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Biomedical subjects

X Fan

Publications and source records attributed to X Fan.

At least 55 records · Page 3Linked to original sources

Osteoclastogenesis is repressed by mechanical strain in an in vitro model.

Functional loading provides a site-specific signal for the regulation of bone mass and morphology. To determine if strain can inhibit the resorptive component of bone remodeling, osteoclast formation was assessed in marrow cultures plated on flexible membranes subjected to 5% strain for 10 cycles/minute, 24 hours per day. Cultures strained during days 2 through 7 inhibited osteoclast formation to 61+/-7% of control cultures (p < 0.05), a degree of inhibition similar to that observed when the cultures were subjected to strains during only days 2 through 4 but also evaluated on day 7 (67+/-4% of control; p < 0.05). In contrast, straining of cultures during days 5 through 7 had little influence on inhibiting the formation of osteoclasts (94+/-5% of control; no significant difference). The nonuniformly strained substrate was subdivided into three concentric rings. and cultures were used to examine the site-specificity of the inhibition caused by strain. Osteoclast formation in the outermost boundary, which was distended from 3.6 to 5%, was 41+/-7% of that observed in outer regions of control wells. The inhibitory potential of mechanical strain was reduced within the middle ring (73+/-6% of control osteoclasts: p < 0.01), where the strain ranged from 0.2 to 3.6%. The central region, which experienced strains equivalent to those in the middle ring (0.2 to -4% strain), showed inhibition of osteoclast formation to a similar degree (75+/-6% of control). Media harvested from strained cultures failed to inhibit osteoclast formation in unstrained cultures; this implies that the inhibitory effect of strain depended on the direct interaction of the cell with the substrate rather than by a humoral factor. A second device, where a uniform strain was delivered at 1.8% throughout the entire plate, inhibited osteoclast recruitment to 48+/-3.6%, emphasizing that uniform strain in the absence of shear stress constrains osteoclast recruitment. These in vitro experiments can but model the complex environment generated by in vivo mechanical strains: however, they provide the first direct evidence that strain must be considered as inhibitory to osteoclast recruitment.

Animals↗

Ultrasound power deposition model for the chest wall.

An ultrasound power deposition model for the chest wall was developed based on secondary-source and plane-wave theories. The anatomic model consisted of a muscle-ribs-lung volume, accounted for wave reflection and refraction at muscle-rib and muscle-lung interfaces, and computed power deposition due to the propagation of both reflected and transmitted waves. Lung tissue was assumed to be air-equivalent. The parts of the theory and numerical program dealing with reflection were experimentally evaluated by comparing simulations with acoustic field measurements using several pertinent reflecting materials. Satisfactory agreement was found. A series of simulations were performed to study the influence of angle of incidence of the beam, frequency, and thickness of muscle tissue overlying the ribs on power deposition distributions that may be expected during superficial ultrasound (US) hyperthermia of chest wall recurrences. Both reflection at major interfaces and attenuation in bone were the determining factors affecting power deposition, the dominance of one vs. the other depending on the angle of incidence of the beam. Sufficient energy is reflected by these interfaces to suggest that improvements in thermal doses to overlying tissues are possible with adequate manipulation of the sound field (advances in ultrasonic heating devices) and prospective treatment planning.

Computer Simulation↗

The value of assessing pulmonary venous flow velocity for predicting severity of mitral regurgitation: A quantitative assessment integrating left ventricular function.

Although alteration in pulmonary venous flow has been reported to relate to mitral regurgitant severity, it is also known to vary with left ventricular (LV) systolic and diastolic dysfunction. There are few data relating pulmonary venous flow to quantitative indexes of mitral regurgitation (MR). The object of this study was to assess quantitatively the accuracy of pulmonary venous flow for predicting MR severity by using transesophageal echocardiographic measurement in patients with variable LV dysfunction. This study consisted of 73 patients undergoing heart surgery with mild to severe MR. Regurgitant orifice area (ROA), regurgitant stroke volume (RSV), and regurgitant fraction (RF) were obtained by quantitative transesophageal echocardiography and proximal isovelocity surface area. Both left and right upper pulmonary venous flow velocities were recorded and their patterns classified by the ratio of systolic to diastolic velocity: normal (>/=1), blunted (<1), and systolic reversal (<0). Twenty-three percent of patients had discordant patterns between the left and right veins. When the most abnormal patterns either in the left or right vein were used for analysis, the ratio of peak systolic to diastolic flow velocity was negatively correlated with ROA (r = -0.74, P <.001), RSV (r = -0.70, P <.001), and RF (r = -0.66, P <.001) calculated by the Doppler thermodilution method; values were r = -0.70, r = -0.67, and r = -0.57, respectively (all P <.001), for indexes calculated by the proximal isovelocity surface area method. The sensitivity, specificity, and predictive values of the reversed pulmonary venous flow pattern for detecting a large ROA (>0.3 cm(2)) were 69%, 98%, and 97%, respectively. The sensitivity, specificity, and predictive values of the normal pulmonary venous flow pattern for detecting a small ROA (<0.3 cm(2)) were 60%, 96%, and 94%, respectively. However, the blunted pattern had low sensitivity (22%), specificity (61%), and predictive values (30%) for detecting ROA of greater than 0.3 cm(2) with significant overlap with the reversed and normal patterns. Among patients with the blunted pattern, the correlation between the systolic to diastolic velocity ratio was worse in those with LV dysfunction (ejection fraction <50%, r = 0.23, P >.05) than in those with normal LV function (r = -0.57, P <.05). Stepwise linear regression analysis showed that the peak systolic to diastolic velocity ratio was independently correlated with RF (P <.001) and effective stroke volume (P <.01), with a multiple correlation coefficient of 0.71 (P <.001). In conclusion, reversed pulmonary venous flow in systole is a highly specific and reliable marker of moderately severe or severe MR with an ROA greater than 0.3 cm(2), whereas the normal pattern accurately predicts mild to moderate MR. Blunted pulmonary venous flow can be seen in all grades of MR with low predictive value for severity of MR, especially in the presence of LV dysfunction. The blunted pulmonary venous flow pattern must therefore be interpreted cautiously in clinical practice as a marker for severity of MR.

Aged↗

Development of apple superficial scald, soft scald, core flush, and greasiness is reduced by MCP.

1-Methylcyclopropene (MCP) was used to evaluate the role of ethylene in development of apple (Malus x domestica Borkh.) physiological disorders during storage. Granny Smith, Red Chief Delicious, and Fuji apple fruit were treated with MCP at a concentration of 1 microL L(-)(1) for 12 h at 20 degrees C. For all varieties stored at 0 degrees C, ethylene production and respiration rates were reduced for several months following MCP treatment, and firmness and titratable acidity of treated fruit were higher compared to controls. Apples treated with MCP did not develop superficial scald or peel greasiness through 6 months storage plus ripening at 20 degrees C for 7 days. Core flush was not observed in MCP-treated fruit until 6 months after treatment when the incidence was still lower compared to control fruit. MCP delayed the rise in production of alpha-farnesene and reduced accumulation of its oxidation products.

Cyclopropanes↗

Impact of 1-methylcyclopropene and methyl jasmonate on apple volatile production.

Climacteric Fuji apples were treated with 10 microL x L(-1) MCP (1-methylcyclopropene), 2 mmol x L(-1) MJ (methyl jasmonate), or a combination of 10 microL x L(-1) MCP and 2 mmol x L(-1) MJ. Fruit were kept at 20 degrees C for 15 days after treatment. Production of ethylene and other volatile compounds was measured prior to and 3, 7, 11, and 15 days after treatment. Ethylene production decreased 3 days following MJ treatment and then increased. MCP treatment alone or in combination with MJ inhibited ethylene production. MJ and MCP inhibited production of many volatile alcohols and esters. The production of individual alcohols and esters appears to be differentially inhibited by MJ or MCP. MJ and MCP inhibited not only production of alcohols but also formation of esters from alcohols.

Acetates↗

Cloning and expression of Aplysia carboxypeptidase D, a candidate prohormone-processing enzyme.

Many peptide hormones in a variety of species are produced from larger precursors by limited proteolysis at basic amino acid-containing sites. The marine mollusc Aplysia has homologs of mammalian peptide-processing enzymes, including furin, prohormone convertase 1 (PC1), PC2, and carboxypeptidase E (CPE). A novel neuronal Aplysia enzyme was recently identified that was most closely related to carboxypeptidase D (CPD; Fan and Nagle, DNA Cell Biol. 15, 937-945, 1996), a second carboxypeptidase thought to be present in the secretory pathway and to contribute to peptide hormone processing. We have identified and cloned multiple overlapping bag-cell neuron cDNAs that encode two proteins that are members of the CPD family. Sequence analyses demonstrate that the longer CPD protein (1446 residues) contains an N-terminal signal peptide and four carboxypeptidase-like domains; the third and fourth domains are not predicted to form active enzymes, as several critical residues are absent. The shorter CPD protein is predicted to contain two active carboxypeptidase-like domains. Northern blot analysis identified a major Aplysia CPD mRNA (5.3 kb) and several smaller minor transcripts in central nervous system tissue. The CPD was purified from Aplysia ovotestis using a method previously developed for mammalian CPD. The purified Aplysia CPD binds antisera raised against regions of the protein encoded by the Aplysia cDNA clone, as well as an antiserum raised against duck CPD. The enzymatic properties of purified Aplysia CPD are generally similar to those of mammalian CPD. Aplysia CPD is a candidate prohormone-processing enzyme that may play a role in the processing of Aplysia prohormones in the secretory pathway.

Amino Acid Sequence↗

Comparison of genetic heterogeneity of hepatitis C viral RNA in liver tissue and serum.

OBJECTIVE: Hepatitis C virus (HCV) is known to be heterogeneous and to circulate as a group of closely related quasispecies in individual patients, although hepatic viral genetic characteristics have not been well documented. METHODS: Matched serum and liver samples were tested by reverse transcription polymerase chain reaction amplification and single stranded conformation polymorphism analysis of the hypervariable portion of the E2/NS1 region of the HCV genome. The number of quasispecies was compared with the amount of HCV RNA, HCV genotyping, and infection with the hepatitis G virus. RESULTS: Sixteen of 40 patients had HCV RNA detectable in serum and liver. The HCV genotype was identical in serum and liver of all but one case. HCV RNA levels were approximately 10-fold higher in liver than serum. The number of HCV quasispecies in serum ranged between two and six (median 3.0) and in the liver between 2 and 19 (median 3.5, mean liver/serum ratio 1 to 6.3, median 1.8). The number of quasispecies in liver was equal to or greater than that in serum in all cases. HGV infection was found in 14 cases and did not influence serum or hepatic levels of HCV RNA. CONCLUSIONS: The number of hepatic HCV quasispecies usually exceeds that in serum, independent of the amount of HCV RNA and HCV genotype. This finding is compatible with clearance of some quasispecies from serum, but not liver, by putative neutralizing antibodies.

Adult↗

Experimental assessment of power and temperature penetration depth control with a dual frequency ultrasonic system.

A novel ultrasound applicator for superficial simultaneous thermoradiotherapy consisting of two parallel-opposed linear arrays and a double-sided scanning reflector was constructed and tested for penetration depth control. In this design the arrays operate at different frequencies (1 and 5 MHz, in this study) and the input power to each array element (five 2 X 2 cm2 elements per array) is computer adjustable. The ultrasonic beams from the arrays are aimed at the scanning reflector which in turn deflects them simultaneously and in parallel toward the treatment volume. Relative intensity distributions generated by the prototype were measured in a degassed water phantom using a thermal technique for a selected reflector position; these showed that the ultrasonic intensity distribution can be controlled in the lateral dimensions by varying the input power level to individual array elements. A fixed-perfused canine kidney phantom was employed to demonstrate experimentally that real time penetration depth control is possible by varying the excitation magnitude of one array (frequency) relative to that of the other. It is concluded that the dual-frequency scanned-reflected ultrasound applicator offers a degree of dynamic three-dimensional control of the power deposition pattern of clinical significance.

Animals↗

Existing antilisterial immunity does not inhibit the development of a Listeria monocytogenes-specific primary cytotoxic T-lymphocyte response.

Infection of BALB/c mice with Listeria monocytogenes stimulates an antilisterial immune response evident by the appearance of H2-Kd-restricted CD8(+) cytotoxic T lymphocytes (CTLs) specific for the nanomer peptides amino acids (aa) 91 to 99 of listeriolysin O (LLO 91-99) and aa 217 to 225 of the p60 molecule (p60 217-225). We have introduced point mutations at anchor residues within LLO 91-99 (92F) or p60 217-225 (218F), and BALB/c mice infected with L. monocytogenes strains containing these point mutations do not develop CTLs specific for LLO 91-99 or p60 217-225, respectively. We have used these strains to test whether primary CTL responses against L. monocytogenes-derived determinants can be stimulated within an environment of existing antilisterial immunity. We found that the development of a primary L. monocytogenes-specific CTL response is not altered by existing immunity to L. monocytogenes. For example, primary immunization with the p60 218F strain of L. monocytogenes followed by a secondary immunization with wild-type L. monocytogenes results in stimulation of p60 217-225-specific CTLs at primary response levels and LLO 91-99-specific effectors at levels consistent with a memory CTL response. Similarly, primary immunization with the 92F strain of L. monocytogenes followed by a secondary immunization with wild-type L. monocytogenes results in stimulation of LLO 91-99-specific CTLs at primary response levels and p60 217-225-specific effectors at levels consistent with a memory CTL response. These results provide additional support for the use of L. monocytogenes as a recombinant vaccine vector and show that antivector immunity does not inhibit the development of a primary CTL response when the epitope is delivered by L. monocytogenes as the vaccine strain.

Amino Acid Substitution↗

Structures in Bacillus subtilis are recognized by CD14 in a lipopolysaccharide binding protein-dependent reaction.

The CD14 molecule expressed on monocytes and macrophages is a high-affinity receptor for bacterial lipopolysaccharide (LPS) and hence an important component of the innate immune system. LPS binding protein (LBP) is required to facilitate the binding of LPS to CD14 in vitro and is necessary for the induction of an inflammatory response to LPS in vivo. Here we show that CD14 and LBP can also bind to lipoteichoic acid from the gram-positive bacterium Bacillus subtilis. Although CD14 does not interact with intact B. subtilis organisms, a brief exposure of the bacteria to serum converts them into a form which can bind to CD14 in an LBP-dependent reaction. When serum-pretreated B. subtilis organisms are incubated with the myelomonocytic cell line U937, which expresses CD14, the bacteria are rapidly phagocytosed. The phagocytosis is strictly dependent both on LBP and on CD14. These in vitro results suggest that LBP plays a role in the innate response not only to gram-negative but also to gram-positive infections.

Acute-Phase Proteins↗

Lymphoguanylin: cloning and characterization of a unique member of the guanylin peptide family.

Guanylin and uroguanylin are small peptides containing two disulfide bonds that activate membrane guanylate cyclase-receptors in the intestine, kidney and other epithelia. Hybridization assays with a uroguanylin complementary DNA (cDNA) detected uroguanylin-like messenger RNAs (mRNAs) in the opossum spleen and testis, but these transcripts are larger than uroguanylin mRNAs. RT of RNA from spleen to produce cDNAs for amplification in the PCR followed by cloning and sequencing revealed a novel lymphoid-derived cDNA containing an open reading frame encoding a 109-amino acid polypeptide. This protein shares 84% and 40% of its residues with preprouroguanylin and preproguanylin, respectively. A 15-amino acid, uroguanylin-like peptide occurs at the COOH-terminus of the precursor polypeptide. However, this peptide is unique in having only three cysteine residues. We named the gene and its peptide product lymphoguanylin because the source of the first cDNA isolated was spleen and its mRNA is expressed in all of the lymphoid tissues tested. A 15-amino acid form of lymphoguanylin containing a single disulfide bond was synthesized that activates the guanylate cyclase receptors of human T84 intestinal and opossum kidney (OK) cells, although with less potency than uroguanylin and guanylin. Northern and/or RT-PCR assays detected lymphoguanylin mRNA transcripts in many tissues and organs of opossums, including those within the lymphoid/immune, cardiovascular/renal, reproductive, and central nervous organ systems. Lymphoguanylin joins guanylin and uroguanylin in a growing family of peptide agonists that activate transmembrane guanylate cyclase receptors, thus influencing target cell function via the intracellular second messenger, cGMP.

Amino Acid Sequence↗

A study of early pregnancy factor activity in the sera of women with trophoblastic tumor.

PROBLEM: To detect whether or not the early pregnancy factor (EPF)-like activity, or chaperonin 10, could be in the sera of patients with trophoblastic tumor in order to find another more efficient means to diagnose this kind of tumor. METHOD OF STUDY: The rosette inhibition assay was used to detect EPF-like activity in 216 sera, collected from patients with gestational trophoblastic tumor, including 47 sera of patients with choriocarcinoma, 68 sera of patients bearing invasive mole, and 101 sera of patients with vesicular mole. RESULTS: The accuracy of diagnosing malignant trophoblastic tumor by detecting EPF-like activity is 91.3% (105/115), with a false positive rate of 14.58% and a false negative rate of 4.8% by this method. Furthermore, the rosette inhibition titer (RIT) values have significant difference (P < 0.001) between the sera in patients with malignant trophoblastic tumor before treatment and those after treatment. CONCLUSIONS: This study demonstrated that diagnosis of malignant trophoblastic tumor could be made with an accuracy of 91.3% by detecting EPF-like activity and that EPF-like activity could be used as an indicator to distinguish benign from malignant trophoblastic tumor.

Adult↗

Is hepatitis G/GB virus-C virus hepatotropic? Detection of hepatitis G/GB virus-C viral RNA in liver and serum.

The recently identified hepatitis G virus (HGV, also named GB virus-C, GBV-C) appears to have similarities to hepatitis C virus and other flaviviridae. To better understand its clinical significance and hepatotropism, we collected liver tissue and matched serum samples from 56 patients undergoing liver transplantation. HGV/GBV-C RNA was detected by reverse transcription-nested PCR, using primers from the relatively conserved 5' noncoding region of the genome to detect HGV/GBV-C RNA and the amount was semiquantitatively estimated by serial 10-fold endpoint dilution. The presence and amount of HCV RNA was estimated by the same methodology. Seventeen patients (30%) had HGV/GBV-C RNA detectable either in liver or in serum, including two of three with cryptogenic liver disease. Interestingly, 5 of 17 (29%) patients had HGV/GBV-C RNA in serum but not liver, even with repeated testing of hepatic RNA from different portions of the liver. Furthermore, the titer of HGV/GBV-C RNA was significantly lower in liver than in serum in most samples (mean log titer, 1.33 vs. 2.56, P < 0.05). In contrast, all 21 patients with HCV RNA in serum also had the virus detectable in liver. In five patients coinfected with HCV and HGV/GBV-C, the mean titer of HCV RNA in liver was higher than that in serum (log titer, 2.8 vs. 3.0, P > 0.05). Thus, our results suggest that HGV/GBV-C is probably not hepatotropic and may replicate predominantly in sites other than the liver. These findings brings into question the role of HGV in causing significant liver disease.

5' Untranslated Regions↗

Telephone adaptation of the Modified Mini-Mental State Exam (3MS). The Cache County Study.

OBJECTIVE: To examine the concurrent validity of a newly developed telephone adaptation of the Modified Mini-Mental State Exam. BACKGROUND: Longitudinal studies of cognition may be advantaged by availability of assessment instruments that can be used over the telephone, as well as in person. METHOD: Subjects were 263 noninstitutionalized elderly residents of a rural community in southern Idaho, aged 65 to 93, who had little or no cognitive difficulty. At an average interval of four weeks, we administered the Modified Mini-Mental State Exam (3MS) and the newly adapted Telephone Modified Mini-Mental State Exam (T3MS). Order of administration was randomly assigned. RESULTS: Agreement between scores on the two instruments was good (r = 0.82, p < 0.001). When we applied various cutoff scores to the instruments, thereby generating assignments of individuals to "screen positive" and "screen negative" groups, the percent agreement in screening results ranged from 80% to 96% as we reduced the cutoff scores from 90 to 74 (100 points possible). CONCLUSIONS: At least among subjects without major cognitive syndromes, the Telephone Modified Mini-Mental State Exam provides a reasonable substitute for the more costly in-person 3MS. The telephone instrument should now be tested over a broader range of cognitive abilities in order to assess its validity in more impaired subjects, e.g., by studying an institutionalized sample.

Aged↗

Insecticidal activity of transgenic tobacco plants expressing both Bt and CpTI genes on cotton bollworm (Helicoverpa armigera).

Transgenic tobacco plants expressing both Bacillus thuringiensis (Bt) insecticidal protein and cowpea trypsin inhibitor (CpTI) genes was used to evaluate the insecticidal activity on the cotton bollworm (Helicoverpa armigera) compared with transgenic tobacco with Bt insecticidal protein gene alone. Mortality of first to third instar larvae fed on transgenic two genes tobacco for three days was significantly higher than that fed on transgenic Bt tobacco. First to fourth instar larvae fed on transgenic two genes tobacco continuously could not survive until pupation. Second instar larvae were fed on transgenic tobacco plants for three days and then were transferred to an artificial diet. The efficacy of transgenic two genes tobacco on mortality, larvae weight, percentage of pupation, and time to pupation were significantly higher than that of transgenic tobacco with Bt insecticidal protein gene alone. The results with transgenic tobacco as model plant were valuable for other crops both for the enhancement of insecticidal efficacy and for the delay of insect adaptation to transgenic Bt crops.

Animals↗

Mechanisms of hyaluronan-induced up-regulation of ICAM-1 and VCAM-1 expression by murine kidney tubular epithelial cells: hyaluronan triggers cell adhesion molecule expression through a mechanism involving activation of nuclear factor-kappa B and activating protein-1.

The matrix constituent hyaluronan (HA) markedly accumulates in inflammatory lesions. To gain insight into the biologic significance of this phenomenon we tested the hypothesis that HA could regulate cell adhesion molecule expression in epithelial cells. Using a clonal line of mouse cortical tubular (MCT) cells we found that fragmented intermediate m.w., but not high m.w., HA markedly increased ICAM-1 and VCAM-1 steady state mRNA and cell surface expression. Up-regulation of ICAM-1 and VCAM-1 mRNA by HA was preceded by a marked increase in NF-kappaB and activating protein-1 DNA binding activity in MCT cells. Transcript levels for the NF-kappaB inhibitor IkappaBalpha and for the activating protein-1 constituents c-jun and c-fos also increased in response to HA stimulation of tubular cells. Inhibition of NF-kappaB with the serine protease inhibitor N-tosyl-L-phenylalanine chloromethyl ketone blocked the HA-mediated expression of ICAM-1 and VCAM-1 in MCT cells. In conclusion, HA displays proinflammatory effects by directly stimulating the expression of the cell adhesion molecules ICAM-1 and VCAM-1 in mouse kidney epithelial cells. HA could thereby play an important role in leukocyte adhesion in inflammatory renal diseases.

3T3 Cells↗

The role of C2 domains in Ca2+-activated and Ca2+-independent protein kinase Cs in aplysia.

In the nervous system of the marine mollusk Aplysia there are two protein kinase C (PKC) isoforms, the Ca2+-activated PKC Apl I and the Ca2+-independent PKC Apl II. PKC Apl I, but not PKC Apl II is activated by a short-term application of the neurotransmitter serotonin. This may be explained by the fact that purified PKC Apl II requires a higher mole percentage of phosphatidylserine to stimulate enzyme activity than does PKC Apl I. In order to understand the molecular basis for this difference, we have compared the ability of lipids to interact with the purified kinases and with regulatory domain fusion proteins derived from the kinases using a variety of assays including kinase activity, phorbol dibutyrate binding, and liposome binding. We found that a C2 domain fusion protein derived from PKC Apl I binds to lipids constitutively, while a C2 domain fusion protein derived from PKC Apl II does not. In contrast, fusion proteins containing the C1 domains of PKC Apl I and PKC Apl II showed only small differences in lipid interactions. Thus, while the presence of a C2 domain assists lipid-mediated activation of PKC Apl I, it inhibits activation of PKC Apl II.

Animals↗