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Biomedical subjects

R Song

Publications and source records attributed to R Song.

At least 19 recordsLinked to original sources

Phagosomes are fully competent antigen-processing organelles that mediate the formation of peptide:class II MHC complexes.

During the processing of particulate Ags, it is unclear whether peptide:class II MHC (MHC-II) complexes are formed within phagosomes or within endocytic compartments that receive Ag fragments from phagosomes. Murine macrophages were pulsed with latex beads conjugated with OVA. Flow or Western blot analysis of isolated phagosomes showed extensive acquisition of MHC-II, H-2M, and invariant chain within 30 min, with concurrent degradation of OVA. T hybridoma responses to isolated subcellular fractions demonstrated OVA (323-339):I-Ad complexes in phagosomes and plasma membrane but not within dense late endocytic compartments. Furthermore, when two physically separable sets of phagosomes were present within the same cells, OVA(323-339):I-Ad complexes were demonstrated in latex-OVA phagosomes but not in phagosomes containing latex beads conjugated with another protein. This implies that these complexes were formed specifically within phagosomes and were not formed elsewhere and subsequently transported to phagosomes. In addition, peptide:MHC-II complexes were shown to traffic from phagosomes to the cell surface. In conclusion, phagosomes are fully competent to process Ags and generate peptide:MHC-II complexes that are transported to the cell surface and presented to T cells.

Animals

Antipeptide antibodies reveal interrelationships of MBP 200 and MBP 235: unique apoB-specific receptors for triglyceride-rich lipoproteins on human monocyte-macrophages.

Two human monocyte-macrophage (HMM) membrane binding proteins, (MBP) 200 and 235, are receptor candidates that bind to the apolipoprotein (apo)B-48 domain in triglyceride-rich lipoproteins for uptake independent of apoE. Microsequence analysis of the purified reduced MBP 200R characterized tryptic peptides of MBP 200R. A synthetic peptide mimicking a unique, unambiguous 10-residue sequence (AEGLMVTGGR) induced antipeptide antibodies that specifically recognized MBP 200, 235 and 200R, in 1- and 2-dimensional analyses, indicating 1) the ligand binding protein was sequenced and 2) MBP 200 and 235 yielded MBP 200R upon reduction. These antibodies identified the MBPs in human blood-borne, THP-1, U937 MMs, and endothelial cells (EC) but not in human fibroblasts or Chinese hamster ovary (CHO) cells. Fluorescence activated cell sorting (FACS) analysis located the MBPs on the MM surface as necessary for receptor function. The 10-residue, unambiguous MBP 200-derived sequence is unique, with no matches in extant protein databases. Antipeptide antibodies bind to the MBPs in reticuloendothelial cells that have this receptor activity, but not to proteins in cells that lack this receptor activity. These studies provide the first direct protein sequence and immunochemical data that a new, unique apoB receptor for triglyceride-rich lipoproteins exists in human monocytes, macrophages, and endothelial cells.

Amino Acid Sequence

Expression of multidrug resistance protein/GS-X pump and gamma-glutamylcysteine synthetase genes is regulated by oxidative stress.

Expression of the MRP1 gene encoding the GS-X pump and of the gamma-GCSh gene encoding the heavy (catalytic) subunit of the gamma-glutamylcysteine synthetase is frequently elevated in many drug-resistant cell lines and can be co-induced by many cytotoxic agents. However, mechanisms that regulate the expression of these genes remain to be elucidated. We report here that like gamma-GCSh, the expression of MRP1 can be induced in cultured cells treated with pro-oxidants such as tert-butylhydroquinone, 2,3-dimethoxy-1, 4-naphthoquinone, and menadione. Intracellular reactive oxygen intermediate (ROI) levels were increased in hepatoma cells treated with tert-butylhydroquinone for 2 h as measured by flow cytometry using an ROI-specific probe, dihydrorhodamine 123. Elevated GSH levels in stably gamma-GCSh-transfected cell lines down-regulated endogenous MRP1 and gamma-GCSh expression. ROI levels in these transfected cells were lower than those in the untransfected control. In the cell lines in which depleting cellular GSH pools did not affect the expression of the MRP1 and gamma-GCSh genes, only minor increased intracellular levels of ROIs were observed. These results suggest that intracellular ROI levels play an important role in the regulation of MRP1 and gamma-GCSh expression. Our data also suggest that elevated intracellular GSH levels not only facilitate substrate transport by the MRP1/GS-X pump as previously demonstrated, but also suppress MRP1 and gamma-GCSh expression.

Biological Transport

A new principle for unilateral complete cleft lip repair, the lateral columellar flap method.

This article presents a different method for unilateral complete cleft lip repair. The tissue deficiency of the medial lip segment is filled with a flap from the lateral surface of the columella, which is in continuity with the lip segment. The lateral columellar flap and the medial lip segment are lowered in one piece vertically downward to such a position that the height of the arch of the Cupid's bow is equal on both sides. No lateral advancement is done. The suture line of the medial and lateral lip segments is an uninterrupted straight line, which imitates the natural line of the philtral column. The repaired lip has more fullness than the lips repaired with the conventional methods. The procedure is described in detail.

Cleft Lip

Apolipoprotein B-48 or its apolipoprotein B-100 equivalent mediates the binding of triglyceride-rich lipoproteins to their unique human monocyte-macrophage receptor.

Studies in animals and humans have demonstrated uptake of plasma chylomicrons (triglyceride-rich lipoprotein [TGRLP] of Sf>400) by accessible macrophages in vivo. One potential mechanism is via a unique receptor pathway we previously identified in human blood and THP-1 monocytes and macrophages for the lipoprotein lipase (LpL)- and apolipoprotein (apo) E-independent, high-affinity, specific binding of plasma chylomicrons and hypertriglyceridemic VLDL (HTG-VLDL) to cell-surface membrane-binding proteins (MBP 200, 235; apparent Mr 200, 235 kD on SDS-PAGE) that leads to lipid accumulation in vitro. Competitive binding studies reported here demonstrate that anti-apoB antibodies specifically block the high-affinity binding of TGRLP to this receptor on THP-1 cells and on ligand blots. LpL, which binds to an N-terminal domain of apoB, also inhibits TGRLP binding both to this site on THP-1s and to MBP 200, 235 by binding to apoB. Chylomicrons of Sf>1100 that contain apoB-48, but not apoB-100, bind specifically to MBP 200, 235, and this binding is blocked by anti-apoB IgG. In contrast, lactoferrin and heparin do not inhibit TGRLP binding. We conclude that the receptor-binding domain is within apoB-48 (or an equivalent in apoB-100) near the LpL-binding domain, but not a heparin-binding domain. Uptake of TGRLP by this mechanism could provide essential nutrients or, in HTG, cause excess lipid accumulation and foam cell formation.

Antibodies

Nurses' job satisfaction, absenteeism, and turnover after implementing a special care unit practice model.

The purpose of the study was to compare job satisfaction, absenteeism, and turnover between nurses working in a nurse-managed special care unit (SCU) and those working in traditional intensive care units (ICU). A case management practice model with a shared governance management model and minimal technology was implemented in the SCU while contrasting features of a primary nursing practice model with a bureaucratic management model and high technology already in place in the traditional ICU. Individual nurses' perceptions of and their preferences for the SCU practice model also were examined related to job satisfaction. Using analysis of covariance, greater satisfaction with a lower absenteeism rate was found in nurses working in the SCU. Nurses' perceptions and preferences for the SCU practice model were closely related to their job satisfaction and growth satisfaction. The findings suggest that individual perception and preference should be taken into account before implementing autonomy, authority, and responsibility at the organizational level to lead to the desired nurse outcomes in a given working environment.

Absenteeism

Interleukin-12 promotes enhanced resistance to Listeria monocytogenes infection of lead-exposed mice.

The heavy metal lead (Pb) has been shown to downregulate various parameters of cell-mediated immune (CMI) responses. This inhibition of CMI responses by Pb is exemplified by a higher mortality rate upon infections with sublethal doses of a variety of pathogens. Unlike Pb, which lowers host resistance, interleukin-12 (IL-12) exerts a substantial stimulatory influence on the host response to intracellular bacteria such as Listeria monocytogenes. To explore the influence of IL-12 in mice rendered susceptible to Listerial infection by oral exposure to Pb, we determined bacterial burdens and production of interferon gamma (IFN-gamma). As expected, Pb-exposed mice had increased morbidity due to higher Listerial titers as compared to control mice. However, administration of exogenous IL-12 reversed the Pb-induced inhibition of host defense and boosted the resistance of the non-Pb-treated mice. The enhanced CMI responses observed in both IL-12-treated groups were accompanied with elevations of IFN-gamma in the sera and spleens. Significant reduction in the number of viable Listeria in Pb-exposed mice upon IL-12 administration suggests that the processes downstream of IL-12 production were intact in the Pb-exposed mice and that the inhibition by Pb was due to the lack of functional IL-12. Alternatively, the exogenous IL-12 may have overcome a downstream effect by enhancing an secondary pathway. Support for the former hypothesis is based on the observation that Pb induced elevated levels of p40 splenic messenger RNA since increased p40 expression would result from lack of IL-12 formation. Contrary to the IFN-gamma levels, significantly higher levels of IL-6 and corticosterone were observed in the sera and spleens of Pb-exposed mice upon infection, suggesting heightened stress in the absence of IL-12. Overall, the results suggest that an environmental pollutant such as Pb can enhance the stress response, which naturally occurs during an infection, and can further compromise health by lowering host resistance by altering cytokine levels.

Animals

Interlaboratory and intralaboratory variabilities in the Environmental Lead Proficiency Analytical Testing (ELPAT) Program.

The Environmental Lead Proficiency Analytical Testing (ELPAT) Program evaluates over 400 laboratories that perform lead measurements in paints, soils, and dusts. A previous National Institute for Occupational Safety and Health study, based on the ELPAT data over a 3-year period (1992-1995), found no large biases among common hotplate and microwave digestion techniques, but did detect small consistent bias between two common instrumental methods. This study expands on the earlier study by examining the total sample variability and its variation components (interlaboratory and intralaboratory). A correlation model was used to separate the variation components by estimating a variation ratio. The correlation model leads to a more general approach than a sample pairing technique developed by Youden. This study found no significant evidence that the relative contribution of intralaboratory and interlaboratory variability to total variability changes with lead loading levels. There were no significant differences in the relative contribution of variation components among three most commonly used analytical methods (combinations of sample preparation techniques and instrumental methods). The interlaboratory relative standard deviation is about 1.7 times the intralaboratory relative standard deviation. Both variation components are important parts of total variation although the laboratory-to-laboratory (including analyst-to-analyst) difference is greater than the within laboratory (including sample-to-sample) variation.

Chemistry Techniques, Analytical

Study of boron neutron capture therapy used neutron source with protons bombarding a thick 9Be target.

Neutron sources created by 4-, 3.5-, and 3-MeV protons striking a thick beryllium target were studied via the time-of-flight technique. Protons were accelerated by the Peking University 4.5 MV electrostatic accelerator. Two disk-shaped 9Be targets with thickness 1.5 and 3 mm were used in the measurements. The time-of-flight spectra were observed at zero degrees with respect to the incident proton beam. The analysis to these time-of-flight spectra is given. The time-of-flight spectra were converted to the energy spectra and compared to a neutron spectrum of 7Li(p, n)7 Be reaction with incident energy 2.5 MeV, which was also measured in this work. Restricted by the spectrometer itself, the threshold of the measurements is 400 keV. The results show that by using several MeV protons bombarding a thick beryllium target, reactions other than 9Be(p, n)9B produce significant contributions to the neutron yield with energy less than 1 MeV.

Beryllium

[Dynamic study on the mutagenic activity of organic concentration of water from one branch port of Huangpu River].

From 1991 to 1994, we studied the mutagenicity of drinking water in Shanghai by seasons by Ames test. The results indicated that the mean value of mutagenic activity of organic concentration on TA98 is 162 revertants per liter. Studies on the four branch ports of Huangpu river showed that they contributed 74% of the total mutagenicity, which accounted for the predominant fraction of the mutagenic activity of Shanghai drinking water. The mutagenicity of Hongkou branch was the highest, accounting for 1/3 of the total mutagenic activity of the 4 branches. Studies showed that the mutagenic activity of organic concentration of Hongkou branch water was declining year by year, and factors affecting the mutagenic activity were analysed.

China

[Miniplate osteosynthesis for multiple mandibular fractures].

This retrospective study involved 34 patients with multiple mandibular fractures. Monocortical miniplates were used for fixation of fractures followed by a short term intermaxillary fixation. All the patients were evaluated radiographically and clinically. The results indicated that all fractures healed uneventfully. No malocclusion, delayed union, injury of the nerves or teeth due to the procedure were found. We concluded that monocortical miniplate osteosynthesis is effective and reliable for multiple mandibular fractures, and a short term intermaxillary fixation is helpful for the healing of fracture.

Adolescent

Roles of proteasomes, transporter for antigen presentation (TAP), and beta 2-microglobulin in the processing of bacterial or particulate antigens via an alternate class I MHC processing pathway.

Latex-OVA and bacteria expressing an OVA fusion protein were processed by macrophages via an alternate class I MHC (MHC-I) processing pathway to present OVA(257-264):Kb. This pathway was resistant to dipeptide aldehyde proteasome inhibitors and brefeldin A, unlike the cytosolic MHC-I pathway. TAP1-/- macrophages exhibited decreases in cell surface peptide-receptive MHC-I and binding of extracellular peptide during transient incubations. This may explain an apparent influence of TAP on alternate MHC-I processing. Alternate MHC-I processing by TAP1-/- cells was enhanced by preincubation at 26 degrees C or with beta 2-microglobulin to increase peptide-receptive MHC-I. Thus, peptides may bind to MHC-I within post-Golgi vacuolar organelles accessible to exogenous beta 2-microglobulin or on the cell surface (following peptide regurgitation).

ATP Binding Cassette Transporter, Subfamily B, Mem

Surface thiols of human lymphocytes and their changes after in vitro and in vivo activation.

Cellular thiols have long been known to play a role in cell activation and proliferation; however, the differential expression of surface thiols on the lymphoid subsets had not been described. Neither was it known whether alteration of surface thiols occurs after exposure to mitogens or infectious agents. Herein, an impermeant thiol-specific fluorescent probe was employed for flow cytometric analysis of surface thiols. Quantification of surface thiols on resting lymphocytes revealed that some subsets expressed different concentrations of surface thiols (CD19+ > CD8+ > CD4+). Furthermore, surface thiols increased on all subsets by 8 h after mitogenic activation. This increase was blocked by cycloheximide or monensin but not by actinomycin D or inhibition of glutathione synthesis by buthionine sulfoximine. In addition, bacitracin, an inhibitor of protein disulfide isomerase, inhibited the increase in surface thiols and DNA synthesis. Lymphocytes from HIV-infected individuals displayed increased surface thiols on CD19+ and CD4+ cells but not CD8+ cells. Although cellular thiols in general have been believed to play a role in protection against oxidants, signaling associated with cell growth, and apoptosis, there is now evidence that changes in exofacial thiols appear to be involved in some of these critical cell reactivities. Thus, quantitative and possibly qualitative differences in surface thiols correlate with membrane differences between lymphoid subsets and with their differential sensitivities to oxidative stress, which suggests that the mechanisms by which surface thiols are maintained and modified after activation are important cellular functions that need to be further evaluated.

Antigens, CD19

Summary of the NIOSH guidelines for air sampling and analytical method development and evaluation.

Suggested guidelines for the development and evaluation of sampling and analytical methods for industrial hygiene monitoring have recently been published in a NIOSH technical report. These guidelines are based in part on various published approaches for method development and evaluation and serve as an attempt at a more unified experimental approach. This paper presents some salient features of this unified approach for method development and evaluation. The basic goal of the approach is to determine if the method under study meets the criterion to produce a result that fell within 25% of the true value 95 times out of 100 on average, although other factors of method performance are evaluated. The experiments proposed for the evaluation of method performance include determination of analytical recovery from the sampler, sampler capacity, storage stability of samples and effect of environmental factors. Evaluation criteria for the experimental data and procedures for the calculation of method bias, precision and accuracy are also included.

Air Pollutants, Occupational

Laboratory and analytical method performance of lead measurements in paint chips, soils, and dusts.

The National Lead Laboratory Accreditation Program (NLLAP) recognizes laboratories capable of analyzing lead in paints, soils, and dusts. NLLAP requires successful participation in the Environmental Lead Proficiency Analytical Testing (ELPAT) program. For paint chip analyses, laboratory-to-laboratory variability is about 10% relative standard deviation (RSD) for lead levels near 0.5%, the HUD definition of lead-based paint. For soil analyses, RSDs are about 9 to 10% near relevant federal soil standards and 16% near the lowest state bare soil standard that currently exists. For dust wipe analyses, RSDs range from 10 to 16% for lead levels near relevant HUD standards. Of participating laboratories, 92 to 93% consistently meet ELPAT performance limits. A variety of analytical methods gives similar results. No conclusive significant differences were found among most frequently used hotplate and microwave sample preparation techniques. In addition, several participating laboratories have successfully used ultrasonic extraction methods, a method suitable for use at abatement sites. The three most frequently used instrumental techniques, flame atomic absorption (FAA), inductively coupled plasma-atomic emission spectroscopy (ICP-AES), and graphite furnace atomic absorption show no statistically significant differences in ability to meet ELPAT performance limits. However, small statistically significant biases between these methods sometimes occur. The magnitude of biases is less than 5% of the corresponding laboratory mean near relevant federal standards except for lead levels near the lowest HUD lead wipe standard, where biases can be as high as 8%. Other instrumental methods that have been used successfully include ICP-mass spectroscopy, direct current plasma-atomic emission spectroscopy, dithizone spectrophotometry, and anodic stripping voltametry.

Accreditation

Some statistical procedures for analytical method accuracy tests and estimation.

In 1974 the National Institute for Occupational Safety and Health (NIOSH) and the Occupational Safety and Health Administration joined to complete exposure standards promulgated by federal regulations. In that effort NIOSH scientists developed an accuracy criterion (AC) and a statistical protocol for evaluating fulfillment of that AC by an analytical method. This article extends that foundation and proposes a new approach to accuracy analyses. It concentrates on the case of known bias, but attempts to bridge the procedures from that case to one in which the bias is estimated. The article emphasizes a general and flexible approach to the design and analysis of more informative and effective method accuracy studies. These empower the user/investigator to design and analyze studies to be most useful and informative for specific requirements.

Bias

Corrections to the target and critical values for the National Institute for Occupational Safety and Health validation tests.

In 1974 the National Institute for Occupational Safety and Health (NIOSH) and the Occupational Safety and Health Administration joined to complete exposure standards promulgated by federal regulations. In that effort NIOSH scientists developed an accuracy criterion (AC) and a statistical protocol for evaluating its fulfillment. That AC and those procedures have been widely used ever since. This article presents corrections to the target and critical coefficients of variation published as part of the statistical protocol.

Bias

Evaluation of a chemical spot-test kit for the detection of airborne particulate lead in the workplace.

A commercial rhodizonate-based test kit was evaluated for its potential use in the detection of lead in airborne particulate samples at work sites. Over 350 air samples were collected at abrasive blasting lead paint abatement sites using cellulose ester membrane filters and personal sampling pumps. The filter samples were tested with the chemical spot test and then analyzed by graphite furnace atomic absorption spectrophotometry. No positive readings were recorded for lead masses below 1.3 micrograms Pb/filter, and no negative readings were observed for lead amounts above 8.1 micrograms Pb/filter. Experimental data were statistically molded in an effort to estimate the performance parameters of the spot test kit. The identification limit of the kit was found to be approximately 3.6 microgram/filter sample. For lead mass values above approximately 10 micrograms Pb/filter, 95% confidence of a positive reading was found, while 95% confidence of a negative reading was found for lead masses below approximately 0.6 micrograms Pb/filter. Based on the results of this study the rhodizonate-based test kit for lead demonstrates potential for use in field screening for lead in personal breathing zone and area air samples.

Air Pollution