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Biomedical subjects

W Yu

Publications and source records attributed to W Yu.

At least 37 records · Page 2Linked to original sources

Automated LC-LC-MS-MS platform using binary ion-exchange and gradient reversed-phase chromatography for improved proteomic analyses.

A simple multidimensional liquid chromatography system utilizing an isocratic pump and a HPLC system is described for the comprehensive proteomic analysis of complex peptide digest mixtures by coupled LC-LC-MS-MS techniques. A binary ion-exchange separation was achieved through the use of a strong cation-exchange column followed by a reversed-phase column for data-dependent LC-MS-MS analysis of the unbound analytes, and following salt elution (and concomitant column reequilibration), the bound analytes. Off-line validation of the platform showed near quantitative recovery of fractionated peptides and essentially complete ion-exchange partitioning. In comparative analyses of a highly complex peptide digest mixture a >40% increase in the number of peptide and protein identifications was achieved using this multidimensional platform compared to an unfractionated control.

Amino Acid Sequence↗

Comparison of the prevalence of knee osteoarthritis between the elderly Chinese population in Beijing and whites in the United States: The Beijing Osteoarthritis Study.

OBJECTIVE: To estimate the prevalence of radiographic and symptomatic knee osteoarthritis (OA) in a population-based sample of elderly subjects in Beijing, China and compare it with that reported in the Framingham (Massachusetts) OA Study. METHODS: We recruited a sample of persons age > or = 60, using door-to-door enumeration in randomly selected neighborhoods in Beijing. Subjects completed a home interview including questions on knee symptoms and a hospital examination including knee radiographs obtained during weight bearing. The protocol was identical to that used in the Framingham OA Study. A reader read intermingled Beijing and Framingham Study films to ensure high reliability. We defined a subject as having radiographic knee OA when the Kellgren/Lawrence grade was > or = 2 in at least 1 knee. Symptomatic knee OA was recorded as present when knee pain was reported and the symptomatic knee had radiographic OA. We estimated the prevalence of these entities in elderly subjects in Beijing and compared it with OA prevalence in Framingham, using an age-standardized prevalence ratio. RESULTS: Of 2,180 age-eligible Beijing subjects contacted, knee radiographs were obtained in 1,787 (82.0%). The prevalence of radiographic knee OA was 42.8% in women and 21.5% in men. Symptomatic knee OA occurred in 15.0% of women and 5.6% of men. Compared with women of the same age in Framingham, women in Beijing had a higher prevalence of radiographic knee OA (prevalence ratio 1.45, 95% confidence interval 1.31-1.60) and of symptomatic knee OA (prevalence ratio 1.43, 95% confidence interval 1.16-1.75). The prevalence of knee OA in Chinese men was similar to that in their white US counterparts (for radiographic OA, prevalence ratio 0.90; for symptomatic OA, prevalence ratio 1.02). CONCLUSION: Using identical methods and definitions to evaluate the prevalence of OA across populations, we found, surprisingly, that older Chinese women have a higher prevalence of knee OA than women in Framingham, Massachusetts. The prevalence in men was comparable. Possible explanations for these differences range from genetic differences to heavy physical activity among Chinese.

Age Distribution↗

Towards defining the urinary proteome using liquid chromatography-tandem mass spectrometry. II. Limitations of complex mixture analyses.

With an emphasis on obtaining a multitude of high quality tandem mass spectrometry spectra for protein identification, instrumental parameters are described for the liquid chromatography-tandem mass spectrometry analysis of trypsin digested unfractionated urine using a hybrid quadrupole-time-of-flight (Q-TOF) mass spectrometer. Precursor acquisition rates of up to 20 distinct precursors/minute in a single analysis were obtained through the use of parallel precursor selection (four precursors/survey period) and variable collision induced dissociation integration time (1 to 6 periods summed). Maximal exploitation of the gas phase fractionated ions was obtained through the use of narrow survey scans and iterative data-dependent analyses incorporating dynamic exclusion. The impact on data fidelity as a product of data-dependent selection of precursor ions from a dynamically excluded field is discussed with regards to sample complexity, precursor selection rates, survey scan range and facile chemical modifications. Operational and post-analysis strategies are presented to restore data confidence and reconcile the greatest number of matched spectra.

Amino Acid Sequence↗

Towards defining the urinary proteome using liquid chromatography-tandem mass spectrometry. I. Profiling an unfractionated tryptic digest.

The proteome of normal male urine from a commercial pooled source has been examined using direct liquid chromatography-tandem mass spectrometry (LC-MS/MS). The entire urinary protein mixture was denatured, reduced and enzymatically digested prior to LC-MS/MS analysis using a hybrid-quadrupole time-of-flight mass spectrometer (Q-TOF) to perform data-dependent ion selection and fragmentation. To fragment as many peptides as possible, the mixture was analyzed four separate times, with the mass spectrometer selecting ions for fragmentation from a subset of the entire mass range for each run. This approach requires only an autosampler on the HPLC for automation (i.e, unattended operation). Across these four analyses, 1.450 peptide MS/MS spectra were matched to 751 sequences to identify 124 gene products (proteins and translations of expressed sequence tags). Interestingly, the experimental time for these analyses was less than that required to run a single two-dimensional gel.

Amino Acid Sequence↗

Indications for early postoperative intraperitoneal chemotherapy of advanced gastric cancer: results of a prospective randomized trial.

Previous analysis of this prospective randomized trial and meta-analysis of published randomized trials of adjuvant intraperitoneal chemotherapy demonstrated improved survival in patients with advanced gastric cancer. Simple criteria applicable at the time of surgery for patient selection were sought in this analysis. From 1990 to 1995 a series of 248 patients with biopsy-proven gastric cancer were randomized intraoperatively to receive early postoperative intraperitoneal mitomycin C and 5-fluorouracil (125 patients) versus surgery only (123 patients). Gastric resection plus early postoperative intraperitoneal chemotherapy showed improved overall survival compared to surgery only (54% and 38%, respectively; p = 0.0278). There were statistically significant differences in patients with stage III (57% and 23%, respectively; p = 0.0024) and in those with stage IV (28% and 5%, respectively; p = 0.0098) gastric cancer. The improvement in survival rate was statistically significant for the subgroup of patients with gross serosal invasion (52% and 25%, respectively; p = 0.0004) and patients with lymph node metastasis (46% and 22%, respectively; p = 0.0027). The surgeons' impression about lymph node status was unreliable, but assessment of serosal invasion was accurate in 80% of cases. Gross serosal invasion with or without frozen section evaluation of lymph nodes can be used as the major selection criteria for early postoperative intraperitoneal chemotherapy of advanced gastric cancer.

Antibiotics, Antineoplastic↗

Characterisation and high-resolution distribution of a matrix attachment region-binding protein (MFP1) in proliferating cells of onion.

The first matrix attachment region (MAR)-binding protein sequenced in plants, MFP1, has been characterised in two dicot species. Based on their antigenic relationship, we report here the conservation of MFP1-like proteins in proliferating root cells of onion (Allium cepa L). Two MFP1-like proteins with different molecular masses and solubilities were detected. The most abundant was a 90-kDa basic protein, presenting several separate spots in two-dimensional blots. The MFP1 was partially soluble and, similar to the proliferating cell nuclear antigen (PCNA)-labelled replication factories in the nucleus and nuclear matrix, was localised at discrete foci as detected by confocal microscopy. High-resolution immunolocalisation of MFP1 by electron microscopy identified the foci as nuclear structures, some of them containing PCNA, which are ultrastructurally similar to the replication factories described in animal cells. Our data provide the first report on MFP1-like proteins in the Alliaceae. In addition, we present evidence of the presence of AcMFP1 in the putative replication factories.

Arabidopsis Proteins↗

Possible impact of genetic differences on the development of neuropathic pain-like behaviors after unilateral sciatic nerve ischemic injury in rats.

The development of neuropathic-like behaviors following unilateral ischemic injury to the sciatic nerve was examined and compared in four rat strains: Sprague--Dawley (SD), Wistar--Kyoto (WK), spontaneously hypertensive (SHR) and Dark--Agouti (DA). We have also compared two sub-strains of SD rats supplied from two different vendors (SD-BK and SD-DK). The responses to mechanical, heat or cold stimuli of both hind paws were measured before and regularly after injury for up to 10 weeks. Spontaneous paw lifting and changes in paw posture after nerve injury were also examined. Significant differences in basal sensitivity to mechanical or heat stimulation were seen among the four rat strain studied with SHR and DA rats being less sensitive than the SD and WK rats. All strains of rats developed bilateral mechanical allodynia and ipsilateral heat hyperalgesia after photochemically-induced nerve ischemia, but the time-course and magnitude of the responses were significantly different among the strains. Again, the SHR and DA were found to be least susceptible to the development of abnormal pain-like responses. Cold allodynia occurred only in WK and SD-BK. SD-DK rats on the other hand developed more severe mechanical allodynia than SD-BK. SHR and DA rats showed less deficits in paw posture after nerve injury whereas spontaneous pain lifting, a measure of possible spontaneous pain, was comparable among all strains. Light microscopic study of the injured sciatic nerve showed comparable nerve damage in SHR, WK and two sub-strains of SD rats. The DA rats however exhibited reduced area of intraneural damage. Finally, electronmicroscopic examination revealed that damage to both myelinated and unmyelinated fibers occurred in this model in all strains. These results showed that normal sensitivity and the development of pain-like response after partial nerve injury differ substantially among different strains of rats, supporting the emerging concept that genetic factors affect pain sensitivity under normal conditions and after nerve injury. The apparent resistance of DA rats to nerve ischemia, however, may suggest that genetic factors not directly related to pain modulation also play a role in the diverse outcomes. Our results indicate that sub-strains of rats also showed variable development of neuropathic pain-like behaviors to both the modality and magnitude of the effect. Thus, controlling sub-strains is also important in experimental studies of neuropathic pain in rats.

Animals↗

Symmetrical cationic triglycerides: an efficient synthesis and application to gene transfer.

Some cationic triglycerides 1Aa-1Cb which have a symmetrical structure were effectively synthesized and formulated into cationic liposomes with the co-lipid dioleoylphosphatidylethanolamine (DOPE) and/or dilauroylphosphatidylcholine (DLPC). The plasmid encoding a luciferase was delivered into CHO cells by using these cationic liposomes. Our symmetrical cationic triglycerides showed high transfection activity when DOPE was used as a co-lipid. Among the symmetrical cationic triglycerides synthesized here, 1Ab and 1Ac, which have an oleoyl group at the 1- and 3-position in the glycerol backbone and also have a relatively long linker connecting the 2-hydroxy group in glycerol with the quaternary ammonium head group, were found to be the most suitable for gene delivery into cells. The transfection activity of the symmetrical cationic triglyceride 1Ab was comparable with that of its asymmetrical congener 6 and several times higher than that of Lipofectin.

Animals↗

The epidemiology of hospitalization of elderly Americans for septicemia or bacteremia in 1991-1998. Application of Medicare claims data.

PURPOSE: To describe the epidemiology of hospitalization of elderly Americans for septicemia or bacteremia. METHODS: Medicare claims data for discharges from 1991 through 1998 were used to study 75,920 hospitalizations with the principal diagnosis of septicemia or bacteremia in patients aged 65 years or older. RESULTS: "Unspecified septicemia" was the commonest principal diagnosis, followed by septicemia due to Escherichia coli or staphylococci. From 1991 through 1997, annual discharges for "unspecified septicemia" increased 108%, and those for pneumococcal septicemia increased 310%. Decreases in reported septicemia were seen after increases in the proportion of beneficiaries in Medicare health maintenance organizations. Discharge rates for septicemia principal diagnoses increased steeply with age. Age-specific discharge rates were usually highest for black men and lowest for white women. Exceptions included septicemia due to E. coli, with white men at low risk, and pneumococcal septicemia, without significant differences between races or sexes. The case-fatality rate in hospital ranged from 4.2% with "bacteremia" and 6.9% with E. coli septicemia to 22.2% with "septicemia due to gram-negative organism, unspecified," and 26.8% with "unspecified septicemia." Staphylococcal septicemia, septicemia due to pseudomonas, and septicemia due to anaerobes were the costliest common principal diagnoses in terms of the mean duration of hospital stay. CONCLUSIONS: Unexplained sharp increases were reported in hospitalization for septicemia or bacteremia in elderly Americans. Marked variation by race and sex were evident in discharge rates with these principal diagnoses. Prognosis and average cost of treatment also differed substantially among common rubrics. Further investigation of individual diagnoses should concentrate on explaining secular trends, exploring the basis for variation by race and sex, and elucidating risk factors for poor clinical outcomes.

Aged↗

Risk selection in the Massachusetts State employee health insurance program.

Using the Diagnostic Cost Group (DCG) model developed from a national sample, we examine biased selection among one fee-for-service (FFS) plan, one preferred provider organization, and several health maintenance organizations (HMOs) in Massachusetts. The proportions of enrollees in low-risk groups are higher in the HMO plans and lower in the FFS plan. The average age in the FFS plan is 9 years greater than that in the HMO plans. Actual premiums are not consistent with risk levels among HMO plans, resulting in gains in some HMO plans and losses in others as high as 20% compared to expected expenses as computed by the DCG model.

Costs and Cost Analysis↗

Microtubule reconfiguration during axogenesis.

When cultured on polylysine, rat sympathetic neurons extend modest lamellae which contain a mass of relatively short non-aligned microtubules. Microtubules display movements, but these movements do not result in any obvious alterations in the overall configuration of the array. Application of a mixture of growth factors called matrigel results in a rapid expansion of the lamellae followed by the outgrowth of axons. Microtubules undergo rapid behavioral changes that result in dramatic alterations in the microtubule array. Microtubules become significantly longer, and extend to the periphery of the lamellae where they invade newly-forming axons. The microtubules align with one another and relative to the cell cortex, and draw together into bundles. Microtubules within a bundle move apart as well, particularly at the tips of developing axons. These observations demonstrate a complexity of microtubule behaviors, some of which can be explained by interactions with actin and/or by forces generated by molecular motor proteins.

Actin Cytoskeleton↗

Cholesterol-dependent modulation of tau phosphorylation in cultured neurons.

One of the hallmarks of Alzheimer's disease (AD) is the abnormal state of tau. It is both highly phosphorylated and aggregated into paired helical filaments (PHFs) in neurofibrillary tangles (NFTs). However, the mechanism underlying the hyperphosphorylation of tau in NFTs and neuronal degeneration in AD remains to be elucidated. The fact that hyperphosphorylation of tau in NFTs are also found in the patients with Niemann-Pick disease, type C (NPC), which is a cholesterol storage disease associated with defective intracellular trafficking of exogenous cholesterol, implies that perturbation of cholesterol metabolism may be involved in tau phosphorylation and neurodegeneration. Here, we report that cholesterol deficiency induced by inhibition of cholesterol biosynthesis in cultured neurons results in hyperphosphorylation of tau, accompanied by axonal degeneration associated with microtubule depolymerization. These changes were prevented by concurrent treatment with beta-migrating very low-density lipoprotein (beta-VLDL) or cholesterol. We propose that intracellular cholesterol plays an essential role in the modulation of tau phosphorylation and the maintenance of microtubule stability.

Animals↗

Trends and outcomes in the hospitalization of older Americans for cardiac conduction disorders or arrhythmias, 1991-1998.

OBJECTIVE: To identify epidemiological trends and measure outcomes in elderly patients hospitalized for cardiac conduction disorders or arrhythmias. DESIGN: Review of the standard 5% samples of the Medicare Provider Analysis and Review Files to characterize 144,512 discharges from 1991 through 1998 in which the principal diagnosis was a conduction disorder or arrhythmia, using the corresponding Enrollment Databases for denominator data. SETTING: Short-stay hospitals in the United States. PARTICIPANTS: Medicare beneficiaries age 65 and older in the standard 5% sample. MEASUREMENTS: Diagnosis-specific trends and rates; discharges by year; cumulative age-, race-, and sex-specific discharge rates; mean length of stay in hospital and in intensive care; mean Medicare reimbursement to the hospital; case-fatality rate in hospital; discharge destinations of patients discharged alive. RESULTS: Annual hospitalizations for sinoatrial node dysfunction, atrial flutter, atrial fibrillation, or ventricular fibrillation increased more rapidly than did the elderly Medicare beneficiary population. Hospitalizations with a principal diagnosis of ventricular extrasystoles or asystole showed steep secular declines. Discharge rates for sinoatrial node dysfunction, a group of rhythms with a nonsinus pacemaker, atrial fibrillation, Mobitz I, or complete atrioventricular block all increased steeply and continuously with patient age. In contrast, discharge rates for atrial flutter or ventricular tachycardia or fibrillation peaked among 75- to 84-year-old patients. White men were at uniquely high risk of hospitalization for atrial flutter or ventricular tachycardia or fibrillation, and, among the white majority, men had higher discharge rates than women for nine of the 11 commonest rubrics. Whites, particularly white women, had the highest discharge rates for atrial fibrillation. Blacks, especially black women, were at disproportionate risk for hospitalization for the group of nonsinus pacemaker rhythms. Diagnosis-specific mean resource costs were strongly correlated with each other and with mean Medicare reimbursement but not with case-fatality rate. CONCLUSION: Medicare claims data demonstrated striking differences among and within diagnoses of heart blocks or arrhythmias in terms of the populations at greatest risk for hospitalization. This variation should be explored further to generate and test hypotheses about differential causation or delivery of care.

Black or African American↗

Characterization of secretory and morphologic properties of primary cultured endocrine cells from porcine pancreata.

We characterized morphologic and secretory properties of porcine pancreatic endocrine cells in primary culture obtained by autolytic preparation without any exogenous proteolytic enzymes. The endocrine cells exhibited a neuron-like shape, and insulin granules were accumulated at the terminal of the processes. Thus derived endocrine cells survived in culture medium containing nicotinamide and remained sensitive to glucose for at least 6 weeks after preparations. The cells responded well to physiologic concentrations of glucose, and high K+ depolarization and the antidiabetic sulfonylureas, tolbutamide, and glibenclamide also elicited the release. With high glucose, insulin release was markedly potentiated by forskolin, glucagon, glucagon-like peptide-1, and arginine and inhibited by somatostatin, the Ca2+ channel blocker nitrendipine, and the ATP-sensitive K+ channel opener diazoxide. Epinephrine had dual effects on the release by glucose; enhanced within a low nanomolar range and inhibited at 1 micromol/L. However, the cells were unresponsive to leucine. Such secretory sensitivities to nutrients, hormones, and pharmacologic agents, and long survival rate (as long as 5-6 weeks) of these cells suggest to us therefore that derived endocrine cells may be useful for xenotransplantation of pancreatic beta cells for treatment of insulin-dependent diabetes mellitus.

Animals↗

Bis(2,2-bipyridine-N,N')tetra-mu-chloro-tetracopper(I).

A novel centrosymmetric chair-like dimer, bis(2,2'-bipyridine)-1kappa(2)N,N';3kappa(2)N,N'-tetra-mu-chloro-1:2kappa(2)Cl;2:3kappa(2)Cl;3:4kappa(2)Cl;1:4kappa(2)Cl-tetracopper(I), [Cu(4)Cl(4)(C(10)H(8)N(2))(2)], has been solvothermally synthesized and structurally characterized. The complex self-assembles into a three-dimensional network via C-H.Cl hydrogen bonds, pi-pi stacking and weak Cu.Cl electrostatic interactions.

Journal Article↗

Direct activation of an inwardly rectifying potassium channel by arachidonic acid.

Arachidonic acid (AA) is an important constituent of membrane phospholipids and can be liberated by activation of cellular phospholipases. AA modulates a variety of ion channels via diverse mechanisms, including both direct effects by AA itself and indirect actions through AA metabolites. Here, we report excitatory effects of AA on a cloned human inwardly rectifying K(+) channel, Kir2.3, which is highly expressed in the brain and heart and is critical in regulating cell excitability. AA potently and reversibly increased Kir2.3 current amplitudes in whole-cell and excised macro-patch recordings (maximal whole-cell response to AA was 258 +/- 21% of control, with an EC(50) value of 447 nM at -97 mV). This effect was apparently caused by an action of AA at an extracellular site and was not prevented by inhibitors of protein kinase C, free oxygen radicals, or AA metabolic pathways. Fatty acids that are not substrates for metabolism also potentiated Kir2.3 current. AA had no effect on the currents flowing through Kir2.1, Kir2.2, or Kir2.4 channels. Experiments with Kir2.1/2.3 chimeras suggested that, although AA may bind to both Kir2.1 and Kir2.3, the transmembrane and/or intracellular domains of Kir2.3 were essential for channel potentiation. These results argue for a direct mechanism of AA modulation of Kir2.3.

Animals↗

Effects of alcohol on blood pressure and production of vascular aldosterone and corticosterone.

AIMS: The aims of this study were to search for the role of alcohol in blood pressure regulation in rats and to investigate the effects of alcohol on the production of vascular aldosterone and corticosterone. METHODS: Male Wistar rats received alcohol 0.7 g x kg(-1) x day(-1) (alcohol-treated group 1) or 1.4 g x kg(-1) x day(-1) (alcohol-treated group 2) or 2.1 g x kg(-1) x day(-1) (alcohol-treated group 3), orally, for 3 months, and blood pressure was monitored by a pressure transducer. Systolic blood pressure increased in Wistar rats treated with alcohol compared to control rats. Mesenteric artery perfusion ex vivo was performed and pressor responses to norepinephrine were determined. The pressor responses to norepinephrine in mesenteric arteries treated with alcohol were significantly increased. The perfusate from the mesenteric arteries was collected and applied to a Sep-Pak C 18 cartridge column for reverse-phase high-performance liquid chromatography and aldosterone and corticosterone levels were determined by radioimmunoassay, aldosterone was decreased but corticosterone was increased in the perfusate from arteries treated with alcohol. RESULTS: Reverse transcriptase polymerase chain reaction showed that alcohol inhibited the expression of 11beta-HSD2 and CYP11B2 mRNA in mesenteric arteries. CONCLUSIONS: These results reveal that alcohol is able to induce hypertension and provide evidence that alcohol inhibits the transcriptions of both 11beta-HSD2 and CYP11B2 in the vasculature, leading to lower aldosterone and higher corticosterone production in vessels, and increased vasoconstrictor responses to norepinephrine.

11-beta-Hydroxysteroid Dehydrogenase Type 2↗