A note on the inactivation of influenza A viruses by solar radiation, relative humidity and temperature

We critically investigate the claim put forward by Sagripanti and Lytle ([2007] Photochem. Photobiol. 83, 1278-1282) that inactivation of influenza A virus by solar radiation can explain the seasonality of influenza epidemics. We correct an error in the Sagripanti and Lytle paper and show that changes in relative humidity and temperature affect influenza virus inactivation as strongly as variation in solar radiation. Furthermore, it appears unlikely that transmission in outdoor settings plays an important role during influenza outbreaks, because influenza A virus is sensitive to a wide range of environmental factors.     

Inactivation of a Rhizobium bacteriophage by ultraviolet radiation of different wave-lengths

Abstract— Trypsin inactivated by u.v. radiation, gamma radiation, visible radiation in the presence of sensitizing dyes, and autolysis, was examined by the method of disc electrophoresis. Untreated Worthington twice crystallized salt-free trypsin separated into four bands which moved toward the cathode; the main band, which had the greatest mobility, contained all of the detectable tryptic activity. The next most mobile band has been assumed to be a chymotrypsin contaminant. The other two bands are of unknown nature. A progressive loss of all the bands was observed when the enzyme was inactivated by those procedures which produce a ‘damaged’ class of trypsin molecules, i.e. flavin-sensitized photooxidation, autolysis, and treatment with u.v. and gamma radiation. No loss of the main band was observed during photoinactivation with methylene blue and eosin Y as sensitizers. In this latter case, it is postulated that the trypsin inactivation products must be of such a nature that the net charge and conformation of the protein is not greatly changed, thus permitting all of the protein to remain in the same band during electrophoresis.     

Testing of the influenza virus purification by CIEF

In virological practice, the pre‐concentration, purification and subsequent determination of the purity and concentration of the viruses from the cultural medium and/or from the real sample are required. The conventional techniques used today are equipment demanding, time‐consuming and laborious. In this study, the CIEF of influenza viruses with UV detection has been developed and subsequently used to test the purification of the virus from the biological samples. The equine and swine influenza viruses present in infected allantoic fluid of specific pathogen free embryonated chicken eggs were precipitated by using PEG 6000 and sodium chloride. The precipitated viruses were centrifuged at 14 000×g, and the impurities of different densities were removed by using the sucrose gradients. The efficiency of the virus purification technique was examined by the CIEF and compared to the results of real‐time PCR. The pIs of both influenza viruses were determined. Simultaneously, the CIEF was found to be a suitable method for the rapid testing of the efficiency of the virus purification.     

Inhibition of influenza virus sialidase and anti-influenza virus activity by plant flavonoids

Flavonoids (103 species) were tested for inhibitory activity against influenza virus sialidase using sodium p-nitrophenyl-N-acetyl-alpha-D-neuraminate as substrate. 5,7,4'-Trihydroxy-8-methoxyflavone from the root of Scutellaria baicalensis showed the most potent activity (IC50, 55 microM), and this flavone appeared to be a non-competitive inhibitor of the enzyme. Whereas, negligible or weak inhibitory activities were observed for mouse liver sialidase, beta-galactosidase and alpha-mannosidase as tested. This flavone also inhibited the infection by influenza virus A/PR/8/34 of Madin-Darby canine kidney cells, and replication of the virus in the allantoic sack of embryonated egg. These results suggest that flavone, which has potent influenza virus sialidase inhibitory activity, may have anti-influenza virus activity.     

Inactivation of dengue virus by methylene blue/narrow bandwidth light system

Peracetic acid was one of the most commonly used disinfectants on solid surfaces in hospitals or public places. However, peracetic acid is an environmental toxin. Therefore, safer, alternative disinfectants or disinfectant systems should be developed. Because photodynamic virus inactivation with methylene blue (MB)/light system has proven effective in blood banking, MB was selected as a photosensitizing agent, dengue virus as a model virus for enveloped RNA viruses, and an in-house fabricated narrow bandwidth light system overlapping the absorption spectrum of MB as the light source. Dengue virus was mixed with different concentrations of MB, and illuminated by the narrow bandwidth light system under different illumination distances and times. The amount of dengue virus remaining was evaluated by plaque forming assays. Results showed that the concentration of MB working solution, illumination intensity of light source, illumination distance and time were four key factors affecting efficiency of virus inactivation using the MB/narrow bandwidth light system. Dengue virus could be completely inactivated at 2.5 m in 5 min when MB >/= 1.0 microg/ml. However, when the distance reached 3.0 m, only greater concentrations of MB (2.0 microg/ml) could completely inactivate virus in a reasonably short time (20 min), and smaller concentrations of MB (1.0 microg/ml) could only completely inactivate virus using longer times (25 min). The results of this virus inactivation model indicate that our MB/narrow bandwidth light system provides a powerful, easy way to inactivate dengue viruses.     

Exposure to ultraviolet radiation enhances mortality and pathology associated with influenza virus infection in mice

Ultraviolet radiation (UVR) causes systemic immune suppression, decreasing the delayed type and contact hypersensitivity responses in animals and humans and enhancing certain mycobacterial, parasitic and viral infections in mice. This study tests the hypothesis that prior exposure to UVR enhances influenza infections in mice. BALB/c female mice were exposed to 0-8.2 kJ/m2 of UVR. Exposed and unexposed mice were infected intranasally three days later with 150-300 plaque-forming units/mouse (lethal dose (LD)20-LD40) of mouse-adapted Hong Kong Influenza A/68 (H3N2) virus or sham infected with 50 microL Hanks' balanced salt solution/mouse. Mortality from viral infection ranged from 25-50%. UVR exposure increased virus-associated mortality in a dose-dependent manner (up to a two-fold increase at 8.2 kJ/m2). The increased mortality was not associated with bacterial pneumonia. The highest dose of UVR also accelerated the body weight loss and increased the severity and incidence of thymic atrophy associated with influenza infection. However, UVR treatment had little effect on the increase in lung wet weight seen with viral infection, and, to our surprise, did not cause an increase in virus titers in the lung or dissemination of virus. The mice died 5-6 days after infection, too early for adaptive immune responses to have much impact. Also, UVR did not interfere with the development of protective immunity to influenza, as measured by reinfection with a lethal challenge of virus. Also, cells adoptively transferred from UVR or untreated mice were equally protective of recipient mice challenged with a lethal dose of virus. The mice resemble mice succumbing to endotoxin, and influenza infection increased the levels of tumor necrosis factor alpha (TNF-alpha) in bronchoalveolar lavage fluid and serum cortisol levels; however, UVR preexposure did not increase either of these responses to the virus. The results show that UVR increased the morbidity, mortality and pathogenesis of influenza virus in mice without affecting protective immunity to the virus, as measured by resistance to reinfection. The mechanism of enhanced mortality is uncertain, but the data raises concerns that UVR may exacerbate early responses that contribute to the pathogenesis of a primary viral infection.     

AC electrokinetic immobilization of influenza virus

The use of alternating current (AC) electrokinetic forces, like dielectrophoresis and AC electroosmosis, as a simple and fast method to immobilize sub-micrometer objects onto nanoelectrode arrays is presented. Due to its medical relevance, the influenza virus is chosen as a model organism. One of the outstanding features is that the immobilization of viral material to the electrodes can be achieved permanently, allowing subsequent handling independently from the electrical setup. Thus, by using merely electric fields, we demonstrate that the need of prior chemical surface modification could become obsolete. The accumulation of viral material over time is observed by fluorescence microscopy. The influences of side effects like electrothermal fluid flow, causing a fluid motion above the electrodes and causing an intensity gradient within the electrode array, are discussed. Due to the improved resolution by combining fluorescence microscopy with deconvolution, it is shown that the viral material is mainly drawn to the electrode edge and to a lesser extent to the electrode surface. Finally, areas of application for this functionalization technique are presented.     

A study of solar erythema radiation doses

Abstract —Using semi-empirical analytic formulas for the transmitted and scattered ultraviolet spectral irradiance at the ground (Green, A. E. S., T. Sawada and E. P. Shettle, Photochem. Photobiol. 19 , 251–259, 1974), we calculate erythema dose rates and daily erythema doses. Results are illustrated graphically, and for the purpose of photobiological applications are given in terms of approximate analytic forms, with parameters presented in tabular form. The relative erythema data assembled by W. W. Coblentz and R. Stair (U.S. Bureau of Standards J. Res. 12 , 13–14, 1934), as fit by an analytic form, is taken as a standard spectrum in our calculations. Other forms of erythema spectra are also compared.     

Fingerprinting a killer: surveillance of the influenza virus by mass spectrometry

Influenza is a deadly virus that continues to kill and inflict illness and suffering the world over. Despite a global surveillance strategy, an annual response to vaccine preparation and the development of new anti-viral drugs to treat the virus ahead of, or after, infection, no cure exists. Future pandemics are a very real threat and countries have mobilised efforts to stockpile treatments and prepare for outbreaks. A new surveillance approach in which the structure and antigenicity of the virus can be rapidly screened by mass spectrometry is expected to have a greater role in the characterisation of emerging influenza strains, even at the site of an outbreak.     

Oxidation of guanine in cellular DNA by solar UV radiation: biological role.

The formation of cyclobutane pyrimidine dimers (CPD) and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) was investigated in Chinese hamster ovary cells upon exposure to either UVC, UVB, UVA or simulated sunlight (SSL). Two cell lines were used, namely AT3-2 and UVL9, the latter being deficient in nucleotide excision repair and consequently UV sensitive. For all types of radiation, including UVA, CPD were found to be the predominant lesions quantitatively. At the biologically relevant doses used, UVC, UVB and SSL irradiation yielded 8-oxodGuo at a rather low level, whereas UVA radiation produced relatively higher amounts. The formation of CPD was 10(2) and 10(5) more effective upon UVC than UVB and UVA exposure. These yields of formation followed DNA absorption, even in the UVA range. The calculated relative spectral effectiveness in the production of the two lesions showed that efficient induction of 8-oxodGuo upon UVA irradiation was shifted toward longer wavelengths, in comparison with those for CPD formation, in agreement with a photosensitization mechanism. In addition, after exposure to SSL, about 19% and 20% of 8-oxodGuo were produced between 290-320 nm and 320-340 nm, respectively, whereas CPD were essentially (90%) induced in the UVB region. However, the ratio of CPD to 8-oxodGuo greatly differed from one source of light to the other: it was over 100 for UVB but only a few units for UVA source. The extent of 8-oxodGuo and CPD was also compared to the lethality for the different types of radiation. The involvement of 8-oxodGuo in cell killing by solar UV radiation was clearly ruled out. In addition, our previously reported mutation spectra demonstrated that the contribution of 8-oxodGuo in the overall solar UV mutagenic process is very minor.     

Induction of mycosporine-like amino acids (MAAs) in cyanobacteria by solar ultraviolet-B radiation

Three filamentous and heterocystous N₂-fixing cyanobacteria, Anabaena sp., Nostoc commune and Scytonema sp. were tested for the presence of ultraviolet-absorbing mycosporine-like amino acids (MAAs) and their induction by solar ultraviolet-B (UV-B) radiation. High performance liquid chromatographic (HPLC) studies revealed the presence of only one type of MAAs in all three cyanobacteria, that was identified as shinorine, a bisubstituted MAA containing both glycine and serine groups having an absorption maximum at 334 nm and a retention time of around 2.8 min. There was a circadian induction in the synthesis of MAAs when the cultures were exposed to mid-latitude solar radiation (Playa Unión, Rawson, Chubut, Patagonia, Argentina) for 3 days, 4–6th February, 2000. Solar radiation was measured by an ELDONET (European Light Dosimeter Network) filter radiometer permanently installed on the roof of the Estación de Fotobiología Playa Unión (43°18′ S; 65°03′ W). The maximum irradiances were around 450–500, 45–50 and 1.0–1.2 W m⁻² for PAR (photosynthetic active radiation), UV-A (ultraviolet-A) and UV-B (ultraviolet-B), respectively. PAR and UV-A had no significant impact on MAA induction while UV-B induced the synthesis of shinorine in all three cyanobacteria. Shinorine was found to be induced mostly during the light period. During the dark period the concentration stayed almost constant. In addition to shinorine, another unidentified, water-soluble, brownish compound with an absorption maximum at 315 nm was found to be induced by UV-B only in Scytonema sp. and released into the medium. This substance was neither found in Anabaena sp. nor in Nostoc commune. Judging from the results, the studied cyanobacteria may protect themselves from deleterious short wavelength radiation by their ability to synthesize photoprotective compounds in response to UV-B radiation.     

Peculiarities of coagulation kinetics of influenza virus dispersions

The coagulation kinetics of the dispersions of the A/Mississippi/1/85 influenza virus strain in NaCl solutions was studied at various pH values via the flow ultramicroscopy technique. Results obtained for different preparation procedures of virus dispersion are compared and conditions resulted to “superfast” (faster than according to Smoluchowski’s theory) coagulation are determined. Common generalities inherent to the process of superfast coagulation of the A/Mississippi/1/85 influenza virus strain and strains previously studied are found.     

Plant responses to current solar ultraviolet-B radiation and to supplemented solar ultraviolet-B radiation simulating ozone depletion: an experimental comparison

Field experiments assessing UV-B effects on plants have been conducted using two contrasting techniques: supplementation of solar UV-B with radiation from fluorescent UV lamps and the exclusion of solar UV-B with filters. We compared these two approaches by growing lettuce and oat simultaneously under three conditions: UV-B exclusion, near-ambient UV-B (control) and UV-B supplementation (simulating a 30% ozone depletion). This permitted computation of "solar UV-B" and "supplemental UV-B" effects. Microclimate and photosynthetically active radiation were the same under the two treatments and the control. Excluding UV-B changed total UV-B radiation more than did supplementing UV-B, but the UV-B supplementation contained more "biologically effective" shortwave radiation. For oat, solar UV-B had a greater effect than supplemental UV-B on main shoot leaf area and main shoot mass, but supplemental UV-B had a greater effect on leaf and tiller number and UV-B-absorbing compounds. For lettuce, growth and stomatal density generally responded similarly to both solar UV-B and supplemented UV-B radiation, but UV-absorbing compounds responded more to supplemental UV-B, as in oat. Because of the marked spectral differences between the techniques, experiments using UV-B exclusion are most suited to assessing effects of present-day UV-B radiation, whereas UV-B supplementation experiments are most appropriate for addressing the ozone depletion issue.     

Diffuse component of solar ultraviolet radiation in tree shade

The first set of quantitative data of diffuse erythemal UV and UV-A radiation in tree shade at a sub-tropical Southern Hemisphere latitude is presented. Over the summer, approximately 60% of the erythemal UV radiation in tree shade is due to the diffuse component. Similarly, approximately 56% of the UV-A radiation in tree shade is due to the diffuse component. In tree shade these diffuse UV percentages are relatively constant from the morning to noon to afternoon periods. In comparison, in full sun, there is a decrease in the percentage of diffuse UV from morning to noon to afternoon. The exposures to diffuse UV on a horizontal plane in tree shade between 9:00 EST and 15:00 EST are of the order of 4 MED (minimum erythemal dose) and 14 J cm(-2) for erythemal UV and UV-A, respectively. The high diffuse UV component in the shade may result in high UV exposures not only to unprotected parts of the body on a horizontal plane, but also in equally high UV irradiances to parts of the body, including the eyes and face, that are not UV protected.     

A solar erythemal (sunburn) radiation dosimeter

Measurement of solar erythemal radiation is a technically demanding task because it is accompanied by a vastly greater flux of other solar radiation. An inexpensive solar erythemal radiation dosimeter has been designed which is based on the photocleavage of an alkyl disulphide. The reaction is carried out in a hydrocarbon solvent which can readily donate hydrogen atoms and as a result no polysulphides with absorptions in the erythemal action spectrum were formed. This avoided non-linearities in the dosimeter arising from inner filtering effects. The amount of alkylthiol produced as a result of exposure of the corresponding dialkyl disulphide solution to solar erythemal radiation was linearly related to the dose of radiation received.     

Simplified calibration for broadband solar ultraviolet radiation measurements

Aspects of different calibration procedures for erythemally weighing broadband radiometers are presented in this study. These instruments are common in projects dealing with ultraviolet radiation effects on humans. Many erythemally weighing broadband radiometers are still operated using a single calibration factor (cf) that is provided with the instrument. The individual characteristics of every instrument are strongly dependent on the total ozone amount and the solar elevation. Therefore, a calibration procedure also has to take into account the ozone concentrations and the solar elevation to compensate for the effects of the individual characteristics and to provide comparable measurements. Given the variation of the ozone concentrations and the solar elevation, an individual cf has to be calculated for every measurement. Using a simplified version of the calibration procedure, which is presented in this study, can lessen this effort. Taking into account the relevant meteorological conditions for a measuring site, a single cf is calculated to compensate the individual characteristics of the instruments and therefore deliver comparable measurements with less effort.     

ELDONET--a decade of monitoring solar radiation on five continents

The European light dosimeter network (ELDONET) comprises more than 40 stations in 24 countries on 5 continents. The present report compares solar radiation data in the photosynthetic active radiation, UV-A (315-400 nm) and UV-B (280-315 nm) wavelength ranges for 17 stations at different latitudes on the northern and southern hemispheres for up to 10 years of monitoring. While the maximal irradiances on clear days follow a latitudinal gradient due to the cosine dependence on the solar angle, the total doses strongly depend on the local climate and atmospheric conditions as well as the day-length distribution over the year. UV-B irradiances and doses are strongly influenced by the total column ozone, which is recorded for all covered stations.     

Effects of solar radiation on collagen and chitosan films

Photo-aging and photo-degradation are the deleterious effect of chronic exposure to sun light of many materials made of natural polymers. The resistance of the products on the action of solar radiation is very important for material scientists. The effect of solar radiation on two natural polymers: collagen and chitosan as well as collagen/chitosan blends in the form of thin films has been studied by UV-Vis and FTIR spectroscopy. It was found that UV-Vis spectra, which characterise collagen and collagen/chitosan films, were significantly altered by solar radiation. FTIR spectra of collagen and collagen/chitosan films showed that after solar irradiation the positions of amide A and amide I bands were shifted to lower wavenumbers. There was not any significant alteration of chitosan UV-Vis and FTIR spectra after solar radiation. In the condition of the experiment chitosan films were resistant to the action of solar radiation. The effect of solar UV radiation in comparison to artificial UV radiation has been discussed.     

An estimation of biological hazards due to solar radiation

A spectrum evaluator based on four different dosimeter materials has been employed to estimate the spectral irradiances of solar radiation for exposed humans. The result is used to calculate the biologically effective irradiance using the erythemal action spectrum and a fish melanoma action spectrum. Measurements are made in winter at a sub-tropical site on the chest and shoulder of subjects during normal daily activities. Up to 95% of the total UV exposure received is in the UV-A waveband (320-400 nm). The UV-A waveband is found to contribute approximately 14% of the erythemal UV and 93% of the biologically effective UV for fish melanoma. Extrapolation to humans suggests that exposure to the UV-A band will contribute to photodamage in human skin during exposure to solar radiation.