI. Second, amino acid scanning mutagenesis was used to generate a library of mutagenized genes encoding the bioinsecticidal peptide lycotoxin-1 (Lyt-1) and the library was cloned into the TRP-selectable SUMO vector and placed into INVSc1-XI to give the strain designated INVSc1-XI-Lyt-1. Third, the Yersinia pestis xylulokinase gene was cloned into the LEU-selectable SUMO vector and placed into the INVSc1-XI-Lyt-1 yeast. Yeast strains expressing XI and xylulokinase with or without Lyt-1 showed improved growth on xylose compared to INVSc1-XI yeast.

Gram-positive bacteria have been explored to convert lignocellulosic biomass to biofuel and bioproducts. Our long-term goal is to create genetically engineered lactic acid bacteria (LAB) strains that convert agricultural biomass into ethanol and other value-added products. The immediate approaches toward this goal involve genetic manipulations by either introducing ethanol production pathway genes or inactivating pathways genes that lead to production of undesired byproducts. The widely studied species Lactobacillus plantarum is now considered a model for genetic manipulations of LAB. In this study, L. plantarum TF103 strain, in which two of the chromosomal L-ldh and D-ldh genes are inactivated, was used to introduce additional mutations on the chromosome to eliminate undesired fermentation products. We targeted the acetolactate synthase gene (als) that converts pyruvate to acetolactate, to eliminate the production of acetoin and 2,3-butanodial. A pBluescript derivative containing sections of the als coding region and an erythromycin resistance gene was directly introduced into L. plantarum TF103 cells to create mutations under selection pressure. The resulting erythromycin resistant (Em(r)) TF103 strain appears to have chromosomal mutations of both the als and the adjacent lysP genes as revealed by polymerase chain reaction and Southern blot analyses. Mutations were thus generated via targeted homologous recombination using a Gram-negative cloning vector, eliminating the use of a shuttle vector. This method should facilitate research in targeted inactivation of other genes in LAB.

New methods of safe biological pest control are required as a result of evolution of insect resistance to current biopesticides. Yeast strains being developed for conversion of cellulosic biomass to ethanol are potential host systems for expression of commercially valuable peptides, such as bioinsecticides, to increase the cost-effectiveness of the process. Spider venom is one of many potential sources of novel insect-specific peptide toxins. Libraries of mutants of the small amphipathic peptide lycotoxin-1 from the wolf spider were produced in high throughput using an automated integrated plasmid-based functional proteomic platform and screened for ability to kill fall armyworms, a significant cause of damage to corn (maize) and other crops in the United States. Using amino acid scanning mutagenesis (AASM) we generated a library of mutagenized lycotoxin-1 open reading frames (ORF) in a novel small ubiquitin-like modifier (SUMO) yeast expression system. The SUMO technology enhanced expression and improved generation of active lycotoxins. The mutants were engineered to be expressed at high level inside the yeast and ingested by the insect before being cleaved to the active form (so-called Trojan horse strategy). These yeast strains expressing mutant toxin ORFs were also carrying the xylose isomerase (XI) gene and were capable of aerobic growth on xylose. Yeast cultures expressing the peptide toxins were prepared and fed to armyworm larvae to identify the mutant toxins with greatest lethality. The most lethal mutations appeared to increase the ability of the toxin α-helix to interact with insect cell membranes or to increase its pore-forming ability, leading to cell lysis. The toxin peptides have potential as value-added coproducts to increase the cost-effectiveness of fuel ethanol bioproduction. Copyright © 2008 European Peptide Society and John Wiley & Sons, Ltd.

A cold-sensitive Listeria monocytogenes mutant designated cld-14 was obtained by transposon Tn 917 mutagenesis. The gene interrupted by Tn 917 in cld-14 was the L. monocytogenes LMOf2365_ 1485 homolog, which exhibits 45.7% homology to the Bacillus subtilis yqfF locus. LMOf2365_ 1485 , here designated pgpH , encodes a putative integral membrane protein with a predicted molecular mass of 81 kDa. PgpH is predicted to contain a conserved N-terminal signal peptide sequence, seven transmembrane helices, and a hydrophilic C terminus, which likely extends into the cytosol. The Tn 917 insertion in pgpH is predicted to result in production of a premature polypeptide truncated at the fifth transmembrane domain. The C terminus of PgpH, which is probably absent in cld-14, contains a highly conserved HD domain that belongs to a metal-dependent phosphohydrolase family. Strain cld-14 accumulated higher levels of (p)ppGpp than the wild type accumulated, indicating that the function of PgpH may be to adjust cellular (p)ppGpp levels during low-temperature growth. The cld-14 pgpH + complemented strain was able to grow at a low temperature, like the parent strain, providing direct evidence that the activity of PgpH is important in low-temperature adaptation. Because of its predicted membrane location, PgpH may play a critical role in sensing the environmental temperature and altering cellular (p)ppGpp levels to allow the organism to adapt to low temperatures.

Resonant pitch angle scattering by electromagnetic ion cyclotron (EMIC) waves has been suggested to account for the rapid loss of ring current ions and radiation belt electrons. For the rising tone EMIC wave (classified as triggered EMIC emission), its frequency sweep rate strongly affects the efficiency of pitch-angle scattering. Based on the Cluster observations, we analyze three typical cases of rising tone EMIC waves. Two cases locate at the nightside (22.3 and 22.6 magnetic local time (MLT)) equatorial region and one case locates at the duskside (18MLT) higher magnetic latitude (λ = –9.3°) region. For the three cases, the time-dependent wave amplitude, cold electron density, and cold ion density ratio are derived from satellite data; while the ambient magnetic field, thermal proton perpendicular temperature, and the wave spectral can be directly provided by observation. These parameters are input into the nonlinear wave growth model to simulate the time-frequency evolutions of the rising tones. The simulated results show good agreements with the observations of the rising tones, providing further support for the previous finding that the rising tone EMIC wave is excited through the nonlinear wave growth process.

Lactobacillus plantarum ferments glucose through the Embden-Meyerhof-Parnas pathway: the central metabolite pyruvate is converted into lactate via lactate dehydrogenase (LDH). By substituting LDH with pyruvate decarboxylase (PDC) activity, pyruvate may be redirected toward ethanol production instead of lactic acid fermentation. A PDC gene from the Gram-positive bacterium Sarcina ventriculi (Spdc) was introduced into an LDH-deficient strain, L. plantarum TF103, in which both the ldhL and ldhD genes were inactivated. Four different fusion genes between Spdc and either the S. ventriculi promoter or three Lactococcus lactis promoters in pTRKH2 were introduced into TF103. PDC activity was detected in all four recombinant strains. The engineered strains were examined for production of ethanol and other metabolites in flask fermentations. The recombinant strains grew slightly faster than the parent TF103 and produced 90-130 mM ethanol. Although slightly more ethanol was observed, carbon flow was not significantly improved toward ethanol, suggesting that a further understanding of this organism's metabolism is necessary.

To study the transcriptional regulation of the rabbit cytochrome P450 2C2 gene (CYP2C2), transgenic mice were generated using a DNA fragment containing the CYP2C2 5′-flanking region fused to a luciferase reporter gene. In one of three transgenic lines, this transgene was expressed in the liver and at a lower level in the kidney, similar to the tissue-specific pattern of this gene in rabbits. Unlike expression in rabbits, the transgene was expressed in a male-specific pattern in liver and kidney, and in the brain of both sexes. Expression of the transgene in male mice was repressed after castration, while expression in females was stimulated after testosterone treatment and periodic injection of rat growth hormone (GH). The expression of this transgene was also regulated developmentally in the liver. These results suggest that this 3500 bp of CYP2C2 5′-flanking region contains the basal transcriptional regulation elements for tissue-specific and developmental expression. Although a position effect can not be ruled out, this transgene may also include the cis-regulatory element for sexual dimorphic expression.

To study the regulatory functions of the ON promoter region, a ppG1b1GUS construct, consisting of 1402 bp 5′ flanking sequence ofGlbl, 1919 by GUS coding sequence, and 283 by 3′ NOS terminator, was cloned into a binary vector and introduced into tobacco plants byAgrobacterium-mediated transformation. Histochemical GUS assays of To tobacco mature seeds indicate that theGlbl promoter drives GUS expression in ABA treated seeds. Further GUS assays of the T, seeds at different developmental stages revealed that without ABA treatment, theGibl promoter drives GUS expression in immature seeds. The results from both To and T1 tobacco plants indicated thatGlbl-driven GUS expression in tobacco is embryo specific.