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Biomedical subjects

R Song

Publications and source records attributed to R Song.

At least 73 records · Page 4Linked to original sources

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↗

The superior auricular artery and retroauricular arterial island flaps.

This paper reports the finding of the superior auricular artery, which is a fairly large arterial branch, about 1 mm in diameter, arising from the superficial temporal artery anterior to the auricular helix. It is covered by skin, the anterior auricular muscle, and the fascia of the parotid gland. It runs together with its concomitant vein posteriorly in the groove between the cartilage of the ear and the temporal bone and anastomoses with the posterior auricular artery. This artery has been used successfully to raise large retroauricular arterial island flaps for one case of total nose reconstruction and seven cases of reconstruction of the severely constricted anophthalmic socket. These operative procedures are described in detail.

Anophthalmos↗

A novel cis-acting element is involved in the promoter activity of the rat mdr1b gene.

Multidrug resistance (MDR) is often associated with overexpression of P-glycoprotein, which is encoded by the mdr gene family. Three mdr genes, i.e., mdr1a (mdr3), mdr1b (mdr1), and mdr2 are present in rodents, and the expression of these genes is temporally and tissue specifically regulated. Furthermore, expression of mdr1b is highly elevated during rat hepatocarcinogenesis. To elucidate how mdr1b expression is regulated, we cloned the genomic sequence of the rat mdr1b gene and functionally dissected its 5' promoter region in various cell lines. The transcription start site identified by the primer extension and RNase protection assays is identical to that of the murine mdr1b homologue. Sequence analysis revealed that the proximal region (within -1300 bp) of the rat mdr1b gene also shares striking similarity to that of the mouse mdr1b gene. Transient transfection assays using reporter gene constructs containing various lengths of the 5' mdr1b sequences revealed that the sequence located between-247 to -126 bp was important for the expression of the reporter gene in many different cell lines. Further analyses revealed that at least one regulatory element located at -189 to -167 bp, which contained the palindromic sequence 5'-AGACATGTCT-3' (-189 to -180 bp), is involved in the promoter function. Gel mobility shift assays demonstrated that this palindromic sequence is essential for specific protein binding. UV cross-linking experiments identified that two major proteins with molecular masses of approximately 41 and 49 kDa were associated with this sequence. A Genbank search and gel motility shift assay competition experiment suggested that the specific binding protein(s) appears to be a novel transcription factor involved in the regulation of the rat mdr1b gene expression.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Identification and characterization of a hepatoma cell-specific enhancer in the mouse multidrug resistance mdr1b promoter.

The expression of multidrug resistance/P-glycoprotein genes mdr1b(mdr1) and mdr1a(mdr3) is elevated during hepatocarcinogenesis. To investigate the regulation of mdr1b gene expression, we used transient transfection expression assays of reporter constructs containing various 5'-mdr1b flanking sequences in hepatoma and non-hepatoma cells. We found that nucleotides -233 to -116 preferentially enhanced the expression of reporter gene in mouse hepatoma cell lines in an orientation- and promoter context-independent manner. DNase I footprinting using nuclear extracts prepared from hepatoma and non-hepatoma cells identified four protein binding sites at nucleotides -205 to -186 (site A), -181 to -164 (site B), -153 to -135 (site C), and -128 to -120 (site D). Further analyses revealed that, while site B alone played a major part for the enhancer function, sites A and B combined conferred full enhancer activity. Site-directed mutagenesis results also supported these results. Gel retardation experiments using oligonucleotide competitors revealed that the site B contains a dominant binding protein. This is the first report demonstrating a cell type-specific enhancer in the mdr locus. The role of this enhancer in the activation of mdr1b gene during hepatocarcinogenesis is discussed.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Novel dipeptide aldehydes are proteasome inhibitors and block the MHC-I antigen-processing pathway.

Class I MHC (MHC-I) molecules present peptides derived from Ag that are processed in the cytosol. The proteasome is a multicatalytic protease complex that is present in the cytosol and has been implicated in cytosolic Ag processing. Novel dipeptide aldehydes were designed, synthesized, and demonstrated to specifically inhibit the chymotrypsin-like protease activity of isolated proteasomes, but produced relatively little inhibition of cathepsin B, a vacuolar cysteine protease. The inhibitors were membrane permeable and inhibited intracellular cleavage of a membrane-permeable fluorogenic substrate of the chymotrypsin-like proteasome activity. When a model Ag, OVA, was introduced into the cytoplasm of M12.B6 murine B cells by electroporation, the proteasome inhibitors blocked its processing for subsequent presentation by MHC-I molecules. The inhibitors had little effect on class II MHC processing of exogenous Ag. The potencies of different inhibitors for blockade of MHC-I Ag processing correlated directly with their potencies for inhibition of the chymotrypsin-like proteasome activity. In contrast, conventional inhibitors of vacuolar cysteine proteases (e.g., leupeptin and benzyloxycarbonyl-Phe-Ala-CHN2) had little effect on MHC-I processing or the chymotryspin-like activity of isolated proteasomes. These results directly demonstrate that inhibition of proteasome activity blocks MHC-I Ag processing, confirming a role for proteasomes in this pathway. Moreover, they suggest that the chymotrypsin-like activity of the proteasome may be of major importance to the cytosolic processing of at least some Ag.

Aldehydes↗

Regulation of multidrug resistance gene mdr1b/mdr1 expression in isolated mouse uterine epithelial cells.

The mammalian uterine epithelium (UE) undergoes drastic physiological and morphological changes during pregnancy. Steady-state levels of murine mdr1b mRNA, transcribed from a multidrug resistance gene encoding a membrane protein which functions as a transporter of lipophilic cytotoxic agents, are low in nonpregnant, cycling UE, but drastically increase (about 1,500- to 2,000-fold) at day 8 of gestation. At day 16 of gestation, levels of mdr1b mRNA are 2,500- to 3,000-fold higher than those in the cycling UE cells. Levels of mdr1b mRNA were elevated to levels comparable to those observed during pregnancy, in the UE of ovariectomized mice following 5-8 days of estrogen and progesterone administration. Withdrawal of these hormones resulted in a drastic reduction of mdr1b mRNA within 36 hr. These results suggested that steroid hormones alone can account for increased mdr1b mRNA expression and do not require the presence of other placenta/embryo-derived factors. Moreover, the hormonal effect on uterine mdr1b mRNA biosynthesis during pregnancy apparently is a delayed phenomenon. Nuclear run-on assays demonstrated that the rate of mdr1b transcription in UE cells prepared from 15-day pregnant mice (d-15 UE cells) was about two- to three-fold higher than that in nonpregnant UE cells. This increased transcription rate alone cannot account for mdr1b mRNA accumulation during pregnancy. mdr1b mRNA expression was investigated in primary cultures of d-15 UE cells. mdr1b mRNA levels decayed by 50% within 3-4 hr of culture and reached a steady-state 0.5-2% of initial levels by 24 hr. The rate of mdr1b mRNA decay in primary d-15 UE cells was decreased by treatment with alpha-amanitin or cycloheximide, suggesting that the decay pathway requires both transcription and de novo protein synthesis. Our results suggest that multiple mechanisms are involved in the maintenance of the high levels of mdr1b mRNA in pregnant UE cells. Furthermore, these data suggest that increased mRNA stability may contribute to the accumulation of mdr1b transcript during pregnancy.

Amanitins↗

Unidirectional valve patch for repair of cardiac septal defects with pulmonary hypertension.

BACKGROUND: Congenital septal defects with a large left-to-right shunt often cause pulmonary hypertension, which complicates surgical repair of the defects. METHODS: Twenty-four patients with congenital cardiac septal defects and severe pulmonary hypertension had operation to close the septal defect using a unidirectional valve patch during a 3-year period. The ratio of systolic pulmonary artery pressure to systolic arterial blood pressure was near to or more than 1.0 in all patients. RESULTS: Two patients died in the hospital after operation, and there have been no deaths during intermediate term follow-up. Mean pulmonary artery pressure decreased from 80 +/- 12 mm Hg to 56 +/- 18 mm Hg. The ratio of pulmonary artery pressure to systemic arterial pressure dropped from 1.1 +/- 0.1 mm Hg to 0.7 +/- 0.1 mm Hg. The unidirectional valve patch functioned allowing right to left shunting in 4 patients with a systolic pulmonary artery pressure more than systolic arterial blood pressure immediately after closure of a septal defect. The patch sealed or was effectively closed by the third postoperative day. There was impressive improvement in symptoms and exercise tolerance after operation during the 3-month to 3-year (mean, 1.1 year) follow-up period. CONCLUSIONS: The unidirectional valve patch is useful for management of patients having operation to close cardiac septal defects in the presence of severe pulmonary hypertension.

Adolescent↗

The cost-effectiveness of a special care unit to care for the chronically critically ill.

To assess the relative value of healthcare programs, technologic innovations, and clinical decisions, policymakers are searching for ways to evaluate cost-effectiveness. What constitutes cost-effectiveness and how should it be measured? The authors discuss ways in which the cost-effectiveness of clinical programs can be measured and describes various methods of assessing both costs and effectiveness. Comparison of the cost-effectiveness of a nurse managed special care unit with that of traditional intensive care units illustrates some of these methods.

Chronic Disease↗

Specific cleavage of two target sequences of the tobacco mosaic virus RNA by a di-component ribozyme.

Two di-component ribozymes RZ34 and RZ13, the simplest forms of multi-component ribozymes, by ligating mono-ribozymes RZ3 and RZ4 and RZ1 and RZ3 were constructed, respectively. The in vitro results indicate that each di-component ribozyme is able to cleave two different TMV transcripts, with the specificity and efficiency similar to their mono-ribozyme counterparts. Possible effects on the specificity and efficiency by temperature and the positions of the component-ribozymes in these di-component ribozymes were also studied.

Base Sequence↗

Processing of bacterial antigens for presentation to class I and II MHC-restricted T lymphocytes.

Phagocytosis leads to the destruction of many bacteria and the proteolytic degradation of bacterial antigens within phagolysosomes to produce immunogenic peptides that bind to Class II major histocompatibility (MHC) molecules within vacuolar compartments. On the other hand, Class I MHC molecules bind cytosol-derived peptides, including peptides from bacteria that escape the vacuolar system and penetrate into the cytosol. A recently described pathway may also allow the presentation of peptides from intravacuolar organisms by Class I MHC molecules in some cases. T cell recognition of peptide-MHC complexes then provides the primary basis for specific immunity to protein antigens of bacteria. This article will review the subcellular compartments and mechanisms involved in generating immunogenic peptides, the subcellular localization of MHC molecules that bind these peptides, and bacterial parameters that affect antigen processing.

Animals↗

[The anatomic study of levator veli palatini muscle].

The purpose of this study was to obtain more informations about the levator veli palatini (LVP) muscle through the dissection of head and neck cadaver samples in 20 sides of normal adults, 4 sides of normal newborns and 4 sides of cleft palate newborns and histological sections in 6 normal fetuses. The results shown that the middle part of LVP is 7.9 mm in width, 3.4mm in thickness and 29mm2 on cross-section area. There are two angle relationship between LVP and the soft palate, which are 44.5 degrees angle to midline sagittal section inward and 33.0 degrees angle to coronal section forward. The LVP insertion is between the 40% to 70% of the soft palate length and only 25% insertion of LVP is located at the junction area of palatal aponeurosis and fibromuscular component of palatine raphe. Other parts of LVP insertion are merged into the midline fibromuscular component. The muscle has a close relationship with the medial part of the cartilaginous portion of Eustachian tube. The LVP is supplied by the palatal branches of ascending palatine artery and ascending pharyngeal artery, which are into the LVP at or above the level of the hard palate. The LVP nerve branches coming from the pharyngeal plexus pass through the lateral pharyngeal space. The LVP reconstruction in cleft palate repair is discussed.

Adult↗

[Treatment of ototoxic auditory damage caused by kanamycin with electroacupuncture at different acupoints].

We adopted the technique and method of integrating the morphology with function to select the effective acupoints for treatment of deafness. The results show that: (1) Tinggong (SI 19), Yifeng (TE 17), Waiguan (TE 5), Shenshu (UB 23), Sanyinjiao (SP 6) and Zhubin (KI 9) etc. are the effectine acupoints for the treatment of ototoxic auditory damage caused by drug, especially, the effect of Tinggong, Sanyinjiao and Zhubin etc, is much better; (2) electroacupuncture can promote audibility, improve SDH activity and relieve progressing injury of auditory hair cells, (3) FFR method has an important significance in the determination of ototoxic damage caused by drug.

Animals↗

Phagocytic processing of exogenous particulate antigens by macrophages for presentation by class I MHC molecules.

Exogenous Ags that are processed in vacuolar endocytic compartments are generally presented by class II MHC molecules and not class I MHC (MHC-I) molecules, which conventionally present cytoplasmic or endogenous Ags. Accordingly, i.v. immunization of C57BL/6 mice with soluble OVA did not elicit a CD8 T cell response. However, i.v. immunization with OVA coupled to Latex particles (Latex-OVA) elicited an OVA-specific CD8 T cell response in vivo (particles from 59 to 2000 nm diameter were effective). In vitro, Latex-OVA was processed by H-2b macrophages and presented by Kb at least 100- to 1000-fold more efficiently than was soluble OVA. Inhibition of phagocytosis by cytochalasin D blocked the processing of Latex-OVA, whereas processing was not blocked by Brefeldin A. Latex-OVA was presented directly by H-2b macrophages or after "regurgitation" of processed OVA peptide from viable MHC-disparate macrophages for binding to surface Kb molecules on fixed H-2b macrophages. Peptide regurgitation was observed during processing of both Latex-OVA and Salmonella typhimurium 14028s that express an OVA fusion protein (Crl-OVA). However, the regurgitation pathway was less efficient than direct processing by viable H-2b macrophages. Thus, macrophages express an alternate pathway that allows MHC-I presentation of vacuolar exogenous particulate Ags, including inert synthetic particles without lipid membranes and intravacuolar bacteria. Peptides from these Ags are released from intracellular compartments to bind to surface MHC-I molecules, but peptide-MHC-I complexes also may be generated within intracellular compartments.

Animals↗

Cellular binding site and membrane binding proteins for triglyceride-rich lipoproteins in human monocyte-macrophages and THP-1 monocytic cells.

Triglyceride- and cholesterol-rich foam cells derived from monocyte-macrophages are commonly associated with some forms of hypertriglyceridemia. In this report, direct binding studies at 4 degrees C demonstrate that human monocyte-macrophages (HMM) 1-6 days after isolation from blood and human THP-1 monocytic cells, before and up to 7 days after differentiation with phorbol ester, exhibit a high affinity (Kd 3-6 nM), saturable, specific, and apolipoprotein (apo) E-independent binding site for the uptake and degradation of certain triglyceride-rich lipoproteins (TGRLP). Ligand blotting analysis identified two membrane binding proteins (MBP) of apparent molecular weights of 200 and 235 kDa (MBP 200 and MBP 235) in both cell types that share the same ligand specificity as the cellular site and bind hypertriglyceridemic (HTG) VLDL, trypsinized VLDL devoid of apoE (tryp-VLDL), and dietary plasma chylomicrons from normal subjects but not LDL, acetyl LDL, or normal VLDL with high affinity. Neither lipoprotein lipase nor apoE are required for TGRLP binding to the cells or the isolated MBPs. The cellular binding site and the MBPs are expressed at similar levels at all stages of differentiation, unlike the LDL or the acetyl LDL receptor. TGRLP that bind to the MBPs induce rapid, saturable, cellular triglyceride accumulation in monocytes as well as macrophages; normal VLDL does not. In addition, the cellular high affinity binding site and MBP 200 and 235 are not affected by the media sterol content, unlike the LDL receptor. Taken together, these data indicate that human monocyte-macrophages exhibit a high affinity, saturable, specific, apoE- and lipoprotein lipase-independent binding site and membrane binding proteins for TGRLP that differ in expression, specificity, and molecular size from receptors of the LDL receptor gene family or the acetyl LDL receptor. The shared characteristics of the cellular binding site with MBP 200 and MBP 235 suggest that they are candidates for the receptor-mediated, apoE-independent uptake of HTG-VLDL and chylomicrons by monocytes and macrophages and therefore may be involved in foam cell formation.

Animals↗