A Novel Stratification Method in Linkage Studies to Address Inter- and Intra-Family Heterogeneity in Autism

Most genome linkage scans for autism spectrum disorders (ASDs) have failed to be replicated. Recently, a new ASD phenotypic sub-classification method was developed which employed cluster analyses of severity scores from the Autism Diagnostic Interview-Revised (ADI-R). Here, we performed linkage analysis for each of the four identified ADI-R stratified subgroups. Additional stratification was also applied to reduce intra-family heterogeneity and to investigate the impact of gender. For the purpose of replication, two independent sets of single nucleotide polymorphism markers for 392 families were used in our study. This deep subject stratification protocol resulted in 16 distinct group-specific datasets for linkage analysis. No locus reached significance for the combined non-stratified cohort. However, study-wide significant (P = 0.02) linkage scores were reached for chromosomes 22q11 (LOD = 4.43) and 13q21 (LOD = 4.37) for two subsets representing the most severely language impaired individuals with ASD. Notably, 13q21 has been previously linked to autism with language impairment, and 22q11 has been separately associated with either autism or language disorders. Linkage analysis on chromosome 5p15 for a combination of two stratified female-containing subgroups demonstrated suggestive linkage (LOD = 3.5), which replicates previous linkage result for female-containing pedigrees. A trend was also found for the association of previously reported 5p14-p15 SNPs in the same female-containing cohort. This study demonstrates a novel and effective method to address the heterogeneity in genetic studies of ASD. Moreover, the linkage results for the stratified subgroups provide evidence at the gene scan level for both inter- and intra-family heterogeneity as well as for gender-specific loci.     

A Novel Improved Gram Staining Method Based on the Capillary Tube

In this work, an exploratory study was conducted to examine Gram staining based on the capillary tube. Each Gram staining step for all bacterial strains tested was completed in capillary tubes. The results showed that different Gram staining morphologies were clearly visible in the capillary tubes. The results presented here demonstrated that the improved method could effectively distinguish between Gram-positive and Gram-negative bacteria, and only small volumes of reagents were required in this method. Collectively, this efficient method could rapidly and accurately identify the types of bacteria. Therefore, our findings could be used as a useful reference study for other staining methods.Key words: Gram staining, capillary tube, bacteria, and glass slide

Since Hans Christian Joachim Gram reported a staining method in 1884 (Gram 1884), such a technique has experienced more than a century of development and has become frequently used in bacteriology. From 1940 to 1960, Gram staining’s clinical application reaches its peak (Kass 1987). In recent years, several automated instruments for Gram staining have also been applied for microbiological analysis (Baron et al. 2010; Li et al. 2020). With the development of modern science and technology, some new technologies are expected to replace Gram staining. For example, Sizemore et al. (1990) have developed an alternative Gram staining technique using a fluorescent lectin. Later on, several fluorescent Gram staining methods have been established, and some Gram staining techniques suitable for live bacterial suspension have been described (Mason et al. 1998; Fife et al. 2000; Forster et al. 2002; Kwon et al. 2019). Sharma et al. (2020) have found that acridine orange fluorescent staining is more sensitive than the Gram staining. Besides, Berezin et al. (2017) have established a method for detecting Gram-negative bacteria based on enhanced Raman spectroscopy. Lemozerskii et al. (2020) have also reported a method of bacterial discrimination using an acoustic resonator. However, Gram staining is still an vital detection method in practical application for many microbiologists and clinicians due to its rapidity and simplicity (Thompson et al. 2017; Jahangiri et al. 2018; Li et al. 2018a).Over the years, Gram staining has been modified for many times, such as the Brown-Hopps method, Brown-Brenn method, and Gram-Twort method (Brown and Brenn 1931; Brown and Hopps 1973; Peck and Badrick 2017), and these approaches as mentioned earlier are widely used in anatomical pathology laboratories. Through the comparison of various improved methods, it is found that Gram’s original four-step method is still used, and some researchers have adopted the three-step method, while its basic principle has not been changed. As reported by Huang and Cui (1996), the three-step Gram staining method combines the two steps of alcohol decolorization and re-staining procedure in one step. Although Gram staining is one of the most commonly used detection methods in clinical microbiology laboratories, many clinicians are skeptical of its results due to differences in operators, low control, and standardization (Samuel et al. 2016; Thomson 2016). Researchers have made efforts to improve the Gram staining’s accuracy and reliability over the past few years, such as repeated training and standardization of the staining procedure (Thomson 2016; Siguenza et al. 2019). In this study, we developed a standardized Gram staining procedure for bacterial identification using a capillary tube. A modified Gram staining method based on the capillary tube has not yet been reported to the best of our knowledge. Therefore, we proposed a novel improved Gram staining method to improve the accuracy of the detection results and Gram staining efficiency.Eight bacterial strains, Staphylococcus aureus, Escherichia coli, Bacillus subtilis, Bacillus Licheniformis, Serratia marcescens, Vibrio parahaemolyticus, Lactobacillus delbrueckii ssp. bulgaricus, and Streptococcus thermophilus were provided by the Laboratory of Microbial Engineering, College of Life Science, Luoyang Normal University. L. bulgaricus and S. thermophilus were inoculated into skim milk culture medium and maintained at 37°C for 12 h. S. marcescens, B. Licheniformis, E. coli, B. subtilis, V. parahaemolyticus, and S. aureus were inoculated into beef peptone agar slants and maintained at 37°C for 16 h.Capillary tubes with an internal diameter of 0.5 mm and a length of 100 mm were purchased from the Instrument Factory of West China University of Medical Sciences. Gram staining reagent was obtained from the Anhui Chaohuhongci Medical Equipment Co., Ltd.Procedure: (1) One or two drops of sterile water were placed in the center of a clean glass slide. An inoculating loop was hold in a flame until it was red-hot and then allowed to cool approximately 30 seconds. Subsequently, a loop of culture was transferred to the center of the slide. The sample was spread onto the slide using the inoculating loop, and a small volume of bacterial suspension was automatically transferred into the capillary tube.(2) The capillary tube was then heated by passing over a flame for several times until the liquid was completely evaporated. The capillary tube was naturally cooled in the air for several seconds.(3) One end of the capillary tube was hold upward, and the crystal violet solution was automatically transferred to the capillary tube, followed by standing for 1 minute. The remaining crystal violet solution of the capillary tube was then transferred to absorbent paper. The capillary tube was washed in a gentle and indirect stream of tap water for a few seconds, and samples were dried on absorbent paper.(4) One end of the capillary tube was hold upward, and Gram’s iodine solution was automatically transferred to the capillary tube, followed by standing for 1 minute. Subsequently, the capillary tube was washed using the same procedure as described above.(5) One end of the capillary tube was hold upward, and 95% ethanol was automatically transferred to the capillary tube, followed by standing for 30 seconds. Subsequently, the capillary tube was washed using the same procedure as described above.(6) One end of the capillary tube was hold upward, and the Safranin solution was automatically transferred to the capillary tube, followed by standing for 30 seconds to 1 minute. The subsequent procedure was the same as described above. Besides, conventional Gram staining was carried out according to the instructions from the reagent kit. According to the instructions, Gram-negative cells are in pink to red, and Gram-positive cells show a purple or blue color when observed under a microscope.The Gram staining is always the “first-stage criteria” in the preliminary identification of bacterial species according to their cell walls (Li et al. 2018b). Eight different bacterial species were examined to investigate our approach, and the strains were selected according to the Gram staining pattern. Gram-negative bacteria E. coli, V. parahaemolyticus, and S. marcescens were examined. Gram-positive bacteria S. thermophilus, L. bulgaricus, S. aureus, B. licheniformis and B. subtilis were also assessed. Fig. ​Fig.1,1, ​,2,2, and ​and33 illustrate the results of Gram staining of E. coli, V. parahaemolyticus, and S. marcescens, respectively. Fig. ​Fig.4,4, ​,5,5, ​,6,6, ​,7,7, and ​and88 show the Gram staining results of S. thermophilus, L. bulgaricus, S. aureus, B. subtilis, and B. licheniformis, respectively. These results were compared with those obtained using a glass slide for Gram staining. No matter spherical or rod-shaped or not, all bacterial strains could be differentiated into two classifications: Gram-positive and Gram-negative. Comparing these results, we found that the results obtained by the capillary tube method were consistent with the conventional Gram staining approach. It was worth mentioning that in contrast to direct heat fixation of bacteria on glass slides, heat fixation by passing the capillary tube over a flame should be carried out quickly and carefully. If the capillary tube was overheated, it might cause the capillary tube to rupture, and it is easy to blur the field of vision, making it challenging to observe the staining result (Fig. ​(Fig.9).9). Therefore, before the experiments, it is better to conduct a preliminary experiment and achieve the desired results.Open in a separate windowFig. 1.The Gram staining results of E. coli. A – Capillary sample, B – Glass slide sample.Open in a separate windowFig. 2.The Gram staining results of V. parahaemolyticus. A – Capillary sample, B – Glass slide sample.Open in a separate windowFig. 3.The Gram staining results of S. marcescens. A – Capillary sample, B – Glass slide sample.Open in a separate windowFig. 4.The Gram staining results of S. thermophiles. A – Capillary sample, B – Glass slide sample.Open in a separate windowFig. 5.The Gram staining results of L. bulgaricus. A – Capillary sample, B – Glass slide sample.Open in a separate windowFig. 6.The Gram staining results of S. aureus. A – Capillary sample, B – Glass slide sample.Open in a separate windowFig. 7.The Gram staining results of B. subtilis. A – Capillary sample, B – Glass slide sample.Open in a separate windowFig. 8.The Gram staining results of B. Licheniformis. A – Capillary sample, B – Glass slide sample.Open in a separate windowFig. 9.The microstructure of the overheated capillary tube.Several studies (Chimento et al. 1996; Wada et al. 2012; Li Zhu 2018b) have already pointed out that the property of the bacterial cell wall determines whether the organism will be Gram-positive or Gram-negative, and it plays a role in the choice of antibiotics when infection occurs. Since it has frequently been observed that not all bacteria react in the same manner to such staining procedure (Hale and Bisset 1956), it is necessary to make more tests upon a representative selection of Gram-positive and Gram-negative bacteria in future studies.Molecular biology techniques and high-precision measurement systems have been successfully developed, and they can distinguish bacterial types in clinical samples and improve microbial detection (Klaschik et al. 2002; Dolch et al. 2016; Kim et al. 2018). However, it is still urgently needed to develop a rapid and straightforward Gram staining approach to detect bacteria, especially for those who have only primary experimental conditions. Our results indicated a promising method for bacterial differentiation using the capillary tube as a carrier. Successful differentiation required only small volumes of reagents, and the results were achieved within a few minutes by applying an optical microscope. In addition, the method proposed in this paper had reference value to other staining methods requiring expensive reagents.In the present study, the improved Gram staining method was developed based on the pure cultures, and it was only a comparison of the staining results between known Gram-negative and Gram-positive bacteria in a glass slide and capillary tube. In order to improve the accuracy and stability of the results, future study is necessary to detect more bacterial species. In addition, the modified method was not applicable for direct Gram staining of clinical samples. In the future, it may have a positive effect by developing a special method for processing clinical samples.The experimental results demonstrated that an improved Gram staining method was suitable for differentiating the strains tested in our laboratory. The method could rapidly discriminate Gram-positive and Gram-negative bacteria. Besides, the method only required small volumes of reagents. A much more comfortable and reproducible Gram staining approach can be developed for microbiology research based on our studies.     

A Novel Fractal Coding Method Based on M-J Sets

In this paper, we present a novel fractal coding method with the block classification scheme based on a shared domain block pool. In our method, the domain block pool is called dictionary and is constructed from fractal Julia sets. The image is encoded by searching the best matching domain block with the same BTC (Block Truncation Coding) value in the dictionary. The experimental results show that the scheme is competent both in encoding speed and in reconstruction quality. Particularly for large images, the proposed method can avoid excessive growth of the computational complexity compared with the traditional fractal coding algorithm.     

A Novel Hepatitis C Virus Genotyping Method Based on Liquid Microarray

The strategy used to treat HCV infection depends on the genotype involved. An accurate and reliable genotyping method is therefore of paramount importance. We describe here, for the first time, the use of a liquid microarray for HCV genotyping. This liquid microarray is based on the 5′UTR — the most highly conserved region of HCV — and the variable region NS5B sequence. The simultaneous genotyping of two regions can be used to confirm findings and should detect inter-genotypic recombination. Plasma samples from 78 patients infected with viruses with genotypes and subtypes determined in the Versant™ HCV Genotype Assay LiPA (version I; Siemens Medical Solutions, Diagnostics Division, Fernwald, Germany) were tested with our new liquid microarray method. This method successfully determined the genotypes of 74 of the 78 samples previously genotyped in the Versant™ HCV Genotype Assay LiPA (74/78, 95%). The concordance between the two methods was 100% for genotype determination (74/74). At the subtype level, all 3a and 2b samples gave identical results with both methods (17/17 and 7/7, respectively). Two 2c samples were correctly identified by microarray, but could only be determined to the genotype level with the Versant™ HCV assay. Genotype “1” subtypes (1a and 1b) were correctly identified by the Versant™ HCV assay and the microarray in 68% and 40% of cases, respectively. No genotype discordance was found for any sample. HCV was successfully genotyped with both methods, and this is of prime importance for treatment planning. Liquid microarray assays may therefore be added to the list of methods suitable for HCV genotyping. It provides comparable results and may readily be adapted for the detection of other viruses frequently co-infecting HCV patients. Liquid array technology is thus a reliable and promising platform for HCV genotyping.     

Risk Stratification in Young Patients With Channelopathies

Identifying the young patient at risk of malignant arrhythmias and sudden cardiac death remains a challenge. It is increasingly recognised that sudden death, syncope and aborted cardiac arrest at a young age in patients with a structurally normal heart may be the result of various ion channel disorders - the channelopathies. The approach to risk stratification involves a combination of the clinical presentation, taken in conjunction with the family history, genetic testing, invasive electrophysiological studies or other provocative tests where appropriate and feasible. A logical approach to risk stratification in some of the commoner channelopathies seen in paediatric practice is presented.     

基于快速沃尔什变换的分子子序列识别

提出了一种基于快速沃尔什变换的分子子序列识别的方法。这种方法不仅能快速识别出子序列并确定子序列的位置，而且极大地降低了CPU运行时间和计算复杂度，结合实例对这种方法进行了分析。     

A Novel Dictionary Based Computer Vision Method for the Detection of Cell Nuclei

Cell nuclei detection in fluorescent microscopic images is an important and time consuming task in a wide range of biological applications. Blur, clutter, bleed through and partial occlusion of nuclei make individual nuclei detection a challenging task for automated image analysis. This paper proposes a novel and robust detection method based on the active contour framework. Improvement over conventional approaches is achieved by exploiting prior knowledge of the nucleus shape in order to better detect individual nuclei. This prior knowledge is defined using a dictionary based approach which can be formulated as the optimization of a convex energy function. The proposed method shows accurate detection results for dense clusters of nuclei, for example, an F-measure (a measure for detection accuracy) of 0.96 for the detection of cell nuclei in peripheral blood mononuclear cells, compared to an F-measure of 0.90 achieved by state-of-the-art nuclei detection methods.     

A Novel Method to Predict Genomic Islands Based on Mean Shift Clustering Algorithm

Genomic Islands (GIs) are regions of bacterial genomes that are acquired from other organisms by the phenomenon of horizontal transfer. These regions are often responsible for many important acquired adaptations of the bacteria, with great impact on their evolution and behavior. Nevertheless, these adaptations are usually associated with pathogenicity, antibiotic resistance, degradation and metabolism. Identification of such regions is of medical and industrial interest. For this reason, different approaches for genomic islands prediction have been proposed. However, none of them are capable of predicting precisely the complete repertory of GIs in a genome. The difficulties arise due to the changes in performance of different algorithms in the face of the variety of nucleotide distribution in different species. In this paper, we present a novel method to predict GIs that is built upon mean shift clustering algorithm. It does not require any information regarding the number of clusters, and the bandwidth parameter is automatically calculated based on a heuristic approach. The method was implemented in a new user-friendly tool named MSGIP—Mean Shift Genomic Island Predictor. Genomes of bacteria with GIs discussed in other papers were used to evaluate the proposed method. The application of this tool revealed the same GIs predicted by other methods and also different novel unpredicted islands. A detailed investigation of the different features related to typical GI elements inserted in these new regions confirmed its effectiveness. Stand-alone and user-friendly versions for this new methodology are available at http://msgip.integrativebioinformatics.me.     

A Novel Antibody Humanization Method Based on Epitopes Scanning and Molecular Dynamics Simulation

1-17-2 is a rat anti-human DEC-205 monoclonal antibody that induces internalization and delivers antigen to dendritic cells (DCs). The potentially clinical application of this antibody is limited by its murine origin. Traditional humanization method such as complementarity determining regions (CDRs) graft often leads to a decreased or even lost affinity. Here we have developed a novel antibody humanization method based on computer modeling and bioinformatics analysis. First, we used homology modeling technology to build the precise model of Fab. A novel epitope scanning algorithm was designed to identify antigenic residues in the framework regions (FRs) that need to be mutated to human counterpart in the humanization process. Then virtual mutation and molecular dynamics (MD) simulation were used to assess the conformational impact imposed by all the mutations. By comparing the root-mean-square deviations (RMSDs) of CDRs, we found five key residues whose mutations would destroy the original conformation of CDRs. These residues need to be back-mutated to rescue the antibody binding affinity. Finally we constructed the antibodies in vitro and compared their binding affinity by flow cytometry and surface plasmon resonance (SPR) assay. The binding affinity of the refined humanized antibody was similar to that of the original rat antibody. Our results have established a novel method based on epitopes scanning and MD simulation for antibody humanization.     

Novel Genetic Loci Identified for the Pathophysiology of Childhood Obesity in the Hispanic Population

Genetic variants responsible for susceptibility to obesity and its comorbidities among Hispanic children have not been identified. The VIVA LA FAMILIA Study was designed to genetically map childhood obesity and associated biological processes in the Hispanic population. A genome-wide association study (GWAS) entailed genotyping 1.1 million single nucleotide polymorphisms (SNPs) using the Illumina Infinium technology in 815 children. Measured genotype analysis was performed between genetic markers and obesity-related traits i.e., anthropometry, body composition, growth, metabolites, hormones, inflammation, diet, energy expenditure, substrate utilization and physical activity. Identified genome-wide significant loci: 1) corroborated genes implicated in other studies (MTNR1B, ZNF259/APOA5, XPA/FOXE1 (TTF-2), DARC, CCR3, ABO); 2) localized novel genes in plausible biological pathways (PCSK2, ARHGAP11A, CHRNA3); and 3) revealed novel genes with unknown function in obesity pathogenesis (MATK, COL4A1). Salient findings include a nonsynonymous SNP (rs1056513) in INADL (p = 1.2E-07) for weight; an intronic variant in MTNR1B associated with fasting glucose (p = 3.7E-08); variants in the APOA5-ZNF259 region associated with triglycerides (p = 2.5-4.8E-08); an intronic variant in PCSK2 associated with total antioxidants (p = 7.6E-08); a block of 23 SNPs in XPA/FOXE1 (TTF-2) associated with serum TSH (p = 5.5E-08 to 1.0E-09); a nonsynonymous SNP (p = 1.3E-21), an intronic SNP (p = 3.6E-13) in DARC identified for MCP-1; an intronic variant in ARHGAP11A associated with sleep duration (p = 5.0E-08); and, after adjusting for body weight, variants in MATK for total energy expenditure (p = 2.7E-08) and in CHRNA3 for sleeping energy expenditure (p = 6.0E-08). Unprecedented phenotyping and high-density SNP genotyping enabled localization of novel genetic loci associated with the pathophysiology of childhood obesity.     

A Novel Approach for Studying the Physiology and Pathophysiology of Myelinated and Non-Myelinated Axons in the CNS White Matter

Advances in brain connectomics set the need for detailed knowledge of functional properties of myelinated and non-myelinated (if present) axons in specific white matter pathways. The corpus callosum (CC), a major white matter structure interconnecting brain hemispheres, is extensively used for studying CNS axonal function. Unlike another widely used CNS white matter preparation, the optic nerve where all axons are myelinated, the CC contains also a large population of non-myelinated axons, making it particularly useful for studying both types of axons. Electrophysiological studies of optic nerve use suction electrodes on nerve ends to stimulate and record compound action potentials (CAPs) that adequately represent its axonal population, whereas CC studies use microelectrodes (MEs), recording from a limited area within the CC. Here we introduce a novel robust isolated "whole" CC preparation comparable to optic nerve. Unlike ME recordings where the CC CAP peaks representing myelinated and non-myelinated axons vary broadly in size, "whole" CC CAPs show stable reproducible ratios of these two main peaks, and also reveal a third peak, suggesting a distinct group of smaller caliber non-myelinated axons. We provide detailed characterization of "whole" CC CAPs and conduction velocities of myelinated and non-myelinated axons along the rostro-caudal axis of CC body and show advantages of this preparation for comparing axonal function in wild type and dysmyelinated shiverer mice, studying the effects of temperature dependence, bath-applied drugs and ischemia modeled by oxygen-glucose deprivation. Due to the isolation from gray matter, our approach allows for studying CC axonal function without possible "contamination" by reverberating signals from gray matter. Our analysis of "whole" CC CAPs revealed higher complexity of myelinated and non-myelinated axonal populations, not noticed earlier. This preparation may have a broad range of applications as a robust model for studying myelinated and non-myelinated axons of the CNS in various experimental models.     

Novel Parachlamydia acanthamoebae Quantification Method Based on Coculture with Amoebae

Parachlamydia acanthamoebae, belonging to the order Chlamydiales, is an obligately intracellular bacterium that infects free-living amoebae and is a potential human pathogen. However, no method exists to accurately quantify viable bacterial numbers. We present a novel quantification method for P. acanthamoebae based on coculture with amoebae. P. acanthamoebae was cultured either with Acanthamoeba spp. or with mammalian epithelial HEp-2 or Vero cells. The infection rate of P. acanthamoebae (amoeba-infectious dose [AID]) was determined by DAPI (4′,6-diamidino-2-phenylindole) staining and was confirmed by fluorescent in situ hybridization. AIDs were plotted as logistic sigmoid dilution curves, and P. acanthamoebae numbers, defined as amoeba-infectious units (AIU), were calculated. During culture, amoeba numbers and viabilities did not change, and amoebae did not change from trophozoites to cysts. Eight amoeba strains showed similar levels of P. acanthamoebae growth, and bacterial numbers reached ca. 1,000-fold (10⁹ AIU preculture) after 4 days. In contrast, no increase was observed for P. acanthamoebae in either mammalian cell line. However, aberrant structures in epithelial cells, implying possible persistent infection, were seen by transmission electron microscopy. Thus, our method could monitor numbers of P. acanthamoebae bacteria in host cells and may be useful for understanding chlamydiae present in the natural environment as human pathogens.     

Assessment of Mineral Pathophysiology in Patients with Diabetic Foot Ulcer

Biological Trace Element Research - Chronic non-healing diabetic foot ulcers (DFU) with a recurrence rate of over 50% in 3 years account for more than 1,08000 non-traumatic lower extremity...     

A Novel Method Based on Etiolation for Increasing Efficiency of In Vitro Propagation of Populus tomentosa

This paper describes a novel method based on etiolation treatment for micropropagation of Populus tomentosa. It includes: (1) Antibiotic “cefotaxime” (2.5 ppm in nutrient medium) is used to inhibit bacteria growth and the cultures which have been contaminated after long period subculture are recovered by sterilization with 1/10000 HgCl2(w/v). (2)Low concentration of TDZ (thidiazuron, 0.005 ppm) replaces expensive zeatin for enhancing bud differentiation and multiplication of leaf explants. (3) Shoot elongation is promoted after etiolation treatment, which will increase the number of shoots suitable for rooting, about 50 etiolated shoots are obtained in a 100 ml flask, 3—5 times more than those produced in traditional method. (4) Etiolated shoots or after their greening are used as cuttings for rooting in vivo and over 90% survival rate could be achieved when the medium is sterilized. This method is labour and money saving and high efficient.     

Predilection of Segmental Glomerulosclerosis Lesions for the Glomerulotubular Junction Area in Type 1 Diabetic Patients: A Novel Mapping Method

The location of segmental glomerular lesions in relation to the vascular or tubular pole may have diagnostic or prognostic significance. We have developed a model-based method to estimate the distance from a glomerular lesion to a given landmark (vascular or tubular pole) or the glomerular center and applied this to biopsies from 5 microalbuminuric, 5 normoalbuminuric and 7 proteinuric type 1 diabetic patients and 5 normal controls. The distance from each glomerular adhesion to the glomerulotubular junction was measured and divided by the glomerular radius, allowing comparability among different glomeruli, assuming a spherical shape for Bowman''s capsule, an assumption which was validated. The frequency of adhesions in 6 glomerular zones with equal height (zone I adjacent to the glomerulotubular junction and zones II–VI progressively farther away) was determined: 59% of adhesions were in zone I, 15% in zone II, 16% in zone III, 7% in zone IV and 3% in zone VI (adjacent to the hilus). In glomeruli with only one adhesion, 82% of these were in zone I. This new method accurately localizes segmental lesions within glomeruli and revealed a marked predilection in type 1 diabetic patients for segmental sclerosis to develop at the glomerulotubular junction.     

A Novel RUNX2 Mutation in Cleidocranial Dysplasia Patients

Cleidocranial dysplasia (CCD) is an autosomal-dominant heritable skeletal disease caused by heterozygous mutations in the RUNX2 gene. Here, the RUNX2 gene was analyzed within a CCD family from China, and a novel missense mutation (c. 475G --> C [p.G159R]) was identified. Normal and mutant RUNX2 expression vectors were then constructed and expressed transiently in NIH3T3 cells. Immunofluorescent staining and Western blotting showed that wild-type RUNX2 protein was localized exclusively in the nucleus; however, the mutant protein was found in both the nucleus and the cytoplasm, which demonstrated that transport of the RUNX2 mutant into the nucleus was disturbed by the G159R mutation. Therefore, we suggest that G159 is very important to promote RUNX2 nuclear localization. According to clinical analysis, the patient displays severe dysplasia of bones and relatively low-grade craniofacial abnormality, and we infer that G159 may be vital for normal skeletal development, other than control of tooth number. These findings confirm that mutations in the RUNX2 gene are associated with the pathogenesis of CCD across different ethnic backgrounds.     

A Novel Method for Optimum Global Positioning System Satellite Selection Based on a Modified Genetic Algorithm

In this paper, a novel method for selecting a navigation satellite subset for a global positioning system (GPS) based on a genetic algorithm is presented. This approach is based on minimizing the factors in the geometric dilution of precision (GDOP) using a modified genetic algorithm (MGA) with an elite conservation strategy, adaptive selection, adaptive mutation, and a hybrid genetic algorithm that can select a subset of the satellites represented by specific numbers in the interval (4 ∼ n) while maintaining position accuracy. A comprehensive simulation demonstrates that the MGA-based satellite selection method effectively selects the correct number of optimal satellite subsets using receiver autonomous integrity monitoring (RAIM) or fault detection and exclusion (FDE). This method is more adaptable and flexible for GPS receivers, particularly for those used in handset equipment and mobile phones.     

A Novel Method of Object Detection from a Moving Camera Based on Image Matching and Frame Coupling

A new method based on image matching and frame coupling to handle the problems of object detection caused by a moving camera and object motion is presented in this paper. First, feature points are extracted from each frame. Then, motion parameters can be obtained. Sub-images are extracted from the corresponding frame via these motion parameters. Furthermore, a novel searching method for potential orientations improves efficiency and accuracy. Finally, a method based on frame coupling is adopted, which improves the accuracy of object detection. The results demonstrate the effectiveness and feasibility of our proposed method for a moving object with changing posture and with a moving camera.