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Biomedical subjects

L S Johnson

Publications and source records attributed to L S Johnson.

At least 19 recordsLinked to original sources
PubMed2000

Retinoic acid induces sodium/iodide symporter gene expression and radioiodide uptake in the MCF-7 breast cancer cell line.

The sodium/iodide symporter (NIS) stimulates iodide uptake in normal lactating breast, but is not known to be active in nonlactating breast or breast cancer. We studied NIS gene regulation and iodide uptake in MCF-7 cells, an estrogen receptor (ER)-positive human breast cancer cell line. All-trans retinoic acid (tRA) treatment stimulated iodide uptake in a time- and dose-dependent fashion up to approximately 9.4-fold above baseline. Stimulation with selective retinoid compounds indicated that the induction of iodide uptake was mediated by retinoic acid receptor. Treatment with tRA markedly stimulated NIS mRNA and immunoreactive protein ( approximately 68 kDa). tRA stimulated NIS gene transcription approximately 4-fold, as shown by nuclear run-on assay. No induction of iodide uptake was observed with RA treatment of an ER-negative human breast cancer cell line, MDA-MB 231, or a normal human breast cell line, MCF-12A. The iodide efflux rate of tRA-treated MCF-7 cells was slow (t(1/2) = 24 min), compared with that in FRTL-5 thyroid cells (t(1/2) = 3.9 min), favoring iodide retention in MCF-7 cells. An in vitro clonogenic assay demonstrated selective cytotoxicity with (131)I after tRA stimulation of MCF-7 cells. tRA up-regulates NIS gene expression and iodide uptake in an ER-positive breast cancer cell line. Stimulation of radioiodide uptake after systemic retinoid treatment may be useful for diagnosis and treatment of some differentiated breast cancers.

Alitretinoin↗
PubMed1999

Initial clinical experience with a video-based patient positioning system.

PURPOSE: To report initial clinical experience with an interactive, video-based patient positioning system that is inexpensive, quick, accurate, and easy to use. METHODS AND MATERIALS: System hardware includes two black-and-white CCD cameras, zoom lenses, and a PC equipped with a frame grabber. Custom software is used to acquire and archive video images, as well as to display real-time subtraction images revealing patient misalignment in multiple views. Two studies are described. In the first study, video is used to document the daily setup histories of 5 head and neck patients. Time-lapse cine loops are generated for each patient and used to diagnose and correct common setup errors. In the second study, 6 twice-daily (BID) head and neck patients are positioned according to the following protocol: at AM setups conventional treatment room lasers are used; at PM setups lasers are used initially and then video is used for 1-2 minutes to fine-tune the patient position. Lateral video images and lateral verification films are registered off-line to compare the distribution of setup errors per patient, with and without video assistance. RESULTS: In the first study, video images were used to determine the accuracy of our conventional head and neck setup technique, i.e., alignment of lightcast marks and surface anatomy to treatment room lasers and the light field. For this initial cohort of patients, errors ranged from sigma = 5 to 7 mm and were patient-specific. Time-lapse cine loops of the images revealed sources of the error, and as a result, our localization techniques and immobilization device were modified to improve setup accuracy. After the improvements, conventional setup errors were reduced to sigma = 3 to 5 mm. In the second study, when a stereo pair of live subtraction images were introduced to perform daily "on-line" setup correction, errors were reduced to sigma = 1 to 3 mm. Results depended on patient health and cooperation and the length of time spent fine-tuning the position. CONCLUSION: An interactive, video-based patient positioning system was shown to reduce setup errors to within 1 to 3 mm in head and neck patients, without a significant increase in overall treatment time or labor-intensive procedures. Unlike retrospective portal image analysis, use of two live-video images provides the therapists with immediate feedback and allows for true 3-D positioning and correction of out-of-plane rotation before radiation is delivered. With significant improvement in head and neck alignment and the elimination of setup errors greater than 3 to 5 mm, margins associated with treatment volumes potentially can be reduced, thereby decreasing normal tissue irradiation.

Algorithms↗
PubMed1999

Adolescent sexuality and sexually transmitted diseases: attitudes, beliefs, knowledge, and values.

This study described rural adolescents' attitudes, beliefs, knowledge, and values with regard to sexuality and sexually transmitted diseases (STDs). Rotter's Social Learning Theory (1954) provided the theoretical framework for this descriptive, correlational design. The convenience sample consisted of 170 students from one rural high school. Consistent with past studies, results included the following: participants had more correct than incorrect knowledge related to sexual intercourse and STDs; the majority had positive attitudes toward condom use and believed it was OK for peers to have sex with a "steady;" the value of an exciting life correlated positively with attitudes toward sex; knowledge of sexual intercourse correlated positively with attitudes toward condom use; and the value health correlated positively with knowledge of sex and attitudes toward condom use, and negatively with attitudes toward sex. The findings in this study suggest the need for ongoing research with adolescents in the area of sexuality and STDs. Additionally, the findings support past studies, which revealed that knowledge of sexual intercourse and STDs has little impact on attitudes toward sexual intercourse. With the serious nature of some of the undesired consequences of adolescent sexual behavior, current and accurate information on this population is needed to assist health educators in developing interventions in this area.

Adolescent↗
PubMed1997

Performance of a video-image-subtraction-based patient positioning system.

PURPOSE: We have developed and tested an interactive video system that utilizes image subtraction techniques to enable high precision patient repositioning using surface features. We report quantitative measurements of system performance characteristics. METHODS AND MATERIALS: Video images can provide a high precision, low cost measure of patient position. Image subtraction techniques enable one to incorporate detailed information contained in the image of a carefully verified reference position into real-time images. We have developed a system using video cameras providing orthogonal images of the treatment setup. The images are acquired, processed and viewed using an inexpensive frame grabber and a PC. The subtraction images provide the interactive guidance needed to quickly and accurately place a patient in the same position for each treatment session. We describe the design and implementation of our system, and its quantitative performance, using images both to measure changes in position, and to achieve accurate setup reproducibility. RESULTS: Under clinical conditions (60 cm field of view, 3.6 m object distance), the position of static, high contrast objects could be measured with a resolution of 0.04 mm (rms) in each of two dimensions. The two-dimensional position could be reproduced using the real-time image display with a resolution of 0.15 mm (rms). Two-dimensional measurement resolution of the head of a patient undergoing treatment for head and neck cancer was 0.1 mm (rms), using a lateral view, measuring the variation in position of the nose and the ear over the course of a single radiation treatment. Three-dimensional repositioning accuracy of the head of a healthy volunteer using orthogonal camera views was less than 0.7 mm (systematic error) with an rms variation of 1.2 mm. Setup adjustments based on the video images were typically performed within a few minutes. The higher precision achieved using the system to measure objects than to reposition them suggests that the variability in repositioning is dominated by the ability of the therapist to make small, controlled changes in the position of the patient. CONCLUSION: Using affordable, off-the-shelf technology, we have developed a patient positioning system that achieves repositioning accuracy normally associated with fractionated stereotactic systems. The technique provides real-time guidance and can be used to easily and quickly correct patient setup before every treatment, thus significantly reducing overall random positioning error. This improved positioning capability provides the precision required to realize the potential gains of conformal radiotherapy.

Computer Terminals↗
PubMed1997

A diffusible cytotoxin of Haemophilus ducreyi.

Little is known about the virulence mechanisms employed by Haemophilus ducreyi in the production of genital ulcers. This Gram-negative bacterium previously has been shown to produce a soluble cytotoxic activity that kills HeLa and HEp-2 cells. We have now identified a cluster of three H. ducreyi genes that encode this cytotoxic activity. The predicted proteins encoded by these genes are most similar to the products of the Escherichia coli cdtABC genes that comprise the cytolethal distending toxin (CDT) of this enteric pathogen. Eleven of 12 H. ducreyi strains were shown to possess this gene cluster and culture supernatants from these strains readily killed HeLa cells. The culture supernatant from a single strain of H. ducreyi that lacked these genes was unable to kill HeLa cells. When the H. ducreyi cdtABC gene cluster was cloned into E. coli, culture supernatant from the recombinant E. coli clone killed HeLa cells. A monoclonal antibody that neutralized this soluble cytotoxic activity of H. ducreyi was shown to bind to the H. ducreyi cdtC gene product. This soluble H. ducreyi cytotoxin may play a role in the development or persistence of the ulcerative lesions characteristic of chancroid.

Amino Acid Sequence↗
PubMed1995

Beta-particle dosimetry in radiation synovectomy.

Beta-particle dosimetry of various radionuclides used in the treatment of rheumatoid arthritis was estimated using Monte Carlo radiation transport simulation coupled with experiments using reactor-produced radionuclides and radiachromic film dosimeters inserted into joint phantoms and the knees of cadavers. Results are presented as absorbed dose factors (cGy-cm2/MBq-s) versus depth in a mathematical model of the rheumatoid joint which includes regions of bone, articular cartilage, joint capsule, and tissue (synovium) found in all synovial joints. The factors can be used to estimate absorbed dose and dose rate distributions in treated joints. In particular, guidance is provided for those interested in (a) a given radionuclide's therapeutic range, (b) the amount of radioactivity to administer on a case-by-case basis, (c) the expected therapeutic dose to synovium, and (d) the radiation dose imparted to other, nontarget components in the joint, including bone and articular cartilage.

Arthritis, Rheumatoid↗
PubMed1994

A comparison of the effects of sodium saccharin in NBR rats and in intact and castrated male F344 rats.

High doses of sodium saccharin (NaSac) increase proliferation in the bladder of the rat, with a male preponderance. The possibility that alpha 2u-globulin is involved in its mechanism of action was evaluated by feeding it at 7.5% of the diet to NCI-Black-Reiter (NBR) male rats, which do not synthesize liver-derived alpha 2u-globulin. NaSac affected urinary parameters similarly in F344 and NBR male rats, but NBR rats consumed more water leading to greater urinary volume. NaSac produced less proliferation in NBR than in intact F344 rats, with intermediate changes in castrated F344 males, which had intermediate urinary alpha 2u-globulin levels.

Alpha-Globulins↗
PubMed1994

Slowed receptor trafficking in mutant CHO lines of the End1 and End2 complementation groups.

A mutant Chinese hamster ovary cell line with nonconditional kinetic defects in receptor internalization and recycling was isolated, based on selection for resistance to a transferrin-diphtheria toxin conjugate and screening for aberrant receptor trafficking. The 12-4 cell line internalizes transferrin at approximately 75% of the parental rate and recycles transferrin back to the cell surface at approximately 55% of the parental rate. Internalization of low density lipoprotein is also reduced to approximately 70% of the parental cell rate, demonstrating that the mutant phenotype affects the trafficking of multiple receptors. Characterization of somatic cell hybrids indicated that the 12-4 phenotype is recessive, and complementation analysis determined that the 12-4 cell line is a member of the End2 complementation group. End2 mutants have previously been described as defective in endosomal acidification but have not been known to be defective in receptor trafficking. We have found similar defects in another End2 mutant cell line, suggesting that slowed receptor trafficking is characteristic of End2 mutants. Interestingly, transferrin receptor recycling and internalization are also slowed in another complementation group of mutants, End1, that is also defective in endosomal acidification. This study demonstrates altered receptor trafficking in End1 and End2 cell lines, a novel aspect of the mutant phenotypes. These findings provide evidence, based on a cellular genetic approach, that proper endosome acidification is necessary for maintenance of normal receptor recycling.

Animals↗
PubMed1994

Mitogenic effects of propoxur on male rat bladder urothelium.

Propoxur produces bladder tumors in rats, but not other species. The hyperplastic and tumorigenic effects do not occur if urinary pH is lowered by administering propoxur in a semi-synthetic diet or co-administering it with ammonium chloride (NH4Cl). We fed propoxur at 8000 p.p.m. in Altromin 1321 diet to male Wistar rats for 4 weeks, with or without NH4Cl as 10,000 p.p.m. of the diet. The urine of rats fed control diet with or without propoxur had a relatively high urinary pH (approximately 8); the addition of NH4Cl lowered the urinary pH by approximately 0.5-1.0 units. There was no evidence of urinary calculi or amorphous precipitate nor was there an increase in microcrystals or formation of different crystals than occur in normal rat urine. Propoxur produced hyperplasia of the urothelium, as observed by light and scanning electron microscopy, and increased the labeling index for proliferating cell nuclear antigen. These effects were significantly inhibited by co-administration with NH4Cl. There was no evidence of urothelial necrosis. Thus, the hyperplasia appears to result from a direct mitogenic effect of propoxur or a metabolite on the urothelium, rather than from toxicity and consequent regeneration. Based on the present study and previous investigations, the urothelial effects of propoxur in the rat are dependent on high urinary pH and high administered doses, factors which need to be incorporated into any mechanistic model for the chemical and into any extrapolation to potential effects in humans.

Animals↗
PubMed1993

Effects of dietary iron and folate supplementation on the physiological changes produced in weanling rats by sodium saccharin exposure.

Exposure of rats to high dietary levels of sodium saccharin (NaSac) started in utero produce physiological effects at 30 days post-birth that are similar to those found in pups of iron-deficient dams. These similarities suggest that some of the changes due to NaSac are secondary to iron deficiency. The present experiment investigated whether the effects of 7.5% dietary NaSac in the newborn rat could be prevented by dietary iron and/or folate supplementation. The NaSac-related effects prevented by iron supplementation included anaemia, decreased serum iron and folate, increased serum cholesterol and triglyceride and increased serum vitamin E. Folate supplementation prevented NaSac-induced depression of serum folate and increase in serum vitamin E. Although bladder hyperplasia was increased by dietary iron and/or folate supplementation, the majority of the urinary chemistry changes associated with NaSac treatment were not affected. The results show that some physiological changes associated with NaSac treatment in the newborn rat may occur as a consequence of iron deficiency rather than a direct effect of NaSac treatment. These changes may be independent of the urinary and bladder effects, which are not reversed by iron supplementation.

Animals↗
PubMed1993

Measurement of true absorption, endogenous fecal excretion, urinary excretion, and retention of calcium in term infants by using a dual-tracer, stable-isotope method.

A dual-tracer, stable-isotope method was used to measure the percentage of true calcium absorption (alpha), true calcium absorption rate (Va), endogenous fecal calcium excretion rate (Vf), urinary calcium excretion rate (Vu), and calcium retention rate (Vr). Twenty-seven infants with a mean gestation of 30.6 +/- 1.7 weeks and a mean birth weight of 1.4 +/- 0.21 kg were studied at 2 or 3 weeks of age, or both, during feedings of human milk (HM), fortified human milk (HMF), and commercially available formula (20 or 24 calories per ounce) for premature infants (EPF-20/780 and EPF-24/940) (part 1 of our study). Of 13 additional infants with a mean (+/- SD) birth weight of 1.26 +/- 0.25 kg and gestation of 29.6 +/- 2.5 weeks, 11 completed a crossover-design study at 2 and 3 weeks of age, receiving two identical formulas containing calcium, 940 mg/L, and phosphorus, 470 mg/L (EPF-24/940 formula) or calcium 1340 mg/L, and phosphorus, 680 mg/L (EPF-24/1340 formula) (part 2 of our study). The alpha value was higher in infants receiving HM (76.4 +/- 15.1%) or HMF (68.0 +/- 7.8%) than in those receiving EPF-20/750 formula (54.1% +/- 5.6%) or in previously reported infants fed standard formula (47.1% +/- 11.5%); those given EPF-24/940 formula had intermediate values (63.9% +/- 13.9%, part 1; 56.1% +/- 16.5%, part 2). No significant differences existed among groups for either Vu or Vf per kilogram. In the crossover study (part 2), no significant differences were seen between formulas for alpha and for Va, Vf, or Vr per kilogram. However, Vu per kilogram was significantly decreased in infants receiving the higher mineral formula (EPF-24/940: 3.6 +/- 2.3; EPF-24/1340: 2.9 +/- 2.3 mg/kg per day; p = < 0.005). With all feedings, alpha, Vu per kilogram, and Vf per kilogram were not related to gestational age, age at study, calcium intake, or each other. However, Vf per kilogram was inversely related to birth weight. Thus, alpha, Vu, and Vf appear to be independent and may be differentially affected by factors altering calcium dynamics. We conclude that increasing formula mineral content does not ensure increased retention; careful monitoring of individual infants remains indicated.

Calcium↗
PubMed1993

The End2 mutation in CHO cells slows the exit of transferrin receptors from the recycling compartment but bulk membrane recycling is unaffected.

We have characterized a new CHO cell line (12-4) derived from a parental line, TRVb-1, that expresses the human transferrin receptor. This mutant belongs to the end2 complementation group of endocytosis mutants. Like other end2 mutants, the endosomes in 12-4 cells show a partial acidification defect. These cells internalize LDL and transferrin at 70% of the rate of parental cells and externalize transferrin at 55% of the parental rate (Johnson, L. S., J. F. Presley, J. C. Park, and T. E. McGraw. J. Cell Physiol. 1993). In this report, we have used fluorescence microscopy to determine which step in receptor trafficking is affected in the mutants. Transferrin is sorted from LDL and is delivered to a peri-centriolar recycling compartment at rates similar to parental cells. However, the rate constant for exit of transferrin from the recycling compartment in mutant cells is 0.025 min-1 vs 0.062 min-1 in the parental line. We also measured the trafficking of a bulk membrane marker, 6-[N-[7-nitrobenzo-2-oxa-1,3-diazol-4-yl]-amino]hexanoyl- sphingosylphosphorylcholine (C6-NBD-SM) that labels the exofacial side of the plasma membrane. C6-NBD-SM enters the same recycling compartment as transferrin, and it exits the recycling compartment at a rate of 0.060-0.065 min-1 in both parental and 12-4 cells. We conclude that bulk membrane flow in the recycling pathway of 12-4 cells is normal, but exit of transferrin from the recycling compartment is slowed due to retention in this compartment. Thus, in the mutant cell line the recycling compartment carries out a sorting function, retaining transferrin over bulk membrane.

4-Chloro-7-nitrobenzofurazan↗
PubMed1993

Endosome acidification and receptor trafficking: bafilomycin A1 slows receptor externalization by a mechanism involving the receptor's internalization motif.

To examine the relationship between endosome acidification and receptor trafficking, transferrin receptor trafficking was characterized in Chinese hamster ovary cells in which endosome acidification was blocked by treatment with the specific inhibitor of the vacuolar H(+)-ATPase, bafilomycin A1. Elevating endosome pH slowed the receptor externalization rate to approximately one-half of control but did not affect receptor internalization kinetics. The slowed receptor externalization required the receptor's cytoplasmic domain and was largely eliminated by substitutions replacing either of two aromatic amino acids within the receptor's cytoplasmic YTRF internalization motif. These results confirm, using a specific inhibitor of the vacuolar proton pump, that proper endosome acidification is necessary to maintain rapid recycling of intracellular receptors back to the plasma membrane. Moreover, receptor return to the plasma membrane is slowed in the absence of proper endosome acidification by a signal-dependent mechanism involving the receptor's cytoplasmic tyrosine-containing internalization motif. These results, in conjunction with results from other studies, suggest that the mechanism for clustering receptors in plasma membrane clathrin-coated pits may be an example of a more general mechanism that determines the dynamic distribution of membrane proteins among various compartments with luminal acidification playing a crucial role in this process.

Adaptor Proteins, Vesicular Transport↗
PubMed1992

Acute urinary tract toxicity of tetraethylorthosilicate in rats.

Acute stomach, kidney, and bladder toxicity was evaluated in F344 rats after gastric gavage of tetraethylorthosilicate (TES) at daily doses of 0, 0.111, 0.223, and 0.333 g. Five rats of each sex at each dose were sacrificed after 1, 2, and 4 days. In TES-treated groups, silicate accumulated in the stomach glands and the muscle layer of the forestomach and glandular stomach. Serum chemistries demonstrated acute onset of renal failure. In the kidneys, acute tubular necrosis, accumulation of silicates, and superficial necrotizing papillitis were observed. In the renal pelvis and bladder, there was urothelial simple hyperplasia, focal erosion of the mucosa, edema, and inflammation. These acute toxic changes were dose and time dependent, but significant sex differences were not observed. The microscopic changes in the urothelium were similar to those observed following administration of high doses of sodium saccharin to male rats in which urinary silicate precipitate and crystals form.

Animals↗
PubMed1991

Absorbed dose profiles for radionuclides of frequent use in radiation synovectomy.

Absorbed dose profiles are presented for radionuclides of frequent or potential use in radiation synovectomy, an intraarticular technique aimed at treating rheumatoid arthritis through direct and highly selective irradiation of inflamed synovium. The radionuclides investigated were 198Au, 165Dy, 32P, 186Re, 90Y, and 166Ho. These profiles reveal the absorbed dose imparted per unit activity of injected radionuclide (Gy/mCi) as a function of penetration distance (mm) to major components of the arthritic synovial joint. Their usefulness is twofold: they can be employed to select the radionuclide best suited to achieving the proper depth of penetration in tissue (approximating the thickness of the inflamed synovium) and to determine, a priori, the necessary dose of radioactivity (delivering an absorbed dose sufficient to effectively treat all the diseased tissue). The extent of radiation of other synovial joint components, such as bone and articular cartilage, and how the advancing rheumatoid arthritis can be expected to alter the extent and pattern of absorbed dose penetration in the joint are also discussed.

Arthritis, Rheumatoid↗
PubMed1990

Immunological and structural characterization of a high affinity anti-fluorescein single-chain antibody.

Single-chain antibody of the (NH2) VL-linker-VH (COOH) design, was constructed based on prototype high affinity anti-fluorescein monoclonal antibody (mAb) 4-4-20. Purified single-chain antibody (SCA) 4-4-20/212 was studied relative to Ig mAb 4-4-20 in terms of ligand binding, kinetics, idiotypy, metatypy, and stability in denaturing agents. Ligand-binding data correlated with metatypic relatedness of the liganded site. Anti-metatypic reagents reacted preferentially with the liganded conformer of the 4-4-20 antibody active site and were unreactive with free ligand and the non-liganded (idiotypic) state. All results were consistent with the conclusion that SCA 4-4-20/212, with a 14-amino acid linker folded into a native conformational state that closely simulated the prototypical mAb. Furthermore, GndHCl unfolding and refolding studies demonstrated H and L chain variable domain intrinsic stability between SCA 4-4-20/212 and a 50 kDa antigen-binding fragment were nearly identical. This suggested CH1 and CL domain interactions may be more prevalent in V region molecular dynamics than structure.

Amino Acid Sequence↗
PubMed1989

Variable region primary structures of a high affinity anti-fluorescein immunoglobulin M cryoglobulin exhibiting oxazolone cross-reactivity.

Previous studies of murine IgM hybridoma protein 18-2-3, derived from an (NZB/NZW)F1 mouse following hyperimmunization with fluorescein (Fl)-conjugated keyhole limpet hemocyanin, demonstrated a high affinity for Fl (Ka = 2.9 x 10(10) M-1) and cryoprecipitation that was abrogated upon Fl binding to the antibody-combining site. V region sequences of 18-2-3 were determined by Edman degradation and nucleotide sequence analysis. The VH region of 18-2-3 was encoded by a gene VHI(B) of the Q52 VH family with 96% homology to anti-oxazolone antibody NQ7.5.3 but utilized a larger D region (DQ52 plus N region). The V kappa region of 18-2-3 was encoded by a gene V kappa IV with an amino acid sequence 97% homologous to that of anti-oxazolone antibody NQ11.1.18. Although monoclonal anti-Fl antibodies 18-2-3 and 4-4-20 possessed similar binding affinities and quenched bound fluorescein to the same extent (Qmax greater than 96%), they utilized different VH, D, V kappa, and J kappa genes, but the same JH gene segment (JH4). Solid-phase analyses showed that 18-2-3 was not idiotypically related to 4-4-20 and 9-40, prototypic anti-Fl antibodies. Fine specificity binding patterns of Fl analogues by 18-2-3 IgM and IgMs were distinct from other anti-Fl antibodies. Monoclonal antibody 18-2-3 bound phenyloxazolone bovine serum albumin with a lower affinity than for Fl-bovine serum albumin. The first hypervariable region of the 18-2-3 light chain showed homology to human cryoglobulins. This is the first variable region sequence of a murine IgM which self-aggregates at low temperature.

Amino Acid Sequence↗