3T3 fibroblasts transfected with a cDNA for mitochondrial aspartate aminotransferase express plasma membrane fatty acid-binding protein and saturable fatty acid uptake.

To explore the relationship between mitochondrial aspartate aminotransferase (mAspAT; EC 2.6.1.1) and plasma membrane fatty acid-binding protein (FABPpm) and their role in cellular fatty acid uptake, 3T3 fibroblasts were cotransfected with plasmid pMAAT2, containing a full-length mAspAT cDNA downstream of a Zn(2+)-inducible metallothionein promoter, and pFR400, which conveys methotrexate resistance. Transfectants were selected in methotrexate, cloned, and exposed to increasing methotrexate concentrations to induce gene amplification. Stably transfected clones were characterized by Southern blotting; those with highest copy numbers of pFR400 alone (pFR400) or pFR400 and pMAAT2 (pFR400/pMAAT2) were expanded for further study. [3H]Oleate uptake was measured in medium containing 500 microM bovine serum albumin and 125-1000 microM total oleate (unbound oleate, 18-420 nM) and consisted of saturable and nonsaturable components. pFR400/pMAAT2 cells exhibited no increase in the rate constant for nonsaturable oleate uptake or in the uptake rate of [14C]octanoate under any conditions. By contrast, Vmax (fmol/sec per 50,000 cells) of the saturable oleate uptake component increased 3.5-fold in pFR400/pMAAT2 cells compared to pFR400, with a further 3.2-fold increase in the presence of Zn2+. Zn2+ had no effect in pFR400 controls (P > 0.5). The overall increase in Vmax between pFR400 and pFR400/pMAAT2 in the presence of Zn2+ was 10.4-fold (P < 0.01) and was highly correlated (r = 0.99) with expression of FABPpm in plasma membranes as determined by Western blotting. Neither untransfected 3T3 nor pFR400 cells expressed cell surface FABPpm detectable by immunofluorescence. By contrast, plasma membrane immunofluorescence was detected in pFR400/pMAAT2 cells, especially if cultured in 100 microM Zn2+. The data support the dual hypotheses that mAspAT and FABPpm are identical and mediate saturable long-chain free fatty acid uptake.

Screen-printed electrode-based electrochemical detector coupled with in-situ ionic-liquid-assisted dispersive liquid–liquid microextraction for determination of 2,4,6-trinitrotoluene

A novel method is reported, whereby screen-printed electrodes (SPELs) are combined with dispersive liquid–liquid microextraction. In-situ ionic liquid (IL) formation was used as an extractant phase in the microextraction technique and proved to be a simple, fast and inexpensive analytical method. This approach uses miniaturized systems both in sample preparation and in the detection stage, helping to develop environmentally friendly analytical methods and portable devices to enable rapid and onsite measurement. The microextraction method is based on a simple metathesis reaction, in which a water-immiscible IL (1-hexyl-3-methylimidazolium bis[(trifluoromethyl)sulfonyl]imide, [Hmim][NTf2]) is formed from a water-miscible IL (1-hexyl-3-methylimidazolium chloride, [Hmim][Cl]) and an ion-exchange reagent (lithium bis[(trifluoromethyl)sulfonyl]imide, LiNTf2) in sample solutions. The explosive 2,4,6-trinitrotoluene (TNT) was used as a model analyte to develop the method. The electrochemical behavior of TNT in [Hmim][NTf2] has been studied in SPELs. The extraction method was first optimized by use of a two-step multivariate optimization strategy, using Plackett–Burman and central composite designs. The method was then evaluated under optimum conditions and a good level of linearity was obtained, with a correlation coefficient of 0.9990. Limits of detection and quantification were 7 μg L−1 and 9 μg L−1, respectively. The repeatability of the proposed method was evaluated at two different spiking levels (20 and 50 μg L−1), and coefficients of variation of 7 % and 5 % (n = 5) were obtained. Tap water and industrial wastewater were selected as real-world water samples to assess the applicability of the method.

Role of the interfacial water structure on electrocatalysis: Oxygen reduction on Pt(1 1 1) in methanesulfonic acid

Most of electrocatalytic reactions occur in an aqueous environment. Understanding the influence of water structure on reaction dynamics is fundamental in electrocatalysis. In this work, the role of liquid water structure on the oxygen reduction at Pt(1 1 1) electrode is analyzed in methanesulfonic (MTSA) and perchloric acids. This is because these different anions can exert a different influence on liquid water structure. Results reveal a lower ORR electrode activity in MTSA than in HClO4 solutions and they are discussed in light of anion's influence on water structural ordering. From them, the existence of an outer-sphere, rate determining, step in the ORR mechanism is suggested.

Dinoflagellate cysts in surface sediment samples of METEOR cruises M20/2, M23/1, M34/1, M34/2, M34/3 and M34/4

To enhance the limited information available about the palaeo-ecological significance of calcareous dinoflagellates, we have studied their lateral distribution in surface sediments of the equatorial and south Atlantic between 13°N and 36°S. Calcareous dinoflagellate cysts appear to be widely distributed throughout the studied area. In the surface sediments, concentrations (cyst per gram dry sediment) of the vegetative stage Thoracosphaera heimii are generally higher than that of the (presumably) calcareous resting cysts. Distribution patterns in surface sediments of Orthopithonella granifera (Fütterer) Keupp and Versteegh, Rhabdothorax spp. Kamptner., Sphaerodinella albatrosiana (Kamptner) Keupp and Versteegh S. albatrosiana praratabulated, Sphaerodinella tuberosa var. 1 (Kamptner) Keupp and Versteegh and S. tuberosa var. 2 and the ratios between these species have been compared with temperature, salinity, density and stratification gradients in the upper water column. Rhabdothorax spp. is characteristically present in sediments of more temperate regions characterized by high seasonality. Dinoflagellates producing these cysts are able to tolerate high nutrient concentrations, and mixing of the water column. S. albatrosiana is abundant in regions characterized by high sea surface temperatures and oligotrophic surface water conditions. In contrast, the distribution of S. tuberosa var. 2 is negatively related to temperature. The other cyst species did not show a characteristic pattern in relation to the studied environmental gradients. The ratio of Sphaerodinella tuberosa var. 2 to Orthopithonella granifera can be used for reconstructing the presence of stratification in the upper 50 m of the water column, whereas the ratios of S. tuberosa var. 2 to Sphaerodinella albatrosiana and of O. granifera to Rhabdothorax spp. might be used for palaeotemperature reconstructions. Calcareous dinoflagellate cysts are abundant in oligotrophic areas and may be useful for the reconstruction of palaeoenvironmental conditions.

Pami a tniki starinnoi russkoi literatury, izdavaemye grafom Grigoriem Kushelevym -Bezborodko. vyp. 1-4; 1860-1862.

Vols. 1-2, 4, edited by N.I. Kostomarov; v. 3, by A.N. Pypin.

Direct support and general support level : truck, cargo, 1-1/4 ton, 6X6, M561 (NSN 2320-00-873-5407), truck, ambulance, 1-1/4 ton, 6X6, M792 (NSN 2310-00-832-9907).

Includes index.

Unit maintenance manual for truck, cargo, tactical, 1-1/4 ton, 4x4, M1008 (2320-01-123-6827), truck, cargo, tactical, 1-1/4 ton, 4x4, M1008A1 (2320-01-123-2671), truck, utility, tactical, 3/4 ton, 4x4, M1009 (2320-01-123-2665), truck, ambulance, tactical, 1-1/4 ton, 4x4, M1010 (2310-01-123-2666) ....

Includes index.

Intermediate direct support/general support maintenance manual for truck, cargo, tactical, 1-1/4 ton, 4x4, M1008, (2320-01-123-6827), truck, cargo, tactical, 1-1/4 ton, 4x4, M1008A1, (2320-01-123-2671) ... truck, chassis, tactical, 1-1/4 ton, 4x4, M1031, (2320-01-133-5368).

Includes index.

Organizational maintenance repair parts and special tools lists : loader, scoop type, diesel engine, 4-wheel drive, rear wheel steerable pneumatic tires, 2 1/2 cu yd, w/ multi-segment bucket, (Frank G. Hough model H-90CM) FSN 3805-995-3236.

Includes index.

Direct and general support and depot maintenance repair parts and special tools lists [microform] : loader, scoop type, diesel engine, 4 wheel drive, rear wheel steerable pneumatic tires, 2 1/2 cu yd, w/multi-segment bucket (Frank G. Hough model H-90CM), FSN 3805-995-3236.

"November 1967."

Intermediate (field) (direct and general support) and depot level maintenance manual : generator set, diesel engine driven, tactical skid MTD 5KW, 1 phase-2 wire, 1 phase-3 wire, 3 phase-4 wire, 120, 120/240 and 120/208 volts, DoD model MEP-002A, class utility, hertz 60, NSN 6115-00-465-1044.

"July 1977."

Intermediate (field) (direct and general support) and depot level maintenance manual : generator set, diesel engine driven, tactical skid MTD 10KW, 1 phase-2 wire, 1 phase-3 wire, 3 phase-4 wire, 120, 120/240 and 120/208 volts : DoD model MEP-003A, class utility, hertz 60, NSN 6115-00-465-1030.

"July 1977."