Stability of vancomycin in an extemporaneously compounded ophthalmic solution

The stability of vancomycin 31 mg/mL (as the hydrochloride) in an artificial tears solution at -10, 4, 25, and 40 degrees C was studied. Vancomycin power was reconstituted with sterile water for injection to a concentration of 50 mg/mL. Artificial tears solution containing 0.3% hydroxypropyl methylcellulose, 0.1% dextran 70, 0.01% benzalkonium chloride, and 0.05% edetate disodium was used to produce a final concentration of 31 mg/mL. Triplicate solutions for each storage temperature and sampling time were prepared. The solutions were stored at -10, 4, 25, and 40 degrees C. Samples were taken initially and at 3, 7, 10, 21, 30, 45, and 60 days for visual inspection and analysis by high-performance liquid chromatography. All solutions remained clear and colorless at -10, 4, and 25 degrees C throughout the study period. By day 3, crystalline particles formed in the solutions stored at 40 degrees C. No substantial change in pH was observed at any time. At -10 degrees C, the solutions retained more than 90% of their initial vancomycin concentrations throughout the study period. The solutions retained a mean of at least 90% of the initial drug concentration for 21 days at 4 degrees C and for 7 days at 25 degrees C. For the solutions stored at 25 or 40 degrees C, less than 85% of the initial vancomycin concentration remained after 10 and 3 days, respectively. Vancomycin 31 mg/mL (as the hydrochloride) in an artificial tears solution was stable for 45 days at -10 degrees C, 10 days at 4 degrees C, and 7 days at 25 degrees C in the tears solution's original container.     

Degradation of histamine solutions used for bronchoprovocation

STUDY OBJECTIVES: To determine optimal storage conditions for histamine diphosphate (HDP) solutions used for bronchoprovocation. DESIGN: HDP was dissolved in buffered saline solution to concentrations of 0.125 to 16 mg/mL and stored in 3-mL unit dose syringes at different temperatures for varying lengths of time, with and without protection from fluorescent light. SETTING: Dark freezer (-20 degrees C), dark refrigerator (4 degrees C), and laboratory counter top (20 degrees C) illuminated by fluorescent light (375 foot-candles). MEASUREMENTS: HDP concentrations were measured after the solutions were prepared and during storage by a high-performance liquid chromatographic assay that differentiates histamine from its break down products. RESULTS: All dilutions were sterile after preparation and contained 97 to 110% of the labeled amount of HDP. Solutions constantly exposed to fluorescent light (375 foot-candles) and room temperature (20 degrees C) contained only 20 to 37% of the initial concentrations after 7 days. The same dilutions stored at room temperature, but protected from light, contained 83 to 94% of the initial concentrations. Dilutions stored in the dark in a refrigerator (4 degrees C) retained 97% of the initial concentrations after 8 weeks, while dilutions stored in the dark freezer (-20 degrees C) were stable for 12 months. CONCLUSIONS: Exposure to fluorescent light at room temperature results in degradation of histamine solutions used for bronchoprovocation. Dilutions stored in unit dose syringes and protected from light are stable for at least 8 weeks in the refrigerator and up to 12 months frozen. Once removed from the refrigerator or freezer, the solutions should be used within 6 h or discarded.     

Influence of culture duration and ciliogenesis on the relationship between ciliary beat frequency and temperature in nasal epithelial cells

Human nasal epithelial cells from excised mucosal specimens were cultured directly in suspension and sequentially on monolayer and in suspension. Ciliary beat frequency (CBF) was measured by fast Fourier transform analysis of computerized microscopic photometry. In biopsy material CBF increased in an approximately linear fashion at 0.6 Hz/degree C between 20 degrees C and 35 degrees C. Above 35 degrees C the increase was lower and was 0.25 Hz/degree C between 40 degrees C and 44 degrees C. CBF increased more rapidly in suspension culture between 25 and 35 degrees C (1 Hz/degree C) but reached a plateau at approximately 40 degrees C and decreased with further temperature elevation. Up to 44 degrees C all changes were reversible, while irreversible slowing and deterioration occurred above 45 degrees C. Values found after 3 weeks' initial suspension culture were similar to those after 6 weeks' sequential monolayer suspension culture. After 3 weeks of ciliogenesis in sequential suspension culture, all values up to 41 degrees C were statistically significantly higher than those under the other conditions. Ciliary activity was maintained and expressed in culture. CBF was higher than in biopsy material and a reversible decrease was observed at high temperature.     

Transient expression of beta-galactosidase in differentiating sporozoites of Eimeria tenella

A cocktail of seven Listeria monocytogenes isolates of food, human and environmental origin was used to assess the antilisterial activity of the bacteriocins nisin and ALTA 2341 in combination with various atmospheres: air, 100% N2, 40% CO2:60% N2, or 100% CO2. Buffered tryptone soya broth (pH 6.0) was used as the growth medium and incubation was at 4 degrees C (21 days) or 12 degrees C (7 days), or when temperature fluctuated between these values for defined periods. It was observed that atmosphere alone influenced the growth rate of L. monocytogenes, with 100% CO2 exerting the greatest inhibition. A 5 log population increase was observed in all atmospheres after 7 days at 12 degrees C. At 4 degrees C a 4-5 log population increase was observed in air, 100% N2 and 40% CO2:60% N2 within 21 days. Growth was prevented by 100% CO2. In the presence of nisin (400 IU/ml), an increase in the lag phase was observed before growth (5 log population increase after 7 days) in all atmospheres at 12 degrees C. This effect was enhanced at 4 degrees C where a maximum 2 log population increase was observed in all atmospheres except 100% CO2, in which growth was prevented. Increasing the concentration of nisin to 1250 IU/ml prevented L. monocytogenes growth in all atmosphere combinations at 4 and 12 degrees C. Two concentrations of ALTA 2341 were also tested. In the presence of 0.1% ALTA 2341 and at 12 degrees C, a 3-5 log population increase was observed in all atmospheres with the exception of 100% CO2, which prevented L. monocytogenes growth. At 4 degrees C, growth was observed in the combination of 0.1% ALTA 2341 and 100% N2 only (3 log population increase). Use of a higher concentration of ALTA 2341 (1.0%) resulted in a population decrease below the detection level within 24 h in all atmosphere/temperature combinations. Re-growth occurred in the presence of 1.0% ALTA 2341 in all atmospheres at 12 degrees C, and in combination with air or 100% N2 at 4 C. When the effectiveness of either nisin or ALTA 2341 and atmosphere was tested against L. monocytogenes as temperature fluctuated for periods between 4 and 12 degrees C, only the combination of 100% CO2 and 1.0% ALTA 2341 prevented growth. Cells surviving exposure to nisin or ALTA 2341 were recovered from 28 of the 32 combinations tested that contained bacteriocin. Nisin survivors remained sensitive to the bacteriocin. ALTA 2341 survivors had become resistant to the bacteriocin.     

Effects of exogenous nitric oxide on neutrophil oxidative function and viability

Lipid extracts of Spodoptera littoralis pheromone glands submitted to acid methanolysis using: (i) sulfuric acid/methanol/benzene (0.1:4:2, by vol) at 90 degrees C for 1 h; (ii) 12 N HCI/methanol (1:2, vol/vol) at 90 degrees C for 1 h, or (iii) 14% BF3-MeOH at 90 degrees C for 1 h did not reveal the presence of either 11- or 12-hydroxytetradecanoic acid in the extracts, as concluded from the gas chromatography-mass spectrometry analyses. Under the above methanolysis conditions, a synthetic sample of methyl (14, 14, 14-2H3) 12-hydroxytetradecanoate remained unaltered. These results may indicate that formation of (E)-11-tetradecenoic acid from tetradecanoic acid does not occur in the pheromone gland by dehydration of an intermediate hydroxyacid. Acid methanolysis of a lipidic extract using BF3-MeOH led to the formation of a mixture of methoxy fatty acid methyl esters, identified by gas chromatography-mass spectrometry. These methoxy derivatives should arise from BF3-catalyzed addition of methanol to the double bond of the natural monounsaturated fatty acyl derivatives present in the gland. Thus, under the same conditions, a synthetic sample of methyl (Z)-11-tetradecenoate was partially transformed into methyl 11-methoxytetradecanoate and methyl 12-methoxytetradecanoate. This reaction might be a useful alternative procedure to obtain methoxy derivatives of olefins, which are very helpful for the structural characterization of the parent alkenes.     

Glucose polymer ingestion--effect on fluid balance and glycemic state during a 4-d march

The effect of glucose-polymer solution on physical performance has been extensively studied under controlled laboratory conditions. The present study was conducted to investigate the influence of such beverages on fluid balance and on glycemic state during a moderate, prolonged field exercise. Forty-eight endurance trained, male subjects participated in the study. The maneuver consisted of a 4-d march; 29, 39, 36, 30 km.d-1, at a speed of 5-6 km.h-1. The subjects covered a total distance of 134 km at an estimated exercise intensity of approximately 40% VO2max, under hot climate conditions (ambient temperature, 32-41 degrees C; relative humidity, 60-14%). Subjects were randomly assigned to one of two groups: glucose polymer-electrolyte beverage (GP; N = 24) and tap water (TW; N = 24). Each group was then divided into two subgroups consuming fluid ad libitum (TWa, GPa) or instructed to consume 900 ml.h-1 (TWb, GPb). The mean daily fluid consumption of all subgroups was similar (5252 +/- 229 and 4640 +/- 67 ml in TWa and TWb; 5257 +/- 317 and 5253 +/- 216 ml in GPa and GPb, respectively). Weight loss, reflecting the degree of dehydration, was 1.2 +/- 0.1% and 1.9 +/- 0.3% of initial body weight in TW and GP, respectively. On day 1, plasma volume changed by +0.4% and -1.8% in the TW and GP groups, respectively. On the days 2-4 changes in both groups were similar.(ABSTRACT TRUNCATED AT 250 WORDS)     

Pyridyloxobutyl adduct O6-[4-oxo-4-(3-pyridyl)butyl]guanine is present in 4-(acetoxymethylnitrosamino)-1-(3-pyridyl)-1-butanone-treated DNA and is a substrate for O6-alkylguanine-DNA alkyltransferase

The lung carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is activated to reactive metabolites that methylate or pyridyloxobutylate DNA. Previous studies demonstrated that pyridyloxobutylated DNA interferes with the repair of O6-methylguanine (O6-mG) by O6-alkylguanine-DNA alkyltransferase (AGT). The AGT reactivity of pyridyloxobutylated DNA was attributed to (pyridyloxobutyl)guanine adducts. One potential AGT substrate adduct, 2'-deoxy-O6-[4-oxo-4-(3-pyridyl)butyl]guanosine (O6-pobdG), was prepared. This adduct was stable at pH 7.0 for greater than 13 days and to neutral thermal hydrolysis conditions (pH 7.0, 100 degrees C, 30 min). Under mild acid hydrolysis conditions (0.1 N HCl, 80 degrees C), O6-pobdG was depurinated to yield O6-[4-oxo-4-(3-pyridyl)butyl]guanine (O6-pobG). O6-pobdG was hydrolyzed to 4-hydroxy-1-(3-pyridyl)-1-butanone and guanine under strong acid hydrolysis conditions (0.8 N HCl, 80 degrees C). O6-pobG was detected in 0.1 N HCl hydrolysates of DNA alkylated with the model pyridyloxobutylating agent 4-(acetoxymethylnitrosamino)-1-(3-[5-3H]pyridyl)-1-butanone ([5-3H]NNKOAc). When [5-3H]NNKOAc-treated DNA was incubated with either rat liver or recombinant human AGT, O6-pobG was removed, presumably a result of transfer of the pyridyloxobutyl group from the O6-position of guanine to AGT's active site.     

Improved strategies for postoligomerization synthesis of oligodeoxynucleotides bearing structurally defined adducts at the N2 position of deoxyguanosine

Improved methodology has been developed for preparation of oligodeoxynucleotides bearing adducts on the N2 position of guanine in which the adduction reaction is carried out in homogeneous solution rather than while the oligonucleotide is immobilized on a solid matrix. The methodology utilizes a new synthon, 2-fluoro-O6-(trimethylsilylethyl)-2'-deoxyinosine (3). Nucleoside 3 is stable to the conditions of oligonucleotide synthesis, but the O6 protection is eliminated under very mild conditions following displacement of the 2-fluoro group by amine nucleophiles. Oligonucleotides containing 3 could be removed from the solid support by treatment with 0.1 M NaOH (8 h, rt) without disruption of 3. Reaction of the crude, partially deprotected oligonucleotide with (R)-2-amino-2-phenylethanol in homogeneous solution, followed by removal of the remaining protective groups with NH4OH (60 degrees C, 8 h) and then 0.1% acetic acid, gave the adducted oligonucleotide in good purity and yield. Alternatively, fully deprotected oligonucleotide containing 3 could be prepared by use of labile phenoxyacetyl-type protecting groups on the exocyclic amino groups.     

Ultrastructural changes in rabbit ciliary body after extraocular mitomycin C

PURPOSE: To study the ultrastructural changes in ciliary body epithelium of the rabbit eye after subconjunctival injections of mitomycin C. METHODS: One eye of six New Zealand white rabbits was given a subconjunctival injection at the 12-o'clock position with 0.005, 0.02, 0.08, 0.1, 0.12, or 0.16 mg mitomycin C. The fellow eye was given a subconjunctival injection of balanced salt solution. Two weeks after treatment, the eyes were enucleated, and the ciliary body was exposed and submerged in fresh 4% paraformaldehyde/2% glutaraldehyde in 0.1 M phosphate buffer, pH 7.4, at 4 degrees C. Electron microscopy of the ciliary body was performed at two sites: the injection site (12-o'clock position) and 180 degrees away (6-o'clock position). RESULTS: At dosages of 0.1 mg and higher, ciliary body epithelial cells beneath the injection site were thinned. There were vacuoles and expansion of intracellular and intercellular spaces. Plasma membrane infoldings were disrupted, and the apical membrane was thinned. Mitochondria and nuclei were normal. Ciliary body epithelium at 6-o'clock position showed only mild architectural distortion of the plasma membrane infoldings. Eyes that received lower doses of mitomycin C (0.005 mg, 0.02 mg, and 0.08 mg) and balanced salt solution showed normal ciliary body epithelium at the injection site and 180 degrees away. CONCLUSIONS: Subconjunctival injection of mitomycin C in the rabbit produces dose-dependent localized ultrastructural changes of the ciliary body epithelium.     

The LETR-Principle: a novel method to assess electrode-tissue contact in radiofrequency ablation

INTRODUCTION: Stable electrode-tissue contact is crucial for successful radiofrequency ablation of cardiac tachyarrhythmias. In this in vitro study, a custom-made radiofrequency generator was used to evaluate the correlation between tip temperature response to a minimal radiofrequency power delivery (Low Energy Temperature Response: LETR-Principle) and electrode-tissue contact as well as lesion size. METHODS AND RESULTS: A battery-powered radiofrequency generator (LETR-Box, 500 kHz, 0.1 to 0.3 W) could measure the temperature increase at the tip electrode with 0.01 degrees C accuracy. The device was tested in vitro using isolated porcine ventricular tissue. For various electrode-tissue settings (i.e., 0 to 0.89 N contact force), the temperature increase (deltaT) due to 0.1-W power delivery for 10 seconds was recorded. Subsequently, for the same electrode-tissue contact, a temperature-controlled radiofrequency ablation was performed (70 degrees C target temperature, 50-W maximum output, 30 sec). Thereafter, the lesion size was measured histologically. To prove the safety of the applied LETR-Principle, the tissue was inspected microscopically after continuous radiofrequency power delivery of 0.3 W for 1 hour with high contact pressure (1.33 N). The delivery of 0.1-W radiofrequency power resulted in an average deltaT of 0.18 degrees +/- 0.13 degrees C. During temperature-controlled radiofrequency ablation, the tip temperature was 59 degrees +/- 8.5 degrees C, resulting in a lesion depth of 4.8+/-0.6 mm. The correlation coefficient between deltaT and contact force was 0.97 and 0.81, respectively, for lesion depth. No lesion was microscopically visible after power delivery of 0.3 W for 1 hour with 1.33 N contact pressure. CONCLUSION: The LETR-Principle safely indicates electrode-tissue contact and lesion depth under in vitro conditions and can be useful for catheter positioning during radiofrequency ablation procedures.     

Effects of cytochalasin H on the morphogenetic movements of the embryos of Microhyla ornata

The intensity of radioautographic reactions in Ilford L4, Sakura NR-H2 and Kodak NTE emulsions was compared after exposure in either dry air or dry helium gas at 4 degrees C to test the stability of latent images in the presence or absence of oxygen. A light proof container is described in which slides bearing radioactive sections coated with the three emulsions were exposed in dry helium at a constant pressure of approximately 0.5 atm. The comparison of air and helium atmospheres during exposure of radioautographs was estimated qualitatively for 125I-labeled thyroid sections stored for several years and, in addition, quantitative data was derived from 3H-labeled methacrylate sections stored from 21 days to 1 year. With the three emulsions under study, the background fog remains low under both exposure conditions at 4 degrees C for as long as several years duration. Using L4 emulsion, similar high grain densities are obtained in air and helium, and therefore, the latent images in L4 emulsion remain stable in the presence of oxygen. In the case of NTE and NR-H2 emulsions, as the exposure time increases, substantially lower reaction intensities are observed in air than in helium. This difference in reaction intensity is evident by 3 weeks with NTE and after 4 weeks with NR-H2. Hence, there is fading of the latent images in the latter emulsions in the presence of oxygen. It is concluded that reliable results may be obtained with the L4 emulsion by exposure of radioautographs in dry air, whereas with the NR-H2 and NTE emulsions, exposure should be in an oxygen-free medium, such as is provided by a dry helium atmosphere.     

Study of 14 denitrifying soil bacteria of the "pseudomonas stutzeri" group isolated by enrichment culture in the presence of nitrous oxide (author's transl)

The strains were isolated from soil by enrichment in a liquid minimal medium containing ethanol, acetate, succinate, L-malate or tartrate, under an N2O atmosphere at 32 degrees C. All fourteen strains can use the following 25 sources of carbon and energy under aerobic conditions: glycerate, ethanol, propanol, acetate, butyrate, malonate, succinate, glutarate, sebacate, glycollate, L-lactate, D-lactate, L-malate, DL-3-hydroxybutyrate, pyruvate, fumarate, itaconate, mesaconate, crotonate, L-alpha-alanine, D-alpha-alanine, L-leucine, asparagine, L-tyrosine, and L-proline. They hydrolyze Tween 80 but not gelatin. Nitrate is used as nitrogen source. Nitrate reductase A and respiratory nitrite reductase are present. Four of the strains are clearly and easily distinguishable from the others on the basis of six characters: special morphology of colonies; in ability to use isovalerate and DL-valine, inability to use glucose, absence of exocellular amylase, and high level of metapyrocatechase. Their G + C content is 66-67%. One of the strains is distinct from the others by the yellow pigmentation of its colonies, its ability to use D-glucuronate, trehalose, D-sorbitol and citraconate, ability to grow at 4 degrees but not at 40 degrees, and a lower G + C content: 63%. One strain accumulates poly-beta-hydroxybutyrate. This work confirms the well-known, wide variability of the bacteria belonging to the P. stutzeri group. Denitrification by two of the strains was quantitatively studied using cell suspensions. Cells from NO-3-containing anaerobic cultures reduce NO-3, NO-2 and NO to N2O and N2; they reduce slowly N2O to N2. Cells grown in anaerobic cultures under N2O also reduce NO-3, NO-2 and NO to N2O and N2 but they reduce N2O rapidly to N2.     

A mutation in guanylate cyclase activator 1A (GUCA1A) in an autosomal dominant cone dystrophy pedigree mapping to a new locus on chromosome 6p21.1

Corticosteroids containing a C21 primary hydroxyl group were derivatised with 9-anthroyl cyanide. The reagent was prepared as a solution in acetonitrile, containing 0.1% triethylamine, at a concentration of 2 mg/ml. Approximately 1 microg of corticosteroid was reacted with 100 microl of this reagent, at 45 degrees C for 2 h. The fluorescent derivatives were separated by HPLC on a silica column, 250x4.6 mm I.D., by stepwise elution, with a mobile phase of 2-propanol-hexane (2:98) for 20 min, followed by 2-propanol-hexane (7:93) from 20 to 40 min. The fluorescence detector was set to 370-nm excitation and 470-nm emission. The relatively low temperature for derivatisation avoided reaction with secondary hydroxyl groups and also prevented thermal degradation of the corticosteroids.     

Atopic eczema: a clinical psychiatric study

The method of Deutsch and Weeks was modified to provide a reliable and reasonably quick method for assaying the L-ascorbic acid content of culture medium. The modified method was used to determine the decay of L-ascorbic acid under various conditions of culture and the concentration of the vitamin in commercially prepared media. The half-life of L-ascorbic acid in a modified New circulator gassed with 95% O2 + 5% CO2 was 1.5 hr.; and when gassed with 20% O2 + 5% CO2 + 75% N2, about 2 hr. In Petri dishes gassed with 20% O2 + 5% CO2 + 75% N2, the half-life of L-ascorbic acid was 0.9 hr. About 4% of the L-ascorbic acid was lost per day when medium was stored at 0 degrees C and about 9% per day when stored at 5 degrees C. When medium with an initial content of 300 microng per ml was stored at room temperature, the half-life was found to be 15.5 hr. The L-ascorbic acid in five commercially available media, which contain the vitamin in their formulations, was assayed immediately after their delivery to the laboratory. The values of L-ascorbic acid measured in these media were in all cases far lower than prescribed. A continuous-flow organ culture system has been designed which allows the provision of a relatively constant level of L-ascorbic acid to explant by taking advantage of the slow oxidation of L-ascorbic acid at 0 degrees C.     

Chemical ablation of the gallbladder: evaluation of multiple agents in vitro

PURPOSE: To find a more effective chemical regimen for transcatheter ablation of the gallbladder in an in vitro model. MATERIALS AND METHODS: Sectioned and whole pig gallbladders were exposed in vitro to 12 different chemical solutions at varying conditions of exposure time, pH, and temperature. RESULTS: In the in vitro studies, 0.1 N and 1.0 N solutions of sodium hydroxide in water or ethanol and 3% hydrogen peroxide were the most effective sclerosant agents. Ethanol and hydrochloric acid failed to completely eliminate the epithelium from the gallbladder sections. Increasing exposure time from 10 to 20 minutes or increasing the temperature of the solutions from 37 degrees C to 50 degrees C did not alter these results. Sequential 15-minute exposures to 0.1 N sodium hydroxide in ethanol followed by peroxide completely eliminated the epithelium from whole gallbladders in vitro. CONCLUSION: Alkaline solutions and hydrogen peroxide are more effective than ethanol alone, acids, or detergents in eliminating gallbladder epithelium in this model. Further evaluation of these agents in vivo is merited.     

Cryopreservation of expanded mouse blastocysts by vitrification in ethylene glycol-based solutions

Experiments were conducted to find optimal conditions for obtaining high survival of expanded mouse blastocysts after vitrification. The vitrification solutions used were designated EFS20, EFS30 and EFS40, and contained 20%, 30% and 40% ethylene glycol, respectively, diluted in PB1 medium containing 30% Ficoll plus 0.5 mol sucrose l-1. In the toxicity test of the solutions and each cryoprotectant, ethylene glycol was found to be toxic to embryos. For vitrification, expanded blastocysts were exposed to the vitrification solutions at 10, 20 or 25 degrees C for various periods; they were then cooled rapidly in liquid nitrogen, after which they were warmed rapidly. When the embryos were directly exposed to EFS40 at 20 degrees C for 2 min before vitrification, 66% of them re-expanded during 48 h of post-warming culture. The re-expansion rates decreased when exposure time was shortened (0.5 min), when exposure temperature was lowered (10 degrees C), or when embryos were vitrified in EFS20 and EFS30, although these conditions should be less toxic. When embryos had been pretreated in a dilute (10-20%) ethylene glycol solution for 5 min, followed by short exposure (0.5 min) to EFS40 at 20 degrees C, post-vitrification survival rate increased to 83-84%; furthermore, the rate reached 94% when the temperature was increased to 25 degrees C. Expanded blastocysts cryopreserved by this two-step method developed into live young as well as control embryos after transfer.(ABSTRACT TRUNCATED AT 250 WORDS)     

T cell engraftment in lymphoid tissues of human peripheral blood lymphocyte reconstituted SCID mice with or without prior activation of cells

A controlled study was undertaken to determine the stability of LSD in pooled urine samples. The concentrations of LSD in urine samples were followed over time at various temperatures, in different types of storage containers, at various exposures to different wavelengths of light, and at varying pH values. LSD concentrations were measured quantitatively by the Abuscreen RIA and by HPLC using a fluorescence detection method. Good correlation was observed between the immunoassay and the fluorescent integrity of the LSD molecule. Thermostability studies were conducted in the dark with various containers. These studies demonstrated no significant loss in LSD concentration at 25 degrees C for up to 4 weeks. After 4 weeks of incubation, a 30% loss in LSD concentration at 37 degrees C and up to a 40% at 45 degrees C were observed. Urine fortified with LSD and stored in amber glass or nontransparent polyethylene containers showed no change in concentration under any light conditions. Stability of LSD in transparent containers under light was dependent on the distance between the light source and the samples, the wavelength of light, exposure time, and the intensity of light. After prolonged exposure to heat in alkaline pH conditions, 10 to 15% of the parent LSD epimerized to iso-LSD. Under acidic conditions, less than 5% of the LSD was converted to iso-LSD. We also demonstrated that trace amounts of metal ions in buffer or urine could catalyze the decomposition of LSD and that this process can be avoided by the addition of EDTA. This study demonstrates the importance of proper storage conditions of LSD in urine in order to insure proper analytical testing results over time.     

Time course of the endocochlear potential in the isolated cochlea of the guinea pig

The temperature of Hanks' solution changed its potential (-0.48 mV/degrees C in Na(+)-Hanks' and -0.23 mV/degrees C in K(+)-Hanks' solution), but the pH had no clear effect on the potential. The time course of the endocochlear potential (EP) in the isolated cochlea of the guinea pig was measured under some different conditions. When the cochlea was moistened with Hanks' solution, negative EP increased gradually toward 0 mV at 124 min after death. When the cochlea was immersed in Hanks' solution, positive EP was obtained but it depended on the oxygen and the circulation of Hanks' solution. Moderate cooling (5 degrees C) of Hanks' solution had no significant effect on the EP of the isolated cochlea immersed in oxygen-saturated Hanks' solution circulated by oxygen bubbles. The space constant and the amplitude of the displacement responses were independent of the EP in the isolated cochlea. Thus, the positive EP might not show the physiological condition of the isolated cochlea.     

Effect of vasoactive drugs on muscle fibre types and capillaries

Photosensitive great tits (Parus major) and willow tits (P. montanus) were exposed to long days (20L:4D) under three different temperature conditions (4+, +10, and +20 degrees C) in early winter. The two species showed significant differences in their LH and testicular reaction patterns to low temperatures. Testes showed pronounced growth cycles under all temperature regimes. For the willow tit, testes in birds kept at +20 degrees C reached maximum size about 2 weeks earlier than testes in birds living under the two lower temperature regimes, whereas in the great tit testes reached maximum size at about the same time in all three groups. Low temperatures delayed the onset of testicular regression in both species. Plasma levels of LH did change with time in both species. However, the patterns of the induced LH-cycles in the three great tit groups differed significantly from each other, whereas this was not the case for the willow tits. The LH cycle was especially pronounced in great tits kept at +20 degrees C. The initial LH peak in great tits kept at +4 and +10 degrees C was about 50% lower than in great tits kept at +20 degrees C. These results are discussed in relation to species differences in winter ecology and establishment of breeding territories.     

Physiopathology of bedsores

Listeria monocytogenes is a food-borne pathogenic bacterium that can be found in soft cheese. At the beginning of cheese ripening, the pH is about 4.85-4.90. The aim of this work was to study the influence of temperature, preincubation temperature (temperature at which the inoculum was cultivated) and initial bacterial concentration on the survival of L. monocytogenes (strain Scott A) at pH 4.8. It was demonstrated in an earlier study that these factors did influence growth kinetics. Survival studies of L. monocytogenes were done in a laboratory broth simulating cheese composition. Four test temperatures (2, 6, 10 and 14 degrees C) and two preincubation temperatures were studied (30 degrees C or the test temperature). Listeria monocytogenes (strain Scott A) was unable to grow at pH 4.8 under all conditions tested. The time for 10% survival was about 11 and 2 d, at 2 degrees C with preincubation at 2 degrees C and 30 degrees C, respectively; 9 d at 6 degrees C with preincubation at 6 degrees C; 4 d at 6 degrees C with preincubation at 30 degrees C; and 1 d at 14 degrees C with preincubation at 14 degrees C or at 30 degrees C. The results show that survival of L. monocytogenes (strain Scott A) at pH 4.8 is not dependent on initial bacterial concentration but on both the test and preincubation temperatures.