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This paper describes reversible "on-off" switching of the photoluminescence (PL) intensity of CdSe quantum dots (QDs), mediated by photochromic furylfulgide carboxylate (FFC) molecules chemisorbed to the surfaces of the QDs. Repeated cycles of UV and visible illumination switch the FFC between "closed" and "open" isomers. Reversible switching of the QDs' PL intensity by80% is enabled by different rates and yields of PL-quenching photoinduced electron transfer (PET) from the QDs to the respective isomers. This difference is consistent with cyclic voltammetry measurements and density functional calculations of the isomers' frontier orbital energies. This work demonstrates fatigue-resistant modulation of the PL of a QD-molecule complex through remote control of PET. Such control potentially enables applications, such as all-optical memory, sensing, and imaging, that benefit from a fast, tunable, and reversible response to light stimuli.

π-Conjugated polymers can serve as active layers in flexible and lightweight electronics and are conventionally synthesized by transition-metal-mediated polycondensation at elevated temperatures. We recently reported a photopolymerization of electron-deficient heteroaryl Grignard monomers that enables the catalyst-free synthesis of n-type π-conjugated polymers. Herein, we describe an experimental and computational investigation into the mechanism of this photopolymerization. Spectroscopic studies performed in situ and after quenching reveal that the propagating chain is a radical anion with halide end groups. DFT calculations for model oligomers suggest a Mg-templated SRN1-type coupling, in which Grignard monomer coordination to the radical anion chain avoids the formation of free sp2 radicals and allows C-C bond formation with very low barriers. We find that light plays an unusual role in the reaction, photoexciting the radical anion chain to shift electron density to the termini and thus enabling productive monomer binding.

Metallaphotoredox catalysis combines the well-established mechanisms of transition-metal-catalyzed cross-coupling reactions with one-electron redox manipulations enabled by light. In most cases, a transition metal or organic dye serves as the photoredox catalyst while a ground-state Pd or Ni catalyst performs the organometallic steps. Cross-coupling mechanisms that rely on direct photoexcitation of a light-absorbing substrate have the potential to access distinct mechanisms and deliver unique selectivity based on the substrate’s excited-state properties. In this report, we describe a photoinduced, Ni-catalyzed Suzuki–Miyaura cross coupling reaction that selectively functionalizes BODIPY chromophores, a versatile class of tunable, bright, photostable fluorophores. Using a bis-iodo BODIPY substrate, the selectivity for mono- vs. bis-arylation was found to be governed by a remote substituent that subtly alters the excited-state properties of the substrate. Consistent with a substrate photoexcitation mechanism, high chemoselectivity is also observed in mixtures of chromophores with distinct excited-state properties. This reaction is compatible with a variety of substituted BODIPY chromophores and boronic acids and esters, enabling the rapid synthesis of unsymmetrically-substituted chromophores.

Current approaches to synthesize π-conjugated polymers (CPs) are dominated by thermally driven, transition-metal-mediated reactions. Herein we show that electron-deficient Grignard monomers readily polymerize under visible-light irradiation at room temperature in the absence of a catalyst. The product distribution can be tuned by the wavelength of irradiation based on the absorption of the polymer. Conversion studies are consistent with an uncontrolled chain-growth process; correspondingly, chain extension produces all-conjugated n-type block copolymers. Preliminary results demonstrate that the polymerization can be expanded to donor-acceptor alternating copolymers. We anticipate that this method can serve as a platform to access new architectures of n-type CPs without the need for transition-metal catalysis.

Despite the discovery of the first N-acyl homoserine lactone (AHL) based quorum sensing (QS) in the marine environment, relatively little is known about the abundance, nature and diversity of AHL QS systems in this diverse ecosystem. Establishing the prevalence and diversity of AHL QS systems and how they may influence population dynamics within the marine ecosystem, may give a greater insight into the evolution of AHLs as signaling molecules in this important and largely unexplored niche. Microbiome profiling of Stelletta normani and BD1268 sponge samples identified several potential QS active genera. Subsequent biosensor-based screening of a library of 650 marine sponge bacterial isolates identified 10 isolates that could activate at least one of three AHL biosensor strains. Each was further validated and profiled by Ultra-High Performance Liquid Chromatography Mass Spectrometry, with AHLs being detected in 8 out of 10 isolate extracts. Co-culture of QS active isolates with S. normani marine sponge samples led to the isolation of genera such as Pseudomonas and Paenibacillus, both of which were low abundance in the S. normani microbiome. Surprisingly however, addition of AHLs to isolates harvested following co-culture did not measurably affect either growth or biofilm of these strains. Addition of supernatants from QS active strains did however impact significantly on biofilm formation of the marine Bacillus sp. CH8a sporeforming strain suggesting a role for QS systems in moderating the microbe-microbe interaction in marine sponges. Genome sequencing and phylogenetic analysis of a QS positive Psychrobacter isolate identified several QS associated systems, although no classical QS synthase gene was identified. The stark contrast between the biodiverse sponge microbiome and the relatively limited diversity that was observed on standard culture media, even in the presence of QS active compounds, serves to underscore the extent of diversity that remains to be brought into culture.

In the developing world, the identification of clean, potable water continues to pose a pervasive challenge, and waterborne diseases due to fecal contamination of water supplies significantly threaten public health. The ability to efficiently monitor local water supplies is key to water safety, yet no low-cost, reliable method exists to detect contamination quickly. We developed an in vitro assay utilizing an odorant-binding protein (OBP), AgamOBP1, from the mosquito, Anopheles gambiae, to test for the presence of a characteristic metabolite, indole, from harmful coliform bacteria. We demonstrated that recombinantly expressed AgamOBP1 binds indole with high sensitivity. Our proof-of-concept assay is fluorescence-based and demonstrates the usefulness of insect OBPs as detector elements in novel biosensors that rapidly detect the presence of bacterial metabolic markers, and thus of coliform bacteria. We further demonstrated that rAgamOBP1 is suitable for use in portable, inexpensive &ldquo, dipstick&rdquo, biosensors that improve upon lateral flow technology since insect OBPs are robust, easily obtainable via recombinant expression, and resist detector &ldquo, fouling.&rdquo, Moreover, due to their wide diversity and ligand selectivity, insect chemosensory proteins have other biosensor applications for various analytes. The techniques presented here therefore represent platform technologies applicable to various future devices.

Having a compact yet robust structurally based identifier or representation system is a key enabling factor for efficient sharing and dissemination of research results within the chemistry community, and such systems lay down the essential foundations for future informatics and data-driven research. While substantial advances have been made for small molecules, the polymer community has struggled in coming up with an efficient representation system. This is because, unlike other disciplines in chemistry, the basic premise that each distinct chemical species corresponds to a well-defined chemical structure does not hold for polymers. Polymers are intrinsically stochastic molecules that are often ensembles with a distribution of chemical structures. This difficulty limits the applicability of all deterministic representations developed for small molecules. In this work, a new representation system that is capable of handling the stochastic nature of polymers is proposed. The new system is based on the popular “simplified molecular-input line-entry system” (SMILES), and it aims to provide representations that can be used as indexing identifiers for entries in polymer databases. As a pilot test, the entries of the standard data set of the glass transition temperature of linear polymers (Bicerano, 2002) were converted into the new BigSMILES language. Furthermore, it is hoped that the proposed system will provide a more effective language for communication within the polymer community and increase cohesion between the researchers within the community., BigSMILES, a line notation that supports intrinsically stochastic molecules on top of the simplified molecular-input line-entry system (SMILES), is presented to pave the way for polymer informatics.

The conformation changes on reaction of maleic anhydride with two tropomyosins (vertebrate and invertebrate) and paramyosin are reported. Maleylation of tropomyosin leads to more than 90% loss of the α-helical conformation and dissociation into subunits. Only one third of the helix is lost on maleylation of paramyosin and it is not dissociated into subunits. The ORD and CD spectra of maleyl tropomyosin are reported and compared with the spectra of other unordered polypeptide chains. The α-helix can be restored after maleylation of tropomyosin by increase of the ionic strength, variation of pH, or addition of helix-forming solvents such as ethanol, 2-chloroethanol, and trifluoroethanol.

Fumaric acid, a Krebs cycle intermediate, is a potential cancer chemoprevention agent. A high performance liquid chromatographic procedure with UV detection for determination of fumaric acid in large numbers of rat plasma, urine and fecal samples was developed. Fumaric acid was extracted from plasma, urine, and fecal samples utilizing solid phase extraction using Clean Up® Quaternary Amine 1 mL (plasma, fecal samples) and 3 mL (urine) extraction columns followed by reverse phase high performance liquid chromatography with UV detection at 215 nm. Standard curves for plasma (1 μg/mL - 200 μg/mL), urine (5 μg/mL - 200 μg/mL), and fecal material (25 μg/g - 500 μg/g) were analyzed and replicate analysis of controls were used to determine intra-day and inter-day variability. Chlorofumaric acid was used as an internal standard for plasma and fecal samples; trans-glutaconic acid for urine samples. Precision and accuracy were studied using control solutions (low and high) prepared in naive rat plasma, uri...

The purpose of this study was to develop a simple and fast analytical method for quantitation of ampicillin in rabbit plasma, suitable for analysis of large numbers of samples collected from experimental animals. The concentration of ampicillin in rabbit plasma was determined utilizing ion-pair reverse-phase high performance liquid chromatography (HPLC) with UV detection and a column switching technique. Plasma samples were treated with a perchloric acid solution to precipitate proteins and centrifuged to pellet the precipitated proteins. Cephalexin was used as an internal standard. The C18 precolumn was placed in the injector loop of the Rheodyne injector. Samples were injected with the injector in the load position and the precolumn was washed free from interfering compounds. When the injector was switched to the inject position, the mobile phase was passed through the precolumn taking relatively pure compounds onto the analytical column. The limit of quantitation was established to be 400 ng mL−1 of plasma. The standard curves were linear over the range of ampicillin concentrations assayed, 400 to 10,000 ng mL−1 of rabbit plasma, and had a mean regression coefficient of 0.9962 (±0.0043). Intra-day variability was determined using six replicates of controls (low and high) analyzed on a single assay. The percent of relative accuracy for low and high controls were 5.67 and 1.12, respectively. Inter-day variability was determined over a four day period analyzing replicates of each control. The percent of relative accuracy for low and high controls were 4.33 and 1.63, respectively.

Beeturia, the passage of pink or red urine after the ingestion of beetroot, is said to occur in 10-14% of the population, and is more common in iron deficiency and malabsorption. A specific HPLC assay for betacyanins, the red beetroot pigments, in biological fluids was developed to study the prevalence of this apparent polymorphism in humans, and to investigate its basis in rats. Two major peaks were observed in chromatograms of extracts of unpickled beetroot. They had identical UV absorption spectra (lambda max = 535 nm) by diode array analysis, and mass spectrometry indicated that one (betacyanin 1) was betanin or its epimer and the other (betacyanin 2) a disaccharide of betacyanin 1. In a population of 100 normal subjects the 0-8 h urinary recoveries after an oral dose of 60 mg beetroot extract were 0.06-0.54% for betacyanin 1 and 0.01-0.6% for betacyanin 2. The distributions of these data were skewed but not clearly bimodal by visual inspection or by kernel density analysis. Four subjects produced visibly red urine and had betacyanin recoveries at the upper end of the population range. Studies using in situ isolated perfused rat jejunum and liver preparations indicated a negligible absorption of the pigments after 1 h and no detectable metabolism or biliary secretion. Intact anaesthetized rats given i.v. bolus doses of beetroot extract cleared both betacyanins from plasma at the rate of 3.3 +/- 0.9 (SD) ml min-1 (n = 5). The total urinary recovery of both pigments amounted to 80% of the dose, and their renal clearances approached their plasma clearances. These data suggest that beeturia does not arise from deficiencies in hepatic metabolism or renal excretion of betacyanins. After oral administration of beetroot extract to rats the betacyanin content of the stomach decreased rapidly with time but neither the intestines nor the bile duct were stained visibly red. These findings together with those showing instability of the betacyanins in acid conditions suggest that variability in the biological fate of beetroot pigments may be determined largely by gastric pH and emptying rate.

Natural plant products such as essential oils have gained interest for use in pest control in place of synthetic pesticides because of their low environmental impact. Essential oils can be effective in controlling parasitic mites that infest honeybee colonies, but effective encapsulants are needed to provide a sustained and targeted delivery that minimizes the amount of active ingredient used. The present study reports the encapsulation of essential oils in porous microspheres that are within the size range of pollen grains and can be easily dispersed. The microspheres were made by pumping an 8% aqueous high-amylose starch gelatinous melt through an atomizing nozzle. The atomized starch droplets were air-classified into two fractions and collected in ethanol. The size range for each fraction was measured using a particle size analyzer. The mean particle size for the largest fraction was approximately 100 microm with a range from 5 microm to over 300 microm. Part of the reason for the large particle size was attributed to the merging of smaller particles that impinged upon each other before they solidified. The smaller fraction of spheres had a mean particle size of approximately 5 microm. The starch-based porous microspheres were loaded with 16.7% (w/w) essential oils including thymol (5-methyl-2-isopropylphenol), clove, origanum, and camphor white oil. The essential oils appeared to be largely sequestered within the pore structure, since the spheres remained a free-flowing powder and exhibited little if any agglomeration in spite of the high loading rate. Furthermore, SEM micrographs verified that the pore structure was stable, as evidenced by the persistence of pores in spheres that had first been loaded with essential oils and then had the oil removed by solvent extraction. Thermal gravimetric analyses were consistent with a loading rate at predicted levels.

The production rate of endogenous L(+)-lactate in the resting human being can be calculated from results of experiments during which isotopic or unlabelled lactate is infused. The merits of the different experimental techniques are discussed, although all published methods have given similar results, namely a resting production rate of about 1.3 mol . (70 kg body wt)-1 . 24 h-1. This result is similar to that found in sheep, but lower than that in dogs and rats. L(+)-lactate production in human subjects is unchanged after fasting for 36-168 h, a finding which contrasts with some animal experiments. It is not known whether this represents a true species difference or a difference in diet. Net splanchnic uptake of L(+)-lactate in the resting human being is about 0.4 mol 24 h-1 although the normal liver has the capacity to metabolize more than this. The kidney is an important site of L(+)-lactate utilization in rats, but there is no information about its role in normal human beings. There are conflicting reports concerning the uptake of L(+)-lactate by resting skeletal muscle, but there is no evidence that resting muscle can metabolize lactate.

Of the two types of lactic acidosis, Type B is the most difficult to explain in terms of mechanisms of lactic acid accumulation because the tissue hypoxia that accompanies Type A is not present except as a terminal event. The methods of pharmacokinetics show that for lactate to accumulate to an extent that would disturb acid-base balance, either lactate synthesis or lactate removal would have to be increased or decreased, respectively, by about 7-10-fold. To produce lactic acidosis within a few hours, synthesis would have to increase at the same time as clearance decreased. At normal contribution of the liver to the total lactate clearance (30%-40%), a total cessation of hepatic lactate clearance would not result in lactic acidosis without a concomitant rise in the rate of lactate synthesis. Again, to produce an acidosis this increase would have to be about 8-fold at a normal fractional hepatic clearance. The control of hepatic lactate uptake is discussed in the light of the relatively low hepatic extraction of lactate and in relation to to two main models of hepatic drug (lactate) clearance; the 'well-stirred' and 'parallel-tube' models.

Coeur d'Alene Lake in northern Idaho is fed by two major rivers: the Coeur d'Alene River from the east and the St. Joe River from the south, with the Spokane River as its outlet to the north. This phosphorus-limited lake has been subjected to decades of mining (primarily for zinc and silver) and other anthropogenic inputs. A 32 full-factorial experimental design was used to examine the interactive effects of free (uncomplexed) zinc ion and dissolved-orthophosphate concentrations on phytoplankton that were isolated from two sites along a longitudinal zinc-concentration gradient in Coeur d'Alene Lake. The two sites displayed different dominanttaxa. Chlorella minutissima, a dominant species near the southern St. Joe River inlet, exhibited greater sensitivity to free Zn ions than Asterionella formosa, collected nearer the Coeur d'Alene River mouth with elevated dissolved-zinc concentrations. Empirical phytoplankton-response models were generated to describe phytoplankton growth in response to remediation strategies in the surrounding watershed. If dissolved Zn can be reduced in the water column from500 nM (i.e., current concentrations near and down stream of the Coeur d'Alene River plume) to3 nM (i.e., concentrations near the southern St. Joe River inlet) such that the lake is truly phosphorus limited, management of phosphorus inputs by surrounding communities will ultimately determine the limnologic state of the lake.

Objectives: To determine the relation of hemostatic markers to cardiovascular fitness and adiposity and the effect of physical training (PT) on these markers. Study design: Seventy-four obese teenagers were randomly assigned to 8 months of lifestyle education (LSE), LSE plus moderate-intensity PT, or LSE plus high-intensity PT. Measures included fibrinogen, plasminogen activator inhibitor-1 (PAI-1), D-dimer, C-reactive protein (CRP), fitness, percent body fat (%BF), and visceral adiposity (VAT). Results: At baseline, fibrinogen and CRP were significantly correlated with %BF and VAT (0.27 ≤ r ≤ 0.51), and fitness ( r = −0.39, r = −0.43, respectively); PAI-1 with %BF ( r = 0.30) and VAT ( r = 0.48); D-dimer with fitness ( r = −0.24). Although PT produced significant changes in fitness and adiposity, there were no significant group differences in the hemostatic markers. Change in PAI-1 was significantly correlated with its baseline value ( r = −0.47) and change in %BF ( r = 0.38). Changes in D-dimer and CRP were significantly correlated with their respective baseline values ( r = −0.68, r = −0.48, respectively). Conclusions: Unfavorable levels of fitness and adiposity were associated with higher levels of hemostatic markers, putting individuals with this profile at greater risk for future cardiovascular disease. No evidence was provided that 8 months of PT had a direct influence on these markers in obese youths. (J Pediatr 2002;141:415-20)

The aim of this study was to determine the response of inflammatory and vasoactive mediators to 3 consecutive days of exercise in African-American women with and without sickle cell anemia (SCA). Circulating inflammatory mediators [C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor alpha (TNFalpha)] were measured before, and vasoactive mediators [endothelin-1 (ET-1), nitric oxide metabolites (NOx)] before and after each exercise bout in ten subjects with SCA and ten controls. Exercise did not affect ET-1, IL-6 or CRP concentrations (p >.05). TNFalpha was higher in SCA than controls (p < or = .0005) at all times; however, the response pattern was similar for the groups: no change from day 1 to day 2, but a decrease from day 2 to day 3 (p < or = .05). NOx increased significantly after exercise (p < or = .0001) but returned to baseline by 24 h afterward. On the 3rd day, NOx increased after exercise in SCA but not in the controls (p < or = .05). In conclusion, exercise did not cause a harmful inflammatory response in these individuals with SCA. However, NOx increased after exercise on all 3 days in SCA but appeared attenuated after 2 days in controls.

Phenethyl isothiocyanate is unstable in aqueous media and at low pH, and rapidly degrades to phenethylamine. Concentrations of phenethylamine, a phenethyl isothiocyanate marker, in dog plasma, were determined utilizing solid-phase extraction and gas chromatography-mass spectrometry with chemical ionization using acetone as the reagent gas. Deuterated d5-amphetamine was used as an internal standard. After extraction, phenethylamine and d5-amphetamine were derivatized using MBHFBA. Ions monitored for d5-amphetamine were m/z 337 and 338; and for phenethylamine were m/z 318 and 319. Precision and accuracy were studied using control solutions prepared in naive dog plasma (80 and 300 ng/ml). Intra-day variability was determined using six replicates of each control solution analyzed on a single day. The relative standard deviation for the 80 ng/ml control was 12.9% and for the 300 ng/ml it was 12.1%. Relative accuracy was 10.9% for the low control and -4.1% for the high control. Inter-day variability was determined over a 6-day period. For the 80 and 300 ng/ml control solutions, the relative standard deviations were 15.8 and 9.1%, respectively, and relative accuracy values were 10.1 and -5.2%, respectively. Standard curves were prepared in naive dog plasma and were linear over the range of phenethylamine assayed (10-500 ng/ml). The results of this study indicate that the proposed method is simple, precise, accurate and sensitive enough for analysis of large numbers of plasma samples.

Mutations of the tumor suppressor gene discs-large (dlg) lead to postsynaptic structural defects. Here, we report that mutations in dlg also result in larger synaptic currents at fly neuromuscular junctions. By selectively targeting DLG protein to either muscles or motorneurons using Gal-4 enhancer trap lines, we were able to rescue substantially the reduced postsynaptic structure in mutants. Rescue of the physiological defect was accomplished by presynaptic, but not postsynaptic targeting, consistent with our finding that miniature excitatory junctional currents were not changed in dlg mutants. These results suggest that DLG functions in the regulation of neurotransmitter release and postsynaptic structure. We propose that DLG is an integral part of a mechanism by which changes in both neurotransmitter release and synapse structure are accomplished during development and plasticity.

For several decades, the main thrust in infrared technology developments in Australia has been in two main sensor technologies: uncooled silicon chip printed bolometric sensors pioneered by DSTO's Kevin Liddiard, and precision engineered high quality Cadmium Mercury Telluride developed at DSTO under the guidance of Dr. Richard Hartley. In late 1993 a low cost infrared imaging device was developed at DSTO as a sensor for guided missiles. The combination of these three innovations made up a unique package that enabled Australian industry to break through the barriers of commercializing infrared technology. The privately owned company, R.J. Optronics Pty Ltd undertook the process of re-engineering a selection of these DSTO developments to be applicable to a wide range of infrared products. The first project was a novel infrared imager based on a Palmer scan (translated circle) mechanism. This device applies a spinning wedge and a single detector, it uses a video processor to convert the image into a standard rectangular format. Originally developed as an imaging seeker for a stand-off weapon, it is producing such high quality images at such a low cost that it is now also being adapted for a wide variety of other military and commercial applications. A technique for electronically stabilizing it has been developed which uses the inertial signals from co-mounted sensors to compensate for platform motions. This enables it to meet the requirements of aircraft, marine vessels and masthead sight applications without the use of gimbals. After tests on a three-axis motion table, several system configurations have now been successfully operated on a number of lightweight platforms, including a Cessna 172 and the Australian made Seabird Seeker aircraft.© (1995) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

The 0-8 hour urinary distributions of the metabolic ratios of sparteine (100 mg), debrisoquine (10 mg) and metoprolol (100 mg) were measured in 165 healthy, unrelated, black Nigerian medical students. There was a weak correlation (rs = 0.51, p < 0.001; n = 82) between the metoprolol/alpha-hydroxymetoprolol (M/HM) and the sparteine/total (2- + 5-) dehydrosparteine (S/DHS) ratios. No significant correlations were found between the debrisoquine/4-hydroxydebrisoquine (D/HD) and M/HM ratios (rs = 0.16, n = 33) and between the D/HD and S/DHS ratios (rs = 0.31, n = 38). Both visual inspection and kernel density analysis of the data suggested the presence of two phenotypic groups for sparteine oxidation, with 4% of the population studied being putative poor metabolizers. In contrast biomodality was not apparent in the distribution of the log10M/HM and log10D/HD ratios. These findings provide evidence for a dissociation in the control of metoprolol, sparteine and debrisoquine oxidation in Nigerians and highlight the difficulties in the interpretation of data from pharmacogenetic studies in different ethnic groups.

The deposition of urate in the form of a tophus is a characteristic feature of gout. The tophus comprises a mass of urate crystals surrounded by acute inflammatory cells, with an outer layer of giant cells and epithelial cells. Larger deposits — nodules — are made up of large masses of urate (hydrated monosodium urate) together with acute inflammatory cells and giant cells. Calcium salts are commonly included along with fat, protein and polysaccharides. The processes that lead up to the deposition of urate in tissues appear, at first consideration, simple. The solubility of monosodium urate in human plasma is saturated at 7 mg/dl (Seegmiller 1965). At concentrations around or above 7 mg/dl, the urate would be expected to precipitate out to form tophi.

Urinary 3-methylhistidine (3MH) excretion has been studied in 20 thyrotoxic patients before, during and after treatment. 3MH excretion was high on diagnosis and fell significantly, when the patients had been rendered euthyroid, to levels measured in a control group. There was a significant linear correlation between 3MH excretion and free thyroxine index. The calculated muscle protein breakdown and 3MH/creatinine molar ratio also fell significantly after treatment of thyrotoxicosis.

Treatment of several Drosophila cell lines with the molting hormone (20-hydroxyecdysone) resulted in biochemical and cellular changes including the morphogenetic process of cell aggregation. Radiolabeling of the cell surface proteins revealed 34 polypeptides that are modulated by the hormone's action. This modulation included both expression of "new" proteins and disappearance of preexisting polypeptides. Whereas most of the hormone-induced proteins were lentil lectin-binding glycoproteins, only one group of disappearing proteins appears to bind lentil lectin. Labeling of the cell surface prior to hormone addition revealed no specific modification of preexisting surface proteins which could account for the protein changes observed with one possible exception. The potential relationship between the modulation in surface proteins and the increase in cell-cell adhesion that occurs during hormone exposure is discussed.

A questionnaire sent to laboratories in the UK and Ireland elicited information on acceptance or otherwise of values assigned to serum-based calibration materials for 14 analytes. Overall, 33% of assigned values were changed by users. The reasons for change, together with the degree of magnitude and direction of change, were studied for each analyte. Unidirectional changes made to assigned values were related to particular batches and manufacturers. Detailed information is presented for protein (Beckman RIIC Ltd.) on Astras, and sodium, bicarbonate and bilirubin on SMACs and SMAs (Technicon Instrument Co. Ltd.)

The sequential reaction of 3β-acetoxypregna-5,16-dien-20-one (1) with methylmagnesium bromide and then methyl iodide has been examined in detail. The major product (85%) was 3β-hydroxy-16α,17α-dimethylpregn-5-en-20-one (2). Other products identified were 3β-hydroxy-16α-methylpregn-5-en-20-one (4); the 21-methyl derivatives [(5) and (10)] of (2) and (4); the 21-(1-hydroxy-1-methylethyl) derivative (11) of (2); and a trace of the 21,21-dimethyl derivative (12) of (2). The scope of the reaction with regard to the range of substituents that can be introduced at positions 16 and 17 has been established by preparing a series of 16α-substituted 17α-methylpregnenolones with a variety of alkyl, aryl, and aralkyl groups. Some of these have been converted into the corresponding 16α,17α-disubstituted progesterones.

The disposition kinetics of ethylene oxide, ethylene glycol, and 2-chloroethanol were studied following their intravenous administration to beagle dogs. Plasma concentration of ethylene oxide was found to decline exponentially with a mean rate constant of 0.024 +/- 0.008 min-1 (mean +/- SD) and total body clearance of 20.0 +/- 5.2 ml/kg X min. Ethylene oxide was found to be metabolized mainly to ethylene glycol, which had a mean plasma half-life of 221.0 +/- 77.7 min and a total body clearance of 2.13 +/- 0.58 ml/kg X min. Between 7 and 24% of intravenously administered ethylene oxide was eliminated in the urine as ethylene glycol within 24 h. The elimination half-life and clearance values for 2-chlorethanol were 40.8 +/- 5.7 min and 10.3 +/- 1.7 ml/kg X min, respectively. The pharmacokinetic data gathered in the present investigation suggest that ethylene glycol rather than 2-chloroethanol is the major metabolite of ethylene oxide in the dog.

Preparation of the four isomeric 3,11-diamino-5α-pregnanes is reported. Three (3β,11α-, 3α,11α-, and 3α,11β-) were prepared from 5α-pregnane-11,20-dione obtained from 20-oxo-5α-pregnane-3β,11α-diyl diacetate. The fourth (3β,11β-) was prepared from 11-oxo-5α-pregnan-3β-yl acetate which is also available from 5α-pregnane-11,20-dione, though for convenience another source was used.The equatorial amino-groups (3β- and 11α-) were formed by alkali-metal reduction of the oximes; the axial 3α-amino-group by reduction of the 3α-azide; and the axial 11β-amino-group by catalytic hydrogenation of the oxime.

Treatment of methyl 3β-acetoxy-16α,17α-dimethyl-5α-androst-9(11)-ene-l7-carboxylic acid (1), or 3β-hydroxy-16α,17α-dimethyl-5α-pregn-9(11)-en-20-one (4), with methylmagnesium halide in refluxing anisole gives the 20-yne (3). Similarly 3β-acetoxy-16α,17α-dimethylpregn-5-en-20-one (5) gives the 20-yne (6). The mechanism of the reaction is discussed in terms of the steric hindrance due to the 16- and 17-methyl groups. Acetylenes (3) and (6) are converted into compounds of potential pharmacological interest.

The cell-surface proteins of Drosophila embryos at gastrula and myoblast fusion stages were characterized by radioiodination and two-dimensional gel electrophoresis. Over 13% of the cell surface proteins detected in gastrula embryos were not found in myoblast fusion stage embryos or in Drosophila embryonic cell line EH34A3 cells. Nearly 18% of the cell-surface proteins detected in myoblast fusion stage embryos were evident only at that stage. Embryonic cell-surface proteins were compared with cell-surface proteins from untreated EH34A3 cells and EH34A3 cells treated with 20-hydroxyecdysone, which induces cell aggregation and the expression of “new” proteins at the cell surface ( D. F. Woods and C. A. Poodry, 1983 , Dev. Biol.96, 23–31). Only one of the proteins induced by ecdysone in EH34A3 cells was detected in the NP-40 soluble fraction of radioiodinated cell lysates, even after fractionation by lectin affinity chromatography and immunoprecipitation to enrich for putative ecdysone induced proteins. However, extraction of the NP-40 insoluble pellet of embryo cells revealed one additional protein that was present both in myoblast fusion stage embryos and hormone-treated culture cells. It was concluded that except for these two proteins, the cell-surface proteins induced in cultured cell lines by treatment with 20-hydroxyecdysone are not present in significant amounts in gastrula or myoblast fusion stage embryos.

Antisera were raised in rabbits using conjugates of albumin with 11-hemisuccinates of 11α-OH E1 and 11α-OH E2. These antisera were compared with antisera to 6-oxo E2 6-CMO-, E2 17-succinyl- and E3 16, 17-disuccinyl-BSA by radioimmunoassay using a statistically designed three-point assay. Sheep anti-(rabbit γ-globulin) coupled to cellulose was used for the separation of antibody-bound and free labeled hapten. Antisera obtained with haptens linked to the carrier at the 17(16) position poorly discriminated between the various estrogens. Antisera obtained with 6- and 11-conjugates showed a much better specificity. In addition the specificity was influenced by using either estrone, estradiol or estriol as tritiated labels. This gives the possibility to determine different parameters by employing different labeled hormones.

The development of a 21-alkylation reaction which proceeds via the lithium 20(21)-enolate is described and its scope demonstrated by the preparation of a variety of 21-alkylpregnane derivatives. Application of this process to 11β-acetoxy-16α,17α-dimethyl-5α-pregnane-3,20-dione (28a) and its 5β-analogue (28b) led to the corresponding 16α,17α,21-trimethyl derivatives. Several routes from these saturated trimethylpregnane-3,20-diones to 11β-hydroxy-16α,17α,21-trimethylpregna-1,4-diene-3,20-dione (Org 6216) were explored. The best method gave Org 6216 in 75% yield.

1. The activity of monoamine oxidase, when assayed with four substrates, was significantly lowered in platelets prepared from the blood of patients with iron-deficiency anaemia. 2. Treatment with oral iron preparations restored platelet monoamine oxidase activity to normal in those patients whose serum iron concentrations also returned to normal. 3. Platelet monoamine oxidase activity remained low if treatment failed to restore serum iron concentration to within normal limits.

Sodium was determined by flame photometry and by direct potentiometry using four different systems (Ektachem DT60, Corning 614, Orion 1020 and Nova-1) in serum samples from patients with normal proteins and paraproteins. Flame photometric and direct potentiometric differences (ISE-flame), showed the expected relationship with total protein whether or not the sample contained paraprotein, an exception being the Ektachem DT60 on normal protein. With the exception of the Ektachem DT60 with samples with normal protein levels, correlation between different ISE-flame estimates was good. Differences between direct potentiometric measurements are probably more attributable to efforts made by manufacturers to relate them to flame photometric measurements in ‘normal samples’, than to the volume occupied by increased protein content. The apparent lack of influence of protein on ISE-flame for the Ektachem DT60 was shown to relate to the original external reference electrode salt bridge used. A second study showed that changing this fluid to an equi-transferrant high salt content caesium chloride based reference fluid partially unmasked the protein effect.

We present a genetic, developmental and molecular analysis of lethal(1)discs large-1[l(1)d.lg-1; Stewart et al., 1972], an oncogene of Drosophila. Mutations in this gene cause the imaginal discs to grow by cell proliferation beyond their normal final size, transform into solid tumors, fuse with one another and the brain, and lose their ability to differentiate. The oncogene represents the only known complementation group between two deficiency breakpoints, and 15 recessive lethal alleles are available. Cloning of the DNA between the two deficiency breakpoints defines a region of 45 +/- 2 kb. The l(1)d.lg-1 transcription unit is identified by both qualitative and quantitative effects of several l(1)d.lg-1 mutations on the RNA transcripts and by the presence of a DNA insert in one of the l(1)d.lg-1 alleles. It gives rise to at least five different transcripts ranging in size from 1.9 to 6.0 kb. Three other transcription units are present within this region, two 5' to the l(1)d.lg-1 gene and one at the 3' end. A near full-length cDNA from one of the larger transcripts of l(1)d.lg-1 has homology to genomic DNA spanning over 20 kb. A developmental profile of l(1)d.lg-1 transcription is presented. We discuss how mutations in this gene could disrupt epithelial structure and how this might be related to the excessive cell proliferation and interdisc fusion that is observed. We also compare this gene with another recessive oncogene of Drosophila, lethal(2)giant larvae, that has been cloned and characterized.

Natural products often do not possess the optimal properties for their use in human or animal medicine. The recent history of natural products would indicate that such optimization is likely. 2-Aryl-carbapenems were synthesized in an effort to improve upon the DHP-I susceptibility of the parent antibiotics. The synthesis of this class of compounds is described and they are indeed more DHP-I stable. However, it has not been possible to obtain meaningful anti-pseudomonal activity in this series.