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Reverse transcription loop-mediated isothermal amplification (RT-LAMP) for rapid detection of viral hemorrhagic septicaemia virus (VHS).

A one step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for detection of viral hemorrhagic septicaemia virus (VHS). A set of six primers were designed, based on the G-protein sequence of the VHS virus serotypes (He, F1, 23.75, Klapmolle and Rindsholm). The assay was optimised to amplify VHS RNA by incubation at 63 degrees C for only 1h, and required only a simple water bath or heating block to provide a constant temperature of 63 degrees C. RT-LAMP amplification products were detected by visual inspection using SYBR Green I stain and had a ladder-like appearance when electrophoresed on an agarose gel. The detection limit of the RT-LAMP assay was found to be similar to the commonly used RT-PCR method: both methods detected VHS RNA at a dilution of 10(6). The assay was evaluated using clinical samples and the results indicated the suitability and simplicity of the test as a rapid, field diagnostic tool for VHS virus.

Animals↗

Recovery of aluminium, nickel-copper alloys and salts from spent fluorescent lamps.

This study explores a combined pyro-hydrometallurgical method to recover pure aluminium, nickel-copper alloy(s), and some valuable salts from spent fluorescent lamps (SFLs). It also examines the safe recycling of clean glass tubes for the fluorescent lamp industry. Spent lamps were decapped under water containing 35% acetone to achieve safe capture of mercury vapour. Cleaned glass tubes, if broken, were cut using a rotating diamond disc to a standard shorter length. Aluminium and copper-nickel alloys in the separated metallic parts were recovered using suitable flux to decrease metal losses going to slag. Operation variables affecting the quality of the products and the extent of recovery with the suggested method were investigated. Results revealed that total loss in the glass tube recycling operation was 2% of the SFLs. Pure aluminium meeting standard specification DIN 1712 was recovered by melting at 800 degrees C under sodium chloride/carbon flux for 20 min. Standard nickel-copper alloys with less than 0.1% tin were prepared by melting at 1250 degrees C using a sodium borate/carbon flux. De-tinning of the molten nickel-copper alloy was carried out using oxygen gas. Tin in the slag as oxide was recovered by reduction using carbon or hydrogen gas at 650-700 degrees C. Different valuable chloride salts were also obtained in good quality. Further research is recommended on the thermodynamics of nickel-copper recovery, yttrium and europium recovery, and process economics.

Alloys↗

Noncontact corneal pachymetry with slit lamp-adapted optical coherence tomography.

PURPOSE: The purpose of this study was to determine the accuracy, the reproducibility, and the limits of agreement of noncontact central corneal thickness measurement with slit lamp-adapted optical coherence tomography (OCT). DESIGN: Nonrandomized comparative clinical trial. METHODS: In a prospective comparative observational study, a total of 108 consecutive patients (108 eyes) with normal corneas (92 eyes) and different corneal alterations (16 eyes) participated. Six sequential measurements of the central corneal thickness with slit lamp-adapted OCT and with ultrasound (US) pachymetry at 1640 ms(-1) were performed. The main outcome measures were accuracy, reproducibility assessed with precision and coefficient of variation (CV), and limits of agreement of central corneal thickness measurement. RESULTS: The mean central corneal thickness values were 541 +/- 43 microm (OCT) and 549 +/- 44 microm (US) with a mean precision of +/- 5.8 microm (CV 1.08%) and of +/- 4.0 microm (CV 0.73%), respectively. The method comparison revealed equivalence (+/- 2SD) in the 5% range with a mean difference between both methods of 7.9 microm (1.45%). The relative error was 8.7 microm (1.6%), which corresponded to limits of agreement (+/- 2SD) ranging from -9.5 microm to 25.3 microm. CONCLUSIONS: Central corneal pachymetry with slit lamp-adapted OCT revealed, for clinical purposes, an excellent accuracy and reproducibility with a high degree of agreement compared with US pachymetry. Thus, the presented OCT system seems to be a promising diagnostic modality to objectively measure corneal thickness in a convenient noncontact mode.

Adult↗

Slit-lamp beamsplitter for two stereoscopic views.

A double binocular beamsplitter, compatible with Zeiss optics, splits the light from each optical train of a slip-lamp microscope to provide an observer the exact stereoscopic view of the examiner. It has advantages in teaching slit-lamp examination techniques. Further, in surgery at the slit lamp, it allows the observer to function as an assistant and the surgeon to have both hands free.

Eye Diseases↗

LAMP-1 in CHO cells is a primary carrier of poly-N-acetyllactosamine chains and is bound preferentially by a mammalian S-type lectin.

Recent studies indicate that some mammalian S-type lectins bind preferentially to oligosaccharides containing the repeating disaccharide [3Gal beta 1,4GlcNAc beta 1]n or poly-N-acetyllactosamine (PL) sequence. We report here our investigation on the distribution of these sequences in glycoproteins in Chinese hamster ovary (CHO) cells and the interaction of glycoproteins containing PL chains with an immobilized S-type lectin (L14) from calf heart tissue. Our results demonstrate that PL chains are carried by a few high molecular weight glycoproteins which are bound by tomato-lectin Sepharose and one of these was precipitated by antibody to LAMP-1 (a lysosomal-associated membrane glycoprotein). More importantly, these high molecular weight glycoproteins, including LAMP-1, were bound with high affinity by L14. These results indicate that mammalian S-type lectins are highly specific in their interactions with glycoproteins and that LAMPs carry important recognition sequences for these lectins.

Animals↗

Analysis of oil used in late Roman oil lamps with different mass spectrometric techniques revealed the presence of predominantly olive oil together with traces of animal fat.

The lipid fraction of residues in ancient oil lamps found at the archaeological site of Sagalassos (south-west Turkey) was analysed by gas chromatography (GC) coupled to mass spectrometry (MS). The identification of plant sterols and long chain alcohols suggested that a vegetable oil was used in these lamps. The lipid sample was also analysed with reversed-phase liquid chromatography (LC) coupled to MS with atmospheric pressure chemical ionization (APCI). The identification of TAG's detected with LC-APCI-MS showed that predominantly olive oil was used as a fuel for the antique oil lamps. The presence of large quantities of multiply unsaturated triacylglycerol (TAG) and traces of saturated TAG indicated that also other oils and animal fat were added. Summarizing, the analysis of TAG's with LC-APCI-MS in lipid extracts of ancient ceramics proved to be a valuable method to reconstitute the original contents.

Animals↗

Isolation of the gene encoding lamp-1, a lysosomal membrane protein, by differential screening in an animal model of status epilepticus.

The present study employed differential library screening to identify genes associated with kainic acid (KA)-mediated selective neuronal death. One of the isolated clones was lamp-1, which encodes a major lysosomal membrane protein that is also present in the cell membrane. Following systemic KA treatment, lamp-1 was induced in vulnerable hippocampal and other limbic regions. This effect was blocked by cycloheximide (CHX) pre-treatment. Northern blot analysis also demonstrated the presence of lamp-1 transcripts in non-neural tissues. These findings suggest a novel role for lysosomal membrane proteins as markers of selective neuronal vulnerability.

Animals↗

Wood's lamp for superficial chemical peels.

BACKGROUND: Superficial chemical peels are being used with increasing frequency in the treatment of photoaging. OBJECTIVE: Our purpose was to develop a marker to indicate when all areas of skin have been treated and avoid reliance on a map or operator recall. METHODS: With a Wood's lamp (maximum output 365 nm) and addition of salicylic acid and fluorescein sodium, solutions that are readily available, the progress of the superficial chemical peel can be visualized. RESULTS: Wood's lamp accentuates epidermal pigment and therefore highlights dyschromia that will respond to a superficial chemical peel. Similarly, salicylic acid and fluorescein sodium fluoresce when viewed under Wood's lamp examination. CONCLUSION: When fluorescent compounds are incorporated into a superficial chemical peel, then fluorescence can accurately locate skip areas, minimize inadvertent overlayering, and thereby avoid deeper penetration.

Chemexfoliation↗

Environmentally acceptable effect of hydrogen peroxide on cave "lamp-flora", calcite speleothems and limestones.

Mosses, algae, and cyanobacteria (lamp-flora) colonize illuminated areas in show caves. This biota is commonly removed by a sodium hypochlorite solution. Because chlorine and other deleterious compounds are released into a cave environment during lamp-flora cleansing, hydrogen peroxide was tested as an alternative agent. In a multidisciplinary study conducted in the Kateinská Cave (Moravian Karst, Czech Republic), 12 algae- and cyanobacteria taxons and 19 moss taxons were detected. The threshold hydrogen peroxide concentration for the destruction of this lamp-flora was found to be 15 vol.%. Based on laboratory experiments in stirred batch reactors, the dissolution rates of limestones and calcite speleothems in water were determined as 3.77 x 10-3 and 1.81 x 10-3 mol m-2 h-1, respectively. In the 15% peroxide solution, the limestone and speleothem dissolution rates were one order of magnitude higher, 2.00 x 10-2 and 2.21 x 10-2 mol m-2 h-1, respectively. So, the peroxide solution was recognised to attack carbonates somewhat more aggressively than karst water. In order to prevent the potential corrosion of limestone and speleothems, the reaching of preliminary peroxide saturation with respect to calcite is recommended, for example, by adding of few limestone fragments into the solution at least 10 h prior to its application.

Cyanobacteria↗

Effect of lamp type and temperature on development, carbon partitioning and yield of soybean.

Soybeans grown in controlled environments are commonly taller than field-grown plants. In controlled environments, including liquid hydroponics, height of the dwarf cultivar "Hoyt" was reduced from 46 to 33 cm when plants were grown under metal halide lamps compared to high pressure sodium lamps at the same photosynthetic photon flux. Metal halide lamps reduced total biomass 14% but did not significantly reduce seed yield. Neither increasing temperature nor altering the difference between day/night temperature affected plant height. Increasing temperature from 21 to 27 degrees C increased yield 32%. High temperature significantly increased carbon partitioning to stems and increased harvest index.

Biomass↗

Contamination of the environment by the current disposal methods of mercury-containing lamps in the state of Minas Gerais, Brazil.

This survey describes the degree of environmental contamination resulting from the current disposal methods of mercury-containing lamps. The territory studied for this purpose is the federal state of Minas Gerais, one of the most populated areas in the Brazilian federation of states. The results of this survey derive in part from answers received to a questionnaire mailed out to industrial firms, commercial business enterprises, hospitals and departments of public work. The sampling technique used was a nonprobabilistic (purposive sampling). Three types of disposal were found to prevail: (1) as an addition to corporate waste disposal (garbage), (2) recycling and (3) disposal by other methods. Overall, our study shows that the majority of mercury lamps are being disposed in the metropolitan area of Belo Horizonte, primarily by the public sector, followed by industrial, commercial and hospital sectors. Although recycling constitutes a relatively high fraction of the disposal methods, we find Federal regulations in Brazil regarding the disposal of hazardous, mercury-containing lamps to be far behind the state of technological achievement. This gap has permitted the adoption of disposal measures that are obsolete, incorrect, and a primary cause for the extensive contamination of the ecosystems with harmful effects to human health.

Brazil↗

Primary LAMP-2 deficiency causes X-linked vacuolar cardiomyopathy and myopathy (Danon disease).

"Lysosomal glycogen storage disease with normal acid maltase" which was originally described by Danon et al., is characterized clinically by cardiomyopathy, myopathy and variable mental retardation. The pathological hallmark of the disease is intracytoplasmic vacuoles containing autophagic material and glycogen in skeletal and cardiac muscle cells. Sarcolemmal proteins and basal lamina are associated with the vacuolar membranes. Here we report ten unrelated patients, including one of the patients from the original case report, who have primary deficiencies of LAMP-2, a principal lysosomal membrane protein. From these results and the finding that LAMP-2-deficient mice manifest a similar vacuolar cardioskeletal myopathy, we conclude that primary LAMP-2 deficiency is the cause of Danon disease. To our knowledge this is the first example of human cardiopathy-myopathy that is caused by mutations in a lysosomal structural protein rather than an enzymatic protein.

Antigens, CD↗

Cup-to-disc ratio: agreement between slit-lamp indirect ophthalmoscopic estimation and stratus optical coherence tomography measurement.

PURPOSE: To determine agreement between slit-lamp indirect ophthalmoscopy and Stratus optical coherence tomography (OCT) when assessing cup-to-disc ratios (CDRs). METHODS: Twenty-five ocular hypertensive subjects and 56 patients with primary open-angle glaucoma were included. Estimation of vertical (VCDR) and horizontal (HCDR) cup-to-disc ratio with slit-lamp ophthalmoscopy was made by three glaucoma specialists along with OCT scanning of optic nerve head. Agreement between OCT and specialists was measured by intraclass correlation coefficients (ICC), Bland and Altman's scatterplots, and a regression coefficient of the average difference. RESULTS: The mean VCDR and HCDR was significantly higher (P<0.001) with OCT than that estimated by the specialists, with the difference ranging from 0.08 to 0.11, and from 0.13 to 0.18, respectively, depending on the specialist. Difference was higher (P<0.001) for cuppings below 0.3, and looses significance for larger VCDR cuppings (above 0.7). ICC for VCDR was 0.87 among specialists, and ranges from 0.82 to 0.75 when comparing OCT and specialists. ICC for HCDR was 0.83 among specialists and 0.74 between OCT and specialists. When data were plotted according to the Bland-Altman method, as the cupping increased, the agreement also increased. CONCLUSIONS: There is very good agreement among the specialists when estimating CDRs by stereoscopic slit-lamp biomicroscopy. OCT shows higher values than the specialists; the greatest differences occurred when assessing small CDRs and the differences diminished as the cupping increased. These two methods of measurement are not interchangeable, and the difference must be considered, especially in discs with smaller CDRs.

Early Diagnosis↗

Gas chromatographic, mass spectrometric and stable carbon isotopic investigations of organic residues of plant oils and animal fats employed as illuminants in archaeological lamps from Egypt.

Man's use of illuminants in lamps or as torches to extend the working day and range of environments accessible to him would have been a major technological advance in human civilisation. The most obvious evidence for this in the archaeological record comes from pottery and stone vessels showing sooting due to the use of a wick in conjunction with a lipid-based fuel or illuminant. A wide range of potential fuels would have been exploited depending upon availability and burning requirements. Reported herein are the results of chemical investigations of a number of lamps recovered from excavations of the site of Qasr Ibrim, Egypt. Gas chromatographic, mass spectrometric and stable carbon isotopic analyses of both free (solvent extractable) and 'bound'(released from solvent extracted pottery by base treatment) lipids have revealed a wide range of saturated fatty acids, hydroxy fatty acids and alpha, omega-dicarboxylic acids. Examination of the distributions of compounds and comparisons with the fatty acid compositions of modern plant oils have allowed a range of fats and oils to be recognised. Specific illuminants identified include Brassicaceae (Cruciferae) seed oil (most likely radish oil, Raphanus sativus), castor oil (from Ricinus communis), animal fat, with less diagnostic distributions and delta(13)C values being consistent with low stearic acid plant oils, such as linseed (Linum usitatissimum) or sesame (Sesamum indicum) oils. The identifications of the various oils and fats are supported by parallel investigations of illuminant residues produced by burning various oils in replica pottery lamps. The findings are entirely consistent with the classical writers including Strabo, Pliny and Theophrastrus.

Archaeology↗

Surface dosimetry in phototherapy: comparison of three ultraviolet B lamps used in the treatment of psoriasis.

The quantitation (dosimetry) of ultraviolet B irradiation (UVB) administered during phototherapy for psoriasis is fundamental to this modality of treatment because only absorbed irradiation can cause a photochemical reaction and, hence, a phototherapeutic effect. Significant differences may exist in surface dosimetry achieved with different UVB irradiators, and this could have important consequences for the observed clinical efficacy of phototherapy as well as practical implications for avoiding burning when transferring patients between systems. Multiple anatomically referenced polysulphone photodosimeters were applied to the skin of five subjects receiving treatment for severe psoriasis. Three subjects were irradiated with both a free-standing hot quartz lamp and a fluorescent cabinet. Two others were irradiated with a flat bank of fluorescent tubes. Topographical maps of surface irradiation were constructed. 'Cold' areas (< 50% of stated dose) included the neck, upper chest, shoulders, and anterolateral thighs for the hot quartz lamp; the medial arms, medial legs, and feet for the cabinet; and the lateral arms, forearms, lower legs, and feet for the bank of tubes. 'Hot' areas (> 100% of stated dose) included the forearms and posterior thighs for the hot quartz lamp; the lateral arms, dorsal forearm, and abdomen for the cabinet: and the central trunk (anterior and posterior) for the bank of tubes. Thus, striking differences in surface irradiation exist among the three UVB irradiators studied, and this has an impact on the results of phototherapy for psoriasis. Knowledge of these differences is of practical importance in achieving maximum therapeutic potential as well as minimizing the risk of burning reactions, particularly if patients are to be transferred between systems.

Equipment Design↗

Evaluation of a district-wide screening programme for diabetic retinopathy utilizing trained optometrists using slit-lamp and Volk lenses.

BACKGROUND: Debate exists about the optimum way to screen for diabetic retinopathy. Cameras produce a permanent record, but offer patients less choice about when and where to be screened. Optometrists offer flexibility but sensitivity and specificity of schemes have varied considerably, perhaps because of variability in screening methodology and that there is frequently no quality assurance programme. AIMS: To audit our district-wide (population 340000) screening programme for diabetic retinopathy against national targets: sensitivity > 80%, specificity > 95% and referral to review < 3 months. METHODS: Trained optometrists performed slit-lamp examination with Volk lenses (78 dioptre) with standardized reporting. Audit was by ophthalmologist with slit-lamp and Volk lenses through dilated pupils. RESULTS: We examined 872 eyes of 439 patients; 64% were normal, 29% background diabetic retinopathy, 7% sight-threatening eye disease (STED). Sixty-three percent of patients were seen within 6 months of the original screen. Of these, sensitivity for any retinopathy was 72%, specificity 77%, positive predictive value (PPV) 53%, negative predictive value (NPV) 88%. For STED, in this group, sensitivity was 87% and specificity 91%, PPV 30%, NPV 99%. Median interval referral to ophthalmological review was 11.5 weeks with 73% reviewed in under the 13-week target. Of those referred 25% received laser therapy. Eleven patients found to have referable eye disease at their initial screen were not referred to an ophthalmologist by their GP. CONCLUSIONS: We conclude that effective district-wide screening for diabetic retinopathy by optometrists using slit-lamp and Volk lenses is possible; however, only 36% of identified people with diabetes in the district were screened over a 4-year period.

Diabetic Retinopathy↗

Effect of the timing of treatment of port-wine stains with the flash-lamp-pumped pulsed-dye laser.

BACKGROUND: Port-wine stains can be treated with a flash-lamp-pumped pulsed-dye laser, but it is uncertain whether this treatment is more effective if administered early in life, when the skin is thinner and the lesion is smaller. METHODS: We prospectively studied 100 patients with a previously untreated port-wine stain of the head or neck. They were treated with the flash-lamp-pumped pulsed-dye laser and divided into four age groups (0 to 5, 6 to 11, 12 to 17, and 18 to 31 years). The outcome measure was lightening of the port-wine stain (reduction in the difference in color between the skin with the stain and contralateral healthy skin) as measured with a colorimeter after an average of five treatments (range, three to seven) of the entire lesion. RESULTS: Of the 100 patients, 11 could not be included in the analysis because they had received fewer than three or more than seven treatments, had an erroneous base-line color measurement, or were lost to follow-up. The sizes, locations, and colors of the port-wine stains were similar among the groups. When all 89 patients were analyzed together, the average reduction in the difference in color between the skin with the port-wine stain and contralateral healthy skin was 40 percent. The differences between age groups in the average reduction in color differences were not significant (P= 0.26). By the end of the study, only 7 of 89 patients had completed laser therapy, and in no case was clearance complete. Treatment was discontinued in all seven because the last three treatments did not lead to further lightening. CONCLUSIONS: We found no evidence that treatment of port-wine stains with the flash-lamp-pumped pulsed-dye laser in early childhood is more effective than treatment at a later age.

Adolescent↗

An evaluation of ultraviolet radiation personnel hazards from selected 400-watt high intensity discharge lamps.

A study of selected mercury and multi-vapor lamps, commonly used for working environment illumination, was underta ken to evaluate the scope of the ultraviolet personnel hazards one may expect under various operating conditions of the lamps. Spectral irradiance measurements were made for operating conditions with and without reflector-type luminaires, glass covers on luminaries and outer glass bulbs on the lamps.

Environmental Exposure↗