High Doses Of Ultraviolet Radiation Research Articles

DNA Polymerase ζ without the C-Terminus of Catalytic Subunit Rev3 Retains Characteristic Activity, but Alters Mutation Specificity of Ultraviolet Radiation in Yeast.

DNA polymerase ζ (pol ζ) plays a central role in replicating damaged genomic DNA. When DNA synthesis stalls at a lesion, it participates in translesion DNA synthesis (TLS), which helps replication proceed. TLS prevents cell death at the expense of new mutations. The current model indicates that pol ζ-dependent TLS events are mediated by Pol31/Pol32 pol ζ subunits, which are shared with replicative polymerase pol δ. Surprisingly, we found that the mutant rev3-ΔC in yeast, which lacks the C-terminal domain (CTD) of the catalytic subunit of pol ζ and, thus, the platform for interaction with Pol31/Pol32, retains most pol ζ functions. To understand the underlying mechanisms, we studied TLS in normal templates or templates with abasic sites in vitro in primer extension reactions with purified four-subunit pol ζ versus pol ζ with Rev3-ΔC. We also examined the specificity of ultraviolet radiation (UVR)-induced mutagenesis in the rev3-ΔC strains. We found that the absence of Rev3 CTD reduces activity levels, but does not alter the basic biochemical properties of pol ζ, and alters the mutation spectrum only at high doses of UVR, alluding to the existence of mechanisms of recruitment of pol ζ to UVR-damaged sites independent of the interaction of Pol31/Pol32 with the CTD of Rev3.

Photoprotective compounds and radioresistance in pigmented and non-pigmented yeasts.

Ultraviolet radiation, continuously reaching our planet's surface, is a type of electromagnetic energy within the wavelength range of 10 to 400 nm. Despite essential for all life on Earth, ultraviolet radiation may have severe adverse cellular effects, including DNA dimerization and production of reactive oxygen species. Radioresistant microorganisms can survive under high doses of ultraviolet radiation, enduring the direct and indirect effects on nucleic acids and other biomolecules. The synthesis and accumulation of photoprotective compounds are among the main strategies employed by radioresistant yeast species to bear the harmful effects of ultraviolet radiation. A correlation between pigments and resistance to ultraviolet radiation has been widely recognized in these microorganisms; however, there is still some debate on this topic, with non-pigmented strains sometimes being more resistant than their pigmented counterparts. In this review, we explore the role of photoprotective compounds-specifically, melanin, carotenoids, and mycosporines-and compare the differences found in resistance between pigmented and non-pigmented yeasts. We also discuss the biotechnological potential of these photoprotective compounds, with special emphasis on those produced by non-pigmented yeast strains, such as phytoene and phytofluene. The use of "-omics" approaches should further unveil the radioresistance mechanisms of non-pigmented yeasts, opening new opportunities for both research and commercial applications. KEY POINTS: • Updated knowledge on photoprotective compounds from radioresistant yeasts. • Differences on radioresistance between pigmented and non-pigmented yeasts. • Future prospects over the study of non-pigmented photoprotective compounds.

The effect of sun elevation, cloudiness, and altitude on the ultraviolet index in Czechia

Ultraviolet (UV) radiation has recently become an important topic in relation to the loss of stratospheric ozone. High doses of UV radiation have a negative effect on many organisms. This paper focuses on the UV index (UVI), which expresses the risk of UV radiation on human health. The aim of the paper is to describe the definition of UVI, and its measurement, and to summarize geographical parameters and meteorological conditions affecting the values of UVI. The effect of sun elevation, cloudiness, and altitude is demonstrated using observed data from the Hradec Králové, Košetice and Labská bouda stations during the period 2011–2017. The results show a strong effect of both sun elevation and cloudiness. The highest values of UVI (up to 8) are generally observed on sunny days around midday from May to July. The reduction of the UVI caused by clouds, fog, and rain is, on average, 85% of values typical for sunny days. The effect of altitude is distinctly weaker; a rise of UVI with increasing altitude is 0.4 per 1 km for clear sky and the surface without snow cover.

Study of skin rigidity variations due to UV radiation using digital holographic interferometry

The skin elasticity is a relevant biomechanical property that can be related to different physiological processes and diseases: hence, it is important to understand and characterize the mechanisms that affect the skin pathologies. In this work, an analysis of the rigidity variations of porcine skin caused by exposure to ultraviolet radiation is presented. For this purpose digital holographic interferometry (dhi) is used as a non-invasive optical technique to study these skin affectations. A new mathematical scheme for the skin rigidity variations was developed and the experimental data was used to study and determine the rigidity coefficients for the samples before and after ultraviolet exposure of the skin. The results after high doses of ultraviolet radiation at 8 mW/cm2 UVA, 8 mW/cm2 UVB and 10 mW/cm2 UVC for periods of 3 min showed that these rigidity coefficients increased above 100% (more than twice) their value after the ultraviolet exposure. The latter suggests a considerable reduction of the porcine skin elasticity when exposed to high doses of ultraviolet radiation, a feature that has been proved to lead to skin melanoma.

Evaluating sunscreen ultraviolet protection using a polychromatic diffuse reflectance device.

BackgroundSun protection factor (SPF) and UVA protection factor (UVA‐PF) are determined using in vivo tests, with high exposures of subjects to ultraviolet (UV) radiation. Hybrid diffuse reflectance spectroscopy (HDRS) enables estimation of both indices using only trace amounts UVB. However, the equipment requires two expensive monochromators that must synchronously scan the spectrum.MethodsAn alternate approach was developed using a polychromatic source that illuminates the skin via a custom light guide array, and the diffuse reflected light is measured with a photomultiplier. The ratio of the diffuse reflectance with and without the sunscreen on the skin determines the polychromatic diffuse reflectance UVA‐PF (PDRS UVA‐PF0). This factor was used to adjust in vitro UV spectroscopy scans of the sunscreen (with and without UV exposure to assess photostability), to calculate SPF and UVA protection factors. Ten sunscreens were evaluated and compared to in vivo SPF and UVA‐PF values.ResultsThe data show an excellent correlation with known in vivo determinations.ConclusionThis polychromatic HDRS approach uses simpler, faster, and less expensive equipment to determine both UVA‐PF and SPFs without high doses of UV radiation to the test subjects.

The Impact of Ultraviolet Radiation on Barrier Function in Human Skin: Molecular Mechanisms and Topical Therapeutics.

A competent epidermal barrier is crucial for terrestrial mammals. This barrier must keep in water and prevent entry of noxious stimuli. Most importantly, the epidermis must also be a barrier to ultraviolet radiation (UVR) from the sunlight. Currently, the effects of ultraviolet radiation on epidermal barrier function are poorly understood. However, studies in mice and more limited work in humans suggest that the epidermal barrier becomes more permeable, as measured by increased transepidermal water loss, in response UVR, at doses sufficiently high to induce erythema. The mechanisms may include disturbance in the organisation of lipids in the stratum corneum (the outermost layer of the epidermis) and reduction in tight junction function in the granular layer (the first living layer of the skin). By contrast, suberythemal doses of UVR appear to have positive effects on epidermal barrier function. Topical sunscreens have direct and indirect protective effects on the barrier through their ability to block UV and also due to their moisturising or occlusive effects, which trap water in the skin, respectively. Some topical agents such as specific botanical extracts have been shown to prevent the loss of water associated with high doses of UVR. In this review, we discuss the current literature and suggest that the biology of UVR-induced barrier dysfunction, and the use of topical products to protect the barrier, are areas worthy of further investigation.

The molecular products and biogeochemical significance of lipid photooxidation in West Antarctic surface waters

The seasonal depletion of stratospheric ozone over the Southern Hemisphere allows abnormally high doses of ultraviolet radiation (UVR) to reach surface waters of the West Antarctic Peninsula (WAP) in the austral spring, creating a natural laboratory for the study of lipid photooxidation in the shallow mixed layer of the marginal ice zone. The photooxidation of lipids under such conditions has been identified as a significant source of stress to microorganisms and short-chain fatty acids altered by photochemical processes have been found in both marine aerosols and sinking marine particle material. However, the biogeochemical impact of lipid photooxidation has not been quantitatively compared at ecosystem scale to the many other biological and abiotic processes that can transform particulate organic matter in the surface ocean. We combined results from field experiments with diverse environmental data, including high-resolution, accurate-mass HPLC-ESI-MS analysis of lipid extracts and in situ measurements of ultraviolet irradiance, to address several unresolved questions about lipid photooxidation in the marine environment. In our experiments, we used liposomes—nonliving, cell-like aggregations of lipids—to examine the photolability of various moieties of the intact polar diacylglycerol (IP-DAG) phosphatidylcholine (PC), a structural component of membranes in a broad range of microorganisms. We observed significant rates of photooxidation only when the molecule contained the polyunsaturated fatty acid (PUFA) docosahexaenoic acid (DHA). As the DHA-containing lipid was oxidized, we observed the steady ingrowth of a diversity of oxylipins and oxidized IP-DAG; our results suggest both the intact IP-DAG the degradation products were amenable to heterotrophic assimilation. To complement our experiments, we used an enhanced version of a new lipidomics discovery software package to identify the lipids in water column samples and in several diatom isolates. The galactolipid digalactosyldiacylglycerol (DGDG), the sulfolipid sulfoquinovosyldiacylglycerol (SQDG) and the phospholipids PC and phosphatidylglycerol (PG) accounted for the majority of IP-DAG in the water column particulate (≥0.2 µm) size fraction; between 3.4 and 5.3% of the IP-DAG contained fatty acids that were both highly polyunsaturated (i.e., each containing ≥5 double bonds). Using a broadband apparent quantum yield (AQY) that accounted for direct and Type I (i.e., radical-mediated) photooxidation of PUFA-containing IP-DAG, we estimated that 0.7 ± 0.2 µmol IP-DAG m−2 d−1 (0.5 ± 0.1 mg C m−2 d−1) were oxidized by photochemical processes in the mixed layer. This rate represented 4.4% (range, 3–21%) of the mean bacterial production rate measured in the same waters immediately following the retreat of the sea ice. Because our liposome experiments were not designed to account for oxidation by Type II photosensitized processes that often dominate in marine phytodetritus, our rate estimates may represent a sizeable underestimate of the true rate of lipid photooxidation in the water column. While production of such diverse oxidized lipids and oxylipins has been previously observed in terrestrial plants and mammals in response to biological stressors such as disease, we show here that a similar suite of molecules can be produced via an abiotic process in the environment and that the effect can be commensurate in magnitude with other ecosystem-scale biogeochemical processes.

The S-layer Protein DR_2577 Binds Deinoxanthin and under Desiccation Conditions Protects against UV-Radiation in Deinococcus radiodurans.

Deinococcus radiodurans has the puzzling ability to withstand over a broad range of extreme conditions including high doses of ultraviolet radiation and deep desiccation. This bacterium is surrounded by a surface layer (S-layer) built of a regular repetition of several proteins, assembled to form a paracrystalline structure. Here we report that the deletion of a main constituent of this S-layer, the gene DR_2577, causes a decrease in the UVC resistance, especially in desiccated cells. Moreover, we show that the DR_2577 protein binds the carotenoid deinoxanthin, a strong protective antioxidant specific of this bacterium. A further spectroscopical characterization of the deinoxanthin-DR_2577 complex revealed features which could suggest a protective role of DR_2577. We propose that, especially under desiccation, the S-layer shields the bacterium from incident ultraviolet light and could behave as a first lane of defense against UV radiation.

Effects of UV radiation on the growth, photosynthetic and photoprotective components, and reproduction of the Caribbean shallow-water coral Porites furcata

Shallow reef corals can frequently be subjected to high doses of ultraviolet radiation [280–400 nm (UVR)] and have developed mechanisms to cope with this. Nevertheless, slight changes in this stressor may impact their physiology and ultimately their survival. Here, we present results on the effects of artificially enhanced UVR on the growth, reproduction, production of photosynthetic pigments and photoprotective compounds of the Caribbean shallow-water branching coral Porites furcata. Corals were randomly located in one of the three different treatments: normal photosynthetically active radiation (PAR) + UVR; normal PAR+ enhanced UVR; normal PAR+ depleted UVR. Growth rates were measured using the Alizarin red staining method, photosynthetic pigments as well as mycosporine-like amino acids (MAAs) were quantified through high-performance liquid chromatography, and fecundity was estimated after histological analyses. Growth and photosynthetic pigment concentration were negatively correlated with increased UVR, compared to controls exposed to normal UVR. A significant increase in MAAs was also found in colonies under enhanced UVR. Based on their respective concentrations, the primary mycosporine-glycine (λmax = 310 nm) and shinorine (λmax = 333 nm) are the main contributors to UVR absorption in this species, while the levels of the secondary MAA palythine (λmax = 320 nm) tripled toward the end of the 128 days of the experimental period. While several physical factors may influence reef coral physiology, the results suggest that slight increases in UVR can debilitate the skeletal constitution and severely reduce the fecundity of corals living in shallow waters.

The Relation between Patterns of Vacation Sun Exposure and the Development of Acquired Melanocytic Nevi in German Children 6-7 Years of Age

Sun exposure is the main environmental risk factor for the development of melanocytic nevi. Although the general association is not disputed, the interplay between intense intermittent and the cumulative amount of sun exposure in defining the promoting effect on melanocytic nevus development is an area of debate. Trained staff members ascertained total body counts of melanocytic nevi in a cross-sectional study of 2,189 children 6-7 years of age who were recruited in two German centers in 2002. Their parents provided information about a variety of exposure factors. The distribution of melanocytic nevi was skewed markedly to the right; therefore, a negative binomial regression model provided the appropriate framework for a multivariable analysis. A steep gradient with respect to the (adjusted) number of melanocytic nevi was apparent only for the frequency of vacation episodes associated with sun exposure in areas with an intense ultraviolet radiation. In contrast, no such gradients were found for the cumulative duration of vacation sun exposure in such areas or for any variable related to vacation sun exposure in areas with a low ultraviolet radiation. This observation supports the hypothesis that intermittent exposure to high doses of ultraviolet radiation plays an especially important role in nevus development.

Impacto de un programa de foto-educación en los conocimientos y hábitos de una población escolar.

Colombians are exposed year round to high doses of ultraviolet radiation. Although no reliable epidemiological data are available for the incidence of skin cancer, it is a frequent pathology not limited to advanced age classes. The current study was done at elementary schools in Cali in zones categorized at socioeconomic levels 1 to 4 to evaluate the impact of a photo-education program for its effect on the knowledge and behavior of a group of elementary school children. Five hundred forty-seven children from kindergarten to grade three were evaluated in 4 schools categorized in socioeconomic levels 1 to 4. The students were interviewed to evaluate their knowledge and behavior with regard to effects of UV exposure. One school rated at level 1-2 and one school at level 3-4 received education on UV protection. Other two schools, rated at levels 1-2 and 3-4 each received no education on UV protection. The questionnaire was repeated in the 4 schools, and a statistical analysis was done to evaluate the impact the education program had on educated and control groups. The children that received the education program showed statistically significant improvement in behavior and knowledge. The education was more effective in low socioeconomic level populations. Follow-up studies are required to evaluate how knowledge persists over time. The need of a public health campaign to educate children in skin cancer prevention is evident.

Effect of high visible and UV irradiance on subtidal Chondrus crispus: stress, photoinhibition and protective mechanisms

Stress and the mechanisms of protection under excessive photosynthetically active (PAR) and ultraviolet radiation (UVR) were examined in the red alga Chondrus crispus Stackh. collected from the subtidal zone (6 m depth) at Helgoland, North Sea, Germany. Three PAR (400–700 nm) treatments without UVR and three UVR-A/B (290–400 nm) treatments supplemented with 30 μmol m −2 s −1 PAR were given twice. All three stages of plant stress, irritation, damage and acclimation, were observed under high light conditions. In the earlier stages of photoinhibitory PAR and UVR treatment, an induction phase occurred. It was characterized by a significant increase in chlorophyll a and carotenoids. The capacity to cope with high light stress in C. crispus increased with increasing duration of the induction phase. Prolonged exposures to high irradiance induced a substantial decline in the potential quantum yield of photosynthesis ( F v/ F m) and progressive pigment destruction that may be responsible for the damage stage of stress. In plants exposed to elevated PAR (3600 μmol m −2 s −1) and UVR (57.3 and 3.6 W m −2 of UVRA and UVRB, respectively), photoinhibition of F v/ F m exceeded 95% of control. Even after 20 h in low irradiance it was similar to values measured immediately after stress, indicating severe photodamage. The 1800 μmol m −2 s −1 PAR treatment also decreased the pigment content and F v/ F m, although damage was not permanent and the tissue gradually recovered. In contrast, naturally high doses of UVR (37.7 and 2.3 W m −2 of UVRA and UVRB, respectively) resulted in less photoinhibition of photosynthesis with rapid recovery of F v/ F m and pigment content, indicating the dynamic nature of photosynthetic acclimation to growth irradiance. Thus, photoinhibitory PAR damage and its repair predominate in subtidal C. crispus during the process of acclimation to a full-sun environment. Comparison between the acclimation conditions suggests that fluorescence quenching is related to the concentration of carotenoids. The increase in concentration, not only in the first but also after repeated exposures to PAR and UVR, was attributed to a protection of the photosynthetic apparatus. Additionally, accumulation of mycosporine-like amino acids (MAAs), mainly during the repeated exposures to UVR, led to substantially less inhibition of photosynthesis than expected given the practically double UVR dose. This suggests incomplete protection of C. crispus by MAAs during UVR shock. Thus, these experiments indicate that a few basic physiological strategies seem to be active during acclimation to high solar irradiance: a prolonged induction phase, dynamic photoinhibition, increased concentration of carotenoids, and accumulation of MAAs, UV-screening substances.

Ultraviolet-induced degradation of Ziegler-Natta and metallocene catalyzed polyethylenes

The metallocene revolution has aroused a storm of interest and associated questions regarding the performance and durability of polyolefins. This new technology has impacted the additives used to stabilize and process polymers. In this work, Ziegler-Natta and metallocene polyethylene (PE) samples were exposed to natural weather conditions under high doses of ultraviolet radiation, high temperature, and increased humidity. Weather-induced degradation of the two sets of PEs was studied using gel permeation chromatography, mechanical properties testing, differential scanning calorimetry, and Fourier transform infrared spectroscopy. The gel permeation chromatography analysis indicated the change in molecular weight distribution and molecular weights of metallocene PE to be more stable than conventional Ziegler-Natta PE. The tensile properties of metallocene PE are known to have higher values than Ziegler-Natta PE. The former exhibited a lower drop rate in mechanical properties when exposed to natural weather. Formations of nonvolatile carbonyl oxidation products, which absorb in the infrared region with a maximum absorbance level at 1742 cm−1 were determined. This indicated a higher rate of photo-oxidative and thermal degradation of Ziegler-Natta PE as compared with metallocene PE. The ultraviolet stabilization of metallocene PE may require different doses and a new kind of stabilizer system that can impart a longer useful lifetime and are cost effective for PE used for outdoor purposes. © 2000 John Wiley & Sons, Inc. J Appl Polym Sci 78: 1591–1596, 2000

P53-Dependent DNA repair and apoptosis respond differently to high- and low-dose ultraviolet radiation

p53 plays an essential part in the maintenance of the cellular genetic stability after a DNA-damaging event such as ultraviolet (UV) radiation. Following UV radiation, the amount of p53 protein is elevated. The increased p53 is believed to induce cell cycle arrest, promote nucleotide excision repair (NER) and apoptosis. To study if cells respond differently to high- and low-dose UV radiation, we examined the DNA repair efficiency and apoptosis rate of human and murine fibroblasts after UV radiation. Using a host cell reactivation assay, we found that NER was increased after low doses but not after high doses of UV radiation. In contrast, apoptosis occurred only after the cells received high doses (over 200 J/m2), but not low doses of UVB. The induction of both NER and apoptosis was observed only in p53+/+ murine fibroblasts, not in p53-/- cells, indicating that both stress response mechanisms are dependent on wild-type p53 function. UV radiation induced the expression of p53 protein in a dose-dependent manner up to 400 J/m2. In contrast, p21waf1/cip1 was induced only after low doses and bax only after high doses of UV radiation, supporting the roles of p21waf1/cip1 and bax in NER and apoptosis, respectively. Taken together, these results indicate that cellular stress response to UV radiation depends on UV dose, DNA repair after low doses and apoptosis after high doses, and that both mechanisms are dependent on wild-type p53 function.

Comet Assay Analysis of Repair of DNA Strand Breaks in Normal and Deficient Human Cells Exposed to Radiations and Chemicals. Evidence for a Repair Pathway Specificity of DNA Ligation

The induction and resealing of DNA strand breaks in a cell line with a proven defect in DNA ligase I, 46BR, and in two Bloom's syndrome cell lines, YBL6 and GM 1492, were compared to those observed in normal human 1BR/3 fibroblasts after treatment with a variety of genotoxic agents whose lesions are processed by different repair pathways. This analysis was performed using the single-cell gel electrophoresis assay. The three types of cells were found to have similar capabilities to recognize and incise ultraviolet photoproducts and also demonstrated similar amounts of DNA breaks immediately after gamma irradiation. During post-treatment incubation, 46BR cells showed a marked DNA re-ligation defect after ultraviolet radiation damage, GM 1492 cells demonstrated a highly reduced DNA joining ability after relatively high doses of ultraviolet radiation, and YBL6 cells were particularly affected in DNA re-ligation after damage by 4-nitroquinoline-1-oxide. The two Bloom's syndrome cell lines and 46BR cells had a nearly normal ability to reseal breaks resulting from gamma irradiation or treatment with xanthine plus xanthine oxidase. These findings suggest that different DNA ligases may be involved in different DNA repair pathways in human cells.

The effects of ultraviolet irradiation on wound contraction in the hairless guinea pig

Ultraviolet radiation has been shown to alter wound tensile strength and evoke a number of intracellular changes in fibroblasts. We examined the effects of relatively high doses of ultraviolet radiation on subsequent wound contraction of circular wounds in the hairless guinea pig model. Female hairless guinea pigs were divided into two experimental groups receiving 80 J/cm 2 or 480 J/cm 2 every other day for 16 weeks. Age-matched unirradiated animals were used as controls. After exposure, all animals had either a 4 mm punch biopsy (80 J/cm 2) or a 2.4 cm diameter ((480 J/cm 2) Groups 3 and 4) circular area excised from the dorsum. The extent of wound enlargement immediately following wounding of the irradiated animals was decreased as compared to the controls. The rate of wound contraction was significantly lower during early stages of wound contraction in each group of irradiated animals, and wound contraction was significantly slower overall in both groups of irradiated animals compared to controls.

Isolation and characterization ofSerratia marcescens mutants defective in prodigiosin biosynthesis

Mutants deficient in the biosynthesis of prodigiosin have been obtained by treatingSerratia marcescens with high doses of ultraviolet radiation. Mutants were selected on the basis of the color characteristics of their colonies when grown on peptone glycerol medium. New types of mutants, with unusual blocks in the biosynthetic pathway of prodigiosin, were obtained. All the mutants were classified under a new scheme on the basis of the syntrophic pigmentation characteristic and infrared spectroscopic analysis of their pigment. By these criteria mutants could be distinguished into eight distinct classes. Classes I to III include mutants of the three classes (M1, B3, and B1) reported previously [Morrison, DA (1966) J Bacteriol 91:1599–1604] and several new ones. Mutants blocked in the methylamylpyrrole (MAP) arm of the bifurcated pathway were assigned to class I. A class II mutant was distinguished by its inability to synthesize methoxybipyrrolecarboxyaldehyde (MBC), but was able to produce norprodigiosin. Class III mutants were deficient in the synthesis of hydroxybipyrrolecarboxaldehyde (HBC). Double mutants were obtained with defects in the expression of both MBC and MAP and were assigned to class IV. Mutants of class V were unable to synthesize HBC and MAP, but could form MBC when furnished with exogenous HBC. Class VI and VII mutants were defective in the synthesis of all three precursors, but differed in their ability to perform the coupling step. Finally, a mutant of class VIII was found to produce the three intermediates, but was deficient in prodigiosin or norprodigiosin biosynthesis, indicative of a defect in the enzymatic condensation of MAP with the bipyrroles MBC and HBC. The anomalous pattern of syntrophism among certain interclass mutants suggests that the physiology of pigment formation inS. marcescens is quite complex.

DNA replication and UV-induced DNA repair synthesis in human fibroblasts are much less sensitive than DNA polymerase alpha to inhibition by butylphenyl-deoxyguanosine triphosphate.

In mammalian cells, both semiconservative DNA replication and the DNA repair patch synthesis induced by high doses of ultraviolet radiation are known to be inhibited by aphidicolin, indicating the involvement in these processes of one or both of the aphidicolin-sensitive DNA polymerases, alpha and/or delta. In this paper, N2-(p-n-butylphenyl)-2'-deoxyguanosine-5'-triphosphate, a strong inhibitor of polymerase alpha and a weak inhibitor of polymerase delta, is used to further characterize the DNA polymerase(s) involved in these two forms of nuclear DNA synthesis. In permeable human fibroblasts, DNA replication and ultraviolet-induced DNA repair synthesis are more resistant to the inhibitor than DNA polymerase alpha by factors of approximately 500 and 3000, respectively. These findings are most consistent with the involvement of DNA polymerase delta in these processes.

YEAST MUTANTS EXCRETING VITAMIN B1AND THEIR USE IN THE PRODUCTION OF THIAMINE RICH BEERS*,†

In certain strains of Saccharomyces cerevisiae and Saccharomyces uvarum (carlsbergensis), high doses of ultraviolet radiation induced stable mutants excreting thiamine from their living cells during the ethanol production. In S. cerevisiae, the first mutagenesis step yielded mutants with the production of 0·3–0·5 mg extracellular thiamine. HCL/litre minimum medium without thiamine and the second one increased the production up to 2·0–2·1 mg thiamine. HCL/litre. In S. uvarum (carlsbergensis), mutants producing up to 0·76 mg thiamine. HCL/litre 10% Plato hopped wort were obtained in the first mutagenesis step combined with mitotic recombinations. The increase up to 0·92 mg/litre was achieved here by repeated selections. Both laboratory and pilot plant fermentations in 10 and 4% Plato hopped worts showed the suitability of selected mutants for the production of thiamine rich beers which fulfilled all quality requirements and contained 0·67–0·80 and 0·22–0·33 mg thiamine. HCI/litre, respectively.

Comparison of the reversibility of locipet23 andlys2 after UV irradiation in the standard and UV-sensitive strains ofSaccharomyces cerevisiae

Reversibility of the respiration-deficient locuspet23 and auxotrophic locuslys2 was followed in the standard (RAD1) and UV sensitive (rad1–2) strains ofSaccharomyces cerevisiae, both after identical doses of UV radiation and at identical survival. When comparing the reversibility after the treatment with identical doses of UV radiation a much higher reversibility of both loci in strainrad1–2 could be detected. When comparing the reversibility of the loci in question at identical survival of both strains it could be found that the reversibility of thepet23 locus is again much higher in strainrad1–2, whereas the reversibility of thelys2 locus is roughly identical in the two strains. Thus, the function of geneRAD1 in repair processes is apparently associated with the “error-free” repair, both at low and high doses of ultraviolet radiation.