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Confirmation of the chromosomal localization of human lamp genes and their exclusion as candidate genes for Salla disease.

Salla disease is an inherited lysosomal storage disorder caused by accumulation of free sialic acid in the lysosomes. Lamp genes, lamp A and lamp B (lysosome associated membrane proteins), are the first known genes encoding for human lysosomal membrane proteins. Absence of linkage in a large group of families shows that lamp genes are not involved in Salla disease. The lamp genes were localized, using Southern hybridization in hamster--human hybrid cell panels, to chromosomes 13 (lamp A) and X (lamp B).

Antigens, CD↗

Inhibitory effect of different UV lamps on the infectivity of taeniid eggs.

The expanding distribution of alveolar echinococcosis in Hokkaido, Japan necessitates a search for control measures, particularly for killing Echinococcus eggs; and UV radiation is a potential tool. To determine the effective UV lamp and its optimum duration of exposure to inhibit the infectivity of taeniid eggs, three UV lamps (UVA, UVB and UVC) with peak emissions at different wavelengths were used to irradiate eggs of Taenia taeniaeformis. Irradiation was done in durations of 30, 90, 270, 810, 2,430 and 7,290 s. Infectivity of the irradiated eggs was tested by oral inoculation into rats. The inhibitory effect of the UV lamps at different exposure durations was determined by the reduction in the larval cyst establishment and by the delay of larval development. The UVC lamp (narrow peak at 254 nm) was found to be the most effective lamp at an exposure duration of more than 2,430 s (2,430 mJ/cm2), attaining 100% reduction. Also, shorter exposure durations produced a significant effect on the development of irradiated eggs in a greater percentage of small cysts (> 1 mm) among the metacestodes. Irradiation delayed the development of some cysts, as observed in histopathological sections. UVA and UVB lamps were not able to attain 100% reduction. It is concluded that the UVC lamp can be used as a tool in sterilizing taeniid eggs and is thus a promising method to control echinococcosis, although it may require a higher intensity of exposure.

Animals↗

Lysosome-associated membrane proteins: characterization of LAMP-1 of macrophage P388 and mouse embryo 3T3 cultured cells.

Lysosome-associated membrane protein (LAMP)-1, a major glycoprotein of mouse embryo 3T3 cells and specifically associated with the lysosomal membrane, has been identified in P388 macrophage cells and compared with the homologous glycoprotein of NIH 3T3 cells. Immunofluorescence microscopy with anit-LAMP-1 monoclonal antibodies shows that the antigen was distributed throughout P388 cells including the ruffled edges or pseudopodia, identical to the pattern of acridine orange accumulation. LAMP-1 was purified from P388 cells by affinity chromatography with 1D4B monoclonal antibody, yielding a homogeneous glycoprotein comprising 0.1% of the total detergent-extracted cell protein. The apparent mass of P388 LAMP-1 was 130,000 to 150,000 compared to the 3T3 glycoprotein of 105,000 to 115,000. Analysis of tryptic peptides indicated that the two purified glycoproteins were highly homologous. Protein synthesis was analyzed in a variety of cell lines by pulse-chase labeling with [35S]methionine; in every case, LAMP-1 was synthesized as a precursor of apparent Mr 92,000, and then converted to heterogeneous mature forms differing in average Mr from 110,000 to 140,000. The basis for these apparent differences in mass was examined by studies of the biosynthesis and oligosaccharide composition of the glycoprotein. Core polypeptides of 45,000 Da were obtained from both HaNIH and P388 cells by treating immunoprecipitates of [35S]methionine pulse-labeled molecules with endoglycosidase H. Cells treated with monensin contained heterogeneous molecules of 80,000 to 85,000 Da. Isoelectric heterogeneity of mature LAMP-1 was markedly reduced by treatment with neuraminidase whereas there was little effect on the apparent molecular weight of the molecules or the differences between the various cell lines. beta-D-Xyloside inhibition of glycosaminoglycan synthesis had little effect on the apparent mass of LAMP-1.

Animals↗

Toxicity and mutagenicity of radiation from fluorescent lamps and a sunlamp in L5178y mouse lymphoma cells.

Unfiltered broad spectrum radiation emitted by black light, cool white, and black light blue fluorescent lamps and a sunlamp, is both toxic and mutagenic to L5178Y mouse lymphoma cells when the cells are irradiated in phosphate-buffered saline. The increase in mutant frequency seen after exposure of the cells is linear throughout the range of exposures tested. The linear increase in mutagenesis is observed even at exposure levels which do not cause significant toxicity. To facilitate comparison of the differing rates of mutagenesis derived from exposure-response curves obtained for each light source, we have defined a parameter, joule-equivalent mutagenesis (jem), equal to mutants per 10(5) survivors per joule per square meter. Jem values are calculated using the integrated irradiance of each lamp. Based on jem values, the relative mutagenicity of the various lamps tested (compared with a germicidal ultraviolet lamp) is 3 x 10(-3) for the sunlamp, 1 x 10(-4) for the black light and cool white lamps, and 3 x 10(-5) for the black light blue lamp. The toxic and mutagenic effects of the lamps are in reasonable agreement with their relative spectral output from 290 to 330 nm.

Cell Line↗

cDNA cloning and structural analysis of the human limbic-system-associated membrane protein (LAMP).

The limbic-system-associated membrane protein (LAMP) is a 64-68-kDa neuronal surface glycoprotein distributed in cortical and subcortical regions of the limbic system. The human LAMP gene was cloned by RT-PCR using human cerebral cortex mRNA and oligodeoxyribonucleotide (oligo) primers derived from the rat lamp cDNA sequence. The human and rat LAMP cDNAs showed 94% identity at the nucleotide (nt) level, and the encoded 338-amino-acid (aa) polypeptides shared 99% sequence identity. All the important features of LAMP were conserved: (i) the deduced aa sequence reflecting a glycosyl-phosphatidylinositol (GPI)-anchor, (ii) eight putative N-linked glycosylation sites, and (iii) conserved pairs of Cys forming three internal repeats characteristic of the immunoglobulin superfamily (IgSF). Northern blot analysis indicated the presence of two mRNA transcripts in the human brain of a size identical to those identified in adult rat brain. These data indicate that LAMP is a highly conserved new member of the IgSF which, together with the opioid-binding cell adhesion molecule (OBCAM) and neurotrimin, comprises a new subfamily that has been designated as IgLONs. With a unique distribution in limbic structures, LAMP may play an important role in limbic system development and function, as suggested by previous in vitro and in vivo functional studies.

Amino Acid Sequence↗

Phototherapy for neonatal jaundice: clinical equivalence of fluorescent green and "special" blue lamps.

The relative efficacy of fluorescent green (Sylvania F20T12/G) and "special" blue (Westinghouse F20T12/BB) lamps in the phototherapy of jaundiced neonates was investigated. Two groups of low birth weight infants with a mean gestational age of 35 weeks and mean birth weight of 1930 gm, who developed hyperbilirubinemia within the first 5 days of life, were given green or blue lamp phototherapy under the same irradiation conditions. No statistically significant difference in plasma bilirubin concentrations was found between the two groups after 24 or 48 hours of treatment. Because recent measurements indicate that green lamps are much less efficient than special blue lamps for the production of Z, E isomers of bilirubin in vitro and in vivo, the clinical equivalence of these two types of lamps seems to support the hypothesis that production of structural photoisomers of bilirubin is the main mechanism of phototherapy in humans. Therefore, fluorescent green lamps provide an alternative to special blue lamps for treatment of neonatal hyperbilirubinemia.

Bilirubin↗

Different biologic effectiveness of blacklight fluorescent lamps available for therapy with psoralens plus ultraviolet A.

In 1976 we reported a change in spectral distribution of "blacklight" fluorescent lamps. It was not possible to determine the spectral composition of these lamps by any codes or packaging materials. Phototherapy booths utilizing standard BL-HO lamps will accept lamps of at least two spectral distributions; both types are commercially available. This study was conducted to determine the biologic efficacy of these two lamp types that we refer to as BL-O and BL-N. The BL-O spectrum had a peak emission at approximately 350 nm with 98% of the energy between 320 and 400 nm. The BL-N spectrum had its peak emission at 365 nm with a range from 340 to 400 nm. The BL-O spectrum was at least 2.5 to 4 times as effective as BL-N in causing minimally perceptible phototoxicity in albino hairless mice given oral doses of 8 mg/kg of 8-methoxypsoralen. Food and Drug Administration (FDA)-approved specifications imply that the BL-O spectrum is to be used for psoralens and ultraviolet A (PUVA) phototherapy. If lamps with the BL-N spectrum are replaced by lamps with the BL-O spectrum, the metered dose must be reduced to no more than one-fourth of the previous dose or the patient may suffer serious phototoxic reactions.

Animals↗

Do tanning lamps cause melanoma? An epidemiologic assessment.

Sun exposure is a known cause of melanoma, and there are several reasons for concern that tanning lamp use may also cause melanoma, especially because of the spectral output of the lamps,the similarity of tanning lamp use to natural sun exposure (particularly sunbathing), the reported association of melanoma with PUVA therapy, and the positive results in several case-control studies of melanoma risk and tanning lamp use. We have reviewed the epidemiologic literature relevant to the relation between tanning lamp use and melanoma and have discussed methodologic issues that should be considered in interpreting these reports and designing improved studies. Although several investigations have found a positive relation between tanning lamp use and melanoma, in some instances including dose-response or duration-response effects, the methodologic limitations preclude any firm conclusion regarding a causative relation. Meta-analysis of existing data may provide better information, but several limitations could only be overcome by new studies collecting more precise exposure data. The popularity of tanning lamp use, especially in young persons, adds urgency to the quest for elucidation of the relation between the use of these devices and melanoma risk.

Adolescent↗

Carbohydrate structures of recombinant soluble lamp-1 and leukosialin containing sialyl Le(x) terminus.

Recombinant soluble lamp-1 and soluble leukosialin can be produced from CHO cells which express sialyl Le(x) structures after stable transfection of fucosyltransferase-III. It was shown previously that those soluble lamp-1 and leukosialin are potent inhibitors for E-selectin-mediated adhesion of human colonic tumor cells (Sawada, R.; Tsuboi, S.; Fukuda, M. J. Biol. Chem., 1994, 269, 1425). In the present study, we have determined the amount of the sialyl Le(x) structure present in recombinant, soluble lamp-1 and soluble leukosialin. CHO cells were metabolically labeled with [3H]-galactose and recombinant soluble lamp-1 and leukosialin were purified from the spent medium. Glycopeptides containing N-glycans derived from lamp-1 were fractionated by sequential lectin affinity chromatography. Similarly, O-glycans released from leukosialin were fractionated by Bio-Gel P-4 gel filtration. The terminal structures of carbohydrate chains were determined by sequential digestion with specific glycosidases. The results clearly indicate that soluble lamp-1 contains much more sialyl Le(x) structure than soluble leukosialin. Considering that soluble leukosialin and lamp-1 are almost equally effective as inhibitors for E-selectin-mediated adhesion, the results strongly suggest that densely clustered O-glycans are better presenters for E-selectin ligands than N-glycans.

Animals↗

Similar dose-response and persistence of erythema with broad-band and narrow-band ultraviolet B lamps.

Psoriasis may be treated with ultraviolet B from lamps that have a broad emission spectrum or, more effectively, with lamps that have a narrow emission spectrum at 311 +/- 2 nm. There are conflicting reports of either greater or lesser burning episodes with narrow-band compared to broad-band ultraviolet B, even when treatments are based on predetermined minimal erythema dose measurements. This suggests that either the characteristics of the dose-response curve for erythema or the time course for erythema may be different for the two lamps. We examined the erythemal response to narrow-band and broad-band ultraviolet B in 12 patients with psoriasis. A geometric series of 10 doses from each lamp type were used on nonlesional skin on the back. Dose-response curves were constructed from reflectance measurements of erythema at 24 h and 72 h after irradiation. No significant difference was found in steepness of the erythema dose-response curve for the two lamps at 24 or 72 h. Persistence of erythema was assessed as the percentage of erythema remaining at 72 h. The mean persistence was 63% for narrow-band and 64% for broad-band lamps (p = 0.94). Therefore, in terms of erythemal response, no evidence has been found for a difference in burning potential for the two lamps.

Adult↗

The lysosomal-associated membrane protein LAMP-1 is a novel differentiation marker for HC11 mouse mammary epithelial cells.

HC11 cells are a model for mammary epithelial cell differentiation. Following treatment with the lactogenic hormones glucocorticoids, insulin and prolactin the HC11 cells synthesize milk proteins. Stereological analysis at the ultrastructural level suggested that lysosomal biogenesis was activated following lactogenic hormone treatment of HC11 cells. Differentiation was also accompanied by an increase in the cellular content of tri- and tetra-antennary oligosaccharides, which were reactive with isolectin L4 from Phaseolus vulgaris (L-PHA). The lysosomal-associated membrane glycoproteins LAMP-1 and LAMP-2 are the major carriers of this glycosylation pattern. An analysis of LAMP-1 and LAMP-2 expression levels showed that there was a dramatic increase in LAMP-1 following lactogenic hormone treatment of HC11 cells. The control of LAMP-1 expression is mainly post-transcriptional since the level of LAMP-1 RNA is not affected by lactogenic hormones. Stereological analysis also showed an increase in intermediate filament control of differentiated cells. Analysis of the cytokeratins expressed in differentiated cells suggests that HC11 cells have characteristics of a mammary-specific stem cell. Increase in lysosomal vesicles and their contents might play a role in intra- and extra-cellular remodeling, which is characteristic of cell differentiation.

Animals↗

Intensified phototherapy using daylight fluorescent lamps.

Jaundice is a common reason for therapeutic intervention in newborn infants and phototherapy is effective treatment if enough light energy is delivered to a skin surface area of sufficient size. Narrow spectrum blue light is superior to white light, but in developing countries fluorescent blue lamps often have to be imported and are much more expensive than white lamps. We developed a phototherapy unit in which seven daylight fluorescent tubes are placed immediately under the floor of a transparent plexiglass crib. The efficacy of this unit, delivering approximately 19 microW/cm2/nm, was compared with that of two conventional phototherapy units using overhead lamps placed 35 cm above the infants. One unit used daylight fluorescent tubes and delivered approximately 4 microW/cm2/nm, the other unit used special blue fluorescent tubes and delivered approximately 22 microW/cm2/nm. Fifty-one infants were included in the analyses, all of them breastfed on demand. Serum bilirubin levels were determined spectrophotometrically at 0, 12 and 24 h. The decrement in serum bilirubin concentrations was significantly greater in infants undergoing phototherapy with the new device or with special blue lamps compared to conventional overhead daylight lamps (p < 0.001 both at 12 and at 24 h). We conclude that highly efficient phototherapy may be delivered with daylight fluorescent lamps placed in very close proximity to the patient. Thus, lack of access to expensive imported special blue lamps does not preclude delivery of effective phototherapy in developing countries.

Circadian Rhythm↗

Different steady state subcellular distributions of the three splice variants of lysosome-associated membrane protein LAMP-2 are determined largely by the COOH-terminal amino acid residue.

The extensively glycosylated lysosome-associated membrane proteins (LAMP)-2a, b, and c are derived from a single gene by alternative splicing that produces proteins with differences in the transmembrane and cytosolic domains. The lysosomal targeting signals reside in the cytosolic domain of these proteins. LAMPs are not restricted to lysosomes but can also be found in endosomes and at the cell surface. We investigated the subcellular distribution of chimeras comprised of the lumenal domain of avian LAMP-1 and the alternatively spliced domains of avian LAMP-2. Chimeras with the LAMP-2c cytosolic domain showed predominantly lysosomal distribution, while higher levels of chimeras with the LAMP-2a or b cytosolic domain were present at the cell surface. The increase in cell surface expression was due to differences in the recognition of the targeting signals and not saturation of intracellular trafficking machinery. Site-directed mutagenesis defined the COOH-terminal residue of the cytosolic tail as critical in governing the distributions of LAMP-2a, b, and c between intracellular compartments and the cell surface.

Alternative Splicing↗

Reduced phenylalanine ammonia-lyase and tyrosine ammonia-lyase activities and lignin synthesis in wheat grown under low pressure sodium lamps.

Wheat (Triticum aestivum L. cv Fremont) grown in hydroponic culture under 24-hour continuous irradiation at 560 to 580 micromoles per square meter per second from either metalhalide (MH), high pressure sodium (HPS), or low pressure sodium (LPS) lamps reached maturity in 70 days. Grain yields were similar under all three lamps, although LPS-grown plants lodged at maturity. Phenylalanine ammonia-lyase (PAL) and a tyrosine ammonia lyase (TAL) with lesser activity were detected in all extracts of leaf, inflorescence, and stem. Ammonia-lyase activities increased with age of the plant, and plants grown under the LPS lamp displayed PAL and TAL activities lower than wheat cultured under MH and HPS radiation. Greenhouse solar-grown wheat had the highest PAL and TAL activities. Lignin content of LPS-grown wheat was also significantly reduced from that of plants grown under MH or HPS lamps or in the greenhouse, showing a correlation with the reduced PAL and TAL activities. Ratios of far red-absorbing phytochrome to total phytochrome were similar for all three lamps, but the data do not yet warrant a conclusion about specific wavelengths missing from the LPS lamps that might have induced PAL and TAL activities in plants under the other lamps.

Ammonia-Lyases↗

Provocation testing in polymorphic light eruption using fluorescent ultraviolet (UV) A and UVB lamps.

BACKGROUND: Provocation testing is frequently performed during investigation of patients with suspected polymorphic light eruption (PLE). Techniques are not standardized between centres. OBJECTIVES: We sought to evaluate the efficacy of different fluorescent ultraviolet (UV) radiation lamps for provocation testing in PLE. METHODS: We analysed results in 68 patients referred consecutively for phototesting in whom a diagnosis of PLE seemed likely based on clinical history. Patients' case notes were reviewed and responses recorded to provocation testing on forearm skin over three consecutive days using broadband UVA, narrowband and broadband UVB lamps. RESULTS: A positive papular response to broadband UVA exposure was seen in 38 patients [56%, estimated 95% population confidence interval (CI) 43-67.9]. Thirty-four patients (50%) had a positive papular response to narrowband UVB exposure (95% CI 37.6-62.4). The probability of a positive provocation test following irradiation with both lamps was 80.9% (95% CI 69.5-89.4). From April 1999, 34 patients also had provocation testing with broadband UVB. Although six patients (18%) had a positive papular response, they all showed a positive response to one or both of the other lamp types. CONCLUSIONS: Provocation testing with fluorescent UVA and UVB lamps is a cheap and readily available method that can be used as a diagnostic aid to investigate patients with suspected PLE. Using both broadband UVA and narrowband UVB lamps for testing increases the likelihood of confirming the diagnosis than if either lamp is used alone.

Adolescent↗

Unifying nomenclature for the isoforms of the lysosomal membrane protein LAMP-2.

The present nomenclature of the splice variants of the lysosome-associated membrane protein type 2 (LAMP-2) is confusing. The LAMP-2a isoform is uniformly named in human, chicken, and mouse, but the LAMP-2b and LAMP-2c isoforms are switched in human as compared with mouse and chicken. We propose to change the nomenclature of the chicken and mouse b and c isoforms to agree with that currently used for the human isoforms. To avoid confusion in the literature, we further propose to adopt the use of capital letters for the updated nomenclature of all the isoforms in all three species: LAMP-2A, LAMP-2B, and LAMP-2C.

Alternative Splicing↗

Correction of emission spectra in microspectrofluorimetry using a reference lamp: computations.

Accurate correction of emission spectra in microspectrofluorimetry, using a reference lamp, may require that a factor for the emissivity of tungsten be introduced. This is only possible provided that the true temperature of the lamp filament is known. A method is given for obtaining the true temperature from the knowledge of the colour temperature. Also, the values of the spectral concentration of the radiance of the black body, either computed according to Planck's equation or taken from available published tables, have to be converted from energetic units to units of quanta since the photomultiplier is linear not to absorbed power but to units of quanta. When the fluorescence spectra to be corrected extend into the far red it is preferable to use a lower temperature (by lowering the supply voltage) than that for which the lamp is certified. It is possible to determine the new temperature (and then the corresponding spectral distribution) by taking a few pairs of measurements at different wavelengths both at the lower voltage and at the voltage for which the lamp is certified and then introducing these values in a non-linear regression soluble on a PC with a curve fitting program. The microscope tungsten halogen lamp can conveniently be used as a reference, thanks to its small size and its steady spectral characteristics. When high accuracy is required, however, the halogen lamp should be calibrated against a certified ribbon filament lamp.

Flow Cytometry↗

The ultraviolet C energy emitted from FS lamps contributes significantly to the induction of human erythema and murine ear edema.

FS ultraviolet (UV) lamps are used extensively to study the biological effects of ultraviolet radiation. Using published data, we investigated the relative contributions of the UVC (250-289 nm), UVB (290-319 nm), and UVA (320-400 nm) portions of the FS lamp emission spectrum to the induction of human erythema and murine ear edema. Many investigators contend that the biological activity of the spectrum emitted from these lamps resides primarily in the UVB region, based on the proportions of the power emission spectrum in this region (53.6%) versus the small relative contribution of UVC (3.2%) and the low biological activity of UVA (43.2%). However, if the biological effectiveness spectrum of FS lamps is calculated by multiplying the power spectrum with different action spectra, the biological effects of UVC are readily observed. For example, 10.4% of the murine ear edema activity and 11.1% (Parrish) or 16.7% (McKinlay-Diffey) of the human erythemal activity is due to the energy emitted in the UVC region. Experimental determination of human erythema and murine ear edema demonstrated that, for an equal amount of energy delivered, radiation from the unfiltered lamps was more potent in causing these responses than radiation from filtered lamps, and the ratio of effectiveness could be predicted by the effectiveness spectra. Thus, the contribution of UVC emitted from FS lamps to biological effectiveness spectra should not be ignored.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗