PubMed Health⌕ Search

Biomedical subjects

J You

Publications and source records attributed to J You.

At least 19 recordsLinked to original sources
PubMed2007

Human amniotic fluid-derived stem cells have characteristics of multipotent stem cells.

OBJECTIVES: To characterize mesenchymal stem cell-like cells isolated from human amniotic fluid for a new source of therapeutic cells. MATERIALS: Fibroblastoid-type cells obtained from amniotic fluid at the time of birth. METHODS: The ability of ex vivo expansion was investigated until senescence, and stem cell-like characteristics were analyzed by examining differentiation potential, messenger RNA expression and immunophenotypes. RESULTS AND CONCLUSIONS: A morphologically homogenous population of fibroblastoid-type (HAFFTs) cells, similar to mesenchymal stem cells from bone marrow (BM-MSCs), was obtained at the third passage. The cells became senescent after 27 passages over a period of 8 months while undergoing 66 population doublings. Under appropriate culture conditions, by the 8th passage they differentiated into adipocytes, osteocytes, chondrocytes and neuronal cells, as revealed by oil red O, von Kossa, Alcian blue and anti-NeuN antibody staining, respectively. Immunophenotype analyses at the 17th passage demonstrated the presence of TRA-1-60; SSEA-3 and-4; collagen types I, II, III, IV and XII; fibronectin; alpha-SMA; vimentin; desmin; CK18; CD44; CD54; CD106; FSP; vWF; CD31; and HLA ABC. Reverse transcriptase-polymerase chain reaction analysis of the HAFFTs from passages 6-20 showed consistent expression of Rex-1, SCF, GATA-4, vimentin, CK18, FGF-5 and HLA ABC genes. Oct-4 gene expression was observed up to the 19th passage but not at the 20th passage. HAFFTs showed telomerase activity at the 5th passage with a decreased level by the 21st passage. Interestingly, BMP-4, AFP, nestin and HNF-4alpha genes showed differential gene expression during ex vivo expansion. Taken together, these observations suggest that HAFFTs are pluripotent stem cells that are less differentiated than BM-MSCs, and that their gene expression profiles vary with passage number during ex vivo expansion.

Amniotic Fluid↗
PubMed2005

Aquatic toxicity due to residential use of pyrethroid insecticides.

Pyrethroids are the active ingredients in most insecticides available to consumers for residential use in the United States. Yet despite their dominance in the marketplace, there has been no attempt to analyze for most of these compounds in watercourses draining residential areas. Roseville, California was selected as a typical suburban development, and several creeks that drain subdivisions of single-family homes were examined. Nearly all creek sediments collected caused toxicity in laboratory exposures to an aquatic species, the amphipod Hyalella azteca, and about half the samples caused nearly complete mortality. This same species was also found as a resident in the system, but its presence was limited to areas where residential influence was least. The pyrethroid bifenthrin is implicated as the primary cause of the toxicity, with additional contributions to toxicity from the pyrethroids cyfluthrin and cypermethrin. The dominant sources of these pyrethroids are structural pest control by professional applicators and/ or homeowner use of insecticides, particularly lawn care products. The suburbs of Roseville are unlikely to be unique, and similar sediment quality degradation is likely in other suburban areas, particularly in dry regions where landscape irrigation can dominate seasonal flow in some water bodies.

Amphipoda↗
PubMed2005

An FDK-like cone-beam SPECT reconstruction algorithm for non-uniform attenuated projections acquired using a circular trajectory.

In this paper, Novikov's inversion formula of the attenuated two-dimensional (2D) Radon transform is applied to the reconstruction of attenuated fan-beam projections acquired with equal detector spacing and of attenuated cone-beam projections acquired with a flat planar detector and circular trajectory. The derivation of the fan-beam algorithm is obtained by transformation from parallel-beam coordinates to fan-beam coordinates. The cone-beam reconstruction algorithm is an extension of the fan-beam reconstruction algorithm using Feldkamp-Davis-Kress's (FDK) method. Computer simulations indicate that the algorithm is efficient and is accurate in reconstructing slices close to the central slice of the cone-beam orbit plane. When the attenuation map is set to zero the implementation is equivalent to the FDK method. Reconstructed images are also shown for noise corrupted projections.

Algorithms↗
PubMed2004

Distribution and toxicity of sediment-associated pesticides in agriculture-dominated water bodies of California's Central Valley.

The agricultural industry and urban pesticide users are increasingly relying upon pyrethroid insecticides and shifting to more potent members of the class, yet little information is available on residues of these substances in aquatic systems under conditions of actual use. Seventy sediment samples were collected over a 10-county area in the agriculture-dominated Central Valley of California, with most sites located in irrigation canals and small creeks dominated by agricultural effluent. The sediments were analyzed for 26 pesticides including five pyrethroids, 20 organochlorines, and one organophosphate. Ten-day sediment toxicity tests were conducted using the amphipod Hyalella azteca and, for some samples, the midge Chironomus tentans. Forty-two percent of the locations sampled caused significant mortality to one test species on at least one occasion. Fourteen percent of the sites (two creeks and four irrigation canals) showed extreme toxicity (>80% mortality) on at least one occasion. Pyrethroid pesticides were detected in 75% of the sediment samples, with permethrin detected most frequently, followed by esfenvalerate > bifenthrin > lambda-cyhalothrin. Based on a toxicity unit analysis, measured pyrethroid concentrations were sufficiently high to have contributed to the toxicity in 40% of samples toxic to C. tentans and nearly 70% of samples toxic to H. azteca. Organochlorine compounds (endrin, endosulfan) may have contributed to the toxicity at a few other sites. This study provides one of the first geographically broad assessments of pyrethroids in areas highly affected by agriculture, and it suggests there is a greater need to examine sediment-associated pesticide residues and their potential for uptake by and toxicity to benthic organisms.

Agriculture↗
PubMed2004

A sonication extraction method for the analysis of pyrethroid, organophosphate, and organochlorine pesticides from sediment by gas chromatography with electron-capture detection.

A method was developed for the simultaneous determination of 5 pyrethroid, 1 organophosphate, and 20 organochlorine pesticides in sediment. Pesticide residues were extracted using sonication with acetone-methylene chloride (1:1 vol/vol) and the extracts were subsequently cleaned with deactivated Florisil (magnesium silicate; U.S. Silica, Berkeley Springs, West Virginia). Gas chromatography with an electroncapture detector was used for analyte determination, and two columns were used for confirmation of the analytes. Four control sediments from different sources were spiked with a pesticide mix and analyzed for method validation. The method detection limits ranged from 0.22 to 0.85 microg/kg dry sediment. Recoveries for spiked samples at four concentrations (1, 5, 20, and 400 microg/kg dry sediment) were 71.9% to 129.8% with relative standard deviations (RSDs) < 11%. Taking the matrix effect into account, 1 microg/kg was chosen for the threshold of detection, but 0.5 microg/kg of spiked control sediment still provided good recoveries and RSDs. This method was validated using field-collected sediment taken from agricultural areas of Fresno County, California.

Chemistry Techniques, Analytical↗
PubMed2004

Evaluation of desulfuration methods for pyrethroid, organophosphate, and organochlorine pesticides in sediment with high sulfur content.

This study investigated different cleanup procedures to eliminate sulfur interference from sediments from a reclaimed mining site in Sparta, Illinois. Sonication extraction with activated copper powder was the most effective method to eliminate elemental sulfur. Extracts were then cleaned with a Florisil column before pyrethroid, organophosphate, and organochlorine pesticides were determined using gas chromatography with electron-capture detection. The method detection limits ranged from 0.03 to 0.68 microg/kg dry sediment. Recoveries for spiked samples were from 82.2% to 132.7% with relative standard deviations < 15%. Fifty-four sediment samples collected from 15 lakes and 1 stream in the proposed Illinois National Guard Armory were analyzed using the desulfuration procedure. Of the 26 target pesticides analyzed in sediment, 20 were detected above the detection threshold of 1 microg/kg. DDT was detected most frequently (found at 50% of the sampling sites), whereas permethrin was found at the greatest concentration.

Chromatography, Gas↗
PubMed2004

The pre-styloid compartment of the parapharyngeal space: a three-dimensional digitized model based on the Chinese Visible Human.

To build a digitized visible model of the parapharyngeal space of the Chinese Visible Human and to provide a sectional anatomic basis for radiological and clinical diagnosis of the parapharyngeal space, sectional anatomy data of the parapharyngeal space were selected from the Chinese Visible Human male and female to compare with MR imaging findings in the axial planes. From these data the parapharyngeal space and surrounding structures were segmented. They were then reconstructed in three dimensions on PC. In the axial planes of the sectional anatomy and MR imaging, the shape, content and relations of the parapharyngeal space were clearly displayed and the dominant plane for showing the parapharyngeal space was elicited. The three-dimensional reconstructed images displayed perfectly the anatomic relationships of the parapharyngeal space, parotid, muscles, mandible and vessels. All reconstructed structures can be displayed singly, in groups or as a whole; any diameter or angle of the reconstructed structures can be easily measured. The Chinese Visible Human male and female data set can provide complete and accurate data. The digitized model of the parapharyngeal space and its surroundings offers unique insights into the complex anatomy of the area, providing morphologic data for imaging diagnosis and surgery of the parapharyngeal space.

Anatomy, Cross-Sectional↗
PubMed2004

Isolation and characterization of endophytic Streptomyces strains from surface-sterilized tomato (Lycopersicon esculentum) roots.

AIMS: To isolate endophytic Streptomyces strains from tomato and examine their antimicrobial activity. METHODS: Endophytic Streptomyces strains were isolated using surface-sterilization methods and identified by morphological characteristics. Antimicrobial activities were measured by the agar plate sensitivity method. Antifungal activity in vivo was measured by seedling mortality in infested soils. RESULTS: Twenty-one per cent of endophytic streptomycete isolates produced antibacterial metabolites and 41% produced antifungal metabolites in S medium. Sixty-five per cent of the most frequently isolated strains inhibited the growth of Rhizoctonia solani by the antibiosis assay but only 32% produced metabolites active against R. solani in S medium. Growth promotion and enhanced disease resistance of seedlings inoculated with Streptomyces sp. strain S30 were observed in tomato but not in cucumber seedlings. CONCLUSIONS: Endophytic Streptomyces spp. strains were successfully isolated using stringent methods and strain S30 promoted growth and enhanced resistance to R. solani in tomato seedlings. SIGNIFICANCE AND IMPACT OF THE STUDY: Endophytic streptomycetes showing antifungal activity in vitro and in vivo may indicate the potential for their use as biocontrol agents particularly of R. solani disease of tomato seedlings.

Anti-Bacterial Agents↗
PubMed2003

Fluid flow-induced prostaglandin E2 response of osteoblastic ROS 17/2.8 cells is gap junction-mediated and independent of cytosolic calcium.

It has been well demonstrated that bone adapts to mechanical loading. To accomplish this at the cellular level, bone cells must be responsive to mechanical loading (mechanoresponsive). This can occur via such mechanisms as direct cell deformation or signal transduction via complex pathways involving chemotransport, hormone response, and/or gene expression, to name a few. Mechanotransduction is the process by which a bone cell senses a biophysical signal and elicits a response. While it has been demonstrated that bone cells can respond to a wide variety of biophysical signals including fluid flow, stretch, and magnetic fields, the exact pathways and mechanisms involved are not clearly understood. We postulated that gap junctions may play an important role in bone cell responsiveness. Gap junctions (GJ) are membrane-spanning channels that physically link cells and support the transport of small molecules and ions in the process of gap junctional intercellular communication (GJIC). In this study we examined the role of GJ and GJIC in mechanically stimulated osteoblastic cells. Following fluid flow stimulation, we quantified prostaglandin E(2) (PGE(2)) (oscillatory flow) and cytosolic calcium (Ca(2+)) (oscillatory and steady flow) responses in ROS 17/2.8 cells and a derivative of these cells expressing antisense cDNA for the gap junction protein connexin 43 (RCx16) possessing significantly different levels of GJIC. We found that the ROS17/2.8 cells possessing increased GJIC also exhibited increased PGE(2) release to the supernatant following oscillatory fluid flow stimulation in comparison to coupling-decreased RCx16 cells. Interestingly, we found that neither osteoblastic cell line responded to oscillatory or steady fluid flow stimulation with an increase in Ca(2+). Thus, our results suggest that GJ and GJIC may be important in the mechanotransduction mechanisms by which PGE(2) is mechanically induced in osteoblastic cells independent of Ca(2+).

Animals↗
PubMed2003

Assessment of sustainable water management for rapidly developing urban regions in Guangzhou City, China.

The Shawan River will be the focal point in the development of the Panyu District, the southern-most district of Guangzhou City in the Guangdong Province of South China. In this research, through the use of two scenarios, the future water quality of the Shawan River was predicted with relation to changes in the water quantity utilized to fuel industrial and domestic development. The worst-case scenario used, simulated the situation if no wastewater treatment was employed, and the best-case scenario simulated the situation if 90% of the pollution load was removed. The period of simulation was for the years 2020 and 2050. Three flowrates were used in the evaluation, those of: the 90% probability of the month of lowest flow (37.2 m3/s); and the range of flowrates within the low flow period, that is, the dry season from November to February (307 and 432 m3/s). Subsequently, two countermeasures (industrial and domestic water savings)--sustainable initiatives--were nested within the two scenarios to ascertain improvements in water quality as a direct result of reduction in water quantity used. The industrial water saving countermeasure showed the greatest improvement in water quality. For the 90% probability of lowest flow for the worst-case scenario, this countermeasure equated to a 63% decrease in BOD. For the low flow period flowrates the background concentration of pollutants was more influential than improvements imparted by the countermeasures to the future predicted water quality. It was recommended that industrial countermeasures be used that take into account water saving, water recycling, the use of brackish water for cooling, and the implementation of economic pricing initiatives. Also that inter-district governmental policy initiatives be introduced to prevent upstream pollution from influencing downstream proposals, further enhancing sustainable water management of the Shawan River.

China↗
PubMed2001

Effect of combined treatment with praziquantel and artemether on Schistosoma japonicum and Schistosoma mansoni in experimentally infected animals.

Praziquantel and artemether are safe and efficacious antischistosomal drugs that act against different developmental stages of the parasite: praziquantel against adult worms and artemether against schistosomula. A combined treatment has been suggested as a strategy for transmission control. Recent laboratory experiments with rabbits with a mixed infection of Schistosoma japonicum parasites of different ages confirmed the effectiveness of a combination therapy. In the present work, we assessed the effect of a combined treatment on adult worms of S. japonicum and found significantly higher worm reduction rates than with a single dose of praziquantel. In a next step, we extended the study of the combined treatment to Schistosoma mansoni. A combined treatment with 75 mg/kg praziquantel and 150 mg/kg artemether was administered to hamsters infected with juvenile and adult S. mansoni. The two drugs, administered simultaneously or spaced by 6 h, 1, 3 or 7 days, resulted in significantly higher worm reduction rates than a single treatment with praziquantel. A combination therapy with increased doses of 100 mg/kg praziquantel and 300 mg/kg artemether showed very high worm reduction rates of 90% and above, however, some hamsters died in five different combined treatment experiments, suggesting that these drug concentrations were too high. We conclude that a combined treatment with praziquantel and artemether at the lower doses is safe and more effective than praziquantel alone, which forms a foundation for designing respective clinical trials in humans.

Administration, Oral↗
PubMed2001

A HECT domain E3 enzyme assembles novel polyubiquitin chains.

Although polyubiquitin chains linked through Lys(29) of ubiquitin have been implicated in the targeting of certain substrates to proteasomes, the signaling properties of these chains are poorly understood. We previously described a ubiquitin-protein isopeptide ligase (E3) from erythroid cells that assembles polyubiquitin chains through either Lys(29) or Lys(48) of ubiquitin (Mastrandrea, L. D., You, J., Niles, E. G., and Pickart, C. M. (1999) J. Biol. Chem. 274, 27299-27306). Here we describe the purification of this E3 based on its affinity for a linear fusion of ubiquitin to the ubiquitin-conjugating enzyme UbcH5A. Among five major polypeptides in the affinity column eluate, the activity of interest was assigned to the product of a previously cloned human cDNA known as KIAA10 (Nomura, N., Miyajima, N., Sazuka, T., Tanaka, A., Kawarabayasi, Y., Sato, S., Nagase, T., Seki, N., Ishikawa, K., and Tabata, S. (1994) DNA Res. 1, 27-35). The KIAA10 protein is a member of the HECT (homologous to E6-AP carboxyl terminus) domain family of E3s. These E3s share a conserved C-terminal (HECT) domain that functions in the catalysis of ubiquitination, while their divergent N-terminal domains function in cognate substrate binding (Huibregtse, J. M., Scheffner, M., Beaudenon, S., and Howley, P. M. (1995) Proc. Natl. Acad. Sci. U. S. A. 92, 2563-2567). Recombinant KIAA10 catalyzed the assembly of both Lys(29)- and Lys(48)-linked polyubiquitin chains. Surprisingly, the C-terminal 428 residues of KIAA10 were both necessary and sufficient for this activity, suggesting that the ability to assemble polyubiquitin chains may be a general property of HECT domains. The N-terminal domain of KIAA10 interacted in vitro with purified 26 S proteasomes and with the isolated S2/Rpn1 subunit of the proteasome's 19 S regulatory complex, suggesting that the N-terminal domains of HECT E3s may function in proteasome binding as well as substrate binding.

Biopolymers↗
PubMed2001

Determination of alcohols by high-performance liquid chromatography with fluorimetric detection after pre-column derivatisation with carbazole-9-N-acetic acid and chromatographic behaviour of alcoholic derivatives.

Alcohols were derivatised to their carbazole-9-N-acetic acid (CRA) esters with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC.HCl) as the dehydrating agent. Studies on derivatisation conditions indicated that the coupling reaction proceeded rapidly and smoothly in the presence of a base catalyst in acetonitrile to give the corresponding sensitively fluorescent derivatives. The retention behaviour of alcohol derivatives was investigated by varying mobile phase compositions (ACN-water and MeOH-water). The parameters from the equation log k'=A-BX were evaluated by retention data of derivatives using an isocratic elution with different mobile phases. The results indicated that the parameters derived allowed computation of retention factors in good agreement with experiments. At the same time, a general equation was derived that makes possible predictions of partition coefficient in binary mobile phases with different proportions of organic solvent to water based on some simple regression analysis. The LC separation for the derivatised alcohols containing higher carbon alcohols showed good reproducibility on a reversed-phase C18 column with gradient elution. The detection limits (excitation at 335 nm, emission at 360 nm) for derivatised alcohols (signal-to-noise ratio=3:1) were in the range of 0.1-0.4 pg per injection.

Alcohols↗
PubMed2001

Osteopontin gene regulation by oscillatory fluid flow via intracellular calcium mobilization and activation of mitogen-activated protein kinase in MC3T3-E1 osteoblasts.

Recently fluid flow has been shown to be a potent physical stimulus in the regulation of bone cell metabolism. However, most investigators have applied steady or pulsing flow profiles rather than oscillatory fluid flow, which occurs in vivo because of mechanical loading. Here oscillatory fluid flow was demonstrated to be a potentially important physical signal for loading-induced changes in bone cell metabolism. We selected three well known biological response variables including intracellular calcium (Ca(2+)i), mitogen-activated protein kinase (MAPK) activity, and osteopontin (OPN) mRNA levels to examine the response of MC3T3-E1 osteoblastic cells to oscillatory fluid flow with shear stresses ranging from 2 to -2 Newtons/m(2) at 1 Hz, which is in the range expected to occur during routine physical activities. Our results showed that within 1 min, oscillatory flow induced cell Ca(2+)i mobilization, whereas two MAPKs (ERK and p38) were activated over a 2-h time frame. However, there was no activation of JNK. Furthermore 2 h of oscillatory fluid flow increased steady-state OPN mRNA expression levels by approximately 4-fold, 24 h after exposure to fluid flow. The presence of both ERK and p38 inhibitors and thapsigargin completely abolished the effect of oscillatory flow on steady-state OPN mRNA levels. In addition, experiments using a variety of pharmacological agents suggest that oscillatory flow induces Ca(2+)i mobilization via the L-type voltage-operated calcium channel and the inositol 1,4,5-trisphosphate pathway.

3T3 Cells↗
PubMed2001

Effect of slip on movement of body center of mass relative to base of support.

OBJECTIVE: The purpose is to investigate the effect of balance conditions and slippery perturbation on the position and velocity of the body's center of mass relative to the body's base of support. DESIGN: Twenty-two young and healthful subjects were investigated while their walk was perturbed by a soap patch applied over a force plate. A safety harness was used to prevent the subject from falling on knee or buttock. BACKGROUND: Appropriate postural response to meet physiological biomechanical requirements is mandatory in restoration of balance upon slip. METHODS: Twenty-two healthy subjects dressed with safety harness walked first without and then with slippery perturbation, guided by a metronome at 120 steps/min and 90 steps/min cadence. Data were collected from a motion analysis system and force plates. RESULTS: For slippery perturbation, the displacement and velocity of center of mass with respect to base of support became smaller from heel strike to contralateral toe off. Subject's balance condition correlated significantly to the displacement of center of mass with respect to base of support (r=-0.51 at 120 steps/min and r=-0.471 at 90 steps/min), as well as the velocity (r=-0.834 at 120 steps/min, r=-0.673 at 90 steps/min) at contralateral toe off. CONCLUSIONS: For slip during walking, smaller excursion and faster velocity of center of mass with respect to base of support were important for subjects regaining balance from heel strike to contralateral toe off. The critical time for subjects regaining stability is the first double support phase of the gait cycle. RELEVANCE: It is confirmed that two variables, the displacement and the velocity of center of mass with respect to base of support, are valuable biomechanical factors and provide quantifiable determination for investigation of the balance condition in slipping.

Accidental Falls↗
PubMed2001

Potentiated endothelium-derived hyperpolarizing factor-mediated dilations in cerebral arteries following mild head injury.

Evidence in the literature suggests that endothelium-derived hyperpolarizing factor (EDHF) may act in a compensatory manner such that during conditions of compromised nitric oxide (NO), EDHF serves as a back-up mechanism. Given that constitutive NO synthase is chronically downregulated after head trauma, we tested the hypothesis that EDHF is potentiated following injury. Male adult rats were subjected to either sham injury (n = 27) or mild controlled cortical impact (CCI) injury (n = 26). Branches of the middle cerebral artery (MCA) directly within the contusion site were harvested either 1 or 24 h later, pressurized to 60 mm Hg in a vessel chamber and allowed to develop spontaneous tone. Relaxation to luminal application of adenosine triphosphate (ATP) was similar in all groups. Relaxation to ATP in the presence of L-NAME (N(G)-nitro-L-arginine methyl ester) and indomethacin was similar in all groups except for vessels isolated at 24 h following mild CCI injury. In this case, L-NAME and indomethacin had no effect on the ATP-mediated dilation. The ATP-mediated dilation in L-NAME and indomethacin-treated MCA branches was inhibited by charybdotoxin, an inhibitor of large conductance Ca2+-sensitive K+ channels. These findings suggest that there is a significant potentiation of the EDHF-mediated dilation to ATP in cerebral arteries isolated at 24 h following mild CCI injury.

Adenosine Triphosphate↗
PubMed2001

Protamine-modified DDAB lipid vesicles promote gene transfer in the presence of serum.

Cationic lipid vesicle-mediated gene transfer has become common for in vitro gene delivery. However, the transfection efficiency is often impaired by serum. DDAB (dimethyldioctadecyl ammonium bromide) lipid vesicle-mediated gene transfer, which we previously reported, has the same problem. To overcome this obstacle, we here report a novel transfection vehicle using protamine-modified DDAB lipid vesicles. While free protamine was simply added to the DNA/lipid complex in the previous study, in the present method the protamine is chemically conjugated to stearic acid and incorporated into DDAB lipid vesicles. Gene transfer was not significantly inhibited in 10% serum-containing medium by this method for the transfection of cultured cells. Protamine-modified DDAB lipid vesicles also enhanced virus transduction efficiency in the presence of serum using a replication-defective retroviral vector. Furthermore, the vesicles allowed efficient gene transfer for avian embryos in vivo. These results indicate that the method is useful for the production of transgenic animals and gene therapy.

3T3 Cells↗