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Interfacial properties play important roles in processes for hydrocarbon resource production, transport, and refining. Biplex liquid crystal domains, with diameters up to 200 μm, have been identified in unreacted heavy fractions of petroleum resources, particularly asphaltenes, worldwide. These liquid crystal domains comprise thin liquid crystal shells surrounding homogeneous cores dispersed in liquid. The identification of species comprising the shells has proven elusive because they are minor constituents of species present. In a recent work, we showed that produced water is enriched in liquid crystal domains relative to a hydrocarbon resource. In this work, gram quantities of liquid-crystal-rich materials were isolated from produced water. The isolation methods are described, and their compositions are analyzed using a combination of elemental analysis, Fourier transform infrared spectroscopy, and negative-ion electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry. The liquid-crystal-rich materials are shown to include humic substances (humic acid, fulvic acid, and humin) among the principal components. These surface-active and lyotropic liquid-crystal-forming materials are the only species known to be present that meet the constraints imposed by the analyses and self-assemble cooperatively. Validation experiments demonstrating the formation of lyotropic liquid crystal domains with solid and liquid hydrocarbon as well as aqueous cores + humic-acid-rich liquid crystalline shells dispersed in aqueous and hydrocarbon liquids were performed. The formation and behavior of liquid crystal domains in humic substance + hydrocarbon + water mixtures are analogous to those of oil–water–surfactant dispersions, where surfactants self-assemble on interfaces, forming a liquid crystal layer. Humic substances, common hydrocarbon reservoir constituents, may contribute to interfacial stability and surface deposition problems, arising in petroleum production, transport, and refining that are commonly attributed to asphaltenes because they appear to comprise a microscopic surphase on the exterior of asphaltene particles.

At first glance, it looked like very good news when David Cameron appointed Justine Greening as Secretary of State for International Development in his September 2012 reshuffle. Greening is an [...]

Distributions of electric fields in two-dimensional arrays of gold nanodisks on a Si3N4membrane, with light incident through the membrane, are modeled with the aim of determining array geometries for effective plasmon-mediated Brillouin light scattering (“surface-enhanced Brillouin scattering”) from phonons or magnons in a specimen placed in contact with such an array. Particular attention is devoted to average intensities and higher-wavevector components of the fields in a plane 2 nm from the circular nanodisk/vacuum interface, which is anticipated to be in the vicinity of the surface of a specimen. For nanodisks with diameters of 50 nm, the average intensity near the circular nanodisk/vacuum interface increases as the angle of the incident light approaches the normal of the Si3N4surface. At low angles of incidence relative to the Si3N4normal, average intensities also increase with decreasing array spacing, primarily because of the corresponding changes in fractional coverage area of the gold. The highest average intensities (with near-normal incidence and 70 nm array periodicity) are found to be ∼3 times that of the incident light. More significantly, higher-wavevector components of the fields are found to have intensities comparable to the incident light. This finding provides evidence for the feasibility of using surface plasmons in nanodisk or nanoline arrays to mediate Brillouin scattering from phonons or magnons with wavelengths of a few tens of nanometers, which would extend the wavevector range of Brillouin-scattering metrology by an order of magnitude.

Developmentally competent oocytes can be collected from xenografted ovarian tissues; however, optimal xenograft conditions need to be established for this technique to be of use in assisted reproduction. In the present study, common wombat ovarian tissue was xenografted under the kidney capsule of nude mice to clarify the role of recipient gonadal status and donor tissue age on graft establishment, follicle development and oocyte recovery. Eighty-nine per cent of all grafts were recovered; of these, 78% contained growing follicles. In female graft recipients, follicle development to the antral stage occurred earlier in ovariectomised recipients compared with intact graft recipients. Similarly, follicle development occurred earlier in recipients of pouch young ovarian tissue grafts when compared with subadult xenografts. Follicle development proceeded to the antral stage in subadult grafts placed under the kidney capsule of male recipient mice, albeit at a slower rate than subadult grafts placed in female recipients. Oocytes were collected from grafts placed in female and male recipients, but no mature oocytes were observed at the time of collection, nor could these oocytes be matured in vitro. The present study demonstrated that common wombat pouch young tissue xenografted to female recipient mice, and subadult ovarian tissue xenografted to male recipient mice, can develop to the antral stage and can therefore facilitate oocyte collection. However, mature oocytes were not obtained using the current protocol.

Bitumen and heavy oil + alkane solvents exhibit complex phase behaviors. For example, organic components of these materials can be solubilized and inorganic solids dispersed into a high-pressure gas phase. Conversely, they can also be partitioned into as many as three bulk phases:  a gas phase, a liquid phase that is largely free of inorganic solids, and a phase comprising essentially all of the inorganic solids and a small fraction of the organic material. The outcome depends on the temperature, pressure, and solvent-to-feed ratio. Mass balance results for a screening survey for Athabasca vacuum bottoms (ABVB) + alkane solvents (pentane, heptane, decane, and dodecane) are reported along with a limited number of phase composition data for ABVB + pentane and dodecane mixtures. Reversible phase behavior and irreversible thermolysis conditions were considered. The key findings are that inorganic solids are readily partitioned from ABVB irrespective of the solvent and operating conditions employed, while key heavy metals, such as vanadium, require a combination of phase behavior and mild thermolysis. Sulfur- and nitrogen-containing species possess low rejection selectivities in this solvent series.

Over the past 30 years many cryopreservation procedures have been applied to oocytes, embryos, sperm, ovarian and testicular tissue. Over this time many, often specialized, terms have been developed for all aspects of these procedures. This can make it difficult for readers who are not familiar with the terminology or protocols to compare and evaluate different procedures. This paper describes the main cryopreservation procedures, the terminology associated with them, and briefly explains the underlying physical, chemical and biological processes. The aim is to help readers understand and interpret other papers on slow cooling, rapid cooling, ultrarapid cooling and vitrification.

Weight-bearing exercise is traditionally recommended for improving bone health in postmenopausal women. Effects of swim exercise were studied as an alternative to weight-bearing exercise in ovariectomized rats. Rats in a swim group (Sw, n= 8) swam for 12 wk, 5 days/wk for 60 min per session. A control group (Con,n= 9) engaged in no structured exercise. Femurs were analyzed for bone mineral density and for bone mineral content by dual energy X-ray absorptiometry, biomechanical properties by three-point bending (Instron), and bone structure and formation by histomorphometry. Food intake did not differ among groups. Final body weights were significantly lower in Sw compared with Con (P< 0.05). Swimmers had significantly greater femoral shaft bone mineral density and content (P < 0.05) compared with Con. Femurs of the Sw group had greater mechanical properties (P< 0.05) compared with Con. Histomorphometric data were significantly better in the Sw group compared with Con after the 12-wk intervention (P < 0.05). In conclusion, data from this study demonstrate some beneficial effects of swim exercise on bone structure, turnover, and strength.

Double packaging, in which an inner straw containing the specimen is inserted into an outer, larger straw (here termed 'straw-in-straw') to prevent the inner straw from coming into direct contact with liquid nitrogen provides a simple strategy for reducing or eliminating the potential contamination risk associated with storage in liquid nitrogen. This approach has in the past been used in conjunction with cryopreservation by slow cooling, but has not previously been tested for use throughout an entire rapid cooling and warming procedure. This study determined whether keeping the straw containing the embryos inside a second protecting container throughout the cryopreservation and storage protocol would compromise embryo viability. We established that a cryoprotectant containing a high polymer concentration (35% dextran or Ficoll) together with 25% ethylene glycol (as the penetrating cryoprotectant) was highly effective for day 2 and day 3 mouse embryos in both single and double straws. The survival and development of all cryopreserved embryos, as assessed both in vitro and in vivo, was not statistically different to their untreated controls. This established that a protein/serum-free cryoprotectant solution supplemented with polymers could provide complete protection of mouse embryos. It also shows, for the first time, that embryos can be cooled by direct immersion in liquid nitrogen and warmed by direct immersion into a waterbath within a double straw arrangement to reduce the likelihood of contamination.

Purpose. Determine if wet milling technology could be used to formulate water insoluble antitumor agents as stabilized nanocrystalline drug suspensions that retain biological effectiveness following intravenous injection.

ABSTRACTA partial phase diagram for the system athabasca bitumen vacuum bottoms (ABVB) (24.6 wt% / 2 mole % ) + dodecane (73.8 wt. % / 47 mole % ) + hydrogen (1.6 wt % / 51 mole % ) was constructed in the temperature range 425 K to 725 K and the pressure range 2 MPa to 7 MPa using an X-ray view cell apparatus. This fluid system is shown to exhibit two phase L1V and three phase L1L2V phase behaviour over parts of this P-T region. The shape of the low temperature boundary between the L1V and L1L2V zones is characteristic of dilute asymmetric mixtures where a heavy liquid phase, L2appears then disappears within the light liquid phase L1on isothermal compression. Such phase behaviour is referred to as unusual retrograde condensation and is of both practical and theoretical interest. Transitions between the multiphase regions were found to be reversible at temperatures less than 660 K., in all cases. At higher temperatures irreversible “ asphaltene precipitation” arose within the L2phase “ Asphaltene precipitation” did not arise in the absence of the L2phase, i.e.: within the L1V region, even at temperatures in excess of 700 K. These data provide a strong link between “ asphaltene precipitation” and multiphase behaviour, and demonstrate a physical rather than kinetic basis for asphaltene precipitation at elevated temperatures

Mouse pronuclear and 4-cell embryos were cryopreserved by slow cooling to -33 degrees C in 1.5 M 1,2-propanediol or 1.5 M ethylene glycol, with or without 0.1 M sucrose. Straws were thawed by immersion into a 37 degrees C water bath, immediately after their removal from liquid nitrogen (protocol 1), or after being held in air for 15 (protocol 2) or 30 s (protocol 3). Others were held in air until the ice melted (protocol 4). Embryos which formed blastocysts that hatched and attached to the Petri dish in vitro (plated) were considered viable. The thawing protocol did not significantly influence the viability of embryos frozen in propanediol with 0.1 M sucrose (52-72% of pronuclear and 69-97% of 4-cell embryos plated). In the other solutions tested, propanediol without sucrose and ethylene glycol with/without sucrose, only protocol 2 resulted in uniformly high development of both pronuclear (45-65% plating) and 4-cell embryos (70-97% plating). Thawing protocol 4 significantly reduced development, in particular for embryos frozen in ethylene glycol (0% 1-cell; 0-25% 4-cell plating). The difference between thawing protocols 2 and 4 was reduced by continuing slow cooling of ethylene glycol solutions to lower temperatures (-41 degrees C). Adding antifreeze proteins type I or III did not improve survival or development. Thus, although mouse pronuclear and 4-cell embryos can be frozen-thawed in either ethylene glycol or propanediol without significant loss of viability, an appropriate thawing protocol is essential for embryos frozen in ethylene glycol or propanediol-sucrose.

A method has been developed for the selective delivery of lipophilic immunomodulators to macrophages, which results in the induction of antitumor activity. This method utilizes exhaustively acetylated low density lipoprotein (acetyl-LDL) to deliver the lipophilic immunomodulator, muramyl tripeptide phosphatidylethanolamine (MTP-PtdEtn; amide composed of N-acetylmuramyl-L-alanyl-D-isoglutamyl-L-alanine and dipalmitoyl phosphatidylethanolamine) to macrophages (M phi) by means of a scavenger lipoprotein receptor pathway. The binding of acetyl-LDL:MTP-PtdEtn to M phi showed specificity since minimal competition was observed in the presence of excess native LDL or phosphatidylcholine/cholesterol liposomes. Additional binding studies showed that acetyl-LDL may serve as a suitable delivery vehicle to a wide variety of M phi in different stages of activation. Cytostatic and tumoricidal activities by thioglycolate-elicited M phi against two tumor cell lines were examined in vitro following incubation with the acetyl-LDL:MTP-PtdEtn complex. Cytostatic activity against B16F10 melanoma cells was induced after the incubation of thioglycolate-elicited M phi with a minimum of 25 micrograms of acetyl-LDL protein containing 2.5 micrograms of bound MTP-PtdEtn (approximately equal to 40 molecules per particle of acetyl-LDL). The induction of cytostasis was not affected by liposome bilayers, which were also endocytosed by the M phi. In addition, tumoricidal activity against P815 mastocytoma cells was demonstrated at a 40:1 effector-to-target ratio using 18 micrograms of the acetyl-LDL:MTP-PtdEtn complex containing 3.6 micrograms of MTP-PtdEtn (approximately equal to 80 molecules per particle). These studies describe a method for the induction of antitumor activity by use of a chemically modified serum component, acetyl-LDL, to direct lipophilic immunomodulators to M phi.

This study retrospectively compared the success of in vitrofertilization (IVF) among patients whose gametes had been incubated either in medium supplemented with freshly prepared pooled serum (331 cases) or in pooled serum which had been stored at -20°C prior to use (728 cases). Frozen stored serum was as effective as fresh serum with regard to the proportion of oocytes which fertilized and embryos which implanted and was not associated with any increased incidence of fetal loss during postimplantation development.

ABSTRACTThe appearance of solid-liquid-liquid-vapour phase equilibria in reservoir fluids is explained as an extension of the phase behaviour of binary asymmetric fluids. The theoretical approach is demonstrated for the model condensate-rich reservoir-fluid ethane (10 mole %) butane (40 mole %) phenanthrene (50 mole %), to which propane is added. Extant solid-liquid-liquid-vapour phase equilibrium data for reservoir fluids, which include asphaltenes, are found to be consistent with the phase behaviour of this model fluid. The implications for the prediction of asphaltene “precipitation” are discussed.

While optimizing the ultrarapid embryo freezing procedure we noted that embryo survival was lowest when gas bubbles formed in the straws. Here we report the influences of gassing the freezing solutions with 5% CO2 in air, degassing the solution, the pH of the medium and straw irradiation on the survival and development in vitro of 2-cell mouse embryos. Embryos were ultrarapidly frozen in medium M2 containing 3 M dimethyl sulphoxide and 0.25 M sucrose. Significantly fewer embryos survived freezing and thawing in gassed solutions. The subsequent development of intact embryos to blastocysts was similar in gassed and non-gassed solutions. Survival and development of embryos frozen in solutions of pH 7.0, 7.6 or 8.0 was similar, but fewer embryos developed to blastocysts after freezing in media at pH 6.0. In straws sterilized by gamma irradiation before use, embryo survival after freezing and thawing was dramatically reduced compared with that seen when non-irradiated straws were used. With non-gassed solutions significantly more gas bubbles formed in irradiated straws than in non-irradiated straws. Although bubble formation was significantly reduced in all straws by using degassed freezing solutions, this did not improve embryo survival. We conclude that under conditions normally used for ultrarapid freezing, dissolved gas and pH have less effect on embryo viability than the use of irradiated straws.

Interaction with excess unilamellar phosphatidylcholine (PC) vesicles resulted in depletion of as much as 90% of the cholesterol from the membrane of intact vesicular stomatitis (VS) virus. The cholesterol depletion was not significantly influenced by the proteolytic removal of virion glycoprotein spikes, but it was temperature dependent. Cholesterol depletion caused substantial reduction in anisotropy of the VS virion membrane as measured by fluorescence depolarization of the lipophilic probe 1,6-diphenyl-1,3,5-hexatriene; residual adsorbed vesicles represent a significant factor in this apparent increase in virion membrane fluidity. Interaction with PC vesicles resulted in a substantial loss of VS viral infectivity as measured by plating efficiency on L-cell monolayers. Reduction in infectivity appeared to be related to temperature-dependent depletion of virion cholesterol by PC vesicles. Interaction of VS virions with cholesterol-containing PC vesicles resulted in significantly less decline in infectivity, but attempts to restore cholesterol and infectivity to depleted VS virions were unsuccessful. Depletion of virion cholesterol apparently results through collision with PC vesicles rather than movement of cholesterol monomers or micelles through the aqueous phase, because PC vesicle-virion interaction in the presence of cholesterol oxidase did not result in substantial oxidation of translocated cholesterol.

Girls and young women who require ovariectomy or cancer therapy may consider having their own eggs, embryos or ovarian tissue stored (cryopreserved) for their own future use. Ovarian tissue is simple to collect and contains large numbers of germ cells. Transplantation of fresh and frozen-thawed ovarian tissue in healthy sheep and mice has resulted in normal live young. Similar techniques may be effective in the human but it is unclear whether ovarian tissue cryopreservation and grafting is suitable for ovaries from individuals with cancer or infections. If cancer cells were present in an ovary at the time of collection and survived cryopreservation and grafting they could establish cancer in the recipient. We therefore performed ovarian cryopreservation and transplantation trials using a mouse lymphoma model. This established that the lymphoma was transmitted by grafts of both fresh and frozen ovarian tissue. The normal healthy recipient mice died 9-43 days after receiving a small piece (1 mm3) of ovarian tissue from a donor with lymphoma. We conclude that ovarian tissue which is collected, cryopreserved and grafted while it contains cancer cells has the potential to spread the cancer to the graft recipient.

Membrane vesicles were isolated from the basolateral domains of pig and normal human colonocytes. The activity of the ouabain-sensitive K(+)-activated phosphatase, the basolateral membrane marker, was enriched 13-fold in these membrane vesicles over the original homogenate. The membranes displayed cross-reactions with antibodies to the (Na+/K+)ATPase and the RLA class I major histocompatibility antigen, both known indicators of the basolateral membrane. There was negligible contamination by other organelles and the luminal membrane, as revealed by marker-enzyme analysis and Western blotting, using an antibody to villin. The vesicles transported D-glucose in a cytochalasin B-inhibitable Na(+)-independent manner, with a Km of 28.1 +/- 0.8 mM and Vmax. of 3.1 +/- 0.4 nmol/s per mg of protein. The transport was inhibited by 2-deoxy-D-glucose and 3-O-methyl-D-glucose, but not by L-glucose or methyl-alpha-D-glucose. Probing the colonocyte basolateral membranes with an antibody against the C-terminus of the human liver GLUT 2 produced a cross-reaction at 52 kDa. These properties indicate the presence of a GLUT 2 isoform on the basolateral membranes of human and pig colonocytes.

A rabbit polyclonal antibody to purified ox kidney branched-chain oxo acid dehydrogenase complex was shown by a variety of techniques to be an antibody to the E2 (acyltransferase) component. Rocket immunoelectrophoresis showed that the antibody does not discriminate between phosphorylated (inactive) or dephosphorylated (active) complex, and the same technique is used to assay total branched-chain complex (sum of active and inactive forms) in rat liver and heart mitochondrial extracts. The values obtained in normal rats fed on normal diet were comparable with those obtained by spectrophotometric assay of the holocomplex reaction after conversion of inactive complex into active complex. The values obtained in liver mitochondria from rats fed on 0%-casein diet or starved for 48 h were comparable with those in rats fed on normal diet, whereas earlier studies using spectrophotometric assay had shown substantial decreases in rats fed on 0%-casein diet or starved for 48 h. It has been shown that conversion of inactive complex into active complex requires prolonged incubation (120 min) in the presence of ketoleucine (4-methyl-2-oxopentanoate; to inhibit branched-chain oxo acid dehydrogenase kinase) to effect complete conversion in mitochondria from rats fed on 0%-casein diet, or starved for 48 h, or made diabetic with alloxan. By this technique, total activity of the complex in rat liver mitochondria was unaffected by diet or diabetes. The effects of diet and diabetes to decrease the activity of branched-chain complex in rat liver are therefore apparently mediated wholly through inactivation of the complex by phosphorylation.

Summary.This paper investigates the effect of straw handling on the viability of 2-cell mouse embryos rapidly frozen in dimethyl sulphoxide (DMSO) solutions. During the brief (3 min) equilibration step, straws were either rotated periodically to keep the embryos in suspension, or kept still to allow the embryos to settle onto the inner surface of the straw. The effects of these straw movements were tested with cryoprotectant solutions containing 1·5, 3·0 or 4·5 m-DMSO. Rapidly cooled straws containing 4·5 m-DMSO vitrify throughout on cooling, but ice forms on warming. The survival and normality of embryos frozen in 4·5 m-DMSO was not influenced by straw handling as 91–92% formed blastocysts in vitro, 77–78% formed normal fetuses, and no chromosomal rearrangements were observed. In solutions containing <4.5 m-DMSO ice formation occurred throughout (1·5 m-DMSO), or in parts (3·0 m-DMSO) of the cryoprotectant during cooling. The viability of embryos frozen in 3·0 or 1·5 m-DMSO solutions was reduced both in vitroand in vivoand structural chromosome aberrations, predominantly tri- and quadri-radial rearrangements, were observed. The reduction in embryo viability, and the chromosomal damage was particularly pronounced in embryos frozen in 3·0 m-DMSO in straws which were rotated during the equilibration step (47% blastocysts, 15% fetuses, 77% chromosome rearrangements). The results indicate that rapid freezing of 2-cell mouse embryos in 4·5 m-DMSO is safe and efficient, whereas freezing at lower DMSO concentrations is associated with severe chromosome damage, and reduced viability in vitroand in vivo.Keywords:dimethyl sulphoxide; rapid freezing; ice formation; handling; chromosome rearrangements; mouse; embryos

[14C]aztreonam was administered intramuscularly (50 mg/kg) to male and female rats. Groups of 10 rats (five male and five female) were sacrificed at 0.25, 2, 6, and 24 h after dosing. Blood and various tissues were removed from six rats (three male and three female) in each group for determination of total radioactivity and unchanged aztreonam by liquid scintillation counting and thin-layer radiochromatography. The remaining rats were prepared for whole-body autoradiography. Radioactivity was distributed rapidly to most tissues and was rapidly eliminated from blood into urine and bile. Concentrations of total radioactivity in kidney, liver, small and large intestines and their contents, and urinary bladder were higher than those in serum at all the times examined. Concentrations of unchanged aztreonam in serum, kidney, liver, and lung declined at about the same rate as did that of total radioactivity in the same tissues. The results of whole-body autoradiography essentially confirmed the results of the distribution of [14C]aztreonam as determined by liquid scintillation counting. No major differences between males and females were observed when concentrations in organs common to both were compared.

[14C]aztreonam was administered as single 25-mg/kg doses to dogs (intravenously and subcutaneously) and monkeys (intramuscularly and intravenously) and as single 50-mg/kg doses (intramuscularly and intravenously) to rats. In rats and dogs, radioactive moieties were excreted primarily in urine; in monkeys, they were excreted about equally in urine and feces. Unchanged aztreonam accounted for 77 to 86% of the radioactivity excreted in the urine of rats, dogs, and monkeys; SQ 26,992, the metabolite resulting from hydrolysis of the monobactam ring, accounted for 10 to 15%; and minor, unidentified metabolites accounted for the remainder. In rats with cannulated bile ducts, about 15% of an intramuscular dose was excreted in bile in 24 h; the bile contained a greater percentage of metabolites than that found in urine. In dogs, the apparent elimination half-life of aztreonam in serum was 0.7 h after intravenous administration. Aztreonam and SQ 26,992 accounted for most of the radioactivity in the sera of dogs and monkeys. Serum protein binding of aztreonam and its metabolites ranged from 28 to 35% in dogs and from 49 to 59% in monkeys. In the three species studied, aztreonam was most extensively metabolized in monkeys; SQ 26,992 and other minor metabolites from monkey urine were tested and found to be devoid of any significant antimicrobial activity.

Subcutaneous administration of [14C]aztreonam (150 mg/kg) to pregnant rats was followed by the appearance of radioactive moieties in fetuses and amniotic fluid. Concentrations of both total radioactivity and unchanged aztreonam in maternal serum declined more rapidly than those in fetuses and amniotic fluid. [14C]aztreonam (150 mg/kg) was also administered subcutaneously to lactating rats. Radioactivity and unchanged aztreonam were found in the suckling pups and in the serum and milk of the dam. Results obtained by whole-body autoradiography were generally consistent with these obtained by measuring radioactivity present in the excised tissues. Autoradiographs of the pups showed detectable amounts of radioactivity in the brain; since no radioactivity was detectable in the brain of the dam, it appears that the blood-brain barrier was not fully developed in 7-day-old pups.

Two systems for measuring embryo development in vitro were evaluated. One was a 1–4 scale based on a subjective evaluation of embryo quality (EQ) from microscopic appearance. In addition, a formula for scoring embryo growth rate in vitro was developed. The embryo development rating (EDR) was based on the ratio between the time at which embryos were observed at a particular stage after insemination and the time at which they would be expected to reach that stage in a hypothetical “ideal” growth rate with a cell cycle length of 11.9 hr. Using this scoring system, “normally” growing embryos scored 100. This approach was aimed at partially normalizing the data and allowed all embryos to be analyzed similarly regardless of the time of observation. Analysis of 1539 embryo replacements resulting in 232 clinical pregnancies showed that both EDR and embryo-quality scores were of value in predicting success, with clinical pregnancy most likely to eventuate from a combination of moderate to good EQ scores (2–4) coupled with average or above-average growth rates (EDR scores from 90 to 129). Poor-quality and very slowly or very rapidly growing embryos were underrepresented in cycles that proceeded to pregnancy. These inferences were based on all embryos transferred (mean, 2.73 per transfer cycle), and they were substantiated by an analysis of 33 pregnancies resulting from replacement of a single embryo and from 18 pregnancies in which all embryos scored the same with both systems. EQ and EDR were significantly associated with each other and together provide a valuable guide in predicting pregnancy, in selecting embryos for freezing, and in monitoring day-to-day performance in the in vitro fertilization (IVF) program.

In this paper we describe a novel method of inducing conductivity in polyaniline photochemically or by electron-beam exposure. This is accomplished with the use of onium salts which are a class of materials that decompose upon irradiation generating protonic acids. We find that the onium salt may be blended with the polyaniline and upon irradiation, the generated acid acts as an in situ dopant for the polymer. Conductivity on the order of ⋍0.1 S/cm has been attained. This system has significant applications in lithography since it allows patterns of conducting lines to be generated. The polyaniline/onium system represents the first electrically conducting photo and electron-beam resist. In addition, we find that this radiation induced doping technique is applicable to polythiophene systems as well.

The Leishmania tarentolae cytochrome oxidase (EC 1.9.3.1) subunit II (COII) and maxicircle unidentified reading frame 3 (MURF3) mRNAs are edited internally by the addition of four and five uridine residues, respectively, which eliminate -1 and +1 reading frameshifts in the gene sequences. The editing events in COII are conserved in three kinetoplastid species, and those in MURF3 are conserved in two species. A primer extension assay showed that the ratio of edited to unedited RNA differed for each gene: 89% of the COII and 36% of the MURF3 transcripts are edited. Preliminary evidence was obtained for translation of the edited COII transcript into protein: antibodies generated against a synthetic peptide with the predicted carboxyl-terminal amino acid sequence reacted with a polypeptide of the correct molecular weight in immunoblots of a mitochondrial lysate.

The effect of protein-free diet to decrease liver activity of branched-chain (-2-oxoacid) dehydrogenase (BCD) complex (active form) and increase BCD kinase activity was unaffected by preparation of hepatocytes, but partially reversed by 25 h of culture of hepatocytes in medium 199. Activation of BCD complex preceded loss of BCD kinase. The effect of culture on BCD complex was completely prevented by omission of branched-chain amino acids and partially prevented by 1 mM-alpha-cyano-4-hydroxycinnamate or 0.2 mM-pyruvate/2 mM-lactate. Protein-free diet decreased plasma branched-chain amino and oxo (‘keto’) acids and increased plasma pyruvate and lactate. It is concluded: (1) that branched-chain amino acids are involved directly in regulation of activities of BCD complex and BCD kinase; (2) that mitochondrial uptake of branched-chain oxo acids is necessary for regulation of BCD complex activity; and (3) that the stable increase in BCD kinase may function as a hysteresis mechanism.

The cytochrome b gene of Trypanosoma brucei has an ATG codon near its 5' end but the cytochrome b genes of the related kinetoplastids Leishmania tarentolae and Crithidia fasciculata lack an ATG. Recent results have shown that 34 uridines that are not encoded in the genome are added within the 5' end of T. brucei cytochrome b transcripts during or after transcription. These additions create an AUG in the transcript that is 20 amino acids upstream of the AUG predicted from the genomic sequence. We report here that the cytochrome b transcripts of L. tarentolae and C. fasciculata also contain added uridines within their 5' ends. The additions occur in similar numbers and positions and an in frame AUG is created at a similar site in all three species. These data strongly suggest that the created AUG functions as the initiation codon for cytochrome b in these species. Since some other kinetoplastid mitochondrial genes also lack conventional initiation codons, creation of initiation codons may be an important function of uridine addition.