Densitometric analysis of rep-PCR data: Insight into genetic variability and transmission of Clostridium perfringens typed with an improved multiplex PCR.

An epidemiological study was conducted in North-East India (part of Indo-Burma biodiversity hotspot) to better understand the distribution, diversity, and transmission of Clostridium perfringens among livestock, pets, wild animals (captive), and humans. A total of 160 C. perfringens isolates were recovered from 642 diarrhoeic faecal samples with an isolation rate of 24.92%. Isolation rate was the highest among captive wild animals (37.5%) followed by dog (34.6%), human (33.8%), pig (32.7%), cattle (20.8%), goat (18.3%) and poultry (9.3%). Isolates were toxin typed using a seven gene multiplex PCR designed for simultaneous detection of cpa, cpb, cpb2, etx, iap, cpe and netB. The majority of isolates, 128 (80%) were of type A, followed by 17 (10.62%), 5 (3.12%), 4 (2.5%), 3 (1.87%), 2 (1.25%) and 1 (0.63%) isolates of type C, D, E, G, F and B, respectively. Beta 2 toxin gene was present in 65 (50%) of type A isolates, followed by 7 (41.2%), 4 (80%), 1(25%), and 1 (100%) of type C, D, G and B isolates, respectively. Beta 2 toxin has a high prevalence among dogs (28.6%), cattle (27.3%), and pig (20.8%) compared to humans, goat, wild animals, and poultry (1.2-14.3%). The prevalence of CPE and NetB toxin-positive strains was low, with only 3 (1.8%) and 5 (3.1%) isolates, respectively. Association of C. perfringens with diarrhoea in Civet Cat, Golden Langur, and Gray Langur has been reported for the first time. The genetic diversity and transmission of isolates were investigated using automated rep-PCR (Diversilab®, bioMérieux) using two densitometry-based matrices: modified Kullback-Leibler (KL) and Pearson's correlation (PC). The PC and modified KL matrices formed three distinct clusters with 59% and 27.2% similarity, respectively. C. perfringens diversity and transmission were best studied using modified KL matrix that placed more emphasis on the presence of bands rather than intensity. However, the PC method was found to be more suitable for differentiating strains within a toxin type, with slightly higher D-values.

Technical note: age estimation by using pubic bone densitometry according to a twofold mode of CT measurement.

In forensic anthropology, age estimation is a major element in the determination of a biological profile and the identification of individuals. Thus, many anatomical structures have been studied, such as the pubic symphysis, which is a source of major interest due to its late maturation. One of the most well-known methods of assessment is the Suchey-Brooks (SB) system based on the morphological characteristics of the pubic symphysis. The aim of this study was to propose linear regression formulae in order to deduce chronological age from bone density, using both Hounsfield unit (HU), and mean bone density (mBD) values of the pubic symphysis. Moreover, we intended to test the reliability and then to explore the feasibility of using HU instead of mBD values for age estimation. We built retrospectively a reference sample of 400 pubic symphyses using computed tomography at a French hospital and a test sample of 120 pubic symphyses. Equations were created to establish linear regression models for age estimation. Inaccuracy and bias were calculated for individuals aged more or less than 40 years. We highlighted homogeneous mean absolute errors for both HU and mBD values, most of them being less than 10 years. Moreover, we reported a moderate overestimation for younger individuals and a very small underestimation for older individuals. This study proposes a correlation between the bone density and age of individuals with a valuable level of reliability. Finally, HU measurements seem to be suitable for linking bone density with the age of individuals in forensic practice.

A field-to-desktop toolchain for X-ray CT densitometry enables tree ring analysis.

BACKGROUND AND AIMS: Disentangling tree growth requires more than ring width data only. Densitometry is considered a valuable proxy, yet laborious wood sample preparation and lack of dedicated software limit the widespread use of density profiling for tree ring analysis. An X-ray computed tomography-based toolchain of tree increment cores is presented, which results in profile data sets suitable for visual exploration as well as density-based pattern matching. METHODS: Two temperate (Quercus petraea, Fagus sylvatica) and one tropical species (Terminalia superba) were used for density profiling using an X-ray computed tomography facility with custom-made sample holders and dedicated processing software. KEY RESULTS: Density-based pattern matching is developed and able to detect anomalies in ring series that can be corrected via interactive software. CONCLUSIONS: A digital workflow allows generation of structure-corrected profiles of large sets of cores in a short time span that provide sufficient intra-annual density information for tree ring analysis. Furthermore, visual exploration of such data sets is of high value. The dated profiles can be used for high-resolution chronologies and also offer opportunities for fast screening of lesser studied tropical tree species.

Photopigment self-screening and the determination of macular pigment absorbance using heterochromatic flicker photometry.

PURPOSE: Heterochromatic flicker photometry (HFP) is commonly used to determine macular pigment optical density (MPOD). Since HFP in this application is a locus comparison method, an identical relative spectral response at each locus is required for a perfect measure. We know this requirement cannot be strictly true since the optical density of photopigments increases as the foveal center is approached. Thus, the self-screening effect would result in an underestimate of MPOD. An earlier study concluded that the underestimate is on the order of 30%. We examined this issue by manipulating photopigment optical density, and consequently the degree of selfscreening. METHODS: A continuously exposed, 470 nm, background bleached cone photopigments over a range from 0 to 80%. MPOD was determined 10' and 30' from the foveal center. Two subjects were used in the main experiment. Five additional subjects were studied with just the 0% and 80% bleach levels. Spectral measures were obtained at 0% and 70% bleach levels for the two primary subjects. RESULTS: Subjects in the main experiment showed MPOD estimates that increased with increasing bleaching. The effect, however, was small: one observer's MPOD increased 0.08 and 0.02 for the 10' and 30' loci, respectively; the other observer's values were 0.04 and 0.01 for the same loci. Comparable values were obtained for the other five subjects using the 0% and 80% bleach conditions. Spectral measures were consistent with the findings of the main experiment. CONCLUSIONS: When self-screening is nearly abolished (80% bleach), a relatively small underestimation is revealed for the unbleached state. For the 1° target we show about 2-3% underestimation. Our 20' target reveals a larger underestimate (8-9%), consistent with longer photoreceptor outer-segments nearer the foveal center. We conclude that HFP yields values essentially independent of self-screening for targets of 1° diameter or greater. Smaller targets are less than 10% underestimated for near-zero bleach conditions.

Integrated protocol for reliable and fast quantification and documentation of electrophoresis gels.

Quantitative analysis of electrophoresis gels is an important part in molecular cloning, as well as in protein expression and purification. Parallel quantifications in yield and purity can be most conveniently obtained from densitometric analysis. This communication reports a comprehensive, reliable and simple protocol for gel quantification and documentation, applicable for single samples and with special features for protein expression screens. As major component of the protocol, the fully annotated code of a proprietary open source computer program for semi-automatic densitometric quantification of digitized electrophoresis gels is disclosed. The program ("GelQuant") is implemented for the C-based macro-language of the widespread integrated development environment of IGOR Pro.

Intraoperative mechanical measurement of bone quality with the DensiProbe.

Reduced bone stock can result in fractures that mostly occur in the spine, distal radius, and proximal femur. In case of operative treatment, osteoporosis is associated with an increased failure rate. To estimate implant anchorage, mechanical methods seem to be promising to measure bone strength intraoperatively. It has been shown that the mechanical peak torque correlates with the local bone mineral density and screw failure load in hip, hindfoot, humerus, and spine in vitro. One device to measure mechanical peak torque is the DensiProbe (AO Research Institute, Davos, Switzerland). The device has shown its effectiveness in mechanical peak torque measurement in mechanical testing setups for the use in hip, hindfoot, and spine. In all studies, the correlation of mechanical torque measurement and local bone mineral density and screw failure load could be shown. It allows the surgeon to judge local bone strength intraoperatively directly at the region of interest and gives valuable information if additional augmentation is needed. We summarize methods of this new technique, its advantages and limitations, and give an overview of actual and possible future applications.

Application of an online-biomass sensor in an optical multisensory platform prototype for growth monitoring of biotechnical relevant microorganism and cell lines in single-use shake flasks.

In the context of this work we evaluated a multisensory, noninvasive prototype platform for shake flask cultivations by monitoring three basic parameters (pH, pO2 and biomass). The focus lies on the evaluation of the biomass sensor based on backward light scattering. The application spectrum was expanded to four new organisms in addition to E. coli K12 and S. cerevisiae [1]. It could be shown that the sensor is appropriate for a wide range of standard microorganisms, e.g., L. zeae, K. pastoris, A. niger and CHO-K1. The biomass sensor signal could successfully be correlated and calibrated with well-known measurement methods like OD600, cell dry weight (CDW) and cell concentration. Logarithmic and Bleasdale-Nelder derived functions were adequate for data fitting. Measurements at low cell concentrations proved to be critical in terms of a high signal to noise ratio, but the integration of a custom made light shade in the shake flask improved these measurements significantly. This sensor based measurement method has a high potential to initiate a new generation of online bioprocess monitoring. Metabolic studies will particularly benefit from the multisensory data acquisition. The sensor is already used in labscale experiments for shake flask cultivations.

TLC--densitometric method for qualitative analysis of betamethasone and its related compounds in pharmacautical preparations.

A new simple and rapid TLC-densitometric procedure for the separation and identification of betamethasone and its related substances, betamethasone-17,21-dipropionate, betamethasone-17-valerate, betamethasone-21-valerate and also betamethasone disodium phosphate was developed. One of the chromatographic systems proposed in this study, which has been satisfactory applied in separation of four pairs of examined compounds was silica gel 60F254 (E. Merck, Art. 1.05554) and a mixture containing chloroform-methanol-acetic acid (99.5%) in volume composition 28:5:0.5. Densitometric measurements were done using densitometer TLC Scanner 3 at 246 nm. The proposed method was checked in terms of its specificity for the determination of betamethasone-17,21-dipropionate and betamethasone disodium phosphate in commercially available products containing both compounds, separately, as active ingredients. The results showed that the method is suitable for qualitative analysis of betamethasone derivatives in simple and combined pharmaceuticals in various dosage forms e.g., lotion and injection solution. It also can be applied in quality control of pharmaceutical formulations of betamethasone and its related compounds in form of salts and esters.

Corneal densitometry evaluation in archipelago keratitis.

To report a case of archipelago keratitis and its improvement as evaluated by corneal densitometry. A review of the patient's record and relevant literature. A 57-year-old man presented with blurred vision associated with photophobia and halos around lights. Slit-lamp examination revealed multiple confluent, nummular subepithelial infiltrates. The infiltrates were unresponsive to topical nonsteroidal anti-inflammatory drops. The treatment was changed to topical steroids and oral acyclovir and a diagnosis of archipelago keratitis was made. The patient showed quick response to the treatment and the infiltrates disappeared within 5 weeks. However, no clear changes were observed initially by slit-lamp but corneal densitometry allowed us to evaluate improvement as the density value decreased from 96.5 to 38.6. After 10 days of treatment all the symptoms were resolved. Archipelago keratitis responded well to topical steroids and oral acyclovir. We describe the use of corneal densitometry complementary to slit-lamp examination in the clinical observation of archipelago keratitis.

Hybrid femtosecond/picosecond rotational coherent anti-Stokes Raman scattering temperature and concentration measurements using two different picosecond-duration probes.

A hybrid fs/ps pure-rotational CARS scheme is characterized in furnace-heated air at temperatures from 290 to 800 K. Impulsive femtosecond excitation is used to prepare a rotational Raman coherence that is probed with a ps-duration beam generated from an initially broadband fs pulse that is bandwidth limited using air-spaced Fabry-Perot etalons. CARS spectra are generated using 1.5- and 7.0-ps duration probe beams with corresponding coarse and narrow spectral widths. The spectra are fitted using a simple phenomenological model for both shot-averaged and single-shot measurements of temperature and oxygen mole fraction. Our single-shot temperature measurements exhibit high levels of precision and accuracy when the spectrally coarse 1.5-ps probe beam is used, demonstrating that high spectral resolution is not required for thermometry. An initial assessment of concentration measurements in air is also provided, with best results obtained using the higher resolution 7.0-ps probe. This systematic assessment of the hybrid CARS technique demonstrates its utility for practical application in low-temperature gas-phase systems.

Imaging system to assess objectively the optical density of the macular pigment in vivo.

This paper presents an optical system called MacPI, which implements a two-color reflectance technique in combination with various hardware and software tools to assess objectively the macular pigment (MP) optical density in vivo. The system consists of a bespoke optical design, a control architecture, driver electronics, a collection of image-processing techniques, and a graphical user interface. The deficiencies of the technique employed and the solutions implemented in the MacPI system to confront those inherent frailties are presented. An overview of the effective interpretation of the acquired data and the techniques employed by MacPI in the acquisition of that data is discussed. The result of a comparison trial with an alternative device is also presented. We suggest that appropriate design of the hardware and an efficient interpretation of the acquired data should produce a system capable of consistent, accurate, and rapid measurements, while retaining the distinction of ease of use, portability, comfort for the subject, and a design that is economic to produce. Its versatility should allow both for a clinical screening application and for further investigation and establishment of the physiological role of the MP in a laboratory-based environment.

Investigation on dynamic calibration for an optical-fiber solids concentration probe in gas-solid two-phase flows.

This paper presents a review and analysis of the research that has been carried out on dynamic calibration for optical-fiber solids concentration probes. An introduction to the optical-fiber solids concentration probe was given. Different calibration methods of optical-fiber solids concentration probes reported in the literature were reviewed. In addition, a reflection-type optical-fiber solids concentration probe was uniquely calibrated at nearly full range of the solids concentration from 0 to packed bed concentration. The effects of particle properties (particle size, sphericity and color) on the calibration results were comprehensively investigated. The results show that the output voltage has a tendency to increase with the decreasing particle size, and the effect of particle color on calibration result is more predominant than that of sphericity.

[Advances in bone densitometry equipment].

Techniques in bone densitometry have made remarkable progress in the past quarter-century and are now widely used in the daily practice of osteoporosis. Since the skeletal site and the method of measurement differ among the type of equipments, several types are adopted to suit the situation of each medical institute. Central DXA is used to provide a thorough examination of osteoporosis in large hospitals, whereas in general clinics the radial DXA or radiographic photodensitometry of metacarpus is mainly used. Quantitative ultrasound of calcaneus is mostly used for screening osteoporosis. DXA is also going to be applied to vertebral fracture assessment, hip structural analysis and micro-architectural analysis of vertebral trabecular bone.

The readout thickness versus the measured thickness for a range of screen film mammography and full-field digital mammography units.

PURPOSE: To establish a simple method to determine breast readout accuracy on mammography units. METHODS: A thickness measuring device (TMD) was used in conjunction with a breast phantom. This phantom had compression characteristics similar to human female breast tissue. The phantom was compressed, and the thickness was measured using TMD and mammography unit readout. Measurements were performed on a range of screen film mammography (SFM) and full-field digital mammography (FFDM) units (8 units in total; 6 different models/manufacturers) for two different sized paddles and two different compression forces (60 and 100 N). RESULTS: The difference between machine readout and TMD for the breast area, when applying 100 N compression force, for nonflexible paddles was largest for GE Senographe DMR+ (24 cm × 30 cm paddle: +14.3%). For flexible paddles the largest difference occurred for Hologic Lorad Selenia (18 cm × 24 cm paddle: +26.0%). CONCLUSIONS: None of the units assessed were found to have perfect correlation between measured and readout thickness. TMD measures and thickness readouts were different for the duplicate units from two different models/manufacturers.

Short- and mid-term repeatability of macular pigment optical density measurements using spectral fundus reflectance.

BACKGROUND: Several methods have been proposed for measuring macular pigment optical density (MPOD). To date, none of the realized techniques can be considered as "gold standard". A key issue for the clinical applicability of a method is its repeatability. In this study, we investigated short- and mid-term repeatability of MPOD measurements using reflectometry. METHODS: A total of 12 healthy young subjects were measured 5 times on 5 consecutive days. Repeatability over 6 months was investigated in patients with AMD. The data in AMD patients were taken from a recently published placebo controlled study investigating the effect of lutein supplementation on MPOD (n = 37; [1]). Four measurements over 6 months were used to calculate repeatability. Spectral fundus reflectance of the fovea was measured in a 2.3° detection field with a custom-built fundus reflectometer. Calculation of MPOD was based on a previously published fundus reflectance model. RESULTS: The coefficients of variation were 6.2 ± 2.4% and 8.0 ± 5.5% in the healthy and AMD group, respectively. Bland-Altman plots indicate that the difference between measurements at day 1 and day 5 in healthy subjects and day 1 and month 6 in AMD patients was small. The maximum deviation in MPOD in a healthy subject was 0.07 (22.6%), and 0.17 (51.5%) in a patient with AMD. CONCLUSIONS: Reflectometry provides adequate short-term and mid-term repeatability for measuring MPOD. Accordingly, the technique makes it possible to monitor MPOD in patients with AMD and to study the influence of supplementation in these subjects. In addition, the variability of the technique is small enough to allow for clinical trials with reasonable sample size.

Radiopacity of different shades of resin-based restorative materials compared to human and bovine teeth.

This study evaluated the radiopacity of different shades of resin-based restorative materials and compared the results to human and bovine dental hard tissues. Disk specimens 6 mm in diameter and 1 mm thick (N = 220, n = 10) were prepared from the following restorative materials: · eight shades of nanofilled composite (Aelite Aesthetic Enamel), · seven shades of nanohybrid composite (Grandio Universal), · six shades of photopolymerized polyacid modified compomer (Glasiosite), and · one shade of hybrid composite (X-tra fil U). Human canine dentin (n = 10), bovine enamel (n = 10), and an aluminum (Al) step wedge were used as references. The optical density values of each material were measured from radiographic images using a transmission densitometer. Al step wedge thickness and optical density values were plotted, and equivalent Al thickness (eq Al) values were determined for radiopacity measurements of each material. The data were analyzed using a non-parametric one-way ANOVA (Kruskal-Wallis), and multiple comparisons were made with a Student-Newman-Keuls post hoc test (a = 0.05). Different shades of resin-based restorative materials tested did not reveal statistically significant differences within each material group (p > 0.05). Radiopacity values of the resin-based restorative materials investigated varied depending on their types; however, within different shades of one material type, radiopacity values were comparable. Every shade of nanocomposite material other than Aelite Aesthetic Enamel Incisal LT Gray showed comparable radiopacity to human dentin. Other materials tested demonstrated higher radiopacity compared to human dentin and bovine enamel.

A calibration methodology of QCT BMD for human vertebral body with registered micro-CT images.

PURPOSE: The accuracy of QCT-based homogenized finite element (FE) models is strongly related to the accuracy of the prediction of bone volume fraction (BV/TV) from bone mineral density (BMD). The goal of this study was to establish a calibration methodology to relate the BMD computed with QCT with the BV/TV computed with micro-CT (microCT) over a wide range of bone mineral densities and to investigate the effect of region size in which BMD and BV/TV are computed. METHODS: Six human vertebral bodies were dissected from the spine of six donors and scanned submerged in water with QCT (voxel size: 0.391 x 0.391 x 0.450 mm3) and microCT (isotropic voxel size: 0.018(3) mm3). The microCT images were segmented with a single level threshold. Afterward, QCT-grayscale, microCT-grayscale, and microCT-segmented images were registered. Two isotropic grids of 1.230 mm (small) and 4.920 mm (large) were superimposed on every image, and QCT(BMD) was compared both with microCT(BMD) and microCT(BV/TV) for each grid cell. RESULTS: The ranges of QCT(BMD) for large and small regions were 9-559 mg/cm3 and -90 to 1006 mg/cm3, respectively. QCT(BMD) was found to overestimate microCT(BMD). No significant differences were found between the QCT(BMD)-microCT(BV/TV) regression parameters of the two grid sizes. However, the R2 was higher, and the standard error of the estimate (SEE) was lower for large regions when compared to small regions. For the pooled data, an extrapolated QCTBMD value equal to 1062 mg/ cm3 was found to correspond to 100% microCT(BV/TV). CONCLUSIONS: A calibration method was defined to evaluate BV/TV from QCTBMD values for cortical and trabecular bone in vitro. The QCT(BMD-microCT(BV/TV) calibration was found to be dependent on the scanned vertebral section but not on the size of the regions. However, the higher SEE computed for small regions suggests that the deleterious effect of QCT image noise on FE modelling increases with decreasing voxel size.

Determination of analytes in medical herbs extracts by SPE coupled with two-dimensional planar chromatography in combination with diode array scanning densitometry and HPLC-diode array detector.

The purpose of this study is to demonstrate an application of 2-D high-performance planar chromatography-diode array detector (DAD) and HPLC-DAD after solid-phase extraction (SPE) for identification and quantitative analysis of pesticides (isoproturon, aziprotryne, hexazinone, flufenoxuron, methabenzthiazuron, procymidone, and α-cypermethrin) in Melissa officinalis L. (Labiatae) samples. The procedure described for the determination of compounds is inexpensive and can be applied to routine analysis of analytes in medical herbs' samples after preliminary cleanup and concentration by SPE. Average recoveries on C18 SPE cartridges of pesticides eluted with 5 mL tetrahydrofuran by the proposed HPLC-DAD method, before and after 2-D-high-performance planar chromatography separation of analytes from M. officinalis L. samples spiked with pesticide at a concentration level of 10 µg/g in plant material are presented. Method validation parameters for the quantification of pesticides by the proposed HPLC-DAD after SPE method are also presented.

[Assessment of volumetric bone mineral density and geometry for hip with clinical CT device].

It is one of the most important challenges for preventing hip fractures to establish an accurate non-invasive assessment of hip fracture risk. Quantitative computed tomography (QCT) is able to analyze not only volumetric bone mineral density (mg/cm3) of trabecular and cortical bone compartment separately, but also geometry and biomechanical parameters in bone such as cross-sectional area, cortical bone thickness, section modulus, buckling ratio etc. The analysis of geometry and biomechanical parameters at hip could provide better prediction of hip fracture risk, better understanding pathogenesis of hip fractures, and thus better selection of drug and/or lifestyle-based interventions.