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Biomedical subjects

T Xu

Publications and source records attributed to T Xu.

At least 19 recordsLinked to original sources

Fracture spacing in layered materials and pattern transition from parallel to polygonal fractures.

We perform three-dimensional simulations of fracture growth in a three-layered plate model with an embedded heterogeneous layer under horizontal biaxial stretch (representing stretch from directional to isotropic) by the finite element approach. The fractures develop under a quasistatical, slowly increasing biaxial strain. The material inhomogeneities are accounted for by assigning each element a failure threshold that is defined by a given statistical distribution. A universal scale law of fracture spacing to biaxial strain in terms of principal stress ratio is well demonstrated in a three-dimensional fashion. The numerically obtained fracture patterns show a continuous pattern transition from parallel fractures, laddering fracture to polygonal fractures, which depends strongly on the far-field loading conditions in terms of principal stress ratio lambda = sigma(2)/sigma(1), from uniaxial (lambda = 0), anisotropic (0 < lambda < 1) to isotropic stretch (lambda = 1). We find that, except for further opening of existing fractures after they are well-developed (saturation), new fractures may also initiate and propagate along the interface between layers, which may serve as another mechanism to accommodate additional strain for fracture saturated layers.

Journal Article↗

Polymerase chain reaction-based denaturing gradient gel electrophoresis in the evaluation of oral microbiota.

BACKGROUND/AIMS: Clinical evaluation of oral microbial reduction after a standard prophylactic treatment has traditionally been based on bacterial cultivation methods. However, not all microbes in saliva or dental plaque can be cultivated. Polymerase chain reaction-based denaturing gradient gel electrophoresis (PCR-DGGE) is a cultivation-independent molecular fingerprinting technique that allows the assessment of the predominant bacterial species present in the oral cavity. This study sought to evaluate the oral microbial changes that occurred after a standard prophylactic treatment with a conventional oral care product using PCR-DGGE. METHODS: Twelve healthy adults participated in the study. Pooled plaque samples were collected at baseline, 24 h after prophylaxis (T1), and 4 days after toothbrushing with fluoride toothpaste (T4). The total microbial genomic DNA of the plaque was isolated. PCR was performed with a set of universal bacterial 16S rDNA primers. The PCR-amplified 16S rDNA fragments were separated by DGGE. The effects of the treatment and of dental brushing were assessed by comparing the PCR-DGGE fingerprinting profiles. RESULTS: The mean numbers of detected PCR amplicons were 22.3 +/- 6.1 for the baseline group, 13.0 +/- 3.1 for the T1 group, and 13.5 +/- 4.3 for the T4 group; the differences among the three groups were statistically significant (P < 0.01). The study also found a significant difference in the mean similarities of microbial profiles between the baseline and the treatment groups (P < 0.001). CONCLUSION: PCR-based DGGE has been shown to be an excellent means of rapidly and accurately assessing oral microbial changes in this clinical study.

Adult↗

Increased calcium deposits and decreased Ca2+-ATPase in right ventricular myocardium of ascitic broiler chickens.

Right ventricular hypertrophy and failure is an important step in the development of ascites syndrome (AS) in broiler chickens. Cytoplasmic calcium concentration is a major regulator of cardiac contractile function and various physiological processes in cardiac muscle cells. The purpose of this study was to measure the right ventricular pressure and investigate the precise ultrastructural location of Ca(2+) and Ca(2+)-ATPase in the right ventricular myocardium of chickens with AS induced by low ambient temperature. The results showed that the right ventricular diastolic pressure of ascitic broilers was significantly higher than that of control broilers (P < 0.01), and the maximum change ratio of right intraventricular pressure (RV +/- dp/dt(max)) of ascitic broilers was significantly lower than that of the controls (P < 0.01). Extensively increased calcium deposits were observed in the right ventricular myocardium of ascitic broilers, whereas in the age-matched control broilers, calcium deposits were much less. The Ca(2+)-ATPase reactive products were obviously found on the sarcoplasmic reticulum and mitochondrial membrane of the control right ventricular myocardium, but rarely observed in the ascitic broilers. The data suggest that in ascitic broilers there is the right ventricular diastolic dysfunction, in which the overload of intracellular calcium and the decreased Ca(2+)-ATPase activity might be the important factors.

Animals↗

Hypoxia induces major effects on cell cycle kinetics and protein expression in Drosophila melanogaster embryos.

Hypoxia induces a stereotypic response in Drosophila melanogaster embryos: depending on the time of hypoxia, embryos arrest cell cycle activity either at metaphase or just before S phase. To understand the mechanisms underlying hypoxia-induced arrest, two kinds of experiments were conducted. First, embryos carrying a kinesin-green fluorescent protein construct, which permits in vivo confocal microscopic visualization of the cell cycle, showed a dose-response relation between O2 level and cell cycle length. For example, mild hypoxia (Po2 approximately 55 Torr) had no apparent effect on cell cycle length, whereas severe hypoxia (Po2 approximately 25-35 Torr) or anoxia (Po2 = 0 Torr) arrested the cell cycle. Second, we utilized Drosophila embryos carrying a heat shock promoter driving the string (cdc25) gene (HS-STG3), which permits synchronization of embryos before the start of mitosis. Under conditions of anoxia, we induced a stabilization or an increase in the expression of several G1/S (e.g., dE2F1, RBF2) and G2/M (e.g., cyclin A, cyclin B, dWee1) proteins. This study suggests that, in fruit fly embryos, 1) there is a dose-dependent relationship between cell cycle length and O2 levels in fruit fly embryos, and 2) stabilized cyclin A and E2F1 are likely to be the mediators of hypoxia-induced arrest at metaphase and pre-S phase.

Animals↗

Abnormal spectra alteration observed in Triton calibration method for measuring [Ca2+]i with fluorescence indicator, fura-2.

We compared two commonly used calibration methods for measuring the concentration of intracellular free calcium ([Ca2+]i) by ratiometric fluorescence dye, fura-2 in mouse neuroblastoma-rat glioma hybrid cells (NG108-15). One calibration method, the Triton method, employs detergent Triton X-100, while the other, the Ionomycin method, uses a calcium-specific ionophore, Ionomycin. In the Triton method, we observed that at excitation 380 nm, the fura-2 fluorescence intensity of steady-state cells abnormally situated beyond the limiting intensity for calibration. By excitation scan, we demonstrated that this abnormality was caused by the change of fura-2 isosbestic points, which in turn was due to cell lysis after the addition of Triton X-100. This problem was resolved in the Ionomycin method by avoidance of cell lysis. Our results showed the correlation between inconsistent isosbestic points and cell lysis. As the basis for [Ca2+]i calibration, the proportionality between the fluorescence intensity and the concentration of dye species was impaired because of inconsistent isosbestic points. This inconsistency can be eliminated by a preliminary experiment of excitation scan to test the feasibility of different calibration methods.

Animals↗

Inkjet printing for high-throughput cell patterning.

The adaptation of inkjet printing technology to the complex fields of tissue engineering and biomaterial development presents the potential to increase progress in these emerging technologies through the implementation of this high-throughput capability via automated processes to enable precise control and repeatability. In this paper, a method of applying high-throughput inkjet printing to control cellular attachment and proliferation by precise, automated deposition of collagen is presented. The results indicate that commercial inkjet printing technology can be used to create viable cellular patterns with a resolution of 350 microm through the deposition of biologically active proteins. This method demonstrates a combination of off-the-shelf inkjet printing and biomaterials and has potential to be adapted to tissue engineering and colony patterning applications. Adapting this method into the three-dimensional construction of cellular structures for eventual high-throughput tissue engineering using a bottom-up approach is possible.

Animals↗

The release of cupric ion in simulated uterine: New material nano-Cu/low-density polyethylene used for intrauterine devices.

With the development of IUDs, a number of copper-bearing devices are now commercially available, including the copper-T, the Multiload and the copper-T in various other forms, so-called "the second-generation" IUDs. In this article, we report on nano-Cu/low-density polyethylene composite as a potential copper carrier in IUD. Two issues for the new material are addressed: the effectiveness of polymers in reducing the initial burst in cupric ion release and the amount and pattern of continuing release. The aim of this study was to investigate copper ion release from this composite as a basis for considering its used in an IUD.

Cations↗

A simple approach to examine early oral microbial biofilm formation and the effects of treatments.

BACKGROUND/AIMS: A simple in vivo approach to examine early dental plaque formation in the human mouth and to determine the effects of common dietary and oral hygiene procedures on biofilm formation is reported. METHODS: A custom designed device that fits securely behind the teeth of the mandibular arch provides a surface for microbial colonization. This device is prepared with denture acrylic and can be repeatedly used by the subject, exposing a large and constant surface area for microbial accumulation. RESULTS: Large numbers of oral bacteria colonized the device by 2 h; these increased significantly by 4 h (P < 0.05). Bacterial colonization increased significantly after rinsing with a sucrose solution (P < 0.05) but remained unaffected after rinsing with water, a commercially available fluoride mouthrinse without antimicrobial agents, or brushing with a fluoride dentifrice (P > 0.05). Rinsing with mouthrinses formulated with chlorhexidine, cetylpyridinium chloride or triclosan/copolymer significantly inhibited colonization (P < 0.05). A dose-dependent inhibition was noted with chlorhexidine rinses (P < 0.05). Brushing with a triclosan/copolymer dentifrice significantly inhibited microbial colonization compared with a control (P < 0.05). CONCLUSION: This simple approach was useful for examining the effects of common dietary and oral hygiene procedures. Significant biofilm inhibitory effects were noted with formulations that demonstrated efficacy in previous clinical studies.

Actinomyces viscosus↗

Biglycan-deficient mice have delayed osteogenesis after marrow ablation.

Biglycan (bgn) is a small proteoglycan in skeletal tissue that binds and regulates collagen and TGF-beta activities. Mice deficient in bgn (bgn-KO) develop age-dependent osteopenia and have multiple metabolic defects in their bone marrow stromal cells including increased apoptosis, reduced numbers of colony-forming units-fibroblastic (CFU-F) and decreased collagen production. In the present study we tested the hypothesis that bone formation capability in response to a physiological stress is compromised in bgn deficiency. We tested this theory using an in vivo bone marrow ablation assay. Ablation was performed on 6-week-old wild type (wt) and bgn-KO mice and bones were analyzed at days 7, 10, and 17 postsurgery. X-ray analysis showed that bone marrow ablation in femora induced vigorous new bone formation within 10 days in both genotypes but appeared greater in the wt compared to the bgn-KO. In order to quantitate the changes in bone formation in the ablated animals, bone densities of the proximal, midshaft, and distal femora were assessed using peripheral quantitative computed tomography (pQCT). The ratio of cancellous bone density at the midshaft (ablated limb/control limb) was significantly higher in wt compared to bgn-KO at day 10 postsurgery. Wt and bgn-KO femora had similar total and cancellous bone densities at days 7 and 17 postsurgery at all three locations indicating that the ablation effects were temporal and limited to the cancellous bone of the mid-shaft region. These data indicate that the absence of bgn directly impeded bone formation. Our results support the concept that bgn is important in controlling osteogenesis following marrow ablation.

Animals↗

Rapid inhibitory effects of corticosterone on calcium influx in rat dorsal root ganglion neurons.

Corticosterone may nongenomically affect cell functions in addition to its well-characterized effects on gene expression. The purpose of this study is to examine if corticosterone has a rapid nongenomic effect on excitability of dorsal root ganglion neurons by using patch-clamp and single-cell Ca(2+) microfluometry techniques. The results show that corticosterone has a dose-dependent rapid inhibitory effect on the voltage-dependent calcium currents in dorsal root ganglion neurons. Moreover, corticosterone inhibits [Ca(2+)](i) elevation induced by 50 mM high K(+) within just 3 s. The inhibitory effects of corticosterone on the voltage-dependent calcium current and high K(+)-induced calcium influx diminish after adding protein kinase C inhibitor or pretreatment with pertussis toxin for 24 h. Our results demonstrate an nongenomic effect of corticosterone on the excitability of dorsal root ganglion neurons and the effect is mediated through a putative pertussis toxin-sensitive G-protein-coupled receptor and activation of protein kinase C.

Animals↗

The tuberous sclerosis complex (TSC) pathway and mechanism of size control.

We have identified three groups of growth-constraint genes using mosaic genetic screens in Drosophila melanogaster, including PTEN (phosphatase and tensin homologue deleted on chromosome 10), and the tuberous sclerosis complex (TSC) genes, Tsc1 and Tsc2. Our studies show that all three groups of genes participate in mechanisms that regulate organ and organism size in animals. We propose that mechanisms of organ size control are critical targets for diseases, such as tumorigenesis, which require an increase in tissue size and total mass, and for evolutionary events that alter the size of organisms. Using genetic and biochemical methods, we have shown that Tsc1 and Tsc2 function in the insulin/phosphoinositide 3-kinase (PI3K)/Akt pathway. We have shown that Akt regulates the Tsc1-Tsc2 complex by directly phosphorylating Tsc2. We have shown further that S6 kinase (S6K) is a downstream component of the PI3K/Akt/TSC pathway and reduction of S6K activity can block TSC defects. Recent studies from many laboratories have now confirmed our findings in mice, rats and human patients, and have shown that drugs that antagonize S6K activities, such as rapamycin, diminish tumours in TSC-deficient mice and rats. Clinical trials based on these findings have begun. Given that other components of the pathway, such as PTEN, are also mutated in a large number of cancer patients and that these components regulate intracellular insulin signalling, therapeutics based on the knowledge of the pathway could have effects beyond the TSC patient population.

Animals↗

Evaluation of beta-amyloid peptide 25-35 on calcium homeostasis in cultured rat dorsal root ganglion neurons.

Accumulation of beta-amyloid (Abeta) protein in brain is an important characteristic for the etiology of Alzheimer's disease. Of all the possible processes generating the neurotoxic effects by Abeta, disruption of intracellular Ca(2+) homeostasis is the primary event. In this process, various intracellular Ca(2+) regulatory mechanisms are reported to be involved. Using patch-clamp techniques, both low and high voltage activated Ca(2+) channel currents were recorded in the cultured dorsal root ganglion (DRG) neurons. Application of Abeta protein fragment, Abeta(25-35) (2 microM), for 30 s increased the amplitude in both currents. The Abeta-triggered facilitation effect of Ca(2+) channel was found in all the depolarized potentials tested, as shown in the current-voltage relationship. Furthermore, after applying single cell Ca(2+) microfluorometric method, it was found that Abeta(25-35) alone could trigger elevations of intracellular Ca(2+) concentration ([Ca(2+)](i)) level in 90% of the cells tested. The elevation diminished completely by cumulatively adding CdCl(2), NiCl(2), thapsigargin (TG), FCCP and Zn(2+) in the normal bath solution. Combining pharmacological approaches, we found that voltage-dependent Ca(2+) channels, Ca(2+) stores and a putative Zn(2+)-sensitive extracellular Ca(2+) entry, respectively, makes 61.0, 25.1, and 13.9% contribution to the [Ca(2+)](i) increase caused by Abeta. When tested in a Ca(2+)-free buffer, mitochondria was found to contribute 41.3% of Abeta produced [Ca(2+)](i) elevation and the remaining 58.7% was attributed to endoplasmic reticulum (ER) release.

Amyloid beta-Peptides↗

Inhibitory effect of synthetic histatin 5 on leukotoxin from Actinobacillus actinomycetemcomitans.

Actinobacillus actinomycetemcomitans is a gram-negative bacterium strongly implicated in the pathogenesis of juvenile periodontitis. This periodontal pathogen synthesizes a leukotoxin that destroys human polymorphonuclear leukocytes (PMNs), and this toxin is thought to be responsible for the virulence of A. actinomycetemcomitans. It was therefore of interest to assess whether major virulence factors of periodontal pathogens were neutralized by salivary components. This study focuses on the effect of histatins, components of the nonimmune oral defense system, on leukotoxin activity. Leukotoxin was extracted with polymyxin B from freshly grown anaerobic cultures of A. actinomycetemcomitans strain Y4. PMNs isolated from blood of healthy human volunteers were incubated in a cytotoxicity assay containing PMNs (10(7) cells/ml) and leukotoxin preparation (0-500 microg/ml) in Hanks' balanced salt solution at 37 degrees C for 0-120 min with or without synthetic histatin 5 (0-500 microM). Cytotoxicity was measured by release of lactate dehydrogenase (LDH) at different time intervals. Histatin 5 neutralized the toxic effect of the leukotoxin preparation in a concentration-dependent manner, with an IC(50) value of 150 microM. When PMNs were preincubated with histatin 5 (300 microM), washed and subsequently exposed to leukotoxin, no protective effect was observed. This observation suggests a mechanism of inhibition whereby histatin 5 either directly neutralizes the leukotoxin or interferes with the leukotoxin-PMN interaction. The inhibitory effect of histatin 5 on leukotoxic activity may suggest a new biological function of histatins in the oral cavity as a naturally occurring secondary antibiotic.

Adult↗

Phenotypic effects of biglycan deficiency are linked to collagen fibril abnormalities, are synergized by decorin deficiency, and mimic Ehlers-Danlos-like changes in bone and other connective tissues.

Decorin (dcn) and biglycan (bgn), two members of the family of small leucine-rich proteoglycans (SLRPs), are the predominant proteoglycans expressed in skin and bone, respectively. Targeted disruption of the dcn gene results in skin laxity and fragility, whereas disruption of the bgn gene results in reduced skeletal growth and bone mass leading to generalized osteopenia, particularly in older animals. Here, we report that bgn deficiency leads to structural abnormality in collagen fibrils in bone, dermis, and tendon, and to a "subclinical" cutaneous phenotype with thinning of the dermis but without overt skin fragility. A comparative ultrastructural study of different tissues from bgn- and dcn-deficient mice revealed that bgn and dcn deficiency have similar effects on collagen fibril structure in the dermis but not in bone. Ultrastructural and phenotypic analysis of newly generated bgn/dcn double-knockout (KO) mice revealed that the effects of dcn and bgn deficiency are additive in the dermis and synergistic in bone. Severe skin fragility and marked osteopenia characterize the phenotype of double-KO animals in which progeroid changes are observed also in the skin. Ultrastructural analysis of bone collagen fibrils in bone of double-KO mice reveals a complete loss of the basic fibril geometry with the emergence of marked "serrated fibril" morphology. The phenotype of the double-KO animal mimics directly the rare progeroid variant of human Ehlers-Danlos syndrome (EDS), in which skin fragility, progeroid changes in the skin (reduced hypodermis), and osteopenia concur as a result of impaired glycosaminoglycan (GAG) linking to bgn and dcn core proteins. Our data show that changes in collagen fibril morphology reminiscent of those occurring in the varied spectrum of human EDS are induced by both bgn deficiency and den deficiency in mice. The effects of an individual SLRP deficiency are tissue specific, and the expression of a gross phenotype depends on multiple variables including level of expression of individual SLRPs in different tissues and synergisms between different SLRPs (and likely other macromolecules) in determining matrix structure and functional properties.

Animals↗

Profile of RGS expression in single rat atrial myocytes.

'Regulators of G protein signaling' (RGS proteins) are members of a large family of GTPase-activating proteins that are differentially expressed in various cell types and accelerate the termination of heterotrimeric G protein signaling. To identify RGS proteins that may affect autonomic regulation of atrial excitability, we screened the expression of nineteen RGS genes (RGS subfamilies A, B, C, and D) in single spontaneously beating rat atrial myocytes maintained in primary culture. Expression profiling by single-cell reverse transcriptase-polymerase chain reaction (RT-PCR) analysis revealed that seven distinct RGS genes are endogenously expressed in atrial myocytes which were also identified in poly(A)(+) mRNA from rat atria (RGS2, RGS3, RGS4, RGS6, RGS10, GAIP, and RGSZ2). Other RGS transcripts were detected in atrial poly(A)(+) mRNA but not single atrial myocytes (RGS5, RGS12, RGS16, and RGS18), and therefore are likely to originate from non-myocyte sources in atrial tissue. The single-cell RT-PCR experiments also led to the identification of putative splice variants for RGS6 and GAIP. Immunocytochemistry using RGS-specific antibodies confirmed the presence of selected RGS proteins in the cultured atrial myocytes. These results demonstrate a rich diversity of RGS expression in atrial myocytes whose specific role in G-protein signaling is yet to be determined. The identification of endogenous RGS proteins in atrial myocytes will facilitate targeted suppression and/or deletion studies to determine how each RGS protein may affect atrial excitability and its short-term and long-term regulation by G-protein signaling events.

Amino Acid Sequence↗

Human homologue of Drosophila lats, LATS1, negatively regulate growth by inducing G(2)/M arrest or apoptosis.

The lats gene encodes a family of proteins conserved from insects to humans. Drosophila carrying lats mutant cells or mice deficient for Lats1 develop tumors in various tissues. The mammalian LATS1 protein was previously shown to bind to CDC2, suggesting that LATS1 may modulate G(2)/M cell cycle progression by affecting CDC2 activity. In this study, we introduced human LATS1 into LATS(-/-) MEF cells by adenovirus-mediated gene transfer. Overexpression of LATS1 causes G(2)/M arrest through inhibition of CDC2 kinase activity. Furthermore, overexpression of LATS1 significantly suppressed the human tumor cell growth in vitro and tumorigenicity in vivo by inducing either cell cycle arrest in G(2)/M or apoptosis. These observations suggest that LATS1 is a potent growth suppressor and, like other tumor suppressors, it suppresses growth by inducing cell cycle arrest or apoptosis.

Adenoviridae↗

Axon-Schwann cell interactions regulate the expression of fibroblast growth factor-5 (FGF-5).

We screened for genes whose expression is significantly up- or downregulated during Wallerian degeneration in adult rat sciatic nerve with cDNA arrays. Fibroblast growth factor-5 (FGF-5) mRNA seemed to be induced. This was confirmed by northern blotting and in situ hybridization, as well as Western blotting for FGF-5 in axotomized nerve. Axon-Schwann cell interactions decreased the steady-state level of FGF-5 mRNA in regenerating sciatic nerves, and forskolin diminished its expression in cultured Schwann cells. We conclude that denervated Schwann cells synthesize FGF-5, which is a secreted, neuronotrophic member of the FGF family.

Animals↗

Li2AlB5O10.

A new compound, dilithium aluminium pentaborate, Li(2)AlB(5)O(10), has been synthesized by solid-state reaction and its structure determined by single-crystal X-ray diffraction. This compound is composed of [B(5)O(10)](5-) groups linked by AlO(4) tetrahedra. The [B(5)O(10)](5-) group consists of two hexagonal B-O rings perpendicular to each other connected by tetracoordinated boron. All the B-O rings in this structure can be divided into two groups, with one group approximately parallel and the other perpendicular to the c axis.

Journal Article↗