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PubMed2003

Application of a modified culture medium for the simultaneous counting of molds and yeasts and detection of aflatoxigenic strains of Aspergillus flavus and Aspergillus parasiticus.

Molds and yeasts from 91 samples of feed and raw materials used in feed formulation were enumerated on a new culture medium to which a beta cyclodextrin (beta-W7M 1.8-cyclodextrin) had been added. This medium was compared with other media normally used in laboratories for the routine analysis of fungi, such as Sabouraud agar, malt agar supplemented with 2% dextrose, and potato dextrose agar. When a t test for paired data (0.05 significance level, 95% confidence interval) was applied, no statistically significant differences between the results obtained with the new culture medium and those obtained with the other media used to enumerate molds and yeasts were found. For the evaluation of contamination due to aflatoxin for all of the samples, Sabouraud agar and yeast extract agar, both supplemented with 0.3% beta-W7M 1.8-cyclodextrin, and APA (aflatoxin-producing ability) medium were used. Aflatoxin was detected in 21% of the feed samples and in 23% of the raw-material samples analyzed, with maximal amounts of 2.8 and 6.0 microg of aflatoxin B1 per kg, respectively, being detected. In any case, the aflatoxin contents found exceeded the legally stipulated limits. The t test for paired data (0.05 significance level, 95% confidence interval) did not show statistically significant differences between the results obtained with the different culture media used for the detection of aflatoxins. The advantage of the new medium developed (Sabouraud agar with 0.3% beta-W7M 1.8-cyclodextrin) is that it allows simultaneous fungal enumeration and determination (under UV light) of the presence of aflatoxin-producing strains without prior isolation and culture procedures involving expensive and/or complex specific media and thus saves work, time, and money.

Aflatoxins↗
PubMed1984

[Methods for testing surface disinfection procedures under current conventional conditions].

A method of testing surface disinfection procedures is described with which the same conditions are maintained as those encountered in practice. The disinfectant is applied to surfaces in a conventional manner at a natural scale. A standardised wet-mopping procedure (two-bucket method) applied to floors of glazed clinker serves as a reference. The test method is quantitative. For comparison the same procedure using water of standardised hardness is used. The test parameters are the reduction of the colony count (CC) both on the surface treated and in the used washing water following certain periods of exposure to the disinfectant. For the suitability test of disinfectants the surface is contaminated with reference strains, Kl. pneumoniae ATCC 27736 and Staph. aureus Wood 46. In some special cases wild strains were used for contamination. When testing surface disinfection under user conditions, the flora settling on the surface used is investigated. The bacterial collector after Thran is employed for the extraction of the germs from the surface. Wet mopping with water of standardised hardness has already reduced the count on the floor of Kl. pneumoniae by 2.8 log on average and the count of Staph. aureus by 2.2 log. By the use of the three types of disinfectants tested the count was additionally reduced by 0.45 to 0.72 log for Kl. pneumoniae and by 1.2 to 2.2 log for Staph. aureus. Even after disinfection a spread of germs can be observed; however, it is slight and corresponds to the spread found after the use of water. After mopping with 17 degrees dH water, practically the total germ count applied is found in the cleaning water. After application of the disinfectant the germ counts for Kl. pneumoniae are reduced by 1.8 to 3.7 log and by 3.1 to 3.7 log for Staph. aureus, depending on the type of disinfectant after an exposition of 5 min in the washing water. With respect to Kl. pneumoniae, reduction of the count continues to amount to 2.7 to 3.8 log after 30 min, so that merely 20 to 30 CFU/100 ml washing water remain. The concentration of the disinfectant mainly influences the reduction of germs in the washing water. Thus the analysis of the washing water constitutes an important parameter for the evaluation of a surface disinfectant. The testing procedure proposed is very flexible as it can be readily adapted to the various disinfection problems and evaluation criteria.

Disinfectants↗
PubMed1981

Cell population kinetics in rat palatal epithelium treated with the carcinogen 4-nitroquinoline N-oxide (4NQO).

The carcinogen 4NQO was applied to the palates of rats for 2 weeks or 2 months. Eight weeks after termination of carcinogen application, animals were injected with colchicine and sacrificed at intervals of 3 h during a 24 h period. Histological sections were produced and the number of arrested metaphases counted in the palatal epithelium. The two groups had the same daily mitotic rate and exhibited a circadian rhythm in mitotic activity similar to that demonstrated in normal oral mucosa of nocturnal animals. Carcinogen was applied to the palates of other groups of rats for 2 weeks or 2 months. Animals treated with saline served as controls. At various time points during and after carcinogen treatment eight rats from each group were killed. Four of these animals were injected with colchicine prior to sacrifice and the number of arrested metaphases counted in the palatal epithelium. In the noncolchicinized rats all mitotic phases were counted. After cessation of carcinogen application the mitotic rates, the mitotic counts and the mitotic durations did not differ from those of control animals, indicating that the carcinogen had failed to induce irreversible changes in the cell population kinetics of the palatal epithelium.

4-Nitroquinoline-1-oxide↗
PubMed1992

Pharmacokinetic profile of [14C]flutrimazole following single topical application in normal and scarified skin of healthy volunteers.

14C-labelled 1-[(2-fluorophenyl)(4-fluorophenyl)phenylmethyl]-1H- imidazole (flutrimazole, UR-4056, CAS 119006-77-8) was administered topically (dermally) in three male volunteers to normal skin and scarified skin, respectively. Blood, urine and faeces were collected and radioassayed by liquid scintillation counting techniques. After topical application to healthy skin area, less than 1% of flutrimazole was absorbed percutaneously from a 1% cream and the amount absorbed was excreted with the urine. Following administration of the drug to scarified skin, the proportions absorbed were very similar to those measured after application to normal skin. About 0.5% of the administered dose is recovered in urine. The absorbed amount of drug leads to very low concentrations in the circulation, lower than a few ng/ml.

Administration, Topical↗
PubMed1997

Can zinc prevent apoptosis of anterior keratocytes after superficial keratectomy?

OBJECTIVE: To determine whether zinc, which blocks apoptosis in many systems, including in leukemic cells and possibly in retinal dystrophies, can prevent the unwanted loss of anterior stromal keratocytes after superficial keratectomy. METHODS: After mechanical central corneal epithelial debridement, the left eyes of nine New Zealand white rabbits were treated with 25 mM zinc chloride (ZnCl2) in Earle's salts minimal essential medium (MEM) either every 30 min (n = 3), every 2 h (n = 3), or every 4 h (n = 3). The left eyes of nine additional animals, divided into three equal groups, were deepithelialized, and each received pure culture medium at one of the same three frequencies. One eye of each of another six rabbits was deepithelialized but received no drops. After 24 h, all 24 animals were sacrificed and the globes were enucleated. The corneas were processed and sections were stained with hematoxylin and eosin. RESULTS: The cell count of MEM-treated corneas exceeded that of untreated corneas (p = 0.03, analysis of variance [ANOVA]), but there was no difference among eyes that received the different frequencies of MEM application (p = 0.36, ANOVA). Cell counts increased with frequency of zinc application, but the differences were not statistically significant (p = 0.09, ANOVA). Only in the group receiving the most frequent zinc applications were superficial keratocytes retained. This group also possessed a greater number of stromal keratocytes than untreated controls and MEM-treated corneas (p = 0.01). CONCLUSIONS: At a significantly high dosage, zinc can prevent loss of superficial keratocytes to a greater extent than culture medium alone can. Moreover, zinc has the advantage of preserving the most anterior layer of keratocytes. Retention of these cells may prevent the reactive overproliferation that constitutes haze after photorefractive surgery.

Administration, Topical↗
PubMed1998

Comparison of 3 methods for counting fruits and vegetables for fourth-grade students in the Minnesota 5 A Day Power Plus Program.

OBJECTIVE: To compare fruit and vegetable servings calculated from 24-hour dietary recall data using 3 methods: a counting scheme developed for the 5 A Day for Better Health study, a method developed by the University of Minnesota Cancer Prevention Research Unit to quantify total consumption of fruits and vegetables, and a counting scheme based on the US Food and Drug Administration's Reference Amounts. The counting methods differ by food items counted and by serving sizes for those items. SUBJECTS/SETTING: Record-assisted 24-hour dietary recalls were collected from 617 randomly selected fourth-grade students (317 girls, 300 boys) from 23 schools in St Paul, Minn, participating in the Minnesota 5 A Day Power Plus Program. DESIGN: The dietary recalls were analyzed using the Minnesota Nutrition Data System (version 2.6/8a/23). Total servings of fruits and vegetables, servings of vegetables, servings of fruits plus juices, servings of fruit juice, and servings of fruit excluding juice were tallied using each counting method. STATISTICAL ANALYSES: A mixed-model Poisson regression analysis was conducted to compare numbers of servings calculated using the 3 methods. RESULTS: Counts of daily total fruits and vegetables averaged 3.9 servings with the 5 A Day method, 4.1 servings using US Food and Drug Administration Reference Amounts, and 5.1 servings with the Minnesota Cancer Prevention Research Unit method (P < .0001). APPLICATIONS: Because the different counting methods yield different tallies of fruit and vegetable intake, it is important for researchers and practitioners interested in fruit and vegetable consumption to be clear about their uses of the data before choosing a counting scheme.

Child↗
PubMed2006

Efficacy of an alcohol-based healthcare hand rub containing synergistic combination of farnesol and benzethonium chloride.

Healthcare workers are required to disinfect the hands several times a day using hand disinfectants, which leads to chronic hand exposure to high levels of antimicrobials contained in the disinfectants, which could compromise the skin integrity. This problem may be addressed by developing hand disinfectants containing synergistic combinations of small amounts of antimicrobials and other agents. The synergistic effect of farnesol and essential oils with several antimicrobials was studied in vitro to select an effective antimicrobial system in preservative concentration for use in healthcare hand rub. Farnesol and lemon oil showed synergistic activity against S. aureus, in combination with benzalkonium chloride and benzethonium chloride, but not with other antimicrobials studied. All essential oils studied showed synergy with benzethonium chloride against Staphylococcus aureus and Escherichia coli. An alcohol-based healthcare hand rub (ZBF hand rub) containing this unique synergistic combination of farnesol and benzethonium chloride was then developed and its efficacy as a healthcare hand rub was evaluated in human volunteers according to the US FDA-TFM protocol using Serratia marcescens as a marker organism. The ZBF hand rub showed a 3.22 log(10) reduction in the microbial count after the first application and a 5.49 log(10) reduction after the tenth application in vivo and exceeds the US FDA-TFM criteria for healthcare hand rub. The ZBF hand rub did not irritate the hands when tested on human volunteers when applied 10 times everyday for five consecutive days. The ZBF hand rub exhibits more than 5.5 log(10) reduction in the microbial count within 15s and more than 2.8 log(10) reduction in the two types of viruses tested within 30s in vitro. When evaluated in an in vitro pig skin model, the ZBF hand rub shows better prolonged activity (20-35 min post-application) against transient bacteria (S. aureus and E. coli) compared to other alcohol-based hand rubs. These findings suggest that the use of the ZBF hand rub amongst health care workers may lower the risk of chronic hand exposure to high levels of antimicrobials without compromising the efficacy.

Animals↗
PubMed1998

Acceptance and feasibility of peripheral stem cell mobilisation compared to bone marrow collection from healthy unrelated donors.

Allogeneic peripheral blood stem cell transplantation leads to an earlier engraftment compared to BMT. The feasibility, acceptance and long-term side-effects of G-CSF mobilisation of PBSC in unrelated healthy donors needs to be evaluated. Forty unrelated healthy donors received G-CSF in a dose of 10 microg/kg bodyweight for 5 days and two aphereses were performed. The donors were monitored prospectively. The data were compared to bone marrow harvests from unrelated donors. Almost all stem cell donors reported some side-effects due to Filgrastim application. Bone pain (32), headache (20), chest pain (two) and night sweats (one) were complained of. By taking analgesics, the pain was relieved in most cases. No donor discontinued the filgrastim application. Bone pain and headache resolved within 2-4 days after termination of Filgrastim application. There was, as expected, a seven-fold increase in the number of total WBCs. There were no significant changes of platelet counts during G-CSF application. After 4 weeks haemoglobin concentration and platelet counts showed no significant differences compared to baseline values. The aphereses were mostly tolerated very well. Eighteen donors reported paraesthesia, one donor developed dizziness, two complained of nausea and vomiting. There was a significant decrease in platelet count (242 before, 98 x 10(9)/l after aphereses). Autologous platelets were transfused after the second aphereses in four donors. These data were compared to data from 245 unrelated bone marrow donors, who had on average, 14 days bone pain and tiredness after donation. The G-CSF mobilisation and apheresis of peripheral blood stem cells is an alternative to traditional bone marrow harvesting in unrelated healthy donors. It is well tolerated and the duration of side-effects on average is shorter than after the surgical procedure. So far no long-term effects have been observed in the follow-up.

Adult↗
PubMed1969

Characteristics of marrow production and reticulocyte maturation in normal man in response to anemia.

Erythropoiesis in normal man was studied during periods of phlebotomy-induced anemia of varying severity. This study permitted a comparison of marrow production measurements over a wide range of marrow production levels. As long as the serum iron remained above 50 mug/100 ml, measurements of plasma iron turnover provided an excellent index of marrow production at all levels of red cell production. In contrast, the absolute reticulocyte count demonstrated a poor correlation with the other measurements. This was shown to be the result of a prolongation of the time required for circulating reticulocytes to lose their reticulum, which correlated with the severity of the anemia. For the clinical application of the reticulocyte count as a measurement of marrow production, an adjustment must be made for this alteration in the circulating reticulocyte maturation time.

Adult↗
PubMed2004

In-use efficacy of a chlorhexidine in alcohol surgical rub: a comparative study.

BACKGROUND: Although full surgical scrubs are performed prior to each case on an operating list, optimum regimens for hand cleaning have yet to be determined, and in-use efficacy evaluations are very limited. METHODS: A crossover study was undertaken comparing a chlorhexidine in detergent/alcohol regimen with povidine-iodine detergent scrub, within an orthopaedic operating environment. Depending on the skin asepsis regimen used, five surgical team members scrubbed or rubbed prior to each case for a complete operating list. Bactericidal efficacy was measured using the 'glove-juice' technique before and after hand asepsis, and at the completion of each case. RESULTS: The chlorhexidine regimen caused substantial and sustained reductions in hand bacterial counts (>50-fold prior to case 1) during surgical cases. Application of alcoholic chlorhexidine prior to each subsequent case reduced bacterial counts to the same level as the original scrub. In contrast, the povidine-iodine scrub reduced counts <3-fold prior to the first case and <2-fold in subsequent cases. The chlorhexidine regimen also resulted in persistent bactericidal effects between cases, as counts prior to application of cases 2 and higher were significantly lower than prior to case 1 (>7-fold for case 2 vs case 1). CONCLUSIONS: The chlorhexidine regimen demonstrated excellent bactericidal efficacy throughout an operating list, and was superior to povidine-iodine scrubbing in all aspects. The alcoholic chlorhexidine regimen is simpler and should have wide surgical application.

2-Propanol↗
PubMed2000

Correlation of leukocyte esterase detection by reagent strips and the presence of neutrophils: a study in BAL fluid.

STUDY OBJECTIVE: In the present study, we evaluated the leukocyte esterase (LE) area of a reagent strip designed for urinalysis for the semiquantitative measurement of the percentage of polymorphonuclear neutrophils (PMNs) in BAL fluid. DESIGN: Prospective. The relative PMN counts (obtained by conventional microscopy and expressed as a percentage of a 500 cell count) of consecutive BAL fluid samples were compared with the corresponding LE categories as read with a urine chemistry reader. LE categories were graded as follows: negative, trace, +, + +, and + + +. RESULTS: A total of 153 BAL fluid samples were included. The mean PMN counts of the negative LE category (4.1 +/- 4.3%; n = 43) and the + + + category (81.8 +/- 16.3%; n = 37) differed significantly from each other and from the mean PMN counts of the other categories. Within the trace, +, and + + categories, a considerable overlap of PMN counts was noted. Assignment of a BAL fluid to the negative LE category consistently predicted a PMN count < 20%. At a threshold value of 50% PMNs, the + + + LE category predicted the BAL fluid samples to the correct group (PMNs > 50% vs < 50%) with a sensitivity of 70.8% and a specificity of 97.1%. CONCLUSIONS: The reagent strips proved to be useful as a rapid test for semiquantitative measurement of the relative PMN counts in BAL fluid. However, the low predictive value for the exclusion of a high PMN count may limit their application.

Alveolitis, Extrinsic Allergic↗
PubMed1997

Applications of multimedia computers and video mixing to neuroethology.

Inexpensive multimedia computers offer new possibilities for mixing video and computer images, videotaping these mixed images, and extracting quantitative data from videotape. In this paper we describe methods for mixing images from a video camera and a Macintosh computer display using chroma keying, and we describe a simple circuit for analog video mixing and frame-counting. We present three applications of these video mixing methods to our neurophysiological and behavioral research with awake, behaving animals. These technologies enhance accuracy, speed, and flexibility during experiments, by allowing us to record an information-rich videotape of the subject and state of the experimental apparatus. After the experiment, these technologies facilitate selecting and extracting quantitative data from the videotape for further analysis. The videotape is especially useful in resolving minor inconsistencies or incomplete information in the notes and data files that often arise during analysis of complex experiments.

Animals↗
PubMed1998

Estimating surface area by the isotropic fakir method from thick slices cut in an arbitrary direction.

The proposed fakir method for estimating surface area is based on counting the intersections between the surface lying within a thick slice, and an isotropic spatial grid consisting of a combination of linear probes called fakir probes. An unbiased procedure using a directly randomized spatial grid rather than sections with randomized directions is presented. The method is applicable if perfectly registered serial sections of the surface are available in a thick slice while the direction of the slice can be arbitrary. The efficiency of the fakir method using different arrangements of orthogonal triplets of fakir probes is evaluated and it is shown that mutually shifted probes are superior to non-shifted ones. The application software for interactive counting of intersections between computer-generated fakir probes and the surface within the stack of digitized images is described and demonstrated by two examples: estimation of the surface area of individual tobacco cell chains using a confocal microscope, and estimation of the total area of exposed surface of mesophyll cells in a barley leaf using a wide-field transmission microscope.

Journal Article↗
PubMed2006

Cannabinoids and neuronal damage: differential effects of THC, AEA and 2-AG on activated microglial cells and degenerating neurons in excitotoxically lesioned rat organotypic hippocampal slice cultures.

Cannabinoids (CBs) are attributed neuroprotective effects in vivo. Here, we determined the neuroprotective potential of CBs during neuronal damage in excitotoxically lesioned organotypic hippocampal slice cultures (OHSCs). OHSCs are the best characterized in vitro model to investigate the function of microglial cells in neuronal damage since blood-borne monocytes and T-lymphocytes are absent and microglial cells represent the only immunocompetent cell type. Excitotoxic neuronal damage was induced by NMDA (50 microM) application for 4 h. Neuroprotective properties of 9-carboxy-11-nor-delta-9-tetrahydrocannabinol (THC), N-arachidonoylethanolamide (AEA) or 2-arachidonoylglycerol (2-AG) in different concentrations were determined after co-application with NMDA by counting degenerating neurons identified by propidium iodide labeling (PI(+)) and microglial cells labeled by isolectin B(4) (IB(4)(+)). All three CBs used significantly decreased the number of IB(4)(+) microglial cells in the dentate gyrus but the number of PI(+) neurons was reduced only after 2-AG treatment. Application of AM630, antagonizing CB2 receptors highly expressed by activated microglial cells, did not counteract neuroprotective effects of 2-AG, but affected THC-mediated reduction of IB(4)(+) microglial cells. Our results indicate that (1) only 2-AG exerts neuroprotective effects in OHSCs; (2) reduction of IB(4)(+) microglial cells is not a neuroprotective event per se and involves other CB receptors than the CB2 receptor; (3) the discrepancy in the neuroprotective effects of CBs observed in vivo and in our in vitro model system may underline the functional relevance of invading monocytes and T-lymphocytes that are absent in OHSCs.

Animals↗
PubMed1991

Computer-aided cell colony counting.

Counting cell colonies is a tedious task when performed with the light microscope. Moreover, unless strict double-blind protocols are adhered to, biased counts are difficult to avoid. Presented here is a computer software application that performs accurate, reproducible cell colony counts with a minimum of user generated bias. The application is based upon the Apple IICX computer system with Image software and AppleScan. Colonies are grown on 24-well plates and prepared in such a way as to permit good quality scanning. The scans are then transferred to Image and the individual colonies in each well are counted. Good correlation with counts done by light microscopy is achieved.

Clone Cells↗