Computerized odor psychophysical testing in mice

An automated odor psychophysical procedure was developed andused to determine absolute sensitivity to n–amyl acetate.Mice were trained to initiate a trial by interrupting a photobeamat the rear of the test chamber, then sample an odor port andindicate the presence or absence of odorant by either quicklywithdrawing from the port or by continuing to sample the port.Once the air dilution olfactometer had been adjusted prior toa training or testing session, a microcomputer was used to recordall responses by the animal, to control the delivery of stimulito the odor port and to control all events in the test chamber.Correct reponses on both odor and control trials were reinforcedand incorrect responses on both types of trials were punishedwith a forced ‘time-out’ period. The odor sensitivityof all mice was estimated, using a tracking procedure, and wasthen studied in detail using schedules in which odor concentrationswere presented in ascending, descending and random order. Withall three schedules, thresholds to n-amyl acetate were between1x10^-12 and 1 x10^-13 M. Threshold estimates obtained from twoof these same animals more than 1 year later were within 0.25log units of the original values. This method should prove valuablein future studies of nasal chemoreception in mice.     

Intranasal Odorant Concentrations in Relation to Sniff Behavior

Knowledge on how odorants are transported through the nasal cavity to the olfactory epithelium is limited. One facet of this is how the sniffing behavior affects the abundance of odorants transferred to the olfactory cleft and in turn influences odor perception. A novel system that couples an online mass spectrometer with an odorant pulse delivery olfactometer was employed to characterize intranasal odorant concentrations of butane‐2,3‐dione (or butanedione, commonly known as diacetyl) at the interior naris and the olfactory cleft. Volunteers (n=12) were asked to perform different modes of sniffing in relation to the sniff intensity that were categorized as ‘normal’, ‘rapid’ and ‘forced’. The highest concentrations of butanedione at both positions in the nose were observed during normal sniffing, with the lowest concentrations correlating with periods of forced sniffs. This corresponded to the panelists' ratings that normal sniffing elicited the highest odor intensities. These feasibility assessments pave the way for more in‐depth analyses with a variety of odorants of different chemical classes at various intranasal positions, to investigate the passage and uptake of odorants within the nasal cavity.     

Olfactory bulb output cell temporal response patterns to increasing odor concentrations in freely breathing rats

This study compares the single-unit responses of 74 mitral/tufted cells recorded in freely breathing rats to step increases of the intensity of five odorants from 2 x 10(-4) to 10(-1) of saturated vapor pressure. It reveals a stability of the responses of these olfactory bulb output cells. Olfactory stimulation has frequently been shown to produce a strong patterning of mitral/tufted cell discharges highly correlated with respiration. In this study, cells were generally found to show the same response type to two consecutive concentrations, and only a few cells switched their response from excitation to suppression or vice versa. Their firing peak and/or trough occupied the same position in a high proportion of respiratory cycles recorded during a stimulation, and they remained significantly time-locked to the same respiratory epoch for the next higher concentration. Increasing odor concentration did not cause the mean firing frequency of individual cells during a peak to change appreciably between successive or extreme concentrations. By contrast, it tended to shift their maximum frequency during this peak towards an earlier respiratory cycle after stimulation onset. These results are compared with data reported in other electrophysiological studies and with results given by olfactory bulb models before being discussed for their implications in odor coding.      

Odor detection curves for n-pentanoic acid in dogs and humans*

The ability of dogs and humans to detect vapor concentrationsof n-pentanoic (valeric) acid was measured under directly comparabletesting conditions. The minimum concentrations detected by dogswere 3.3 x 10⁶ molecules/cm³ by two German shepherds, and 8.8x 10⁸ molecules/cm³ by a Fox terrier. The average minimum detectedby humans was 5 x 10¹⁰ molecules/cm³. Detection curves for thetwo best performing human subjects showed double reversals inslope (a "notch") at approximately one-half log₁₀ unit of concentrationabove the minimum detectable level. Two distinct notches, separatedby about 2 log₁₀ units of concentration, occurred in the curvesfor the dogs; the lower notch, present in the curves of thetwo best performers, fell approximately 2 log₁₀ units abovethe lowest minimum concentration detected. Performance differencesproducing these latter notches were statistically significantand, according to one interpretation, could reflect the availabilityof less primary stimulus information at the two concentrationswhere they occur, a possible result of three slightly differentranges of receptor sensitivity.     

Psychophysical methodology in odour pollution research: the measurement of poultry house odour detectability and intensity

The main purpose of the present experiment was to develop amethodology for the measurement of the detectability and intensityof agricultural odorants using established psychological procedures. Twelve odorants from three poultry houses differing amongstothers in their manure treatment systems were bag sampled andinvestigated in a controlled laboratory setting. Using a simpleone port dilution olfactometer, six subjects judged the airdiluted concentrations presented. The detectability of the odorants was studied using a procedurerelated to signal detection theory; the perceived odour intensitywas measured by the method of direct magnitude estimation. Thedetectability and psychophysical functions were constructed,and a rationale was developed for anchoring the psychophysicalfunctions from the twelve odorants investigated. Over the period of investigation the exponents of the psychophysicalfunctions obtained for odorants sampled from the same poultryhouse were very similar. Differences in this respect betweenodorants of different poultry houses appeared to be large. However,it was observed that the estimated intensities of undilutedodorants obtained from the same poultry house varied to someextent. These findings were discussed from a methodologicalpoint of view. ^*Present address to which all correspondence should be sent:Department of Human Nutrition, Agricultural University, De Dreijen11, 6703 BC Wageningen, The Netherlands     

Response patterns of single neurons in the tortoise olfactory epithelium and olfactory bulb

The responses to odor stimulation of 40 single units in the olfactory mucosa and of 18 units in the olfactory bulb of the tortoise (Gopherus polyphemus) were recorded with indium-filled, Pt-black-tipped microelectrodes. The test battery consisted of 27 odorants which were proved effective by recording from small bundles of olfactory nerve. Two concentrations of each odorant were employed. These values were adjusted for response magnitudes equal to those for amyl acetate at –2.5 and –3.5 log concentration in olfactory twig recording. Varying concentrations were generated by an injection-type olfactometer. The mucosal responses were exclusively facilitory with a peak frequency of 16 impulses/sec. 19 mucosal units responded to at least one odorant and each unit was sensitive to a limited number of odorants (1–15). The sensitivity pattern of each unit was highly individual, with no clear-cut types, either chemical or qualitative, emerging. Of the 18 olfactory bulb units sampled, all responded to at least one odorant. The maximum frequency observed during a response was 39 impulses/sec. The bulbar neurons can be classified into two types. There are neurons that respond exclusively with facilitation and others that respond with facilitation to some odorants and with inhibition to others. Qualitatively or chemically similar odorants did not generate similar patterns across bulbar units.     

Olfactory cues modulate facial attractiveness

We report an experiment designed to investigate whether olfactory cues can influence people's judgments of facial attractiveness. Sixteen female participants judged the attractiveness of a series of male faces presented briefly on a computer monitor using a 9-point visual rating scale. While viewing each face, the participants were simultaneously presented with either clean air or else with 1 of 4 odorants (the odor was varied on a trial-by-trial basis) from a custom-built olfactometer. We included 2 pleasant odors (geranium and a male fragrance) and 2 unpleasant odors (rubber and body odor) as confirmed by pilot testing. The results showed that the participants rated the male faces as being significantly less attractive in the presence of an unpleasant odor than when the faces were presented together with a pleasant odor or with clean air (these conditions did not differ significantly). These results demonstrate the cross-modal influence that unpleasant odors can have on people's judgments of facial attractiveness. Interestingly, this pattern of results was unaffected by whether the odors were body relevant (the body odor and the male fragrance) or not (the rubber and geranium odors).     

Specific antagonists at the pyridine receptor: evidence from electrophysiological studies with acetylpyridines

The effect of structural isomers of acetylpyridine on the responseof single pyridine-sensitive cells was investigated electrophysiologically.4-Acetylpyridine was identified as a strong specific inhibitorof the response to pyrazine-carboxamide, the most effectivestimulant. When 4-acetylpyridine was present in the concentrationrange from 5 x 10^-5 to 5 x 10^-2 mol/l, the dose-response curveswere shifted to the right, the slope and the common saturationlevel remaining unchanged. This effect can be interpreted asa competitive inhibition. It was also consistent with competitiveantagonism that the Schild plot of these data was linear witha slope close to 1 (0.90). Linear regression indicated an inhibitionconstant K₁ = 40 µmol. 2-Acetylpyridine had a less potentantagonistic effect and an agonistic effect only at high concentrations(>10^–3 mol/1).     

Partition coefficient and comparative olfactometry

The air/water partition coefficients of many odorants can becalculated from available data on their vapor pressures andsolubilities at 25°. A gas-flow method is described forobtaining equilibrium concentration data to calculate the coefficientsof certain odorants for which theoretical or gas chromatographictechniques are not applicable. It is shown that the odor thresholdsobtained in airdilution olfactometry agree with those measuredby sniff tests on aqueous dilutions, when the air/water partitioncoefficient is taken into consideration. Similar calculationsinvolving the oil/water partition coefficient show close correspondencebetween thresholds measured in water or safflower oil dilution.A conversion factor was obtained for comparing squeeze-bottlepair tests with stoppered flask 2/5 tests in the measurementof olfactory threshold.     

Floodwater Oxygen Content, Ethylene Production and Lenticel Hypertrophy in Flooded Mango (Mangifera indica L.) Trees

A system was developed to test the effects of floodwater O₂concentration on ethylene evolution and stem lenticel hypertrophy,and the effects of exogenous ethylene on stem lenticel hypertrophyin mango (Mangifera indica L.) trees. Dissolved O₂ concentrationsof 1–7x10^–9 m³ m^–3 generally resulted in hypertrophyof stem lenticels within about 6 d of flooding, whereas floodwaterO₂ concentrations of 13–15 x 10^–9 m³ m^–3 delayedhypertrophy until about day 9. After 14d of flooding, therewere more than twice the number of hypertrophied lenticels pertree with floodwater O₂ concentrations of 1–7 x 10^–9m³ m^–3 than with floodwater O₂ concentrations of 15 x10^–9 m³ m^–3. Ethylene evolution from stem tissueimmediately above the floodline increased 4- to 8-fold in treesexposed to floodwater O₂ concentrations of 1–2 x 10^–9m³ m^–3, increased 2-fold for trees exposed to floodwaterO₂ concentrations of 6–7 x 10^–9 m³ m^–3, butremained constant with floodwater O₂ concentrations of 13–15x 10^–9 m³ m^–3. Plants maintained in highly oxygenatedfloodwater (13–15 x 10^–9 m³ m^–3), and givenexogenous ethylene developed many hypertrophied lenticels, whereasplants in highly oxygenated water and not given ethylene developedfewer or nohypertrophied lenticels. These data suggest thatethylene plays a role in promotion of stem lenticel hypertrophyin flooded mango trees, and that floodwater dissolved oxygenconcentration can regulate stem lenticel hypertrophy and ethyleneevolution in this species. Key words: Flooding, hypoxia, hypertrophic cell swelling     

Effect of dilution rate on competitive interactions between the cyanobacterium Microcystis novacekii and the green alga Scenedesmus quadricauda in mixed chemostat cultures

We examined the competition between the cyanobacterium Microcystisnovacekii (Kom.) Comp. and the green alga Scenedesmus quadricauda(Turpin) Brébisson using unialgal and mixed chemostatcultures with various supply rates of culture medium where limited algal growth. In unialgal cultures, bothspecies grew at all of the dilution rates examined (0.1, 0.3and 0.8 day^-1): steady-state cell densities were 1 x 10⁴ to8 x 10⁴ cells mL^-1 for M. novacekii and 0.5 x 10⁵ to 2.1 x 10⁵cells mL^-1 for S. quadricauda. Microcystis novacekii was dominantin mixed cultures at a dilution rate of 0.1 day^-1, where thesteady-state cell density was 1 x 10⁴ to 7 x 10⁴ cells mL^-1for M. novacekii and 1 x 10² to 5 x 10² cells mL^-1 for S. quadricauda.Scenedesmus quadricauda was dominant in mixed cultures at thehigher dilution rates (0.3 and 0.8 day^-1), where the final celldensity was 0.5 x 10² to 6.4 x 10² cells mL^-1 for M. novacekiiand 0.2 x 10⁵ to 7 x 10⁵ cells mL^-1 for S. quadricauda. Thisresult indicates that the dilution rate affects the competitiveinteraction. We conclude that it is necessary to consider waterexchange in the study of mechanisms of cyanobacterial blooms.     

Reaction time for the recognition of odor quality

Psychophysical procedures were used to determine human recognitiontimes to three matched intensity levels of the odorants n-butanol,(+)-limonene and propionic acid. A computer controlled the deliveryof the odorants from an air dilution olfactometer and measuredrecognition times. The mean times recorded with the odorantswere significantly different and ranged between 680–867ms, the shortest times being recorded with the acid. The resultsare discussed in relation to reported detection and recognitiontimes, and their relevance to odor mixture perception, in particularit was concluded that reaction times for recognition do notpredict the occurrence of suppression in mixtures.     

Temporal aspects of information processing in the first two stages of the frog olfactory system: influence of stimulus intensity

To understand information processing in the first two stagesof the olfactory system, neural activities were examined froma temporal point of view. The extracellular unitary activityof receptor cells and bulb neurons in response to four odorsdelivered in a range of concentrations from 1 x 10^–6 to5.62 x 10^–2 as a fraction of saturated vapor was recorded.For each population, unitary activities were pooled by 100-msbins, according to the nature and/or to the concentration ofthe stimulus. The results provided a view of the timing of activityof cell populations and allowed a comparison of the temporalstructures of the primary afferent message with that of thebulbar output message. Temporal patterns of receptor cell populationwere characterized by late and sustained discharges; thus, theprimary input volley did not coincide with the early and briefbulbar output message. In the two cell populations, the temporalresponse patterns elicited depended on the nature of the stimulus.At receptor level, interstimulus differences could be explainedin terms of binding interactions between odorants and receptorsites. In the bulb, while the input and output messages werenot strictly synchronous, some temporal characteristics relativeto the nature of the stimulus were preserved. In receptor cells,the activity occurring within 1 s following the stimulus onsetspecified stimulus intensity. By contrast, in the bulb, theevents which seemed to be involved in intensity coding werethose occurring within the first 500 ms. The convergence ofprimary afferences onto the bulb leads to an amplification ofthe earliest peripheral events setting up a sharp, earlier outputbulb message; in the processing of the input information, theolfactory bulb seems to combine an amplifying role for the earliestevents with a shutting action on later events.     

Effect of abscisic acid on elongation and kinetin-induced tuberization of isolated stolons of Solarium tuberosum L.

The response of isolated stolons cultured in vitro, to abscisicacid (ABA) has been studied in the presence and absence of kinetin(6-furfurylaminopurine). ABA alone in concentrations from 7.5x 10^–4 mM to 7.5 x 10^–2 mM, inhibited stolon elongationbut failed to promote tuber initiation. In the presence of kinetin,ABA at concentrations of 3.0 x 10^–2 and 7.5 x 10^–2mM markedly inhibited kinetin-induced tuber initiation and stolonelongation, but at 7.5 x 10^–4 and 7.5 x 10^–3 mMABA did not prevent tuber initiation. When stolons were incubated on a medium containing kinetin andlater transferred to one containing ABA with or without kinetin,the inhibitory effect of ABA decreased appreciably as the timeof incubation on kinetin is increased. The results are discussed in relation to the role of ABA inthe inhibition of nucleic acid and protein synthesis and theinteraction with cytokinins and the possible effect of ABA onkinetin uptake, transport and accumulation at the locus of action. (Received February 26, 1969; )     

Carbon Dioxide Effects on Auxin Responses of Coleoptile Sections

In the wheat cylinder bioassay technique as previously usedhere 5 sections have been enclosed in a 2 x 38 in, assay tubetogether with 0.5 ml. of the test solution. A method developedfor estimating the amount of carbon dioxide which accumulatesin these tubes through the respiration of the enclosed sectionshas shown that the level can rise to 20 per cent. after 24 hrs.at 25°C. In the presence of a 100 p.p.m. IAA(6x10^-4M.) testsolution, growth of 5 enclosed sections is depressed from 8hrs. onwardas and they eventually shrink, releasing their accumulatedIAA back into the solution. The growth of sections under various gas mixtures of carbondioxide in air has also been followed and these experimentsshow that section length is reduced approximately lineraly withrespect to increasing carbon dioxide concentration up to 20per cent. in air, both in the presence and absence of a 100p.p.m. IAA solution. The slope of the fitted regression line,however, is much steeper when the test solution contains IAA—i.e.there is a large interaction. In the presence of IAA, growth-time data show that a reductionin the growth rate, as compared with that in normal air, canbe detected after only 4 hrs, at the highest carbon dioxideconcentration. In the absence of IAA, high concentrations ofcarbon dioxide accelerate growth during the first 8 hrs. ofthe assay but depress it later. The mechanism of action of this interaction is unknown but itis not shown at very high concentrations of IAA, e.g. 1,000p.p.m. (6x10^-3M.).     

Flavanoids in Pollen and Stigma of Brassica oleracea and Their Effects on Pollen Germination In Vitro

Brassica oleracea pollen was applied to a basic medium of 1.5per cent agar and 15 per cent sucrose to which flavanoids wereadded at three concentrations. Two types of agar were used;with agar 1, quercetin at a concentration of 0.5 x 10^–3per cent gave an increase in percentage germinating grains.With agar 2, an increase in germination occurred with kaempferoland naringin at concentrations of 0.5 x 10^–3 and 0.5 x10^–1 per cent respectively. Increase in pollen tube lengthoccurred with agar 2 and quercetin at a concentration of 0.5x 10^–3 per cent. The stigma tissue of B. oleracea contains at least three andthe pollen at least one glycoside of quercetin. The sugars inthe glycosides were not identified. Pollen germination and pollentube extension were not stimulated exclusively by the flavanoidspresent in the stigma. The flavanoid composition of the stigmadid not vary amongst five different S-allele genotypes, indicatingthat flavanoids are probably not directly involved in the incompatibilityreaction of B. oleracea.     

Role of temporary cysts in the population dynamics of Alexandrium taylori (Dinophyceae)

Although temporary cyst stages are common in dinoflagellates,their role remains unclear. Every year Alexandrium taylori (Dinophyceae)forms dense patches (10⁶ cells l^-1) along La Fosca beach (Spain,northwest Mediterranean), which last for 2 months (July, August).One of the characteristics of the life history of A. tayloriis the shift from a vegetative motile stage to non-motile temporarycysts. Here we present the temporal changes in the abundanceof temporary cysts in sediments and their in situ encystmentand excystment rates. The in situ encystment rate of temporarycysts from the water column to the sediment ranged from 1.8x 10⁶ to 4.4 x 10⁶ cysts m^-2 day^-1, whereas the excystment ratewas between 0.9 x 10⁶ to 2.7 x 10⁶ cysts m^-2 day^-1 during thebloom period. Some of the temporary cysts in the sediment tookmore than 1 day to produce vegetative cells and remained viablefor at least 4 days. We propose that temporary cyst formationin this species is a tool for reducing population losses. Theproduction of temporary cysts can be an advantage since partof the population is stored in the sediments.     

Effects of Ca2+ on electro-olfactogram in the isolated olfactory mucosa of the frog

Reports are conflicting as to whether the presence of Ca²⁺ onthe ciliated surface of the olfactory mucosa suppresses or potentiatesthe response of receptor cells to odorants. To resolve thisissue, electro-olfactograms (EOGs) were recorded from the isolatedolfactory mucosae of the frog while its ciliated surface wasperfused with saline solutions containing differing concentrationsof Ca²⁺. A decrease in Ca²⁺ concentration augmented the EOGamplitude, and the magnitude of the augmentation increased asthe Ca²⁺ concentration decreased progressively. The slow forskolin-inducedchange in potential likewise increased in amplitude with theremoval of Ca²⁺ from the perfusate. Desensitization of EOG duringthe prolonged administration of odorants developed similarly,irrespective of the concentration of Ca²⁺ on the ciliated surface.These observations are consistent with findings of patch-clampand biochemical experiments. The augmentation of EOG at lowCa²⁺ concentrations appeared to result from either an increasein activity of adenylate cyclase or an increase in responsivenessof the channels activated by adenosine 3',5'-cyclic monophosphate,but not from an increased sensitivity of the receptor molecules.