Quantification of Mucosa oxygenation using three discrete spectral bands of visible light

Quantification of the mucosa oxygenation levels during Endoscopic imaging provides useful physiological/diagnostic information. In this work a method for non‐contact quantification of the oxygen saturation index during Endoscopic imaging using three discrete spectral‐band in the blue, the green, and the red parts of the spectrum (RGB bands) has been investigated. The oxygen saturation index (TOI_rgb) was calculated from the three discrete RGB spectral bands using diffusion approximation modeling and least‐square analysis. A parametric study performed to identify the optimum band width for each of the three spectral bands. The quantification algorithm was applied to in vivo images of the endobronchial mucosa to calculate (TOI_rgb) from selected areas within the image view. The results were compared to that obtained from the full visible spectral (470–700 nm, 10 nm) measurements. The analysis showed that a band width of at least 20 nm in the blue and the green is required to obtain best results. The results showed that the method provides accurate estimation of the oxygenation levels with about 90% accuracy compared to that obtained using the full spectra. The results suggest the potential of quantifying the oxygen saturation levels from the three narrow RGB spectral bands/images. (© 2009 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Identification and designation of telocentric chromosomes in barley by means of Giemsa N-banding technique

Summary Seven complete chromosomes and nine telocentric chromosomes in telotrisomics of barley (Hordeum vulgare L.) were identified and designated by an improved Giemsa N-banding technique. Karyotype analysis and Giemsa N-banding patterns of complete and telocentric chromosomes at somatic late prophase, prometaphase and metaphase have shown the following results: Chromosome 1 is a median chromosome with a long arm (Telo 1L) carrying a centromeric band, while short arm (Telo 1S) has a centromeric band and two intercalary bands. Chromosome 2 is the longest in the barley chromosome complement. Both arms show a centromeric band, an intercalary band and two faint dots on each chromatid at middle to distal regions. The banding pattern of Telo 2L (a centromeric and an intercalary band) and Telo 2S (a centromeric, two intercalary and a terminal band) corresponded to the banding pattern of the long and short arm of chromosome 2. Chromosome 3 is a submedian chromosome and its long arm is the second longest in the barley chromosome complement. Telo 3L has a centromeric (fainter than Telo 3S) and an intercalary band. It also shows a faint dot on each chromatid at distal region. Telo 3S shows a dark centromeric band only. Chromosome 4 is the most heavily banded one in barley chromosome complement. Both arms showed a dark centromeric band. Three dark intercalary bands and faint telomeric dot were observed in the long arm (4L), while two dark intercalary bands in the short arm (4S) were arranged very close to each other and appeared as a single large band in metaphase chromosomes. A faint dot was observed in each chromatid at the distal region in the 4S. Chromosome 5 is the smallest chromosome, which carries a centromeric band and an intercalary band on the long arm. Telo 5L, with a faint centromeric band and an intercalary band, is similar to the long arm. Chromosomes 6 and 7 are satellited chromosomes showing mainly centromeric bands. Telo 6S is identical to the short arm of chromosome 6 with a centromeric band. Telo 3L and Telo 4L were previously designated as Telo 3S and Telo 4S based on the genetic/linkage analysis. However, from the Giemsa banding pattern it is evident that these telocentric chromosomes are not correctly identified and the linkage map for chromosome 3 and 4 should be reversed. One out of ten triple 2S plants studied showed about 50% deficiency in the distal portion of the short arm. Telo 4L also showed a deletion of the distal euchromatic region of the long arm. This deletion (32%) may complicate genetic analysis, as genes located on the deficient segment would show a disomic ratio. It has been clearly demonstrated that the telocentric chromosomes of barley carry half of the centromere. Banding pattern polymorphism was attributed, at least partly, to the mitotic stages and differences in techniques.Contribution from the Department of Agronomy and published with the approval of the Director of the Colorado State University Experiment Station as Scientific Series Paper No. 2730. This research was supported in part by the USDA/SEA Competitive Research Grant 5901-0410-9-0334-0, USDA/ SEA-CSU Cooperative Research Grant 12-14-5001-265 and Colorado State University Hatch Project. This paper was presented partly at the Fourth International Barley Genetics Symposium, Edinburgh, Scotland, July 22–29, 1981     

基于图像融合与混合像元分解的城市植被盖度提取

城市植被盖度提取对于开展城市绿色空间保护和城市规划具有重要意义。随着遥感技术的发展,混合像元分解模型被广泛用于从中等分辨率的多光谱影像提取城市植被盖度,但较低的影像空间分辨率限制了该模型的应用领域。为此,以杭州市为例,首先引入Gram-Schmidt(GS)方法对Landsat ETM+的多光谱波段和全色波段进行融合,再通过混合像元分解模型从ETM+融合影像上提取城市植被盖度,最后利用SPOT影像进行精度检验。结果发现,采用GS方法对影像进行融合后,标准差、信息熵、平均梯度提高,相对偏差小于0.07,说明在保留多光谱信息的基础上提高了其空间分辨率。与SPOT影像相比,在融合影像上75%以上样本的植被盖度值相似,误差较大的区域是市区植被特别稀疏或茂盛的像元。与源影像相比,从融合影像上提取的植被盖度的均方根误差和系统误差降低了0.01。该方法在降低城市植被监测成本、提高监测精度方面具有潜力。     

High-resolution analysis of DNA replication domain organization across an R/G-band boundary.

Establishing how mammalian chromosome replication is regulated and how groups of replication origins are organized into replication bands will significantly increase our understanding of chromosome organization. Replication time bands in mammalian chromosomes show overall congruency with structural R- and G-banding patterns as revealed by different chromosome banding techniques. Thus, chromosome bands reflect variations in the longitudinal structure and function of the chromosome, but little is known about the structural basis of the metaphase chromosome banding pattern. At the microscopic level, both structural R and G bands and replication bands occupy discrete domains along chromosomes, suggesting separation by distinct boundaries. The purpose of this study was to determine replication timing differences encompassing a boundary between differentially replicating chromosomal bands. Using competitive PCR on replicated DNA from flow-sorted cell cycle fractions, we have analyzed the replication timing of markers spanning roughly 5 Mb of human chromosome 13q14.3/q21.1. This is only the second report of high-resolution analysis of replication timing differences across an R/G-band boundary. In contrast to previous work, however, we find that band boundaries are defined by a gradient in replication timing rather than by a sharp boundary separating R and G bands into functionally distinct chromatin compartments. These findings indicate that topographical band boundaries are not defined by specific sequences or structures.     

Chromosome ideograms of the laboratory rat (Rattus norvegicus) based on high-resolution banding, and anchoring of the cytogenetic map to the DNA sequence by FISH in sample chromosomes

A detailed banded ideogram representation of the rat chromosomes was constructed based on actual G-banded prometaphase chromosomes. The approach yielded 535 individual bands, a significant increase compared to previously presented ideograms. The new ideogram was adapted to the existing band nomenclature. The gene locus positions in the rat draft DNA sequence were compared to the chromosomal positions as determined by dual-color FISH, using rat (RNO) chromosomes 6 and 15 and a segment of RNO4 as sample regions. It was found that there was generally an excellent correlation in the chromosome regions tested between the relative gene position in the DNA molecules and the sub-chromosomal localization by FISH and subsequent information transfer on ideograms from measurements of chromosomal images. However, in the metacentric chromosome (RNO15), the correlation was much better in the short arm than in the long arm, suggesting that the centromeric region may distort the linear relationship between the chromosomal image and the corresponding DNA molecule.     

Use Reference Bands to Accurately Estimate ISCN Band Levels 400, 550, and 850

In their 2002 Guidelines for chromosome analysis of peripheral blood, the American College of Medical Genetics states that "The 550-band stage should be the goal of all constitutional studies..." The College of American Pathologists requires that the average case be analyzed at the 400-band level of resolution for routine work, and that the 550-band level be achieved in appropriate blood samples. The challenge is how to identify the 400, 550, and 850-band levels confidently and consistently. In this study, our objectives were to develop simple and reliable criteria to estimate band level, and to evaluate our laboratory's performance with respect to those criteria. Using the ISCN(1995) ideogram as a reference, candidate bands were selected for the three band levels: 400, 550 and 850. A pilot and two follow-up studies were conducted and a set of candidate bands were validated against the Vancouver method of evaluating band level so that band level scores were similar using either method. The final set of reference bands were the presence of 9q32 and 20q13.2 for the 400-band level; 5q33.2 and 10q22.2 for the 550-band level; and 3p26.1, 18q22.3 and 20q13.32 for the 850-band level. Cell selection improved after each technologist was provided a composite image of chromosomes with reference bands highlighted. The band level criteria presented here involve no band counting, appear to be objective, can help to improve quality and consistency among technologists, and can ensure compliance with regulatory agencies.     

Increasing the resolution of chromosome analysis using pyrido[1,2-a]benzimidazoles

We studied the influence of three derivatives of pyrido[1,2-a]benzimidazoles (PBIs), which have DNA-intercalating properties, on plant mitotic chromosome condensation, in order to increase the resolution of chromosome analysis. The efficiency of the influence of these agents was assessed using the median chromosome length on chromosome slides, as well as by the number and size of chromosome DAPI bands. We used the third chromosome of Linum grandiflorum Desf. in these experiments. The chromosome was identified on the slides using its DAPI band pattern and a molecular marker, viz., the 5S rDNA site, which is located in the proximal region of the long arm of the chromosome. The influence of the well-known 9-aminoacridine (9-AMA) DNA intercalator, which is widely used in karyotype studies of short-chromosome organisms, was used as a control in all of the experiments. It was found that the influence of each of the three PBIs in the study on the root meristem of L. grandiflorum resulted in an increase in the median length of the third chromosome, the linear centromeric DAPI band size, and the number of intercalary DAPI bands. All three PBIs acted more efficiently than 9-AMA. The median chromosome length was increased by 15?C40% and the number of intercalary bands increased by 1.5?C3 times after PBI treatment, as compared to 9-AMA treatment. At the same time, 7-CF₃-PBI, in a similar manner to 9-AMA, did not change the relative size of the centromeric DAPI band, while 7-NH₂-PBI and 9-NH₂-7-CF₃-PBI gradually increased this parameter. It is concluded that these substances can be used as intercalating agents in cytogenetic studies in order to increase the resolution of chromosome analysis.     

Equivalence of the total constitutive heterochromatin content by an interchromosomal compensation in the C band sizes of chromosomes 1, 9, 16, and Y in Caucasian and Japanese individuals

A quantitative analysis of C bands by densitometric measurements in chromosomes 1, 9, 16, and Y was conducted in Caucasians and Japanese living in Brazil. Sixty normal unrelated subjects (30 males and 30 females) were studied in each racial group. Caucasians presented C bands of chromosomes 1, 9, and 16 larger than Japanese, but, on average, only the difference for C bands of chromosome 9 was statistically significant. In the Japanese, the C band sizes of chromosomes Y were, on average, significantly larger than in the Caucasians. The mean C band size of chromosome 9 and the sum of the three pairs were significantly larger in Caucasian than in Japanese males. The total values of constitutive heterochromatin, sigma (1qh,9qh,16qh,Yq12), did not show significant difference between Caucasian and Japanese males. The relative C band sizes of chromosomes 1, 9, and 16 were, on average, similar in Caucasians and Japanese. No sex difference was found in both racial groups. As regards the heteromorphism, only the values of C bands of chromosome 9 were, on average, significantly larger in Caucasians than in Japanese. Partial inversions were detected only among the Caucasians.     

Fc receptor gene translocation in a t(1;19) pre-B ALL cell line

We have recently mapped the human FCGR2 gene to chromosome 1 bands q23-q24. In situ hybridization of FCGR2 cDNA with a cell line containing a t(1;19)(g23;p13) derived from a patient with pre-B ALL has allowed a more accurate localization of this gene to chromosome 1 band q23. Furthermore, this study indicated a splitting of the FCGR2 gene or gene cluster by the t(1;19). However, Southern analysis showed no genetic rearrangement when compared with a karyotypically normal Epstein-Barr virus (EBV)-transformed cell line from the same patient. This suggests that the translocation breakpoint does not occur within the coding region of this gene.     

The application of AFLP fingerprinting to construct a YAC contig containing ADH2 and MTP on sheep chromosome 6.

The low density of genetic markers on livestock maps limits progress in positional cloning projects. We demonstrate a strategy of combining comparative mapping with AFLP fingerprinting to develop physical maps in a defined region of the sheep genome. Sequence tagged sites for alcohol dehydrogenase 2 (ADH2) and microsomal triglyceride transfer protein (MTP) were developed and used to screen a sheep yeast artificial chromosome (YAC) library. Nine YACs were identified containing the microsatellite marker BM1329 and either ADH2 or MTP. Additional markers in the region were not available, and AFLP analysis was developed to identify sheep-specific bands within the YACs to determine their degree of overlap. Fourteen bands common to more than one YAC were analysed and provided the markers necessary to develop a YAC contig containing the three STS markers. One YAC (yac260B5) containing all three markers (ADH2, MTP, and BM1329) was mapped to sheep chromosome 6q1.6-->q1.8 by FISH analysis.     

Iterative image reconstruction using modified non-local means filtering for limited-angle computed tomography

PurposeLimited-angle CT imaging is an effective technique to reduce radiation. However, existing image reconstruction methods can effectively reduce streak artifacts but fail to suppress those artifacts around edges due to incomplete projection data. Thus, a modified NLM (mNLM) based reconstruction method is proposed.MethodsSince the artifacts around edges mainly exist in local position, it is possible to restore the true pixels in artifacts using pixels located in artifacts-free regions. In each iteration, mNLM is performed on image reconstructed by ART followed by positivity constraint. To solve the problem caused by ART-mNLM that there is undesirable information that may appear in the image, ART-TV is then utilized in the following iterative process after ART-mNLM iterates for a number of iterations. The proposed algorithm is named as ART-mNLM/TV.ResultsSimulation experiments are performed to validate the feasibility of algorithm. When the scanning range is [0, 150°], our algorithm outperforms the ART-NLM and ART-TV with more than 40% and 29% improvement in terms of SNR and with more than 58% and 49% reduction in terms of MAE. Consistently, reconstructed images from real projection data also demonstrate the effectiveness of presented algorithm.ConclusionThis paper uses mNLM which benefits from redundancy of information across the whole image, to recover the true value of pixels in artifacts region by utilizing pixels from artifact-free regions, and artifacts around the edges can be mitigated effectively. Experiments show that the proposed ART-mNLM/TV is able to achieve better performances compared to traditional methods.     

Generation of a complete set of human telomeric band painting probes by chromosome microdissection

Chromosomal rearrangements involving telomeric bands have been frequently detected in many malignancies and congenital diseases. To develop a useful tool to study chromosomal rearrangements within the telomeric band effectively and accurately, a whole set of telomeric band painting probes (TBP) has been generated by chromosome microdissection. The intensity and specificity of these TBPs have been tested by fluorescence in situ hybridization and all TBPs showed strong and specific signals to target regions. TBPs of 6q and 17p were successfully used to detect the loss of the terminal band of 6q in a hepatocellular carcinoma cell line and a complex translocation involving the 17p terminal band in a melanoma cell line. Meanwhile, the TBP of 21q was used to detect a de novo translocation, t(12;21), and the breakpoint at 21q was located at 21q22.2. Further application of these TBPs should greatly facilitate the cytogenetic analysis of complex chromosome rearrangements involving telomeric bands.     

Demonstration of two different regions of lateral asymmetry in human Y chromosomes

Summary Two differently stained regions of lateral asymmetry were observed in the long arm of the human Y chromosome, following FPG staining. The first asymmetry was confined to band q12 of the long arm. The second asymmetrically stained region was located at the junction between bands q11 and q12. In the non-fluorescent Y chromosomes only one region of lateral asymmetry was found at the end of the long arm and its staining properties were similar to the region situated at the junction between q11 and q12 bands in the fluorescent Ys. The two morphologically distinguishable regions of lateral asymmetry are presumed to indicate sites containing different satellite DNAs in the human Y chromosome.     

Chromosome-band-specific painting: chromosome in situ suppression hybridization using PCR products from a microdissected chromosome band as a probe pool

Summary We describe a chromosome-band-specific painting method that involves (1) microdissection of the chromosome, chromosomal region or band, (2) amplification of a variety of chromosome/region/band-specific DNA fragments with the polymerase chain reaction (PCR), and (3) chromosome in situ suppression hybridization (CISS) with the direct use of the PCR products as a probe pool. With this method, it was possible 1) to paint an entire X or Y chromosome, a distal one-fourth of 2q, and only a band at 8q24.1, 2) to identify the origin of a minute marker chromosome in a mentally retarded patient, 3) to detect an X;Y translocation in another patient, and 4) to identify one human chromosome 2 in a human-mouse hybrid cell line. This method allows us to identify not only structural chromosome abnormalities at the band level, but also the origin of cytogenetically unidentifiable marker chromosomes. It will also be useful in studies of evolutionary cytogenetics.     

Stylized chromosome images

Stylized chromosome images 1) serve as a format to test effects of preprocessing algorithms used in automated karyotyping; 2) enhance the ability of humans to perform quantitative analysis of chromosomal aberrations; 3) provide an alternative format for karyotype hard copies produced by automated systems. Stylized chromosomes are two-dimensional computer-generated images based on information extracted from one-dimensional width and density profiles. These profiles correspond to what cytogeneticists observe through the microscope as the shape and banding patterns of stained chromosomes. Stylized presentation sharpens chromosome band boundaries and perimeters, reduces "noise," and enhances gray level variations, which are difficult to distinguish by humans on photographic or computer generated karyotypes. Karyotyping accuracy using stylized images was used to detect difficult areas for automated chromosome identification. Landmark bands sufficient to classify chromosomes were identified; shapes of chromosomes reflected in width profiles were said to aid classification. A two-step automated karyotyping strategy proposed is: 1) classify chromosomes by landmarks, minimum information needed for identification; 2) subsequently employ the full banding pattern with maximum resolution to detect aberrations. Stylized images of abnormal chromosomes have potential for testing hypothesis regarding breakpoints and quantitative analysis, but improvements are needed in homologue normalization and definition of termini of chromosomes.     

Color standardization method and system for whole slide imaging based on spectral sensing

In the field of whole slide imaging, the imaging device or staining process cause color variations for each slide that affect the result of image analysis made by pathologist. In order to stabilize the analysis, we developed a color standardization method and system as described below. (1) Color standardization method based on RGB imaging and multi spectral sensing, which utilize less band (16 bands) than conventional method (60 bands). (2) High speed spectral sensing module. As a result, we confirmed the following effect. (1) We confirmed the performance improvement of nucleus detection by the color standardization. And we can conduct without training data set which is needed in conventional method. (2) We can get detection performance of H&E component equivalent to conventional method (60 bands). And measurement process is more than 255 times faster.     

BrdU处理的鱼类染色体高分辨G-带带型分析

本文应用鱼类染色体高分辨G-带技术,重点将黄鳝培养细胞具不同长度染色体的正中期分裂相做成G-带核型加以比较分析。随着染色体长度的增加,带纹数目也增加。但增加是有限度的。染色体带纹数目的增加,明显地表现在深染带再分为若干亚带。当染色体从前期向中、后期过渡收缩变短时,一些亚带融合为原来数目的带。染色体上各个带的收缩程度、收缩时间是不均等的。实验证明大剂量的BrdU不仅能阻断鱼类细胞于中S期,也可使染色体伸长、小剂量的伸长作用不明显。最后讨论了BrdU处理与G-显带的关系、染色体带纹数目相对恒定以及染色体伸长缩短问题。     

Raman spectroscopic investigations of sarcoplasmic reticulum membranes.

Raman spectra are presented for sarcoplasmic reticulum membranes. Interpretation of the 1000-1130 cm-1 region of the spectrum indicates that the sarcoplasmic reticulum membrane may be more fluid than erythrocyte membranes that have been examined by the I portion of the membrane spectrum with a strong 1658 cm-1 band characteristic of C=C stretching in hydrocarbon side chains exhibiting cis conformation. This band is unaltered in intensity and position in H2O and in 2H2O thus obscuring amide I protein conformation. Of particular interest is the appearance of strong, resonantly enhanced bands at 1160 and 1527 cm-1 attributable to membrane-associated carotenoids.     

Methylation imprinting was observed of mouse mo-2 macrosatellite on the pseudoautosomal region but not on chromosome 9

Mouse mo-2 macrosatellites consisting of 31-bp tandem repeat units are mainly located at two loci in the C57BL/6 genome, one being at the centromere-distal telomeric region of chromosome 9 and the other at the pseudoautosomal (PA) region of chromosomes X and Y. The two clustes constitute approximately 300 kb and 150 kb, respectively. Southern analysis of a methylation-sensitive enzyme, HpaII-digested DNA showed that the mo-2 macrosatellites are detected as more than 30 polymorphic bands. Comparison of those bands between reciprocally crossed F₁ mice revealed that approximately 20% of the allele-specific fragments exhibit different band intensities depending on the sex of the parent of origin. The differential methylation is observed in the mo-2 macrosatellite on the PA region but not in that on chromosome 9. Several fragments including the 3.4-kb fragment without internal HpaII site are more clearly detected when paternally derived, suggesting that the male-derived macrosatellite is undermethylated. Interestingly the difference is much more remarkable in inter-subspecific F₁ mice between C57BL/6 and MSM than F₁ between C57BL/6 and C3H/He. This suggests the presence of a modifier(s) that affect(s) the methylation of mo-2 in the MSM genome.