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Optical Density (optical + density)
Kinds of Optical Density Terms modified by Optical Density Selected AbstractsElectrochromic Films Composed of MnO2 Nanosheets with Controlled Optical Density and High Coloration Efficiency.CHEMINFORM, Issue 11 2006Nobuyuki Sakai Abstract ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 200 leading journals. To access a ChemInform Abstract, please click on HTML or PDF. [source] Spectral Imaging Microscopy Can Define Hyperchromasia in Small Lymphoid CellsJOURNAL OF CUTANEOUS PATHOLOGY, Issue 1 2005N.S. McNutt Spectral Imaging Microscopy (SIM) is a technique for performing image-enabled spectroscopic analysis using a standard light microscope. We employed a prototype to a current, commercially available, spectral imaging system (NuanceTM, CRI, Inc.) that electro-optically selects narrow bands of light to be collected by a cooled, monochrome CCD camera. For each sample, images were taken every 10 nm from 420 nm to 720 nm to create a spectral image "stack." Analysis of the images with a prototype to the Nuance TM Analyze software (CRI, Inc.) gave both standard color (RGB) and spectral information at each pixel of the images. A representative 40x field was spectrally imaged from H&E slides of 104 biopsy specimens of mycosis fungoides (MF). Optical densities and spectral properties of small lymphocytes were determined. Hyperchromatic cells were pseudocolored to show their distribution. In addition to the important architectural features of MF, SIM can define whether hyperchromatic or normochromatic small lymphocytes are epidermotropic in the lesion by setting appropriate threshold values. Hyperchromasia can be attributed to increased nuclear eosinophilia or basophilia. These differences are difficult to estimate by eye. Since lymphoblasts have enlarged nuclei with dispersed chromatin, a combination of spectral and spatial data is required for their analysis. [source] An in-vitro investigation of the antibacterial effect of nisin in root canals and canal wall radicular dentineINTERNATIONAL ENDODONTIC JOURNAL, Issue 10 2004S. R. Turner Abstract Aim, To determine whether nisin, a bacteriocin, would be effective at killing Enterococcus faecalis and Streptococcus gordonii cells in solution and within the root canal system. Methodology, Bacterial isolates of E. faecalis and S. gordonii were grown from glycerol stocks in closed tubes containing BHY broth at 37 °C. The minimum bactericidal concentration (MBC) of nisin for both bacterial species was determined by a microdilution method. Extracted human teeth were decoronated to produce roots of equal length with a single canal and divided into six groups of 10 roots. The canals were prepared to a master apical size 30 file using 0.04 taper Ni-Ti rotary instruments. Bacterial samples of each species were inoculated into three groups of prepared roots and incubated in closed tubes at 37 °C for 21 days. The root canals in each group were then medicated with water (control), calcium hydroxide powder mixed with sterile water [Ca(OH)2], or nisin and incubated for a further 7 days. Rotary Ni-Ti files were used to take radicular dentine samples from the walls of each canal which were then incubated in BHY broth for 24 h. Optical density (OD600) readings were taken as a measure of bacterial growth. Results, The MBC of nisin for E. faecalis and S. gordonii was 70 and 20 mg mL,1 respectively. Calcium hydroxide and nisin medication eradicated infection within the root canal while cells remained viable in the control group. Mean optical density (OD600) readings from canal wall dentine shavings infected with E. faecalis were 1.32 ± 0.98, 0.73 ± 0.27 and 0.69 ± 0.38 for the control, Ca(OH)2 and nisin samples respectively. Corresponding mean readings for S. gordonii were 1.19 ± 0.18, 0.73 ± 0.15 and 0.60 ± 0.29. The Ca(OH)2 and nisin group readings were significantly (P < 0.01) lower than the control for each species as tested by Student's t -test and Mann,Whitney U statistical analysis. Values for Ca(OH)2 and nisin were not significantly (P > 0.01) different. Conclusion, Nisin was effective at eradicating E. faecalis and S. gordonii cells in pure culture and was comparable with Ca(OH)2 in the elimination of these species from within the root canal system. [source] Measurement of gp210 autoantibodies in sera of patients with primary biliary cirrhosisJOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 4 2007Alicja Bauer Abstract Primary biliary cirrhosis (PBC) is an autoimmune liver disease with unknown etiology. Patients with PBC have antimitochondrial autoantibodies (AMA) and additionally 50% of them have antinuclear antibodies (ANA). A 15,amino acid fragment (DRKASPPSGLWSPAY) from the C-terminal part of the nuclear envelope glycoprotein gp210 has been proposed as one of the antigenic targets for ANA. The aim of this work was to develop an immunoenzymatic assay for determination of gp210 autoantibodies using for its binding a synthetic pentadecapeptide derived from the gp210 amino acid sequence and to determine level of these autoantibodies in sera of patients with PBC and other autoimmune liver diseases from Poland. Polystyrene microtitration plates coated with the synthetic peptide were consecutively incubated with diluted sera, anti-human immunoglobulin G (IgG) antibodies conjugated with horseradish peroxidase, and with tetramethylobenzidine. Optical density (OD) was read at 450 nm. The mean values of the intra- and interassay of variation coefficients of the test were 4.1 and 10.2%, respectively. Anti-gp210 was detected in 44% of PBC patients and in 6% of patients with PSC. The results were negative for healthy blood donors and other controls. The specificity of the test was 99%, so the anti-gp-210 autoantibodies estimated on DRKASPPSGLWSPAY can be a reliable marker of PBC. J. Clin. Lab. Anal. 21:227,231, 2007. © 2007 Wiley-Liss, Inc. [source] Hen Egg Yolk Prevents Bacterial Adherence: A Novel Function for a Familiar FoodJOURNAL OF FOOD SCIENCE, Issue 1 2001T. Deignan ABSTRACT: The aim of this study was to evaluate the ability of unfractionated hen egg yolk to prevent the attachment of Salmonella typhimurium to murine intestinal epithelial cells in vitro. Inhibition of adherence was examined microscopically and by an ELISA. Both methods showed that egg yolk from immunized and unimmunized hens significantly reduced bacterial adherence. Optical density (OD) readings ± SD of 0.825 ± 0.016 for untreated bacteria were reduced to 0.335 ± 0.024 and 0.267 ± 0.021 for bacteria pretreated with egg yolk from immunized hens and unimmunized hens, respectively. Microscopic analysis showed that egg yolk from either immunized or unimmunized hens reduced bacterial adherence from 17 ± 2.7 bacteria per epithelial cell to less than 4 bacteria per epithelial cell. These results show that hen egg yolk significantly inhibits adherence of S. typhimurium to intestinal epithelial cells in vitro. Boosting of specific antibody levels does not enhance this antiadhesive effect, suggesting that egg yolk contains novel antiadhesive factors. [source] Habitual use of the primate forelimb is reflected in the material properties of subchondral bone in the distal radiusJOURNAL OF ANATOMY, Issue 6 2006Kristian J. Carlson Abstract Bone mineral density is directly proportional to compressive strength, which affords an opportunity to estimate in vivo joint load history from the subchondral cortical plate of articular surfaces in isolated skeletal elements. Subchondral bone experiencing greater compressive loads should be of relatively greater density than subchondral bone experiencing less compressive loading. Distribution of the densest areas, either concentrated or diffuse, also may be influenced by the extent of habitual compressive loading. We evaluated subchondral bone in the distal radius of several primates whose locomotion could be characterized in one of three general ways (quadrupedal, suspensory or bipedal), each exemplifying a different manner of habitual forelimb loading (i.e. compression, tension or non-weight-bearing, respectively). We employed computed tomography osteoabsorptiometry (CT-OAM) to acquire optical densities from which false-colour maps were constructed. The false-colour maps were used to evaluate patterns in subchondral density (i.e. apparent density). Suspensory apes and bipedal humans had both smaller percentage areas and less well-defined concentrations of regions of high apparent density relative to quadrupedal primates. Quadrupedal primates exhibited a positive allometric effect of articular surface size on high-density area, whereas suspensory primates exhibited an isometric effect and bipedal humans exhibited no significant relationship between the two. A significant difference between groups characterized by predominantly compressive forelimb loading regimes vs. tensile or non-weight-bearing regimes indicates that subchondral apparent density in the distal radial articular surface distinguishes modes of habitually supporting of body mass. [source] Evaluation of hepatitis C antibody testing in saliva specimens collected by two different systems in comparison with HCV antibody and HCV rna in serum,JOURNAL OF MEDICAL VIROLOGY, Issue 1 2001G.J.J. van Doornum Abstract Two different ELISA assays, the Ortho HCV 3.0 ELISA (Ortho Diagnostics Systems) and the Mono-Lisa anti-HCV Plus (Sanofi Diagnostics Pasteur) were evaluated for the detection of hepatitis C virus (HCV) antibody in saliva samples. Specimens were collected from 152 individuals who participated in a longitudinal cohort study on HIV infection, and who used illicit drugs. Saliva specimens were collected using two different systems: Salivette (Sarstedt) and Omni-Sal (Saliva Diagnostic Systems). Saliva specimens were tested following modified protocols by both ELISAs, and the results were compared with serum specimens that were tested according to the instructions of the manufacturer. Serum samples of 102 (67%) participants were positive by both assays, and 50 persons were negative for HCV antibody. A total of 99 of the 102 serum specimens were confirmed as positive using Ortho Riba HCV 3.0 (Ortho Diagnostics System) and Deciscan HCV (Sanofi Diagnostics Pasteur), and 3 yielded discrepant results. As no cut-off level is known for testing saliva samples by ELISA, 3 different levels were chosen: mean (M),+,1 standard deviation (SD), M,+,2 SD, and M,+,3 SD of the optical densities of saliva tests of the 50 HCV serum antibody negative persons. At a level of M,+,1 SD and M,+,2 SD the Salivette/Mono-Lisa combination gave the greatest proportion of HCV antibody positive saliva specimens obtained from the 102 HCV serum antibody positive participants, 88% and 79%, respectively. Differences between the various collection systems and assay combinations were not significant statistically. In 76 of the 102 persons with HCV antibodies in serum, HCV RNA was detected in serum. Salivary presence of HCV RNA, however, could not be demonstrated. The results show that the assays compared are unsuitable for diagnostic use, but the sensitivities of the assays are acceptable for use in epidemiological studies. J. Med. Virol. 64:13,20, 2001. © 2001 Wiley-Liss, Inc. [source] Production of Shiga toxin by Shiga toxin-expressing Escherichia coli (STEC) in broth media: from divergence to definitionLETTERS IN APPLIED MICROBIOLOGY, Issue 4 2007L.B. Rocha Abstract Aims:, To determine the suitability of eight different commercial broth media for Shiga toxin (Stx) production. Methods and Results:, Shiga toxin-producing Escherichia coli (STEC) strains producing Stx1 or Stx2 were grown at 37°C (250 rev min,1) for 24 h in brain heart infusion broth, E. coli broth, Evans medium, Luria-Bertani broth, Penassay broth, buffered-peptone water, syncase broth and trypticase soy broth. Toxin production was measured by enzyme-linked immunosorbent assay (ELISA) in polymyxin-treated cell pellets and/or supernatants of cultures, ELISA optical densities reached 1 when isolates were grown for 2,4 h in E. coli broth in the presence of antibiotic. Besides, a collection of STEC-expressing Stx strains was evaluated and the Stx production was assayed in the supernatants and in polymyxin-treated pellets of bacterial growth after 4 h of cultivation in E. coli broth in the presence of antibiotic. Conclusions:, The most suitable medium for Stx production was E. coli broth when the bacterial isolates were grown for 4 h in the presence of ciprofloxacin and the Stx production is detected in the supernatant. Significance and Impact of the Study:, This study presents the first comprehensive comparison of different broth media with regard to Stx production to establish optimal culture conditions for STEC detection in routine diagnostic laboratories. [source] Endotoxin Level Measurement in Hemodialysis Biofilm Using "The Whole Blood Assay"ARTIFICIAL ORGANS, Issue 6 2005Karine Marion-Ferey Abstract:, Biofilms have been found on the inner surface of silicone tubing inside dialysis machines. Endotoxin releasing from those biofilms increases the bioincompatibility of dialysis liquids and leads to long-term inflammatory complications among dialysis patients. Endotoxin measurement is recommended for the control of dialysis liquids. This article describes the use of a new method, the Whole Blood Assay (WBA), for endotoxin quantification in dialysis biofilms. Biofilms were suspended in sterile water by scraping the tubing samples. Diluted blood samples from healthy donors were stimulated overnight with the contaminated suspension. Stimulated mononuclear cells released IL-1, in response to endotoxins. IL-1, level was then measured using an ultrasensitive ELISA method. We demonstrated a semilogarithmic model in which the optical densities measured after the ELISA assay increases linearly with the levels of endotoxin. This model allowed the determination of the amount of endotoxins in biofilm samples with a detection limit of 0.032 EU/mL. Most of the time, the amounts of endotoxin measured by the WBA were higher than those measured by the Limulus Amoebocyte Lysate (LAL) assay. This study suggested the presence of "endotoxin-like" compounds different from the lipopolysaccharides that are not detected by the LAL assay. We concluded that the LAL is necessary but insufficient to have a representative quantification of endotoxins that could be hazardous to patient health. [source] 3123: Non-invasive measurement of retinal oxygenation: principles and expectationsACTA OPHTHALMOLOGICA, Issue 2010M HAMMER Purpose To determine oxygen saturation (SO2) of blood inside retinal vessels which is an essential measure for the estimation of oxygen supply to the tissue as well as its oxygen consumption. Methods Two-, four-, and multiple - wavelength approaches to the non-invasive measurement of SO2 will be discussed. The dual wavelength technique, imaging the fundus at 548 and 610 nm, showed to be most appropriate for clinical routine investigations. The SO2 of the hemoglobin in retinal arterioles and venules is calculated from the ratio of the optical densities of the vessels at both wavelengths. Results From a healthy control population, mean arterial and venous SO2 were measured to be 98±10.1% and 65±11.7% with reproducibility of 2.52% and 3.25% respectively. In a cohort of 41 patients (mean age: 65±12.3 years) with diabetic retinopathy (DR), we found an increase of the venous SO2 with the severity of DR: Mild non-proliferative DR 69±7%, moderate non-proliferative DR 70±5%, severe non-proliferative DR, 75±5%, and proliferative DR 75±8%. Measurements of SO2 in accordance with vessel diameters revealed a correlation of the venous SO2 with arterial as well as venous diameters in 159 diabetic patients (mean age: 55.8±13.9 years) with no or non , proliferative DR. Increased venous SO2 is an indicator of insufficient oxygen supply to the retinal tissue. The correlation of the vessel diameters with venous SO2 may point to compensatory mechanisms of retinal blood flow regulation. Conclusion Accurate retinal vessel oximetry is possible by non , invasive optical methods. Combined with measurements characterising the retinal blood flow, it is a powerful tool for the estimation of retinal oxygen supply and consumption. Commercial interest [source] High-frequency stimuli preferentially release large dense-core vesicles located in the proximity of nonspecialized zones of the presynaptic membrane in sympathetic gangliaDEVELOPMENTAL NEUROBIOLOGY, Issue 4 2008F. Cifuentes Abstract We characterized the effect of a brief high-frequency stimulus on the number, distribution, and optical density of large dense-core vesicles (LDCVs) in the nerve terminals of the rat superior cervical ganglia. From 4.21 ± 0.37 LDCVs/bouton detected in control nerve terminals, a stimulus of 40 Hz for 1 min released 41% of LDCVs, decreasing their number to 2.48 ± 0.14 LDCVs/bouton (p = 0.0009). In control ganglia, most dense vesicles were located close to the plasma membrane (at ,100 nm); in contrast, in stimulated ganglia they were broadly distributed with respect to the active zone. The mean distance of LDCVs to membrane and active zones was 95 ± 8 nm and 473 ± 15 nm, respectively. The analysis of the core density showed that both groups had a similar asymmetric distribution with the same average. Stimulation preferentially released those vesicles located ,100 nm from the plasma membrane that had no apparent relationship with the active zone. After the stimulus, the average distances of LDCVs to the plasma membrane and active zone did not change, suggesting that the stimulus also caused the relocation of inner LDCVs. Interestingly, optical density analysis showed that the released vesicles had low range densities, and suggested that LDCVs release their entire content. We conclude that LDCV exocytosis mainly involves those vesicles located ,100 nm from the plasma membrane and occurs in regions of synaptic boutons presumed to be nonspecialized. These results agree with the characteristics of the classical model that proposes full content release. © 2008 Wiley Periodicals, Inc. Develop Neurobiol, 2008. [source] Recruitment pattern of muscle fibre type during flat and sloped treadmill running in Thoroughbred horsesEQUINE VETERINARY JOURNAL, Issue S36 2006D. ETO Summary Reasons for performing the study: There is little information about the muscle fibre recruitment pattern during sloped and flat track running in Thoroughbred horses. Objectives: To examine the glycogen depletion pattern of each muscle fibre type during running on a flat and sloped treadmill. Methods: Thirteen Thoroughbred horses (3,9 years old) were used. They were initially subjected to incremental exercise tests on a treadmill at 10 and 0% inclines in each horse to determine running speed at 90 and 60% VO2max. Needle biopsy samples were obtained from the middle gluteal muscle immediately after the running at 90% VO2max for 4 min and 60% VO2max for 12 min on 10% and 0% inclines treadmill. Four muscle fibre types (Types I, IIA, IIA/IIX, and IIX) were immunohistochemically identified, and optical density of Periodic Acid Schiff staining (OD-PAS) in each fibre type and the glycogen content of the muscle sample were determined by quantitative histochemical and biochemical procedures. Results: The changes in OD-PAS showed that the recruitment of all fibre types were identical after each exercise bout, i.e., 4 min running at 90% VO2max (8.4,9.4 m/sec on 10%, 13.9,14.1 m/sec on 0%), and 12 min running at 60% VO2max (5.4,6.0 m/sec on 10%, 7.9,11.2 m/sec on 0%). No significant differences were found in the recruitment patterns of each muscle fibre type between 10 and 0% inclined exercise bouts at the same exercise intensity. Conclusions: The recruitment pattern of muscle fibre type is mainly determined by exercise intensity (%VO2max) and duration, but not by running speed. Potential relevance: The results of this study indicate the possibility that up-hill running results in the same training effect as faster running on a flat track. [source] Surface podzolization in Cambisols under deciduous forest in the Belgian loess beltEUROPEAN JOURNAL OF SOIL SCIENCE, Issue 1 2000V. Brahy Summary Surface podzolization involves the migration of metal,humus complexes to a depth of a few centimetres. In acid soils derived from loess, this process has been diagnosed mainly by morphological observation. We investigated this process in a toposequence of Luvisols and Cambisols on loess using selective extraction and mineralogical data as well as characteristics of the leaf litter. The humus type (O and OAh horizons) is a moder in the three Luvisols and one of the Cambisols, whereas it is a fibrimor in the two other Cambisols. The contents in total alkaline and alkaline-earth cations range from 35 to 60 cmolc kg,1 in the fibrimor and from 40 to 90 cmolc kg,1 in the moder humus. In the two Cambisols with fibrimor smectite occurs in the clay fraction of the Ah horizon; Fe,humus complexes seem to have moved, but no more than 9 cm, from the Ah to the AB horizon beneath. Relative to the Ah horizon, the upper part of the AB has larger tetraborate-extractable Fe/Al ratio and optical density of the oxalate extract. Such features converge to diagnose surface podzolization in the Cambisols with fibrimor. However, they were not detected in the Cambisol and Luvisols with moder. In the two Cambisols with fibrimor, surface podzolization is consistent with (i) their smaller iron content, (ii) their more advanced weathering stage and (iii) their lower acid neutralizing capacity. [source] Immunological quantification of the nematode parasitic bacterium Pasteuria penetrans in soilFEMS MICROBIOLOGY ECOLOGY, Issue 3 2001S Fould Abstract Currently, the abundance of Pasteuria penetrans in soils, an unculturable bacterial parasite of root-knot nematodes (Meloidogyne spp.), is estimated by the percentage of nematode juveniles infected with bacteria and the number of spores attached to their cuticle. Indirect immunofluorescence led to detection of free spores directly in soil suspensions using UV light and polyclonal antibodies raised against two P. penetrans populations (ORS-21414-Sen and PP1). Three extraction methods were compared in order to improve spore recovery. A gentle shaking/sieving method recovered more than 90% of the spores inoculated in soils and was more efficient and simple than aqueous two-phase partitioning and polyethylene glycol extractions. All the spores inoculated in sandy or sandy,clay soils were detected with immunofluorescence microscopy. The quantification of the spores was improved using an ELISA technique that showed a good correlation between optical density and spore concentration in inoculated soils. Specific antibodies provide a suitable method to quantify P. penetrans and may be used to follow the evolution of the real pool of bacteria either in native suppressive soils or in inoculated ones. [source] An in-vitro investigation of the antibacterial effect of nisin in root canals and canal wall radicular dentineINTERNATIONAL ENDODONTIC JOURNAL, Issue 10 2004S. R. Turner Abstract Aim, To determine whether nisin, a bacteriocin, would be effective at killing Enterococcus faecalis and Streptococcus gordonii cells in solution and within the root canal system. Methodology, Bacterial isolates of E. faecalis and S. gordonii were grown from glycerol stocks in closed tubes containing BHY broth at 37 °C. The minimum bactericidal concentration (MBC) of nisin for both bacterial species was determined by a microdilution method. Extracted human teeth were decoronated to produce roots of equal length with a single canal and divided into six groups of 10 roots. The canals were prepared to a master apical size 30 file using 0.04 taper Ni-Ti rotary instruments. Bacterial samples of each species were inoculated into three groups of prepared roots and incubated in closed tubes at 37 °C for 21 days. The root canals in each group were then medicated with water (control), calcium hydroxide powder mixed with sterile water [Ca(OH)2], or nisin and incubated for a further 7 days. Rotary Ni-Ti files were used to take radicular dentine samples from the walls of each canal which were then incubated in BHY broth for 24 h. Optical density (OD600) readings were taken as a measure of bacterial growth. Results, The MBC of nisin for E. faecalis and S. gordonii was 70 and 20 mg mL,1 respectively. Calcium hydroxide and nisin medication eradicated infection within the root canal while cells remained viable in the control group. Mean optical density (OD600) readings from canal wall dentine shavings infected with E. faecalis were 1.32 ± 0.98, 0.73 ± 0.27 and 0.69 ± 0.38 for the control, Ca(OH)2 and nisin samples respectively. Corresponding mean readings for S. gordonii were 1.19 ± 0.18, 0.73 ± 0.15 and 0.60 ± 0.29. The Ca(OH)2 and nisin group readings were significantly (P < 0.01) lower than the control for each species as tested by Student's t -test and Mann,Whitney U statistical analysis. Values for Ca(OH)2 and nisin were not significantly (P > 0.01) different. Conclusion, Nisin was effective at eradicating E. faecalis and S. gordonii cells in pure culture and was comparable with Ca(OH)2 in the elimination of these species from within the root canal system. [source] Decreasing myelin density reflected increasing white matter pathology in Alzheimer's disease,a neuropathological studyINTERNATIONAL JOURNAL OF GERIATRIC PSYCHIATRY, Issue 10 2005Martin Sjöbeck Abstract Background White matter disease (WMD) is frequently seen in Alzheimer's disease (AD) at neuropathological examination. It is defined as a subtotal tissue loss with a reduction of myelin, axons and oligodendrocytes as well as astrocytosis. Studies quantitatively defining the myelin loss in AD are scarce. The aim was to develop a method that could provide numerical values of myelin density in AD. The purpose was to compare the myelin contents in increasing grades of pathology of WMD, with age and cortical AD pathology as well as in different regions of the brain in AD. Material and methods Sixteen cases with AD and concomitant WMD were investigated with an in-house developed image analysis technique to determine the myelin attenuation with optical density (OD) in frontoparietal, parietal, temporal and occipital white matter on whole brain coronal sections stained for myelin with Luxol Fast Blue (LFB). The OD values in LFB were compared grouped according to Haematoxylin/Eosin (HE) evaluated mild, moderate and severe WMD or normal tissue. The OD values were also correlated with age and cortical AD pathology and compared between the different studied white matter regions. Results Increasing severity of WMD was associated with a statistically significant OD reduction. No correlation was seen between age and OD or overall cortical AD pathology. The OD values were significantly lower in frontoparietal-compared to occipital white matter. Conclusions Myelin loss in AD with WMD is a marked morphologic component of the disease and it is possible to determine the reduction objectively in neuropathological specimens with quantitative measures. This may be of use for clinical diagnostics including brain imaging. Copyright © 2005 John Wiley & Sons, Ltd. [source] Effect of recombinant Rv1009 protein on promoting the growth of Mycobacterium tuberculosisJOURNAL OF APPLIED MICROBIOLOGY, Issue 4 2008X. Wu Abstract Aims:, To determine whether resuscitation-promoting factor (RPF) from Mycobacterium tuberculosis can promote mycobacterial growth and shorten culture time. Method and Results:, We cloned, expressed and purified an RPF from M. tuberculosis, Rv1009 protein and subsequently studied the biological activity of the recombinant Rv1009 (rRv1009) in liquid and on solid media. Our results indicate that the molecular weight of rRv1009 protein expressed in Escherichia coli BL21 was approximately 39 kDa. At picomolar and micromolar concentrations, rRv1009 protein could increase the optical density of freeze-dried Mycobacterium bovis BCG three to fivefold in Middlebrook 7H9 medium, stimulate the growth of viable mycobacteria on solid medium, and shorten positive growth detection time of a small number of M. tuberculosis in BACTEC 960 medium. Conclusions:, The rRv1009 could promote proliferation of mycobacteria. It may be useful for culture of mycobacteria presented in clinical samples. Significance and Impact of the Study:, rRv1009 protein can be used as a growth-promoting reagent of mycobacteria in the medium to shorten the time of culture. [source] In vitro assessment of antimicrobial activity of carvacrol, thymol and cinnamaldehyde towards Salmonella serotype Typhimurium DT104: effects of pig diets and emulsification in hydrocolloidsJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2006W. Si Abstract Aims:, To determine the effect of pig diets in vitro on the antimicrobial activity of carvacrol, thymol and cinnamaldehyde, and to identify an emulsifier/stabilizer that can stabilize the essential oil (EO) components in aqueous solution and retain their antimicrobial activity in the presence of the diets. Methods and Results:, Emulsification of essential oil components with hydrocolloid solution was achieved by blending with a Polytron. Antimicrobial activity was measured through in vitro assays to determine the inhibition of bacterial growth by measuring the optical density at 600 nm or plating on nutrition agar after incubation of the mixtures of an EO component with the culture of Salmonella serotype Typhimurium DT104 in the presence or absence of pig diets. The results generated through the in vitro assays indicated that pig diets were able to abolish the antimicrobial activity of EOs. Xanthan, fenugreek and yellow mustard gums were the best in forming stable emulsions of five different EO components among ten different plant polysaccharides and surfactants examined. Emulsification of all the EO components in the fenugreek gum solution did not alter their antimicrobial activity. However, the antimicrobial activity of geraniol was significantly reduced when emulsified with other polysaccharides and surfactants. Both fenugreek and xanthan gum solutions were unable to protect the antimicrobial activity of carvacrol and thymol when mixed with the diets. Although cinnamaldehyde required no emulsification, but a high concentration (equivalent to at least three times of minimum bactericidal concentration for cinnamon oil) to inhibit Salmonella growth significantly in the presence of the diets, emulsification in fenugreek gum appeared to be essential for cinnamaldehyde solution to retain its antimicrobial activity during storage. Conclusions:, The diets for newly weaned pigs were a significant factor limiting the antimicrobial activity of EOs and their components. Cinnamaldehyde required a high concentration to retain its antimicrobial activity in the diets, in addition to its requirement for emulsification to stabilize its activity during the storage. Significance and Impact of the Study:, The assay with the diets used in this study for measuring the antimicrobial activity can be used in vitro for rapid and effective screening of potential antimicrobials for swine production. This study has identified polysaccharides that are able to stabilize EO component solutions. It has also identified cinnamaldehyde for further in vivo studies that may have potential in future application in controlling Salmonella and possibly other enteric pathogens in swine production. [source] Semiquantitative analysis of urinary low protein levels using silver dot blot assayJOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 4 2001Kazuyuki Matsuda Abstract We designed a semiquantitative analysis of urinary low protein levels using silver dot blot assay. In this method, 3 ,l of urine are blotted to one dot onto a cellulose acetate membrane, which is stained by a colloidal silver staining reagent, and the optical density of the silver stained urinary protein is measured at 500 nm using a densitometer. There was a good linearity between 2.5 mg/L and 100 mg/L and a gentle linearity between 100 mg/L and 200 mg/L, and the minimum sensitivity was 2.5 mg/L. This method is suitable for semiquantitative analysis of urinary protein levels less than 300 mg/L, which can not be determined precisely by dipstick. J. Clin. Lab. Anal. 15:171,174, 2001. © 2001 Wiley-Liss, Inc. [source] A methodology using subjective and objective measures to compare plaque inhibition by toothpastesJOURNAL OF CLINICAL PERIODONTOLOGY, Issue 12 2004N. Claydon Abstract Objectives: Plaque scoring usually employs subjective indices. The aim was to compare plaque inhibition of three toothpastes using two objective and one subjective measures of plaque. Material and Methods: Formulations were: (1) an experimental anti-plaque paste (test); (2) an experimental paste (minus active or negative control); and (3) a proprietary anti-plaque toothpaste product (positive control). The study was a blind, randomised crossover design using a 4-day, no tooth brushing, plaque regrowth model and involving 22 healthy subjects. After baseline plaque removal, subjects rinsed twice a day with slurries of the allocated paste. On day 5, plaque was scored by index, wet weight and optical density of extracted disclosing solution from the plaque (stain intensity). Results: All data showed the same pattern. There were highly significant subject and treatment effects but not period effects. The positive control was highly significantly more effective in plaque control than the test and minus active experimental formulations, which in turn were not significantly different from each other. There were strong and significant correlations between pairs of scoring methods particularly wet weight and stain intensity. Conclusion: The use of objective methods of plaque alongside conventional subjective indices provided convincing evidence for increased discriminatory power in a study comparing plaque inhibition by toothpastes. [source] DESIGN, CONSTRUCTION AND VALIDATION OF A SANITARY GLOVE BOX PACKAGING SYSTEM FOR PRODUCT SHELF-LIFE STUDIESJOURNAL OF FOOD PROCESSING AND PRESERVATION, Issue 3 2001ZEHRA AYHAN A glove box has been constructed as pan of an integrated pilot plant scale pulsed electric field processing and packaging system to facilitate studies of product shelf-life with selected packaging materials. The glove box was sanitized using combination of hydrogen peroxide and germicidal UV light. A HEPA air filter provided positive pressure of bacteria-free air. Nonselective nutrient broth was sterilized and filled into presanitized bottles inside the glove box. Negative and positive controls were included in the experiment. All bottles were incubated at 22C and 37C for two weeks and checked for rnicrobial growth by measuring optical density at 600 nm using a spectrophotometer and by plating on plate count agar and potato dextrose agar for total aerobic and, yeast and mold counts, respectively. No turbidity or microbial growth was observed in the media filled in the sanitized bottles using the sanitized glove box at 22 and 37C. PEF processed orange juice using this system had a shelf-life of more than 16 weeks at 4C. [source] Fermentation Capabilities of Bifidobacteria Using Nondigestible Oligosaccharides, and Their Viability as Probiotics in Commercial Powder Infant FormulaJOURNAL OF FOOD SCIENCE, Issue 6 2005Darío Pérez-Conesa ABSTRACT The species Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum biotype infantis (Spanish type culture collection), and Bifidobacterium longum (Morinaga nutritional foods) were evaluated in vitro in the presence of 4 commercial nondigestible oligosaccharides (NDO) (short-chain fructooligosaccharides [SCFOS] [degree of polymerization, DP: 2,3], inulin [DP: 10,0], oligofructose [DP: 2,0] and 4,-galactosyllactose [4,-GOS] [DP: 3,]). Each species was incubated anaerobically in tryptone phytone yeast (TPY) broth for 7 d with NDO. Every 24 h, bifidobacteria growth was evaluated by means of broth turbidity as optical density at 600 nm. Moreover, another sample was collected for pH culture measurement. Results showed that inulin was the substrate with the least effect on the stimulation of bifidobacteria growth and pH decrease. On the last day of incubation, the substrate 4,-GOS stimulated bacterial growth more strongly and produced a larger decrease in culture broth pH than the other substrates. On the other hand, B. bifidum and B. longum showed a greater growth with 4,-GOS. In a 2nd study, these 2 bifidobacteria species were added to a powder follow-on probiotic infant formula. The viability of the bifidobacteria during the formula's period of consumption was evaluated in 2 studies of 6 and 14 d. Both corresponded to the minimum and maximum time of consumption of the formula according to the manufacturer's directions. It was found that, although in both studies bifidobacteria counts decreased significantly (P < 0.05) with time, they were always above the recommended addition level (106 colony-forming units [CFU]/g) at the time of sale for dairy products by the Intl. Standard of Fédération Internationale de Laiterie/International Dairy Federation (FIL/IDF). Moreover, because the pH of the reconstituted formula was always close to neutrality (from 6.74 to 7.06), the number of bacteria did not drop below the recommended level. [source] Participation of the secreted dipeptidyl and tripeptidyl aminopeptidases in asaccharolytic growth of Porphyromonas gingivalisJOURNAL OF PERIODONTAL RESEARCH, Issue 3 2009H. Oda Background and Objective:,Porphyromonas gingivalis secretes gingipains, endopeptidases essential for the asaccharolytic growth of this bacterium. P. gingivalis also secretes dipeptidyl aminopeptidases (DPPIV and DPP-7) and a tripeptidyl aminopeptidase (PTP-A), although their role in asaccharolytic growth is unclear. The present study was carried out to elucidate the role of these dipeptidyl/tripeptidyl aminopeptidases on the asaccharolytic growth of P. gingivalis. Material and Methods:, Knockout mutants for the DPPIV (dpp), dpp7 and/or PTP-A genes were constructed. Brain,heart infusion medium supplemented with sterile hemin and menadione (BHIHM) was used as a complex medium, and the minimal medium used was GA, in which the sole energy source was a mixture of immunoglobulin G and bovine serum albumin. Growth of P. gingivalis was monitored by measuring the optical density of the culture. Results:, All knockout mutants for DPPIV, dpp7 and PTP-A grew as well as strain W83 in BHIHM. In GA, growth of single-knockout and double-knockout mutants was similar to that of W83, whereas growth of a triple-knockout mutant (83-47A) was reduced. We purified recombinant DPPIV and recombinant PTP-A from recombinant Escherichia coli overproducers, and purified DPP-7 from the triple-knockout mutant 83-4A. GA supplemented with the three purified dipeptidyl/tripeptidyl aminopeptidases supported the growth of 83-47A. Conclusion:, DPPIV, DPP-7 and PTP-A contribute to the normal growth of P. gingivalis by cleaving substrate peptides into short-chain polypeptides that are efficient energy sources for P. gingivalis. [source] Accounting for co-extractable compounds (blank correction) in spectrophotometric measurement of extractable and total-bound proanthocyanidin in Leucaena sppJOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 8 2002Scott A Dalzell Abstract Methods to account for the spectral interference of co-extractable compounds (blank correction) in the spectrophotometric analysis of both extractable and bound proanthocyanidin (PA) using the proanthocyanidin (butanol/HCl) assay were evaluated. Crude extractable and bound PA sample matrices of PA-free Leucaena magnifica were used. Extractable PA blanks generated in heated 95% butanol/5% H2O reagent underestimated the optical density (absorbance) of co-extractable compounds by 24% (P,<,0.01), whereas unheated 95% butanol/5% HCl blanks, incubated at room temperature, accurately measured the absorbance of the background matrix (P,<,0.01). Current procedures that estimate bound PA concentrations using the proanthocyanidin assay produce intensely coloured background matrices. Recovery measurements from total-bound PA extracts spiked with 1071 and 2142,µg anthocyanidin per tube indicated that existing analytical procedures that do not account for the spectral interference of co-extractable compounds overestimated (P,<,0.01) bound PA concentrations by 69 and 38% respectively. An innovative technique that generated an internal correction factor for each sample, using wavelength-scanning spectrophotometry and non-linear curve-fitting computer software, was developed. This procedure recovered 100% of added anthocyanidins from bound PA matrices. © 2002 Society of Chemical Industry [source] Quantitative interpretation of optical density measurements using PF4-dependent enzyme-immunoassaysJOURNAL OF THROMBOSIS AND HAEMOSTASIS, Issue 8 2008T. E. WARKENTIN Summary.,Background:,Many laboratories test for heparin-induced thrombocytopenia (HIT) using a PF4-dependent enzyme-immunoassay (EIA). An advantage of the EIA is its simplicity; a disadvantage is that it only indirectly detects heparin-dependent, platelet-activating antibodies (,HIT antibodies'). Objectives:,To determine whether the magnitude of a positive EIA result, expressed in optical density (OD) units, predicts risk of HIT antibodies, defined as a strong-positive platelet serotonin-release assay (SRA) result (,50% serotonin release). Patients/methods:,We determined the risk of a strong-positive SRA result for five categories of OD reactivity (<0.40, 0.40,<1.00, 1.00,<1.40, 1.40,<2.00, and ,2.00 OD units) using two EIAs (commercial anti-PF4/polyanion IgG/A/M and in-house anti-PF4/heparin,IgG). Results:,For patient sera investigated for HIT antibodies, a weak-positive result (0.40,<1.00 OD units) in either EIA indicated a low probability (,5%) of a strong-positive SRA; the risk increased to ,90% with an OD , 2.00 units. Quantifying the EIA,SRA relationship for 1553 referred patient sera, we found that for every increase of 0.50 OD units in the EIA,IgG, the risk of a strong-positive SRA result increased by OR = 6.39 [95% confidence interval (CI), 5.13, 7.95; P < 0.0001]. For every increase of 1.00 OD units in the EIA,IgG, the risk increased by OR = 40.81 (95% CI, 26.35, 63.20; P < 0.0001). Conclusions:,The probability of HIT antibodies (strong-positive SRA result) inferred by a positive PF4-dependent EIA varies considerably in relation to the magnitude of the EIA result, expressed as OD values. In our laboratory, the probability of HIT antibodies being present reached ,50% only when the OD level was ,1.40 units. [source] IgG classification of anti-PF4/heparin antibodies to identify patients with heparin-induced thrombocytopenia during mechanical circulatory supportJOURNAL OF THROMBOSIS AND HAEMOSTASIS, Issue 2 2007S. SCHENK Summary., Commercial immunoassays frequently detect anti-PF4/heparin antibodies during mechanical circulatory support (MCS), but only a small minority of patients develops heparin-induced thrombocytopenia (HIT). Whereas platelet functional tests can distinguish between platelet-activating and non-platelet-activating antibodies, commercial PF4-dependent immunoassays do not. Between 2003 and 2004, 113 patients were placed on MCS. Blood samples were obtained on postimplant day 5,7 for analyses by antibody assays and the functional heparin-induced platelet activation (HIPA) assay. Three distinct groups of patient sera were identified: platelet-activating anti-PF4/heparin antibodies (n = 10), non-platelet-activating anti-PF4/heparin antibodies (n = 53), and anti-PF4/heparin antibody negative (n = 50). Patients with platelet-activating antibodies had the highest risk for thromboembolic events (P < 0.005), whereas those with non-platelet-activating antibodies did not differ from antibody negative patients (P = 0.369). The enzyme-immunoassay and column agglutination assays, which cover all immunoglobulin classes, demonstrated adequate sensitivity and negative predictive value; yet, both lacked specificity with respect to the platelet-activating antibodies. If all antibody positive patients were further classified by an IgG-specific anti-PF4/heparin enzyme-immuno assay, specificity for platelet-activating antibodies increased. Whereas IgG-specific optical density (OD) values below 1.0 were likely for non-platelet-activating anti-PF4/heparin antibodies, higher values were progressively predictive for pathogenic platelet activation. The probability of the development of clinical HIT also increased steeply. In conclusion, platelet-activating anti-PF4/heparin antibodies are relatively common (about 9%) in patients on MCS and are associated with significantly higher thrombotic event rates. Low IgG-specific OD values (< 1.0) in the enzyme-immunoassay indicate low likelihood for the presence of platelet-activating antibodies. These results justify further validation so that anticoagulation during MCS becomes safer and adequate. [source] Effect of the proton motive force inhibitor carbonyl cyanide- m -chlorophenylhydrazone (CCCP) on Pseudomonas aeruginosa biofilm developmentLETTERS IN APPLIED MICROBIOLOGY, Issue 4 2008A. Ikonomidis Abstract Aims:, Proton motive force (PMF) inhibition enhances the intracellular accumulation of autoinducers possibly interfering with biofilm formation. We evaluated the effect of the PMF inhibitor carbonyl cyanide- m -chlorophenylhydrazone (CCCP) on Pseudomonas aeruginosa biofilm development. Methods and Results:, Four epidemiologically unrelated P. aeruginosa isolates were studied. A MexAB-oprM overproducing strain was used as control. Expression of gene mexB was examined and biofilm formation after incubation with 0, 12·5 and 25 ,mol l,1 of CCCP was investigated. Mean values of optical density were analysed with one-way analysis of variance and t -test. Two isolates subexpressed mexB gene and only 25 ,mol l,1 of CCCP affected biofilm formation. Biofilms of the other two isolates and control strain PA140 exhibited significantly lower absorbance (P ranging from <0·01 to <0·05) with either 12·5 or 25 ,mol l,1 of CCCP. Conclusions:, The PMF inhibitor CCCP effect was correlated with the expression of MexAB-OprM efflux system and found to compromise biofilm formation in P. aeruginosa. Significance and Impact of the Study:, These data suggest that inhibition of PMF-dependent trasporters might decrease biofilm formation in P. aeruginosa. [source] In vitro studies on the effects of herbicides on the growth of rhizobiaLETTERS IN APPLIED MICROBIOLOGY, Issue 1 2002G. Singh Aims: ,To study the possible adverse effect of herbicides on nodulation and nitrogen fixation in legumes by affecting the nitrogen-fixing rhizobia. Methods and Results: ,Experiments were conducted to study the effect of four herbicides (terbutryn/terbuthylazine, trietazine/simazine, prometryn and bentazone) on the growth of nitrogen-fixing pea rhizobia ( Rhizobium leguminosarum ) in vitro by measuring optical density. Terbutryn/terbuthylazine, trietazine/simazine and prometryn had an adverse effect on the growth of rhizobia whereas bentazone was safe to rhizobia. Conclusions: ,The above herbicides could be used in pea at the recommended rates. Significance and Impact of the Study: ,The adverse effects of herbicides on rhizobia were observed at concentrations not normally expected to occur under field conditions. Further, previously observed adverse effects of these herbicides on nodulation and nitrogen fixation of peas were, possibly, not due to their effects in rhizobia but to their adverse effects on the plant growth itself. [source] Automutanolysin disrupts clinical isolates of cariogenic streptococci in biofilms and planktonic cellsMOLECULAR ORAL MICROBIOLOGY, Issue 6 2009P. Thanyasrisung Introduction:, Dental caries remains one of the most common chronic infectious diseases throughout the world. The formation of dental plaque is one of the caries risk factors. As a consequence, the removal of plaque may reduce the incidence of caries development. We identified an autolysin produced by Streptococcus mutans named auto-mutanolysin (Aml). Aml selectively lyses S. mutans and Streptococcus sobrinus. The specificity towards these cariogenic bacteria suggests that Aml may be used to prevent dental caries. Here, with the aim towards therapeutic application, we investigated the lytic activity of Aml against clinical isolates of S. mutans and S. sobrinus using planktonic cells and biofilms. Methods:, Planktonic cell suspensions and biofilms of clinically isolated streptococci were treated with Aml in the absence or the presence of Triton X-100. The lytic activity of Aml was monitored as the change in turbidity. The disruption of biofilms was evaluated by detecting the released DNA by polymerase chain reaction and observing the alteration of optical density of treated biofilms. Results:, Triton X-100 enhances the lytic ability of Aml. Using planktonic cells, Aml had various lysis levels against clinical strains. Repeated Aml treatment showed disruption of the biofilm using the representative clinical strains. Conclusion:, Our study demonstrates that Aml has an ability to lyse planktonic and biofilm cells of clinically isolated mutans streptococci in the presence of Triton X-100. These results suggest the possibility of using Aml as an alternative or additional approach for caries prevention. [source] Stress and the periodontal diseases: growth responses of periodontal bacteria to Escherichia coli stress-associated autoinducer and exogenous FeMOLECULAR ORAL MICROBIOLOGY, Issue 3 2005A. Roberts Psychological stress is known to increase the circulating levels of the catecholamine hormones noradrenaline and adrenaline, which have been shown to influence the growth of a large number of bacterial species by acting in a siderophore-like manner or by inducing the production of novel autoinducers of growth. As we have previously demonstrated that periodontal organisms display differing growth responses to noradrenaline and adrenaline, the aim of this study was to determine whether these growth effects were based upon either siderophore-like or autoinducer mechanisms. Initial inocula of 43 microbial organisms normally found within the subgingival biofilm were established under anaerobic conditions (35°C). Each strain was re-inoculated into a serum-based minimal medium and growth was assessed by optical density (OD600nm) with test and control cultures performed in triplicate. Test cultures were supplemented with either 50 ,m ferric nitrate or a previously described Escherichia coli autoinducer of growth. Significant growth effects for supplementation with ferric nitrate (13 species responding positively) and E. coli autoinducer (24 species responding positively) were observed, with differences in growth response within bacterial species and within microbial complexes. When data for all organisms were compared with published responses to catecholamines there were only weak correlations with Fe (r = 0.28) and E. coli autoinducer (r = 0.34) responses. However, large positive responses (> 25% increase) to free Fe and/or E. coli autoinducer were significantly more prevalent in the group of organisms (n = 12) known to exhibit similar responses to catecholamine hormones (P < 0.01; ,2 = 4.56). The results support the view that catecholamines may exert their effects on subgingival organisms by initiating autoinducer production, or simply by acting in a siderophore-like manner, scavenging bound iron from the local environment. It is possible that autoinducer mechanisms may play an important role in the response of oral microorganisms to stress hormones, thereby contributing to the clinical course of stress-associated periodontal diseases. [source] | ||||||||||||||||||||||||||||||||||