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At least 1,387 records · Page 77Linked to original sources
PubMed1986

2,3,5-Triphenyltetrazolium chloride as a novel tool in germicide dynamics.

A novel, colorimetric method using 2,3,5-triphenyltetrazolium chloride (TTC) in tandem with membrane filtration is described for the determination of death rates of Escherichia coli (E. coli) due to microbiocides. This method enables results to be obtained on the same day in contrast to the 18 h required by the accepted aerobic plate count method. The microbiocides investigated were the preservatives 2-bromo-2-nitro-1,3-propanediol (Bronopol), N-(hydroxymethyl)-N-(1,3-dihydroxymethyl-2,5- dioxo-4-imidazolidinyl)-N'-(hydroxymethyl)-urea (Germall II), phenethyl alcohol, and benzyl alcohol. D values (time required per log reduction of E coli) were determined by this method, and equations relating the D values to preservative concentrations were derived [i.e., eta values (the logarithmic values relating changes in rates of kill for specified changes in concentration) and A values (extrapolated D values at 1% concentration) were determined]. these equations are compared with those previously published using the accepted aerobic plate count method. The potential advantages of this method are that it has a broad range of application as TTC is reduced by a wide variety of microbes; the test is easily done; results can be achieved in one day; dead cells do not cause interference; test sensitivity can be increased by increasing the length of incubation time or by using membrane filtration in tandem with TTC reduction; and preservative inactivation may be achieved by filtration and flushing with an inactivator, or by adding neutralizers to the TTC broth.

Colorimetry↗
PubMed1983

Indole metabolism in the pineal organ of the pigeon with special reference to melatonin-synthesizing cells. In vitro study combining thin layer chromatography, autoradiography and pharmacological treatment.

By means of radioimmunoassay a clear-cut peak of melatonin concentration was found in the pineal organ of the pigeon at the middle of the scotophase (Coisin et al. 1982a). The aim of the present study was to identify the cell type responsible for the nocturnal indole metabolism, including melatonin synthesis, in the pineal of this avian species. After a short-term incubation or organ culture in the presence of [3H]-indolic precursors, [3H]-5-hydroxytryptophan or [3H]-5-hydroxytryptamine, the relative amounts of deaminated and acetylated products occurring in the pineal organ were measured by the use of thin layer chromatography and liquid-scintillation counting. It was possible to modify the relative amounts of deaminated and acetylated indoles by the application of some inhibitors of monoamine oxidase and cyclic nucleotide phosphodiesterase. Irrespective of the experimental conditions, high-resolution autoradiography combined with the above-mentioned radiochemical experiments showed that the cells of the receptor line (modified photoreceptor cells) are responsible for indole storage and metabolism, and very probably also for melatonin biosynthesis. The other cell types of the pineal parenchyma did not display significant labeling.

4-(3-Butoxy-4-methoxybenzyl)-2-imidazolidinone↗
PubMed2006

Ketamine and N-acetylaspartylglutamate peptidase inhibitor exert analgesia in bone cancer pain.

PURPOSE: Not all bone cancer pain can be effectively treated with current therapies. In the present study, the effects of ip administration of alpha-2 agonists (dexmedetomidine and clonidine), N-methyl-D-aspartate (NMDA) antagonists (MK-801 and ketamine), an N-acetylaspartylglutamate peptidase inhibitor (ZJ-43), and morphine were examined in a mouse bone cancer pain model. METHODS: A bone cancer pain model was produced by injection of murine sarcoma cells into the medullary cavity of the distal femur. To estimate the level of bone cancer pain, the number of pain-related behaviours induced by repeated applications of a von Frey monofilament (0.166 g) to the site of tumour cells implantation was counted. Drugs were administered two weeks after the implantation. RESULTS: Morphine produced a significant analgesic effect (P < 0.001). The alpha-2 agonists produced analgesic effects (P < 0.001) with an efficacy similar to that of morphine, but only at doses that produced severe sedation. MK-801 had only limited analgesic effects, while ketamine produced an analgesic effect (P < 0.001) with the same efficacy as morphine. ZJ-43 (100 mg.kg(-1)) had a significant analgesic effect (P < 0.05) and the effect of ZJ-43 was antagonized by the selective group II metabotropic glutamate receptor (mGluR) antagonist. CONCLUSION: These data suggest that alpha-2 agonists produce an analgesic effect only at a sedative dose and that ketamine, but not MK-801, is associated with an analgesic response without overt side effects. The effect of ZJ-43 is mediated by activating group II mGluRs.

Adrenergic alpha-Agonists↗
PubMed2005

Comparing recovering efficiency of immunomagnetic separation and centrifugation of mycobacteria in metalworking fluids.

The accurate detection and enumeration of Mycobacterium immunogenum in metalworking fluids (MWFs) is imperative from an occupational health and industrial fluids management perspective. We report here a comparison of immunomagnetic separation (IMS) coupled to flow-cytometric enumeration, with traditional centrifugation techniques for mycobacteria in a semisynthetic MWF. This immunolabeling involves the coating of laboratory-synthesized nanometer-scale magnetic particles with protein A, to conjugate a primary antibody (Ab), specific to Mycobacterium spp. By using magnetic separation and flow-cytometric quantification, this approach enabled much higher recovery efficiency and fluorescent light intensities in comparison to the widely applied centrifugation technique. This IMS technique increased the cell recovery efficiency by one order of magnitude, and improved the fluorescence intensity of the secondary Ab conjugate by 2-fold, as compared with traditional techniques. By employing nanometer-scale magnetic particles, IMS was found to be compatible with flow cytometry (FCM), thereby increasing cell detection and enumeration speed by up to two orders of magnitude over microscopic techniques. Moreover, the use of primary Ab conjugated magnetic nanoparticles showed better correlation between epifluorescent microscopy counts and FCM analysis than that achieved using traditional centrifugation techniques. The results strongly support the applicability of the flow-cytometric IMS for microbial detection in complex matrices.

Centrifugation↗
PubMed1982

Preparation of anti-GM4 antiserum and its assay by a solid-phase radioimmunoassay.

Anti-GM4 antiserum was prepared from rabbits by immunization with pure human brain GM4 ganglioside in complete Freund's adjuvant and methylated bovine serum albumin. None of the immunized animals developed any clinically apparent neurological dysfunction. The antiserum titer and specificity were analyzed by double immunodiffusion and a novel solid-phase radioimmunoassay (RIA). In the latter procedure, microtiter plate wells were coated first with the glycolipid antigen, followed by sequential application of antiserum and [125I]-Staphylococcal Protein A. The absorbed radioactivity in the well was then counted. Employing the RIA procedure, anti-GM4 antibody achieved a titer of 1:1600. The antiserum also exhibited a high degree of specificity to GM4; cross-reactivity with glycolipids of similar structure was negligible. The production of highly specific antiserum to GM4 and the feasibility of detecting antibodies to glycolipid antigens by a convenient solid-phase RIA should be useful to further study the biological and immunological roles of GM4 and other glycolipids in the central nervous system.

Animals↗
PubMed2005

Buffer system for the separation of neutral and charged small molecules using micellar electrokinetic chromatography with mass spectrometric detection.

An organic buffer system will be discussed that is suitable for the separation of neutral as well as charged molecules be means of micellar electrokinetic chromatography (MEKC). The buffers are based on the combination of a long chain alkyl acid, such as lauric acid with ammonium hydroxide or an organic base such as tris-hydroxymethylaminomethane (Tris). The resulting buffer system is able to separate neutral compounds based on its micellar properties. These buffers exhibit much reduced conductivity compared to traditional MEKC buffers, such as sodium dodecylsulfate (SDS), which contain inorganic salts. They also have inherent buffer capacity at high pH resulting from the basic buffer component, which in our studies had pK values from about 8-11. The separations that were observed showed high efficiency with plate counts in many cases above 500,000 plates per meter. The reduced conductivity allowed for the application of much higher electric fields, resulting in very fast analysis times. Alternatively, an increase in detection sensitivity could be achieved, as the reduced conductivity allowed for the use of capillaries with lager internal diameters. Combinations of different alkyl acids and organic bases provided for significant flexibility in selectivity tuning. Finally, the fact that the organic micellar buffer systems discussed here do not contain inorganic ions, allows for coupling with mass spectrometric (MS) detection. The possibility of MS detection combined with the high speed in analysis that can be obtained using these organic buffer systems, could make this approach an interesting option for high throughput analysis of combinatorial libraries.

Buffers↗
PubMed2000

Dose finding in the Ames Salmonella assay.

Threshold dose/concentration values, such as the lowest effective dose, minimum effective dose or the lowest effective concentration (LED, MED or LEC, respectively) are in use as an alternative to the mutagen potency measures based on the 'rate' measurements (e.g., the slope of the initial part of the dose-response curve). In this respect, several statistical procedures for the corresponding so-called 'dose finding' were proposed during the last decades. However, most of them disregard the discrete nature of responses such as the plate colony count in the Ames Salmonella assay. When the plate counts agree with the Poisson assumption, two procedures considered here seem to be appropriate for the dose finding. One is based on the stepwise collapsing of the homogeneous control and dose counts; another consists of constructing the confidence limits for the mutation induction factor (MIF). When the dose and control counts are non-overlapping, the simple 'visual' non-parametric estimation of LED is possible. Applicability and validity of the methods is demonstrated with the two data sets on the mutagenicity of the beta-carboline alkaloid, harmine, and one of the oxidation products of apomorphine.

Dose-Response Relationship, Drug↗
PubMed2000

Topical dinitrochlorobenzene therapy in the treatment of refractory atopic dermatitis: systemic immunotherapy.

BACKGROUND: Repeated dinitrochlorobenzene (DNCB) application has been proposed as a systemic immunotherapy on the basis of its ability to stimulate T helper 1 (T(H)1) responses, such as those for systemic lupus erythematosus and HIV infection. OBJECTIVE: We report the effect of topical DNCB therapy in an open trial in patients with refractory atopic dermatitis (AD). METHODS: Eight patients with refractory AD received weekly application of 0.2% to 1% DNCB to a 2. 5-cm(2) area on the upper arm after sensitization with 5% DNCB; the position was rotated at each application. Disease activity was monitored by pruritus score, percentage of body involvement, clinical severity score, eosinophil counts, serum IgE levels, and serum soluble interleukin 2 receptor levels. RESULTS: Six of 8 patients (patients 1-6) showed apparent improvement both on clinical scores and laboratory data until 16 weeks after DNCB therapy (week 16). The clinical severity scores of patients 1 to 6 were significantly correlated with eosinophil counts, IgE levels, and serum soluble interleukin 2 receptor levels. One patient did not show clear improvement, and another (patient 8) showed deterioration. DNCB therapy was discontinued at week 12 for patient 8. CONCLUSION: Topical DNCB may systemically stimulate T(H)1 cell responses of patients with AD, resulting in restoration of the T(H)1/T(H)2 imbalance and possible clinical improvement. These results, however, should be interpreted with caution until additional documentation is obtained.

Administration, Topical↗
PubMed1989

Improved synchronized accumulating radioisotope detector for gas chromatography.

A synchronized accumulating radioisotope detector for radio gas chromatography was developed. It comprised seven gas-flow proportional counters each with an inner volume of 10 ml. Every counter tube was connected by a mutual anti-coincidence circuit to reduce the background. The transit time of gas particles in one counter tube could be set to an optimal value between 1 and 4 s by regulating the flow-rate of the counting gas, according to analytical requirements. The improved detector maintained high chromatographic resolution, which suggested the applicability of the apparatus to capillary gas chromatography.

Chromatography, Gas↗
PubMed1991

Surface properties of intraocular lens materials and their influence on in vitro cell adhesion.

An in vitro model to assess lens epithelial cell adhesion to a variety of intraocular lens materials was developed. Rabbit anterior lens capsules were isolated and cultured in serum-containing medium. Test surfaces included poly(methyl methacrylate), two new silicones (SLM-1/UV, SLM-2/UV), two hydrogels (HEMA, Lidofilcon A), and polytetrafluoroethylene (PTFE). Following the application and culturing of cells on the test surfaces, adherent cells were removed by trypsinization and counted at eight and 24 hours. The material surfaces were characterized by electron spectroscopy for chemical analysis and scanning electron microscopy. The captive bubble technique was also used to assess interfacial free energy. More cells adhered to PMMA than to the other materials tested (P less than .01). The two silicones, HEMA, and PTFE did not differ significantly from each other; Lidofilcon A had the lowest cell adhesion of all materials tested. Cell adhesion results were related to the interfacial free energy of each material. Materials of low (less than 5 ergs/cm2) or high (greater than 40 ergs/cm2) interfacial free energies had lower cell adhesion than materials of intermediate free energies (5 to 40 ergs/cm2) which exhibited the highest cell adhesion.

Animals↗
PubMed2004

Sputum tests in the diagnosis and monitoring of asthma.

OBJECTIVE: To review the techniques of sputum analysis with relevance to the diagnosis and monitoring of asthma. DATA SOURCES: MEDLINE databases were searched to identify all publications involving sputum studies related to the diagnosis and treatment of asthma from 1990 to October 2003. We also used internal reference files related to sputum and searched bibliographies of relevant articles. The review was limited to human data in English-language publications. STUDY SELECTION: Studies were selected by the expert opinions of the authors for quality and relevance to the evaluation of asthmatic inflammation by induced sputum. RESULTS: During the past 10 years, there have been an increasing number of publications concerning the diagnosis and treatment of asthma using sputum analysis. Analysis of induced sputum provides similar data to secretions obtained through bronchial wash, bronchoalveolar lavage, and, to some extent, bronchial biopsy. The techniques of cellular counting and immunochemical analysis are described along with potential problems and pitfalls of these methods. Clinical application of sputum analysis is discussed as it pertains to the diagnosis and monitoring of asthma and asthma-related conditions. CONCLUSIONS: Analysis of induced sputum is increasingly being considered as a noninvasive means of evaluating airway inflammation and may provide useful information with regard to the diagnosis and monitoring of asthma in select individuals.

Asthma↗
PubMed2004

Relationship between physical activity and diet among African-American girls.

OBJECTIVE: To examine the cross-sectional relationships between physical activity and dietary behaviors among 8- to 10-year-old African-American girls. RESEARCH METHODS AND PROCEDURES: Two hundred ten 8- to 10-year-old African-American girls from four field centers participated. Computer Science and Applications (CSA) activity monitors were worn for 3 days. CSA data were expressed as mean CSA counts per minute, mean minutes of moderate to vigorous activity per day, and mean metabolic equivalents (METS) per minute. Two nonconsecutive 24-hour dietary recalls were analyzed for kilocalories; percent kilocalories from fat; daily servings of fruit, 100% fruit juice, and vegetables; sweetened beverages; and water consumption. Height and weight were measured, and information on household income, material possessions, and participant age were obtained. RESULTS: All three expressions of physical activity were significantly negatively associated with percentage calories from fat (r = -0.147 to -0.177, p < 0.01), and mean METS per minute were significantly positively associated with percentage calories from carbohydrate (r = 0.149, p < 0.05) after controlling for household income, material possessions, field center, and total caloric intake. Income was inversely associated with percentage calories from fat. DISCUSSION: Physical activity and dietary fat consumption were inversely related among African-American girls. Efforts to prevent obesity in preadolescent African-American girls should focus on increasing physical activity and lowering dietary fat consumption.

Black or African American↗
PubMed2005

Assisted procreation in cases of hepatitis B, hepatitis C or human immunodeficiency virus infection of the male partner.

Guidelines for assisted procreation impose a special responsibility upon physicians for the health of the expected child because of their active role in inducing pregnancy. Therefore, careful clinical evaluation of both partners has to precede every application of these methods. Risks for the mother's health or the development of the child count as a relative contraindication for a treatment. To balance these relative contraindications, the existing risk factors have to be recognized through screening examination. If a chronic infection occurs in the male partner, prevention for the female partner is theoretically possible by using a condom. As this inhibits a pregnancy, at least in cases of human immunodeficiency virus and hepatitis C virus infections, realization of a pregnancy requires assisted procreation. The main question in these cases is whether infectious particles can be eliminated by sperm processing to ensure the safe treatment of the healthy female partner.

Female↗
PubMed2001

Protegrin-1 enhances bacterial killing in thermally injured skin.

OBJECTIVE: Septic complications and the emergence of drug-resistant microbes represent serious risks to patients. Recently, naturally occurring peptides have been discovered that possess potent and broad-spectrum antimicrobial activity. Protegrin-1 is particularly attractive for clinical use in human wounds because, unlike defensins, protegrin-1 retains broad antimicrobial and antifungal activity at physiologic salt concentration and in the presence of serum. The objective of this study was to examine the efficacy of protegrin-1 in killing multiple drug-resistant microbes isolated from human burn patients. DESIGN: For thein vitroexperiment, bilayer radial diffusion was performed comparing standard antibiotics with protegrin-1 on multiple-drug-resistant microbial organisms isolated from infected burn wounds. In vivo, rats received a 20% total body surface area partial-thickness burn by immersion in 60 degrees C water for 20 secs followed by wound seeding with 106 colony forming units of Silvadene-resistant Pseudomonas aeruginosa. SETTING: University of Michigan research laboratory. SUBJECTS: Adult, male Sprague-Dawley rats. INTERVENTIONS: Rats were randomized into three groups: those receiving synthetic protegrin-1, acetic acid (carrier), or gentamicin (positive control). Protegrin-1 was administered by topical application or intradermal injection. Wound tissues were harvested aseptically at different time points for quantitative bacterial counts. MEASUREMENTS AND MAIN RESULTS: In vivo and in vitro experiments revealed rapid and significant decreases in bacterial counts for protegrin-1-treated groups compared with controls. CONCLUSIONS: This study shows that protegrin-1 potentially may be used as an alternative or adjunct therapy to standard agents used to treat wound infections.

Administration, Topical↗
PubMed1982

Iontophoretically applied microtubule inhibitors induce transganglionic degenerative atrophy of primary central nociceptive terminals and abolish chronic autochtonous pain.

Transcutaneous iontophoresis of microtubule inhibitors (Vinblastin, Vincristin, Formyl-Leurosin) in rats induces depletion of fluoride-resistant acid phosphatase (FRAP) and transganglionic degenerative atrophy (trggl. deg. atr.) of the central terminals of primary nociceptive neurons, probably via blockade of axoplasmic transport in the peripheral sensory nerves. Radiochemical experiments prove that about 0.2% of the microtubule inhibitors applied iontophoretically at the skin reach the level of nociceptive axon terminals. 40 out of 48 patients suffering from chronic intractable pain of diverse etiology (postherpetic, paresthetic, ischaemic and trigeminal neuralgia, alcoholic and diabetic polyneuropathy, meralgia, brachialgia, discopathia, arthropathia and terminal pain) were successfully treated with Vinblastin or Vincristin iontophoresis. Iontophoretically applied microtubule inhibitors do not affect the blood cell count, have no side-effects and do not impair the skin at the site of application.

Acid Phosphatase↗
PubMed1975

Comparison of agents producing a neutrophilic leukocytosis in man. Hydrocortisone, prednisone, endotoxin, and etiocholanolone.

To study the potential application of glucocorticosteroid administration for the measurement of the bone marrow neutrophil reserve response, blood neutrophil count changes were measured in normal subjects after the administration of intravenous hydrocortisone (25, 50, 100, 200, and 400 mg) and oral prednisone (5, 10, 20, 40, and 80 mg). The upper three doses of both steroids increased the blood neutrophil count by approximately 4,000 cells/mm3. The neutrophilia occurring after hydrocortisone (200 mg) and/or prednisone (40 mg) was compared with that observed after endotoxin (0.8 ng/kg) and etiocholanolone (0.1 mg/kg) in 14 normal subjects, 7 patients with Wegener's granulomatosis on cyclophosphamide therapy and 10 patients with chronic idiopathic neutropenia. The normal responses (mean increase of blood neutrophils/mm3 above base line +/- 1 SEM) were: hydrocortisone 4,220 +/- 320, prednisone 4,610 +/- 360, endotoxin 6,060 +/- 880, and etiocholanolone 3,780 +/- 440. In the patient studies, etiocholanolone gave the smallest mean responses, but, in general, the results were similar for all agents. These data indicate that these glucocorticosteroids can be used as equivalent agents to endotoxin and etiocholanolone for measuring the neutrophil reserve response.

Adult↗
PubMed2001

Evaluation of the petrifilm plate method for the enumeration of aerobic microorganisms and coliforms in retailed meat samples.

This study was designed to compare the effectiveness and applicability of the Petrifilm plate method with the Association of Official Analytical Chemists' (AOAC) standard aerobic count method and violet red bile agar method for meat products. The comparison was carried out using 303 meat samples collected from various retailers: 110 pork samples, 87 chicken samples, and 107 beef samples. In the comparison of the correlation coefficient (R) between the conventional method and the Petrifilm plate method by a linear regression analysis, the correlation coefficient in total microorganisms was 0.99, 0.95, and 0.94 in pork, beef, and chicken samples, respectively. The correlation coefficient in coliform count was 0.83, 0.96, and 0.81 in pork, beef, and chicken samples, respectively. Based on the high correlation in the total microorganism count, it might be possible to replace the conventional methods with the Petrifilm plate method. For coliform counts, the Petrifilm plate method also showed a generally high correlation coefficient, except for pork samples, which are more subject to contamination. The Petrifilm plate method was simpler and less time-consuming in sample preparation and, in procedures, faster than the conventional method. These results suggested that the 3M Petrifilm plate method could replace the conventional methods in the analysis of microorganism contamination measurement in meat products.

Agar↗
PubMed2001

Monitoring of microbial hazards at farms, slaughterhouses, and processing lines of swine in Korea.

This study was executed to investigate microbiological hazards at swine farms, slaughterhouses, dressing operations, and local markets for the application of the hazard analysis critical control point system in Korea by analyzing total aerobic plate count (APC) and presence of pathogens. Six integrated pig farms and meat packers were selected from six different provinces, and samples were collected from pig carcasses by swabbing and excision methods at the slaughterhouses, processing rooms, and local markets, respectively. APCs of water in water tanks were relatively low, 1.9 to 3.1 log10 CFU/ml; however, they were increased to 4.6 to 6.9 log10 CFU/ml when sampled from water nipples in the pigpen. APCs of feeds in the feed bins and in the pigpens were 4.4 to 5.4 and 5.2 to 6.7 log10 CFU/g, respectively. Salmonella spp., Staphylococcus aureus, and Clostridium perfringens were detected from water and feed sampled in pigpens and pigpen floors. S. aureus was the most frequently detected pathogenic bacteria in slaughterhouses and processing rooms. Listeria monocytogenes and Yersinia enterocolitica were also detected from the processing rooms of the Kyonggi, Kyongsang, and Cheju provinces. Even though APCs were maintained at the low level of 3.0 log10 CFU/g during slaughtering and processing steps, those of final pork products produced by the same companies showed relatively high numbers when purchased from the local market. These results indicated that the cold chain system for transporting and merchandising of pork products was deficient in Korea. Water supply and feed bins in swine farms and individual operations can be identified as critical control points to reduce microbiological hazards in swine farms, slaughterhouses, and processing plants.

Abattoirs↗