Methanogenium frigidum sp. nov., a psychrophilic, H2-using methanogen from Ace Lake, Antarctica

Methanogenium frigidum sp. nov. was isolated from the perennially cold, anoxic hypolimnion of Ace Lake in the Vesfold Hills of Antarctica. The cells were psychrophilic, exhibiting most rapid growth at 15 degrees C and no growth at temperatures above 18 to 20 degrees C. The cells were irregular, nonmotile coccoids (diameter, 1.2 to 2.5 microns) that occurred singly and grew by CO2 reduction by using H2 as a reductant. Formate could replace H2, but growth was slower. Acetate, methanol, and trimethylamine were not catabolized. Cells grew with acetate as the only organic compounds in the culture medium, but growth was much faster in medium also supplemented with peptones and yeast extract. The cells were slightly halophilic; good growth occurred in medium supplemented with 350 to 600 mM Na+, but no growth occurred with 100 or 850 mM Na+. The pH range for growth was 6.5 to 7.9; no growth occurred at pH 6.0 or 8.5. Growth was slow (maximum specific growth rate, 0.24 day-1; doubling time, 2.9 days). This is the first report of a psychrophilic methanogen growing by CO2 reduction.     

Distribution circuit state estimation using a probabilisticapproach

Past work on distribution circuit state estimation has focused on the adoption of a transmission state estimator approach, without necessarily accounting for the specific requirement of a distribution circuit-based analysis. On distribution circuits, typically, there are very few available real-time measurements, and thus, researchers have treated customer load demand estimates as pseudo-measurements in a weighted-least-squares formulation. This can lead to convergence problems and also, the approach effectively assumes that all bus load demands are normally distributed (Gaussian) which may not be valid on distribution circuits. This paper presents an alternative approach to distribution circuit state estimation using a probabilistic extension of the radial load flow algorithm while accounting for real-time measurements as solution constraint. The algorithm which takes advantage of the radial nature of distribution circuits also accounts for other issues specific to distribution circuits. Namely, the algorithm accounts for nonnormally distributed loads, incorporates the concept of load diversity (load correlation) and can interact with a load allocation routine. The effectiveness of the algorithm is illustrated through comparisons made with Monte Carlo simulations     

Caecal disease in equids

OBJECTIVE: To review the breed, age, gender, clinical and laboratory findings, treatment and outcome of horses with caecal disease presented to a referral centre. DESIGN: Retrospective study of 96 cases. PROCEDURE: The breed, age, and gender of the study population were compared with the corresponding hospital population for the same period. The means (+/- SD) for clinical and laboratory findings were recorded for each caecal disorder. Treatment was categorised as medical or surgical, and outcome was recorded. RESULTS: Caecal diseases included impaction (40% of total cases), rupture associated with concurrent unrelated disease (13%), rupture with parturition (9%), rupture with no associated disease (5%), infarction (11%), torsion (9%), abscess or adhesion (7%), tumour (3%), and miscellaneous conditions (3%). The breed or gender of affected horses did not differ from the hospital population, although horses > 15 years were more frequently represented (P < 0.05). This age group was specifically more predisposed to caecal impaction (P < 0.05), as were Arabian, Morgan, and Appaloosa breeds (P < 0.05). In horses with caecal impaction transrectal examination was the most useful diagnostic procedure; 90% of affected horses treated medically were discharged while horses treated by typhlotomy alone, or typhlotomy and blind end ileocolostomy, had survival rates to discharge of 71% and 86%, respectively. Horses with caecal rupture associated with concurrent un-related disease showed no signs of impending rupture; all were receiving phenylbutazone, all were euthanased, and 50% had caecal ulceration at necropsy. Of horses with caecal rupture with parturition 56% had prior dystocia; in two-thirds the site of rupture was the ileocaecal junction and all were euthanased. Horses with caecal rupture with no associated disease died or were euthanased; rupture was idiopathic. Horses with caecal infarction usually had signs of abdominal pain and abdominal fluid changes consistent with peritonitis; transrectal examination was nonspecific, and typhlectomy was successful in seven of eight horses. Horses with caecal torsion had signs of severe, acute abdominal pain and typhlectomy was successful in three of five horses. Diagnosis of caecal adhesion or abscess was assisted by transrectal palpation in two of seven horses and surgical treatment was successful in two of five horses. A caecal tumour was diagnosed in three horses aged 20 years or older that presented with chronic weight loss. Other caecal diseases were uncommon. CONCLUSION: Caecal disease is uncommon in equids but some specific features of the history and physical findings can alert the veterinarian to the possibility of caecal involvement in horses with gastrointestinal dysfunction. Medical or surgical therapy can be effective in horses where caecal rupture has not occurred.     

An enhancer that directs lineage-specific expression of CD8 in positively selected thymocytes and mature T cells

Positive selection of CD4+CD8+ T cells to the CD4+CD8- helper and CD4- CD8+ cytotoxic lineages is a multistep process that involves complex regulation of coreceptor gene expression. By analyzing expression of a reporter gene in transgenic mice, we have identified a DNA segment, located between the murine CD8beta and CD8alpha genes, that has enhancer activity restricted to CD8 lineage cells. Remarkably, this enhancer functions in thymocytes undergoing positive selection to the CD4-CD8+ phenotype but not in immature double-positive thymocytes. The enhancer also functions in gut intraepithelial lymphocytes that express CD8alpha but not CD8beta, suggesting that it is specific for CD8alpha expression. The tight correlation between activation of this enhancer and the final step in positive selection has important implications for understanding the mechanism of lineage commitment in thymocytes.     

Dynamic kinetic asymmetric cycloadditions of isocyanates to vinylaziridines

The first examples of the use of racemic vinylaziridines in a Pd-catalyzed dynamic kinetic asymmetric transformation have been examined. Optimization studies of the Pd-catalyzed addition of vinylaziridines to isocyanates revealed that the chiral ligand between trans-1,2-diaminocyclohexane and 2-diphenylphosphino-1-naphthoic acid is superior to that involving 2-diphenylphosphino benzoic acid. Surprisingly, high ee's required the use of an acid whose pKa was about 4.7 +/- 0.1 as a cocatalyst. Both acetic acid and hydroxybenzotriazole meet this requirement. Less electrophilic isocyanates (e.g., benzyl, p-methoxyphenyl) gave higher ee's than more electrophilic ones (phenyl or benzoyl). Both N-benzyl and N-arylaziridines react well to give good yields and ee's, whereas N-tosylaziridines gave lower ee's. A 1,1-disubstituted aziridine led to the formation of a tertiary C-N bond with ee's comparable to the formation of the secondary C-N bond. The products were easily reduced almost quantitatively to the sensitive imidazolidines which can be readily hydrolyzed to the vicinal diamines. The reactivity pattern is consistent with a Curtin-Hammett situation wherein the enantiodiscriminating event is the cyclization of a rapidly equilibrating dynamic pi-allyl palladium intermediate.     

2-Hydroxypropyl-beta-cyclodextrin enhances phorbol ester effects on glucose transport and/or protein kinase C-beta translocation to the plasma membrane in rat adipocytes and soleus muscles

In rat adipocytes and soleus muscles, 2-hydroxypropyl-beta-cyclodextrin (CD) was found to have a relatively small or no effect on basal or insulin-stimulated hexose uptake, but markedly enhanced hexose uptake effects of phorbol esters and/or diacylglycerol. In rat adipocytes, the CD-induced enhancement of hexose uptake during concurrent phorbol ester treatment was not associated with an increase in GLUT4 glucose transporter translocation to the plasma membrane, which was stimulated comparably by insulin and phorbol esters. Moreover, CD appeared to activate or facilitate the activation of glucose transporters subsequent to their translocation to the plasma membrane during ongoing phorbol ester treatment. In rat adipocytes, CD also enhanced the translocation of protein kinase C (PKC)-beta to the plasma membrane during the action of phorbol esters, which alone had little or no effect on this specific PKC translocation. Although it is uncertain how CD alters the function of plasma membranes to enhance the translocation of PKC-beta to, and the activation of glucose transporters within, this subcellular fraction during phorbol ester treatment, our findings provide direct support for a two-step model in the activation of glucose transport. In addition, it seems clear that, at least in some cell types, simple phorbol ester treatment does not necessarily serve as a ubiquitous activator of all activable PKC pools and all potential PKC-mediated responses.     

Plant genetics: fast flowering

Bacillus cereus hemolysin III activity was tested in crude extracts, from Escherichia coli carrying the hly-III gene. It was concluded that hemolysin III is a pore-forming hemolysin with functional pore diameter of about 3-3.5 nm. Hemolysis occurs in at least three steps: (i) the temperature-dependent binding of the Hly-III monomers to the erythrocyte membrane; (ii) the temperature-dependent formation of the transmembrane oligomeric pore; (iii) the temperature-independent erythrocyte lysis.     

Mass spectrometric analysis of 2-deoxyribonucleoside and 2'-deoxyribonucleotide adducts with aldehydes derived from lipid peroxidation

An important emerging issue in chemical carcinogenesis is the role that products of endogenous metabolism play in formation of covalently modified DNA. One example is the formation of alpha, beta-unsaturated aldehydes as a result of endogenous and drug-stimulated lipid peroxidation. Malondialdehyde (MDA), crotonaldehyde (CR), 2-hexenal (HX), and 4-hydroxy-2-nonenal (HNE) react covalently with 2'-deoxyguanosine (dG) and 2'-deoxyadenosine (dA) residues on DNA to form promutagenic cyclic adducts that may be important in the etiology of cancer in humans and animals. The accurate quantification of such adducts provides a powerful tool in molecular epidemiology for assessing carcinogenic risks from various lifestyle choices (e.g. diet, drug use) in humans. 32P-Postlabeling is recognized as one of the most sensitive methods available for detection of DNA adducts in human tissues, but without adequate validation such methodology can yield inaccurate quantitative measurements. We have used LC separations in conjunction with electrospray ionization MS and tandem MS (triple quadrupole and hybrid quadrupole-orthogonal acceleration time of flight analyzers) to characterize MDA-, CR-, HX- and HNE-modified dG and nucleotide (3'- and 5'-monophosphate; 3',5'-bisphosphate) adducts. These data have been used to validate 32P-postlabeling methods for quantification of low level MDA-dG adducts formed in DNA of human and animal tissues. Availability of reliable methods for quantification of endogenous DNA damage in humans and animals is essential for determining unknown etiologies of cancer and for the assessment of cancer risks in humans.