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Biomedical subjects

Gang Lin

Publications and source records attributed to Gang Lin.

14 recordsLinked to original sources

RPK118, a PX domain-containing protein, interacts with peroxiredoxin-3 through pseudo-kinase domains.

RPK118 is a sphingosine kinase-1-binding protein that has been implicated in sphingosine 1 phosphate-mediated signaling. It contains a PX (phox homology) domain and two pseudo-kinase domains, and co-localizes with sphingosine kinase-1 on early endosomes. In this study we identified a novel RPK118-binding protein, PRDX3 (peroxiredoxin-3), by yeast two-hybrid screening. The interaction between these proteins was confirmed by pull-down assays and co-immunoprecipitation experiments. Deletion studies showed that RPK118 interacted with PRDX3 through its pseudokinase domains, and with early endosomes through its PX domain. Double immunofluorescence experiments demonstrated that PRDX3 co-localized with RPK118 on early endosomes in COS7 cells. PRDX3 is a member of the antioxidant family of proteins synthesized in the cytoplasm and functioning in mitochondria. Our findings indicate that RPK118 is a PRDX3-binding protein that may be involved in transporting PRDX3 from the cytoplasm to its mitochondrial site of function or to other membrane structures via endosome trafficking.

Animals↗

Hierarchical, model-based merging of multiple fragments for improved three-dimensional segmentation of nuclei.

BACKGROUND: Automated segmentation of fluorescently labeled cell nuclei in three-dimensional confocal images is essential for numerous studies, e.g., spatiotemporal fluorescence in situ hybridization quantification of immediate early gene transcription. High accuracy and automation levels are required in high-throughput and large-scale studies. Common sources of segmentation error include tight clustering and fragmentation of nuclei. Previous region-based methods are limited because they perform merging of two nuclear fragments at a time. To achieve higher accuracy without sacrificing scale, more sophisticated yet computationally efficient algorithms are needed. METHODS: A recursive tree-based algorithm that can consider multiple object fragments simultaneously is described. Starting with oversegmented data, it searches efficiently for the optimal merging pattern guided by a quantitative scoring criterion based on object modeling. Computation is bounded by limiting the depth of the merging tree. RESULTS: The proposed method was found to perform consistently better, achieving merging accuracy in the range of 92% to 100% compared with our previous algorithm, which varied in the range of 75% to 97%, even with a modest merging tree depth of 3. The overall average accuracy improved from 90% to 96%, with roughly the same computational cost for a set of representative images drawn from the CA1, CA3, and parietal cortex regions of the rat hippocampus. CONCLUSION: Hierarchical tree model-based algorithms significantly improve the accuracy of automated nuclear segmentation without sacrificing speed.

Algorithms↗

Characterization of a Mycobacterium tuberculosis proteasomal ATPase homologue.

A screen for Mycobacterium tuberculosis (Mtb) mutants sensitive to reactive nitrogen intermediates identified transposon insertions in the presumptive proteasomal ATPase gene mpa (mycobacterium proteasome ATPase; Rv2115c). mpa mutants are attenuated in both wild type and nitric oxide synthase 2 deficient mice. In this work, we show that attenuation of mpa mutants is severe, and that Mpa is an ATPase associated with various cellular activities (AAA) ATPase that forms hexameric rings resembling the eukaryotic complex p97/valosin-containing protein (VCP). Point mutations in the conserved Walker box ATPase motifs of Mpa greatly reduced or abolished ATPase activity in vitro and abrogated protection of Mtb against acidified nitrite. A mutant Mpa protein missing only its last two amino acids retained ATPase activity, yet failed to protect Mtb against nitrite. The corresponding strain was attenuated in mice. Thus, Mpa is an ATPase whose enzymatic activity is necessary but not sufficient to protect against reactive nitrogen intermediates.

Adenosine Triphosphatases↗

3D-catFISH: a system for automated quantitative three-dimensional compartmental analysis of temporal gene transcription activity imaged by fluorescence in situ hybridization.

Fluorescence in situ hybridization (FISH) of neural activity-regulated, immediate-early gene (IEG) expression provides a method of functional brain imaging with cellular resolution. This enables the identification, in one brain, of which specific principal neurons were active during each of two distinct behavioral epochs. The unprecedented potential of this differential method for large-scale analysis of functional neural circuits is limited, however, by the time-intensive nature of manual image analysis. A comprehensive software tool for processing three-dimensional, multi-spectral confocal image stacks is described which supports the automation of this analysis. Nuclei counterstained with conventional DNA dyes and FISH signals indicating the sub-cellular distribution of specific, IEG RNA species are imaged using different spectral channels. The DNA channel data are segmented into individual nuclei by a three-dimensional multi-step algorithm that corrects for depth-dependent attenuation, non-isotropic voxels, and imaging noise. Intra-nuclear and cytoplasmic FISH signals are associated spatially with the nuclear segmentation results to generate a detailed tabular/database and graphic representation. Here we present a comprehensive validation of data generated by the automated software against manual quantification by human experts on hippocampal and parietal cortical regions (96.5% concordance with multi-expert consensus). The high degree of reliability and accuracy suggests that the software will generalize well to multiple brain areas and eventually to large-scale brain analysis.

Algorithms↗

The synthesis, distribution, and anti-hepatic cancer activity of YSL.

YSL was prepared stepwise from C terminal to N terminal with the side chain un-protective amino acids, Boc-Leu-OMe, Boc-Ser-OH, and Boc-Tyr-OH, as the starting materials in 39.5% total yield (31.2g/per batch). With the side chain un-protective Boc-(3,5-dibromo)-Tyr-OH and HCl.Ser-Leu-OMe as the starting materials (3,5-(3)H-Tyr)-Ser-Leu-OH was obtained in 29% yield. The determination of radioactive quantity in the urine and feces indicated that even after the administration for 130 h only 8.4% (5.35% in urine and 3.05% in feces) of total radioactive quantity from the metabolite of [3,5-(3)H-Tyr]-Ser-Leu-OH were monitored. The distribution study revealed the relative accumulation level of the individual tissue was arranged in the sequence of spleen>liver>kidney>lung>heart>muscle>brain. Selecting hepatic cancer as the target YSL significantly increased the survival time of H22 tumor cells implanted mice.

Animals↗

Predictive scheduling algorithms for real-time feature extraction and spatial referencing: application to retinal image sequences.

Real-time spatial referencing is an important alternative to tracking for designing spatially aware ophthalmic instrumentation for procedures such as laser photocoagulation and perimetry. It requires independent, fast registration of each image frame from a digital video stream (1024 x 1024 pixels) to a spatial map of the retina. Recently, we have introduced a spatial referencing algorithm that works in three primary steps: 1) tracing the retinal vasculature to extract image feature (landmarks); 2) invariant indexing to generate hypothesized landmark correspondences and initial transformations; and 3) alignment and verification steps to robustly estimate a 12-parameter quadratic spatial transformation between the image frame and the map. The goal of this paper is to introduce techniques to minimize the amount of computation for successful spatial referencing. The fundamental driving idea is to make feature extraction subservient to registration and, therefore, only produce the information needed for verified, accurate transformations. To this end, the image is analyzed along one-dimensional, vertical and horizontal grid lines to produce a regular sampling of the vasculature, needed for step 3) and to initiate step 1). Tracing of the vascular is then prioritized hierarchically to quickly extract landmarks and groups (constellations) of landmarks for indexing. Finally, the tracing and spatial referencing computations are integrated so that landmark constellations found by tracing are tested immediately. The resulting implementation is an order-of-magnitude faster with the same success rate. The average total computation time is 31.2 ms per image on a 2.2-GHz Pentium Xeon processor.

Algorithms↗

Kinetically controlled synthesis of wurtzite ZnS nanorods through mild thermolysis of a covalent organic-inorganic network.

The high-temperature (over 1020 degrees C) polymorph of ZnS, wurtzite ZnS, has been successfully prepared through a low-temperature (180 degrees C) hydrothermal synthesis route in the presence of ethylenediamine (en). The effects of en concentrations, reactant concentrations, reaction temperatures, and reaction times on crystal structures and shapes of ZnS have been investigated. We have demonstrated that the wurtzite ZnS showing rodlike morphology can be kinetically stabilized in the presence of en, especially at a high reactant concentration under appropriate hydrothermal conditions. Besides phase evolution of ZnS from hexagonal to cubic, morphological transformation from nanorods to nanograins has also been observed in the present investigation. Nanograins of phase-pure cubic ZnS, the thermodynamically stable polymorph, are easily prepared, and no hexagonal ZnS nanorods are detected in "pure" water, i.e., in the absence of en molecules. The above investigations indicate that the controlled fabrication of wurtzite ZnS nanorods is due to a mediated generation of the lamellar phase, ZnS.0.5en, a covalent organic-inorganic network based on ZnS slabs, and to its subsequent thermolysis in aqueous solution. The controlled growth of wurtzite ZnS nanorods and sphalerite ZnS nanograins provides us an opportunity to structurally modulate physical properties. These wurtzite ZnS nanorods display narrower and stronger blue emission than sphalerite ZnS nanograins.

Journal Article↗

A hybrid 3D watershed algorithm incorporating gradient cues and object models for automatic segmentation of nuclei in confocal image stacks.

BACKGROUND: Automated segmentation of fluorescently-labeled cell nuclei in 3D confocal microscope images is essential to many studies involving morphological and functional analysis. A common source of segmentation error is tight clustering of nuclei. There is a compelling need to minimize these errors for constructing highly automated scoring systems. METHODS: A combination of two approaches is presented. First, an improved distance transform combining intensity gradients and geometric distance is used for the watershed step. Second, an explicit mathematical model for the anatomic characteristics of cell nuclei such as size and shape measures is incorporated. This model is constructed automatically from the data. Deliberate initial over-segmentation of the image data is performed, followed by statistical model-based merging. A confidence score is computed for each detected nucleus, measuring how well the nucleus fits the model. This is used in combination with the intensity gradient to control the merge decisions. RESULTS: Experimental validation on a set of rodent brain cell images showed 97% concordance with the human observer and significant improvement over prior methods. CONCLUSIONS: Combining a gradient-weighted distance transform with a richer morphometric model significantly improves the accuracy of automated segmentation and FISH analysis.

Algorithms↗

Studies on the large scale synthesis and anti-tumor activity of YSL.

H-Tyr-Ser-Leu-OH (YSL), isolated from the degradation products of porcine spleen, was synthesized by a solution method on a large scale via two synthetic strategies. Its structure was confirmed by spectroscopic and related analyses. The in vivo anti-tumor activity was evaluated with murine H22 tumor models. The result indicated that, with the administration of YSL, the survival time of the murine H22 implanted mice was significantly prolonged.

Amino Acid Sequence↗

[Enhance of reproduction of Phanerochaete chrysosporium by adding natural lixiviums in liquid medium].

Great promotion to the reproduction of white rot fungus (Phanerochaete chrysosporium) by adding natural lixiviums such as from wood, maize core and potato in liquid medium was found in this research. Incubated in the liquid medium contained 10 g/L glucose as carbon source with natural lixiviums for three days, the production of mycelium pellet reached more than 80 g/L, which was 5 times of without natural lixiviums. Incubated in the liquid medium contained 5 mg/L glucose as carbon source with natural lixiviums for three days, the production of mycelium pellet can reach 69.5 g/L, while the production of medium without natural lixiviums was very low. When the basic medium contained 1 g/L-20 g/L glucose as carbon source, the production of mycelium pellet in 3 days can only reach 12.5 g/L to 14.5 g/L. The media with potato lixiviums were easily contaminated and the media with maize core lixiviums were easily bulking, while the media with wood lixiviums were not easily contaminated and bulking. Medium with wood lixiviums can produce more pellet than other medium, endure contamination and keep better sedimentation capacity. So, wood lixivium is better additive to the culture of white rot fungi in liquid medium. Addition of the mixture of wood, maize core and potato lixiviums is of advantage to the production of mycelium pellet. The difference of the production in the media with different amount of wood lixiviums showed little in the first 3 days, while it expanded after 3 days. Wood lixiviums stimulate the growth of P. chrysosporium instead of supply organics which fungi need.

Culture Media↗

[Analysis of alkaline phosphatase staining results in 238 cases of malignant lymphoma].

OBJECTIVE: To examine the significance of alkaline phosphatase (ALP) staining in clinical characterization of malignant lymphoma cases. METHODS: Fresh peripheral blood smear obtained from 238 cases of malignant lymphoma, including 23 cases with bone marrow involvement (BMI) 189 cases without BMI(NBMI), and 26 cases of lymphoma cell leukemia (LCL), were examined by ALP staining, and the results were compared with those of 40 normal control subjects. RESULTS: With the gradual progression of lymphoma, the patients tended to have higher ALP scores, and the percentages of the subjects with a score higher than 250 in the control, NBMI, BMI and LCL were 0, 4.8%, 39.1%, 53.9% respectively, showing significant differences between the four groups. CONCLUSIONS: Increased ALP activity can be indirectly indicative of lymphoma progression, and ALP staining results may provide insight into the clinical staging and prognosis of malignant lymphoma.

Adolescent↗

To enhance the reproduction of Phanerochaete chrysosporium by adding natural lixiviums in liquid medium.

Great promotion to the reproduction of white rot fungus Phanerochaete chrysosporium by adding natural lixiviums such as from wood, maize core and potato in liquid medium was found in this research. Incubated in the liquid medium contained 10 mg/L glucose as carbon source with natural lixiviums for three days, the production of mycelium pellet reaches more than 80 g/L, which is 5 times more than that of without natural lixiviums. Incubated in the liquid medium contained 5 mg/L glucose as carbon source with natural lixiviums for three days, the production of mycelium pellet can reach 69.5 g/L, while the production in the medium without natural lixiviums is very low. When the liquid medium contained 1-20 g/L glucose as carbon source, the production of mycelium pellet in 3 d can only reach 12.5 g/L to 14.5 g/L. The fungus in the medium with potato lixiviums are easily contaminated by other microorganisms and in the medium with maize core lixiviums are easily bulking, while in the medium with wood lixiviums are neither easily contaminated nor bulking. Medium with wood lixiviums can produce more pellet than other medium, endure contamination and keep better sedimentation capacity. So that, wood lixivium is better additive to the culture of white rot fungi in liquid medium. Addition of the mixture of wood, maize core and potato lixiviums is of advantage to the production of mycelium pellet. The difference of the production in the medium with different amount of wood lixiviums showed little in the first 3 d, while it expanded after 3 d. Wood lixiviums stimulate the growth of P. chrysosporium instead of supply organics which fungi need.

Culture Media↗

[Super-selective pump-insertion into the target artery and regional intraarterial infusion chemotherapeutant and immunizator in treatment of the latter gastrointestinal cancer].

OBJECTIVE: To evaluate the effect of operative selective pump-insertion into the tumorous target artery, postoperative regional infusion chemotherapeutant and immunizator for treatment the latter gastrointestinal cancer. METHODS: The effect of operative super-selective pump-insertion into the tumorous nutritious artery, postoperative regional infusion chemotherapeutant and immunizator for treatment 88 cases patients suffering from irremovable gastrointestinal cancer was observed. Of them, 45 cases were gastric cancer, 31 cases were rectal cancer, 11cases were colic cancer. RESULTS: Complete response 2 case; Part response 77 cases, 11 cases patients had received secondary resection after intraarterial chemotherapy. Non chang 9 cases; effective rates reach to 89.8%. One, two and three years survival rates were 86.4%, 30.7% and 10.2%. Average survival period were 21.5 mouths. CONCLUSION: Super-selective pump-insertion into the artery and regional intraarterial chemotherapy is an efficient way in treatment of the latter gastrointestinal cancer, which can delay the survival period of patients with tumor, and increase the resectable rate.

Adolescent↗