A Compressed-Sensing Based Blind Deconvolution Method for Image Deblurring in Dental Cone-Beam Computed Tomography.

In cone-beam computed tomography (CBCT), reconstructed images are inherently degraded, restricting its image performance, due mainly to imperfections in the imaging process resulting from detector resolution, noise, X-ray tube's focal spot, and reconstruction procedure as well. Thus, the recovery of CBCT images from their degraded version is essential for improving image quality. In this study, we investigated a compressed-sensing (CS)-based blind deconvolution method to solve the blurring problem in CBCT where both the image to be recovered and the blur kernel (or point-spread function) of the imaging system are simultaneously recursively identified. We implemented the proposed algorithm and performed a systematic simulation and experiment to demonstrate the feasibility of using the algorithm for image deblurring in dental CBCT. In the experiment, we used a commercially available dental CBCT system that consisted of an X-ray tube, which was operated at 90 kVp and 5 mA, and a CMOS flat-panel detector with a 200-µm pixel size. The image characteristics were quantitatively investigated in terms of the image intensity, the root-mean-square error, the contrast-to-noise ratio, and the noise power spectrum. The results indicate that our proposed method effectively reduced the image blur in dental CBCT, excluding repetitious measurement of the system's blur kernel.

Eliminating artifacts in single-grid phase-contrast x-ray imaging for improving image quality.

In this study, we propose a modification to a single-grid phase-contrast x-ray imaging (PCXI) system using a Fourier domain analysis technique to extract absorption, scattering, and differential phase-contrast images. The proposed modification is to rotate the x-ray grid in the image plane to achieve spectral separation between the desired information and the moiré artifact, which is introduced by the superposition of the periodic image of the grid shadow and the periodic sampling by the detector. In addition, we performed some system optimization by adjusting distances between source, object, grid, and detector to further improve image quality. This optimization aimed to increase the spectral spacing between the primary spectrum (lower frequency) and the harmonics of the spectrum (higher frequency) used to extract the various image contrasts. The table-top setup used in the experiment consisted of a focused-linear grid with a 200-lines/inch strip density, a microfocus x-ray tube with a 55-µm focal spot size, and a CMOS flat-panel detector with a 49.5-µm pixel size. The x-ray grid was rotated at 27.8° with respect to the detector and the sample was placed as close as possible to the x-ray tube. Our results indicated that the proposed method effectively eliminated the PCXI artifacts, thus improving image quality.

3D reconstruction based on compressed-sensing (CS)-based framework by using a dental panoramic detector.

In this work, we propose a practical method that can combine the two functionalities of dental panoramic and cone-beam CT (CBCT) features in one by using a single panoramic detector. We implemented a CS-based reconstruction algorithm for the proposed method and performed a systematic simulation to demonstrate its viability for 3D dental X-ray imaging. We successfully reconstructed volumetric images of considerably high accuracy by using a panoramic detector having an active area of 198.4 mm × 6.4 mm and evaluated the reconstruction quality as a function of the pitch (p) and the angle step (Δθ). Our simulation results indicate that the CS-based reconstruction almost completely recovered the phantom structures, as in CBCT, for p≤2.0 and θ≤6°, indicating that it seems very promising for accurate image reconstruction even for large-pitch and few-view data. We expect the proposed method to be applicable to developing a cost-effective, volumetric dental X-ray imaging system.

Structure and expression of the cytosolic aldehyde dehydrogenase gene in cyclophosphamide-resistant murine leukemia L1210 cells.

These investigations were performed to clarify the molecular basis for the enhanced expression of cytosolic aldehyde dehydrogenase (ALDH-1) enzymatic activity in the cyclophosphamide-resistant L1210/CPA murine leukemia cell line, as compared to the parental L1210/O strain. Western immunoblot analysis was performed using a 15-fold greater quantity of cytosolic protein from the L1210/O as compared to the L1210/CPA cell line. Nevertheless, ALDH-1 immunoreactive protein could be detected only in the L1210/CPA cells. Northern analyses, performed using total cellular and polyadenylated RNA, again demonstrated ALDH-1-specific transcripts only in the L1210/CPA cell line. This transcript was identical in size to the ALDH-1 message expressed by normal murine hepatocytes. On Southern analysis, no evidence of gene amplification, gene rearrangement, or significant mutations of length was detected. These studies suggest that the ALDH-1 protein produced by the L1210/CPA cell line is structurally normal. Moreover, overexpression of the gene does not appear to have arisen as a result of an incremental process, such as gene amplification. Rather, a qualitative abnormality in the regulation of this gene appears to exist in the L1210/CPA cells, which distinguishes them from L1210/O cells and from normal murine lymphocytes.

Rapid distinction between Leptospira interrogans and Leptospira biflexa by PCR amplification of 23S ribosomal DNA.

Bacterial specific primers were used to amplify 23S rRNA genes from a representative strain from each of the 23 serogroups of the pathogenic Leptospira interrogans and 8 strains from 6 serogroups of the non-pathogenic Leptospira biflexa. Only regions of extreme variability, which had been identified on the basis of homology-based search of all the 23S rRNA sequences available in GenBank database, were sequenced from the amplified products. PCR primers that had the potential to distinguish L. interrogans from L. biflexa species were designed from the derived sequences and a sensitive PCR protocol developed. The PCR method enabled the differentiation of the 59 strains of the 23 serogroups of L. interrogans from the 8 strains of 6 serogroups of L. biflexa. Further investigation by 16S rDNA sequencing of two strains of L. interrogans, which gave unexpected PCR results, provided evidence that they had been misclassified and hence we propose to reassign them to L. biflexa.

Rapid distinction between Leptonema and Leptospira by PCR amplification of 16S-23S ribosomal DNA spacer.

The PCR amplification of the genomic DNA of Leptonema illini strain 3055 using primers directed against conserved regions of the rRNA operon provided evidence that the 16S and 23S rRNA genes were linked via an intergenic spacer region. The sequencing of the intergenic spacer region indicated that it was 435 nucleotides in length and sequence similarity searches revealed that it bore no homology to any known sequences including tRNA available in databases. Further investigations using Southern blot hybridization revealed that there were two copies of these linked genes in the genome. However, similar PCR studies on a representative strain from each of the 23 serogroups of Leptospira interrogans, which are pathogenic, and eight strains from the 6 serogroups of Leptospira biflexa, which are non-pathogenic, revealed that the 16S and 23S rRNA genes were not linked.

Comparison of two PCR methods for rapid identification of Leptospira genospecies interrogans.

Based on (i) an analysis of Leptospira 16S rDNA sequences determined by us and of those from databases and (ii) a previously published finding that restriction fragment length polymorphisms (RFLPs) within the Leptospira 16S and 23S rDNA were detected by nine restriction enzymes and these RFLPs allowed categorisation of Leptospira into eight genospecies, we predicted that one particular DdeI restriction site polymorphism within 16S rDNA could be independently used for identifications of Leptospira strains belonging to the genospecies interrogans. Two PCR-based methods, namely allele-specific amplification (ASA) and PCR-RFLP, were tested for the rapid detection of the DdeI restriction site polymorphism. One or two representative strains from each of nine genospecies were tested by ASA, whereas 73 strains from nine genospecies and two field isolates were tested by PCR-RFLP. Our experiments showed that the ASA method was not as specific as intended, but the PCR-RFLP method was useful for rapid identifications of the genospecies interrogans. We have not only confirmed a previous finding and extended the number of samples particularly from the genospecies biflexa, weilii, and inadai, but also simplified a previous PCR-RFLP protocol.

Investigation of the epidemiology of hospital isolates of Acinetobacter anitratus by two molecular methods.

Two hundred and two isolates of Acinetobacter anitratus from 126 patients in 36 wards belonging to 11 specialties of a university teaching hospital were compared by ribotyping and restriction enzyme digest analysis (REA) of total DNA. Forty-six groups were defined by both techniques and there was 96 center dot 5% agreement between the methods for the designation of isolates into groups. Only two groups were endemic and circulating in the whole hospital while others were less common. Burns and intensive therapy units had the highest number of isolates and these were mainly of the two endemic groups while renal dialysis and neonatal units had isolates belonging to the less common groups. Of the 32 patients with multiple isolates, 17 were infected or colonized at different sites by two and up to four groups of A. anitratus. Both ribotyping and REA of total DNA are discriminatory methods for typing A. anitratus, however, the latter is a simpler and more rapid method and it can be used in a routine clinical laboratory.

Identification of Leptospira biflexa by real-time homogeneous detection of rapid cycle PCR product.

Sequence analysis of 16S rRNA genes extracted from nucleic acids databases enabled the identification of a Leptospira biflexa (L. biflexa) signature sequence, against which a reverse primer designated L613, was designed. This primer, when used in conjunction with a universal bacterial specific forward primer designated Fd1, enabled the development of a LightCycler-based PCR protocol in which fluorescence emission due to binding of SYBR Green I dye to amplified products could be detected and monitored. A melting temperature (Tm), determined from the melting curve of the amplified product immediately following the termination of thermal cycling, confirmed that the product was that of L. biflexa. Agarose gel electrophoresis therefore was not necessary for identification of PCR products. The PCR protocol was very rapid, and consisted of 30 cycles with a duration of 20 s for each cycle with the monitoring of the melting curve requiring an additional 3 min. The whole protocol was completed in less than 20 min. The PCR protocol was also specific and enabled the identification of 18 strains of L. biflexa, whilst excluding 14 strains of L. interrogans and Leptonema illini. Two examples of its utility in improving work flow of a Leptospira reference laboratory are presented in this article. The use of a simple boiling method for extraction of DNA from all the members of the Leptospiraceae family DNA further simplifies the procedure and makes its use conducive to diagnostic laboratories.

Identification of Leptospira inadai by continuous monitoring of fluorescence during rapid cycle PCR.

Seven new Leptospira isolates from rats, a buffalo, and contaminated media showed either reactive serology against more than 1 serogroup or no reactive serology against a reference panel of 22 serovars in the microscopic agglutination test (MAT). Because of these inconclusive results, the 16S rDNA sequences of these isolates were determined and found to resemble that of the type strain of Leptospira inadai (L. inadai), serovar lyme strain 10, which is considered to be nonpathogenic for humans. Comparative analyses of other Leptospira 16S rDNA sequences from databases revealed a L. inadai-specific signature sequence, against which an amplification primer was designed. This primer when used in conjunction with an universal primer enabled the trial of a rapid PCR protocol in which fluorescence emissions due to binding of SYBR Green I dye to PCR products were continuously monitored during rapid thermal cycling. A melting curve acquired immediately after PCR was used to distinguish the intended product. The thermal cycling and continuous monitoring of fluorescence emission were accomplished by the LightCycler; the whole procedure of 30 PCR cycles and melting curve acquisition required only 20 minutes. The primer achieved the required specificity, as the intended PCR product resulted only from 6 confirmed L. inadai reference strains and 7 field isolates that had been verified as L. inadai by the 16S rDNA sequencing, but not from 16 reference strains of Leptospira belonging to 7 other genospecies. Furthermore, these experiments showed that the PCR protocol was robust because target DNA of different conditions, which were extracted by either 1 of the 4 methods used, could be detected.

An early human ovum.

An early embryo at stage 5 of development is presented. The specimen's estimated age of about ten days makes it one of the youngest human products of conception on record. As such, it provides information on the earliest phases of human development.

Diffuse viral papillomatosis (condyloma) of the uterine cavity.

A case of viral papillomatosis (condyloma) of the uterine cervix is reported, which was followed for 13 years and in which the lesion spread to involve superficially the entire uterine cavity without any change in its histologic appearance. The presence of intranuclear papillomavirus antigen was demonstrated by using the peroxidase-antiperoxidase technique.

Lipogenesis from amino acids in perfused isolated dog skin.

Lipogenesis from amino acids has been studied in isolated perfused dog skin. Uniformly labeled alanine-(14)C, glycine-(14)C, isoleucine-(14)C, leucine-(14)C, phenylalanine-(14)C, and valine-(14)C are all incorporated into the cutaneous lipids, with significant incorporation into most of the isolated lipid fractions. Efficiency of lipogenesis has been expressed relative to the extent of incorporation of acetate under the same experimental conditions. This efficiency was highest for the three branched-chain amino acids. The accuracy, uses, and limitations of the perfusion technique for the study of cutaneous lipogenesis have been evaluated.

Antimicrobial susceptibilities and beta-lactamase production of Hong Kong isolates of gastroenteric salmonellae and Salmonella typhi.

We examined the in-vitro antibiotic susceptibility of 760 gastroenteric salmonellae and 36 strains of Salmonella typhi isolated in Hong Kong between 1985 and 1988. S. typhi remained susceptible to all the antibiotics tested except for one isolate resistant to chloramphenicol, another to kanamycin and co-trimoxazole, and a third to nalidixic acid. In contrast, resistance and multiple resistance has increased significantly in gastroenteric salmonellae over the last ten years. Seventeen percent were resistant to ampicillin, 61% to tetracycline, 23% to chloramphenicol and 8% to gentamicin. Many ampicillin-resistant strains remained resistant to ampicillin even in the presence of sulbactam (69%) or clavulanic acid (25%). More than 50% of isolates were resistant to two or more antibiotics and one isolate was resistant to eleven. Ampicillin-resistance was usually due to the production of TEM-1 or OXA-1 beta-lactamases but a few isolates produced AER-1, PSE-1 or PSE-2. Genetic determinants for these enzymes were usually borne on plasmids ranging in size from 2 to 143.7 Md but half of the OXA-1 genes were chromosomally located.

Identification of pathogenic Leptospira genospecies by continuous monitoring of fluorogenic hybridization probes during rapid-cycle PCR.

Partial sequences of 23S rRNA gene PCR products from 23 strains of 6 pathogenic Leptospira genospecies and from 8 strains of the saprophytic Leptospira biflexa were determined. Sequence analyses enabled Leptospira genus-specific amplification primers and pathogen-specific fluorogenic adjacent hybridization probes to be designed and synthesized. A PCR protocol was developed in which changes in fluorescence emission resulting from specific annealing of fluorogenic adjacent hybridization probes to the target DNA were continuously monitored. Nine strains of the pathogenic Leptospira genospecies could be differentiated from Leptonema illini, Escherichia coli, and eight strains of Leptospira biflexa. The PCR method was rapid, requiring 18 min for the completion of 45 cycles. It was also simple and flexible, as DNA templates prepared by four different methods, including the simple boiling method, could be used without adverse effects. Two hundred copies of target, equivalent to 100 cells, could be detected.