Drosophila alcohol dehydrogenase activity in vitro and in vivo: Effects of acetone feeding

When adult Drosophila are placed on medium containing 0.5% acetone, their level of alcohol dehydrogenase activity drops rapidly. At the same time, the proportion of activity in the various electrophoretic forms of the enzyme shifts; most of the activity becomes localized in what is ordinarily a minor form of the enzyme. Moreover, the loss of enzyme activity occurs in vivo as well, as shown by sensitivity to ethanol poisoning, insensitivity to pentenol treatment, and inability to utilize ethanol as an energy source. These observations are discussed in light of a model advanced for the origin of the multiple forms of alcohol dehydrogenase in Drosophila.This research was supported by NIH Grants GM-18254 and ES-1527 and by DOE Contract No. EY-76-5-02-2965. Publication No. 1016 from the Department of Biology, Johns Hopkins University.     

Single cell recordings from tsetse (Glossina m.morsitans) antennae reveal olfactory, mechano - and cold receptors

Abstract. Action potentials from individual cells and receptor potentials were recorded from antennae (funiculi) of living tsetse flies, Glossina m.morsitans Westwood, using a 'surface-contact' recording technique. Stimuli were pressure of the electrode, changes in temperature and the vapours of l-octen-3–ol, acetone, 3–methylphenol, dichloromethane and CO₂. Two types of mechano-receptive cells were found. One type only fired action potentials when pressure was increased, the other type continued firing when pressure was maintained. Of the 182 cells tested for their sensitivity to temperature change and the odour stimuli, 19% did not respond to any of the stimuli, suggesting that we are still unaware of cues which may be of import to the flies, and 4% responded to temperature only, increasing and decreasing their activity with a decrement and increment in temperature, respectively. Of the 141 cells which were olfactory receptors, 52% responded to 1-octen-3–ol and 40% were exclusively sensitive to this substance. For 3–methylphenol these percentages were 23 and 18, for acetone 13 and 11, for dichloromethane 12 and 7, for CO₂ 13 and 13. No clustering of cell types in certain areas of the funiculi was found, nor was a difference apparent between the sexes. The majority of the olfactory cells responded by increasing their activities on odour stimulation. Inhibition was found in three cells only, in which spontaneous activity was suppressed on stimulation with 3–methylphenol. Spike responses were phasic-tonic and varied between two extremes: Cells showing relatively rapid cessation of spike activity after the end of stimulation, and cells which continued firing for several seconds or minutes after stimulation. Possible behavioural effects of the activities of the various cell types are discussed.     

Perception of breath components by the tropical bont tick,Amblyomma variegatum Fabricius (Ixodidae)

Summary Wall-pore sensilla in the capsule of Haller's organ on foreleg tarsi of the tick, Amblyomma variegatum, show multicellular responses upon stimulation with human and bovine breath. Filtering breath through charcoal removes the stimulant for some of these receptors. Analysis by gas chromatography coupled with olfactory sensillum electrophysiological recordings indicates that an ethanol extract of the breath components trapped on charcoal contains a major stimulant eluting at the same retention time as H₂S. Two types of H₂S-sensitive receptors have been identified. They are housed in separate sensilla, and are called sulfide-receptor 1 and 2.Although, both receptor types are characterized by a high sensitivity to H₂S with an estimated threshold of ca. 0.1 ppb and a response range covering 5–6 log orders of magnitude, their overall response to sulfides and mercaptans is nevertheless dissimilar. The type 1 receptor fires slightly more upon stimulations with H₂S than type 2, whereas ethylmercaptan induces a stronger response from type 2, and dimethyl sulfide activates only receptor 2.In a bioassay, H₂S tested at concentrations of ca. 0.02 ppm and 1 ppm equally arouses 60% of resting ticks. Two-thirds of these ticks quest the air with their first pair of legs, and the remainder start active search. By contrast, H₂S at ca. l ppm in a mixture with CO₂ severely diminishes the locomotor stimulating effect of CO₂.     

In-vivo dosimetry comparison of supraclavicular junction dose for breast and chest-wall patients with and without deep inspiration breath hold (DIBH)

PurposeThe use of deep inspiration breath-hold (DIBH) for patients with left-sided breast cancer reduces cardiac dose, with the aim of reducing the risk of major coronary events. However, this technique has not been universally adopted for patients requiring regional nodal irradiation (RNI) with one concern related to the junction dose. This study evaluates the dose received at the junction for both DIBH and free-breathing patients having tangential breast/chest wall radiation and regional nodal radiation treated with 3D-conformal or hybrid IMRT radiotherapy.MethodsIn-vivo dosimetry measurements utilizing EBT3 GafChromic™ film were performed for 19 patients during three fractions over their course of treatment. The mean junction dose and variability in junction dose were compared between the DIBH and free breathing patients.ResultsOur results show that for voluntary DIBH (v-DIBH) patients the junction dose is more variable between fractions. However, when comparing the average junction dose for DIBH and free breathing patients over the three measurements, the difference was small and not statistically significant. A larger difference was seen when patient measurements were analysed based on treatment linac.ConclusionsThese results show that the mean junction dose is not significantly compromised by the use of v-DIBH. The small possibility of a change in junction dose due to breathing technique should be weighed against the proven increased risks associated with excess cardiac dose received by free-breathing patients. If junction dose is of concern, an in-vivo study, such as this one, could allow cautious introduction of DIBH for patients requiring supraclavicular irradiation.     

Responses of the biting midge Culicoides impunctatus to acetone, CO2 and 1-octen-3-ol in a wind tunnel

Responses of host-seeking female Culicoides impunctatus (Goetghebuer) (Diptera: Ceratopogonidae) to acetone, carbon dioxide and 1-octen-3-ol were measured in a wind tunnel. Carbon dioxide, presented as a filamentous plume, increased upwind flight in a dose-dependent manner, up to 0.09% concentration. A homogenous CO2 plume elicited similar upwind responses at concentrations up to 0.09%, whereas higher plume concentrations (> 0.1%) induced erratic responses with a suppression of upwind flight. Bovine equivalent concentrations of acetone (1.5 x 10(-6)g/l) and 1-octen-3-ol (1.3 x 10(-8)g/) failed to induce any significant upwind response when tested alone. In the presence of CO2, however, 1-octen-3-ol showed highly significant increases in upwind responses at concentrations of 1.3 x 10(-1) - 10(-8)g/l. Mixtures of CO2+ acetone also enhanced upwind flight at 1.5 x 10(-9)g/l. High tunnel concentrations of both 1-octen-3-ol and acetone inhibited upwind responses. These findings are discussed in relation to host finding by C. impunctatus and known mechanisms by which upwind flight is initiated and arrested at high odour concentrations.     

13C-urea breath test to assess Helicobacter pylori bacterial load

BACKGROUND: Some authors have reported, using different protocols, that 13C-urea breath test (13C-UBT) is capable of assessing the intragastric Helicobacter pylori bacterial load, whereas others have not confirmed these data. Our aim is to evaluate the correlation between 13C-UBT values and H. pylori bacterial load. MATERIALS AND METHODS: One hundred ninety-two patients diagnosed H. pylori-positive by rapid urease test, histology, and 13C-UBT were enrolled. H. pylori bacterial load (H. pylori score) and gastritis activity (activity score) were evaluated according to the Updated Sydney System. 13C-UBT was performed according to the European Standard Protocol. Breath samples were obtained every 10 minutes for 60 minutes in 52 patients and at 30 minutes (T30) in 140 patients and analyzed by mass spectrometry. RESULTS: At T30, mean +/- SD excess delta 13CO2 excretion was 17.4 +/- 1.1, 29.9 +/- 2.2, and 48.7 +/- 4.8 in patients with H. pylori scores 1, 2, and 3, respectively. This difference was statistically significant: H. pylori score 1 versus 2, p < .005; score 1 versus 3, p < .05; score 2 versus 3, p < .05. A significant positive correlation (G = 0.59) was found between H. pylori score and activity score of chronic gastritis. At T40 and T50 significant correlation between mean excess delta 13CO2 excretion and bacterial load was achieved only in patients with H. pylori scores 1 and 3. CONCLUSIONS: 13C-UBT European Standard Protocol values correlate with H. pylori bacterial load and the activity of gastritis at T30 breath sampling time.     

Effect of the carbonic anhydrase inhibitor, acetazolamide, on Helicobacter pylori infection in vivo: a pilot study

BACKGROUND: Carbonic anhydrase inhibitors have been successfully used to treat peptic ulcers. Although carbonic anhydrase restriction does not inhibit Helicobacter pylori in vitro, recent studies suggest that carbonic anhydrase inhibition reduces the ability of H. pylori to survive in an acid environment as present in the stomach. METHODS: In a pilot study, we examined the effect of acetazolamide 500 mg as twice a day for 4 days in volunteers with active H. pylori infection. Effectiveness was judged by changes in the results of the urea breath test. RESULTS: Eight H. pylori infected volunteers completed the test. No urea breath test reverted to negative and there was a trend for the urea breath test value to increase [e.g. delta over baseline (DOB) mean +/- SE increased from 50.9 +/- 13 at baseline to 64.9 +/- 13 at day 5] during treatment with acetazolamide. CONCLUSION: The potential effect of carbonic anhydrase inhibitors on acid secretion may prevent effect on H. pylori in vivo and/or the sites of infection at the surface of the stomach may have a pH higher for any postulated acid-dependent effect to have an effect clinically.     

Mechanistic aspects of cyanogenesis from active-site mutant Ser80Ala of hydroxynitrile lyase from Manihot esculenta in complex with acetone cyanohydrin

The structure and function of hydroxynitrile lyase from Manihot esculenta (MeHNL) have been analyzed by X-ray crystallography and site-directed mutagenesis. The crystal structure of the MeHNL-S80A mutant enzyme has been refined to an R-factor of 18.0% against diffraction data to 2.1-A resolution. The three-dimensional structure of the MeHNL-S80A-acetone cyanohydrin complex was determined at 2.2-A resolution and refined to an R-factor of 18.7%. Thr11 and Cys81 involved in substrate binding have been substituted by Ala in site-directed mutagenesis. The kinetic measurements of these mutant enzymes are presented. Combined with structural data, the results support a mechanism for cyanogenesis in which His236 as a general base abstracts a proton from Ser80, thereby allowing proton transfer from the hydroxyl group of acetone cyanohydrin to Ser80. The His236 imidazolium cation then facilitates the leaving of the nitrile group by proton donating.     

Exhaled breath volatile alterations in pregnancy assessed with electronic nose

Context: Pregnancy-linked accelerated metabolism and oxidative stress may alter the exhaled volatile compound pattern (“breathprint”). Electronic noses can distinguish “breathprints” associated with different disorders.

Objective: This is the first study assessing alterations in “breathprint” during gestation.

Material and methods: 130 women participated in our study (78 pregnant vs. 52 non-pregnant). Breath samples were processed by an electronic nose and analyzed using principal component analysis.

Results: Significant differences were found in exhaled breath pattern between pregnant and non-pregnant women (p?=?0.001).

Conclusion: Pregnancy-induced changes in exhaled gases need to be considered when pregnant women with respiratory disorders carry out breath tests.