Use of a commercially available ELISA kit for detection of salmonellas in water

The accuracy and specificity of a commercially available ELISA kit (Locate, Rhone-Poulenc Diagnostics) were assessed when applied to enrichment cultures of naturally contaminated water and sewage. The kit showed 66/180 samples positive by both culture and ELISA. The ELISA showed six positives which could not be confirmed by culture.     

A spectrophotometric assay for lipid peroxides in serum lipoproteins using a commercially available reagent

A method is described for measuring lipid peroxides by means of the color reagent of a commercially available test kit for cholesterol estimation. In principle, this assay makes use of the oxidative capacity of lipid peroxides to convert iodide to iodine, which can be measured photometrically at 365 nm. Calibration curves were obtained using peroxides such as H2O2, t-butyl hydroperoxide, and cumene hydroperoxide. A stoichiometric relationship was observed between the amount of organic peroxides assayed and the concentration of iodine produced. Concentrations of lipid peroxides as small as 1 nmol/ml could be measured. The ability to estimate lipid peroxides of isolated low density lipoprotein was demonstrated.     

Use of commercially available monoclonal antibodies for immunoenzyme double staining

Summary An immunoenzyme double-staining method for the simultaneous detection of two cellular epitopes, using commercially available mouse monoclonal antibodies, is described. The method employs a combination of the suppression of endogenous biotin and two successive indirect techniques with a blocking step in between. The first indirect method involves an unlabelled monoclonal antibody followed by an alkaline phosphatase-conjugated goat anti-mouse immunoglobulin. After a blocking step with normal mouse serum, the second indirect method is applied using a biotinylated monoclonal antibody followed by the visualization of this antibody by avidin-biotinylated peroxidase complex (ABC) or rabbit anti-biotin and peroxidase-conjugated swine anti-rabbit immunoglobulin in successive steps. Using these methods in combination with the introduction of dioctyl sodium sulphosuccinate and tetramethylbenzidine as chromogens for peroxidase activity, two cellular epitopes could be distinguished clearly in tissue sections by the green-and violet-stained peroxidase and alkaline phosphatase activities, respectively. The expression of two epitopes on the same cellular constituent is outlined by the coappearance of both enzyme activities as a bluish-purple colour. This method allows for the simultaneous identification, localization and enumeration of two cellular epitopes. These can serve as parameters for a number of pathological processes.     

Detection of methicillin-resistant Staphylococcus pseudintermedius with commercially available selective media

Aims: Commercially available selective media for methicillin‐resistant Staphylococcus aureus (MRSA) were tested for the detection and isolation of methicillin‐resistant Staphylococcus pseudintermedius (MRSP). Methods and Results: Five different screening agars [mannitol salt agar with oxacillin and BD BBL? Chromagar? MRSA (BD Diagnostics); chromID? MRSA agar (bioMérieux); Oxacillin resistance screening agar base (ORSAB); and Brilliance MRSA agar (Oxoid)] were analysed for the detection of MRSP. Bacteria that may be isolated together with MRSP and may grow on the screening agars were included in the study to determine possible interference with the growth of MRSP. MRSP grew well on all selective media except on BD BBL? Chromagar? MRSA (BD Diagnostics) and chromID? MRSA agar (bioMérieux), on which a low to moderate growth rate was noted. Conclusions: ORSAB (Oxoid) and Brilliance MRSA agar (Oxoid) are most suitable for the detection and isolation of MRSP from clinical material. Significance and Impact of the Study: The importance of MRSP in veterinary medicine is increasing. Diagnostic systems are needed to detect MRSP carrier as soon as possible. This study provides information about selected MRSA screening agars for the detection of MRSP to the clinical microbiologists.     

Comparison of commercially available kits with standard methods for detection of Salmonella strains in foods.

Six commercial kits were compared with the U.S. Food and Drug Administration (USFDA) method and the Japanese standard method for Salmonella isolation in foods. When only Salmonella serovars were tested, many of the methods performed well; however, when foods were artificially inoculated, only the USFDA method and immunomagnetic separation coupled with the xylose-lysine-brilliant green agar method (MS-XLBG) could positively detect Salmonella serovars. All seven wild-type Salmonella serovars were detected by the USFDA method, and the MS-XLBG method detected salmonellae from six samples.     

Improved HPLC method for total plasma homocysteine detection and quantification

Recent clinical research has pointed at hyperhomocysteinemia as an independent risk factor in a number of cardiovascular and neurological diseases. We have improved a chromatographic method of total plasma homocysteine measurements in order to obtain higher sensitivity, reliability and reproducibility. The method demonstrates excellent linearity (R=0.999), range (<2-100 microM), precision (instrumental RSD 0.06 and method RSD 1.17), accuracy (recovery of 99.92 and RSD 1.27), reproducibility, quantification limit and ruggedness (e.g. pH from 2.0 to 2.5). Because even a small increase in homocysteine level can be a significant risk factor of cardiovascular diseases, such a precise method is required. The constructed method allows the measurement of plasma pyridoxal phosphate, PLP, the co-enzyme form of vitamin B(6), on the same column and similar reagents. The developed method has been successfully applied to measure both total plasma and serum homocysteine in a group of acute stroke patients.     

Comparison of commercially available kits with standard methods for the detection of coliforms and Escherichia coli in foods.

Three commercially available kits that were supplemented with substrates for enzyme reactions were evaluated to determine their abilities to detect coliforms and fecal coliforms in foods. Japanese and U.S. Food and Drug Administration standard methods, as well as two agar plate methods, were compared with the three commercial kits. A total of 50 food samples from various retailers were examined. The levels of detection of coliforms were high with the commercial kits (78 to 98%) compared with the levels of detection with the standard methods (80 to 83%) and the agar plate methods (56 to 83%). Among the kits tested, the Colilert kit had highest level of recovery of coliforms (98%), and the level of recovery of Escherichia coli as determined by beta-glucuronidase activity with the Colilert kit (83%) was comparable to the level of recovery obtained by the U.S. Food and Drug Administration method (87%). Isolation of E. coli on the basis of the beta-glucuronidase enzyme reaction was found to be good. Levine's eosine methylene blue agar, which has been widely used in various laboratories to isolate E. coli was compared with 4-methylumbelliferyl-beta-D-glucuronide (MUG)-supplemented agar for isolation of E. coli. Only 47% of the E. coli was detected when eosine methylene blue agar was used; however, when violet red bile (VRB)-MUG agar was used, the E. coli detection rate was twice as high. Of the 200 E. coli strains isolated, only 2 were found to be MUG negative, and the gene responsible for beta-glucuronidase activity (uidA gene) was detected by the PCR method in these 2 strains. Of the 90 false-positive strains isolated that exhibited various E. coli characteristic features, only 2 non-E.coli strains hydrolyzed MUG and produced fluorescent substrate in VRB-MUG agar. However, the PCR did not amplify uidA gene products in these VRB-MUG fluorescence-positive strains.     

Compressive fatigue properties of a commercially available acrylic bone cement for vertebroplasty

Acrylic bone cements are widely used for fixation of joint prostheses as well as for vertebral body augmentation procedures of vertebroplasty and balloon kyphoplasty, with the cement zone(s) being subjected to repeated mechanical loading in each of these applications. Although, in vertebroplasty and balloon kyphoplasty, the cement zone is exposed to mainly cyclical compressive load, the compressive fatigue properties of acrylic bone cements used in these procedures are yet to be determined. The purposes of the present study were to determine the compressive fatigue properties of a commercially available cement brand used in vertebroplasty, including the effect of frequency on these properties; to identify the cement failure modes under compressive cyclical load; and to introduce a screening method that may be used to shorten the lengthy character of the standardized fatigue tests. Osteopal \({^\circledR } \mathrm{V}\) was used as the model cement in this study. The combinations of maximum stress and frequency used were 50.0, 55.0, 60.0, 62.5 and 75.5 MPa at 2 Hz; and of 40.0, 55.0, 60.0, 62.5 or 75.5 MPa at 10 Hz. Through analysis of nominal strain-number of loading cycles results, three cement failure modes were identified. The estimated mean fatigue limit at 2 Hz (55.4 MPa) was significantly higher than that at 10 Hz (41.1 MPa). The estimated fatigue limit at 2 Hz is much higher than stresses commonly found in the spine and also higher than that for other acrylic bone cements tested in a full tension–compression fatigue test, which indicates that tension–compression fatigue testing may substantially underestimate the performance of cements intended for vertebroplasty. A screening method was introduced which may be used to shorten the time spent in performing compressive fatigue tests on specimens of acrylic bone cement for use in vertebral body augmentation procedures.     

A latex agglutination test for the detection of Mycoplasma pneumoniae in respiratory exudates: a comparative study with a commercially available DNA-probe test.

We prepared polyclonal antibody specific to Mycoplasma pneumoniae. Using this antibody, we developed a latex agglutination test (LAT) for detecting the organism in respiratory exudates as rapid diagnosis of M. pneumoniae infection. Further, LAT was compared with DNA-probe test (DP) which was the only commercially available test for the rapid detection of the organism. In LAT, both M. pneumoniae and M. genitalium give positive agglutination, but the titer of M. genitalium was significantly lower than that of M. pneumoniae. The detection limit of LAT was 2 x 10(5) CFU/ml and that of DP was 5 x 10(4) CFU/ml in vitro. It was considered that target molecules in LAT were accumulated in the pharyngeal portion of the patients, because of their long half-life at 37 C. However, ribosomal RNA which was target molecule in DP was destroyed at 37 C much sooner, and the accumulation could not be expected. Actually, positive rate in LAT was higher than that in DP among clinical specimens in which M. pneumoniae was detected by culture method. The procedure of LAT is much easier and more rapid than that of DP in which radioactive isotope is required. LAT could be the choice of test for rapid diagnosis of M. pneumoniae infection.     

A novel resolution of a pharmaceutically important bridged bicyclic ketone intermediate via selective enzymatic reduction with a commercially available ketoreductase

An efficient route to the pharmaceutically important (6S,9R)-11-oxo-5,6,7,8,9,10-hexahydro-6,9-methanobenzocyclooctene intermediate has been demonstrated via kinetic resolution of 11-oxo-5,6,7,8,9,10-hexahydro-6,9-methanobenzocyclooctene using a commercially available ketoreductase. Biocatalytics KRED 101 has been shown to selectively reduce the (6R,9S) enantiomer leaving behind the desired (6S,9R) enantiomer. This novel reaction is the first demonstration of a high yielding (44% versus 50% maximum theoretical yield) highly stereoselective (>99% ee) resolution of a bicyclic ketone, via enzymatic reduction using a commercially available ketoreductase, where the stereochemistry of the substrate is determined by a bridged ring system. Several challenges were overcome, including enhancing the selectivity of the enzyme by controlling temperature and increasing substrate solubility by employing a combination of cyclodextrin and organic co-solvent in the aqueous reaction system.     

Evaluation of a commercially available beneficial insect habitat for management of lepidoptera pests

A field study was conducted in 2003 and 2004 at the Center for Environmental Farming Systems in Goldsboro, NC, to evaluate the effectiveness of a commercially available beneficial insect habitat in decreasing pest caterpillar populations in organically managed tomato, Lycopersicon esculentum Mill., plots. Six pairs of tomato plots were established and a commercial beneficial insect habitat seed mix (Peaceful Valley's Good Bug Blend) transplanted around the perimeter of treatment plots, whereas a brown-top millet, Brachiaria ramose (L.) Stapf., border was planted around control plots. Egg predation, egg parasitism by trichogrammatid wasps, and larval parasitism by braconid wasps was monitored throughout the growing season to determine whether habitat increased their activity. In both years of this study, the density of Helicoverpa zea (Boddie) and Manduca spp. eggs was not significantly different between treatment and control plots. Although parasitism was the most important component of egg mortality (19-49%), parasitism was not significantly different between habitat types. Identifiable predation was a minor component (3-9%) of egg fate; it is possible that unidentified predation may be part of the approximately 35-52% of eggs that met unknown fates. Larval parasitism levels ranged from approximately 10 to 90% but was not significantly influenced by the presence of beneficial insect habitat in either year of the study. These results demonstrate that natural enemy activity in organic tomatoes was not amplified, and pest populations were not reduced by the presence of a commercially available beneficial insect habitat.     

Synthesized effective atomic numbers for commercially available dual-energy CT

PurposeThe objective of this study was to assess synthesized effective atomic number (Z_eff) values with a new developed tissue characteristic phantom and contrast material of varying iodine concentrations using single-source fast kilovoltage switching dual-energy CT (DECT) scanner.MethodsA newly developed multi energy tissue characterisation CT phantom and an acrylic phantom with various iodine concentrations of were scanned using single-source fast kilovoltage switching DECT (GE-DECT) scanner. The difference between the measured and theoretical values of Z_eff were evaluated. Additionally, the difference and coefficient of variation (CV) values of the theoretical and measured values were compared with values obtained with the Canon-DECT scanner that was analysed in our previous study.ResultsThe average Z_eff difference in the Multi-energy phantom was within 4.5%. The average difference of the theoretical and measured Z_eff values for the acrylic phantom with variation of iodine concentration was within 3.3%. Compared to the results for the single-source Canon-DECT scanner used in our previous study, the average difference and CV of the theoretical and measured Z_eff values obtained with the GE-DECT scanner were markedly smaller.ConclusionsThe accuracy of the synthesized Z_eff values with GE-DECT had a good agreement with the theoretical Z_eff values for the Multi-Energy phantom. The GE-DECT could reduce the noise and the accuracy of the Z_eff values than that with Canon-DECT for the varying iodine concentrations of contrast medium.Advances in knowledgeThe accuracy and precision of the Z_eff values of the contrast medium with the GE-DECT could be sufficient with human equivalent materials.     

Pregnancy diagnosis in wild canids using a commercially available relaxin assay

Accurate detection of pregnancy is a useful tool in zoo management and husbandry, conservation breeding programs and research settings. Our study evaluated the ability of a commercial relaxin hormone assay used in domestic dogs (ReproCHEK^TM) to accurately detect pregnancy through plasma analysis in two wolf and two fox species. The relaxin assay detected all of the pregnancies greater than 25 days gestation for island foxes, fennec foxes, gray wolves, and Mexican gray wolves. For island foxes, three negative relaxin results were attributed to using the test earlier postconception than manufacturer recommendation (before day 20). Five other negative results were found for females estimated at 15–25 days gestation, spanning the early and intermediate period (21–30 days gestation) when relaxin may be detected but less reliably. There were no false‐positive results in nonmated negative control animals. Relaxin assay results were highly correlated with ultrasound results and the intra‐assay replicate agreement was 100%. Our results show that the ReproCHEK^TM commercial relaxin assay is a minimally invasive and reliable method for pregnancy detection in these wild species when used after 25 days gestation and might be applied to other canids as well. Furthermore, this assay is easy to run and requires no specialized equipment, making it extremely useful for zoo and field research applications. Zoo Biol 27:406–413, 2008. © 2008 Wiley‐Liss, Inc.     

Mutagenicity of some commercially available nitro compounds for Salmonella typhimurium

Benzoyl chloride and 53 commercially available aromatic, heterocyclic and aliphatic nitro compounds were tested for mutagenicity in Salmonella typhimurium TA98 and TA100. 34 of 53 nitro compounds (64%) were mutagenic, 4 in TA100 only, 15 in TA98 only, and 15 in both strains. 13 of the heterocyclic derivatives of pyridine, indole, indazole, quinoline, and benzimidazole were mutagenic. 21 of 34 mutagenic nitro compounds were bactericidal. Nitromethane was the only aliphatic tested and was not mutagenic. Benzoyl chloride, a human carcinogen, was mutagenic for TA98.     

WNT unrelated activities in commercially available preparations of recombinant WNT3a

WNT signaling pathways play an important role in both development and disease. By analyzing the signaling capabilities of commercially available WNT3a preparations towards the PI3K/AKT/GSK3 signaling pathway, we discovered unexpected inconsistencies from lot to lot of recombinant WNT3a. We provide evidence that: (1) The ability to trigger AKT/GSK3 signaling varies dramatically between different lots of WNT3a, without any variation in their ability to activate the canonical WNT/β‐catenin signaling. (2) sFRP1, a WNT signaling inhibitor, is unable to interfere with the activation of AKT/GSK3 signaling induced by some of the WNT3a lots. (3) Pharmacological inhibition of AKT/GSK3 phosphorylation by PI3K inhibitors fails to affect the stabilization of β‐catenin, the central effector of the canonical WNT/β‐catenin signaling pathway. In summary, while all tested lots of recombinant WNT3a activated WNT/β‐catenin pathway, our results suggest that individual lots of recombinant WNT3a activate the PI3K/AKT/GSK3 pathway in a WNT‐independent manner, hampering thus the analysis of regulation of PI3K/AKT/GSK3 by WNT ligand. J. Cell. Biochem. 111: 1077–1079, 2010. © 2010 Wiley‐Liss, Inc.     

Evaluation of a commercially available automated sleep staging tool in healthy infants

Sleep and Biological Rhythms - The ALICE5 software package provides a commercially available automated sleep staging system designed for infants. This study aims to evaluate the accuracy of this...