Understanding the natural processes that have taphonomic effects on bone is an important part of accurately determining the postmortem interval (PMI). Ultraviolet radiation is one of these various natural processes that weather bone. The present study quantifies the degree to which differential exposure to sunlight affects bone bleaching and weathering. In this study, 140 Sus scrofa long bones were placed in two different microenvironments (grassland and woodland) within a New England setting. Upon the completion of a one-year observation period, 100% of the bones had reached bleaching level 4, and 9.85% (n = 13) of the bones had reached weathering stage 1, with the majority (n = 11) being from the woodland sample. The results indicated that the microhabitat that each sub-sample was deposited within played a statistically significant role in the degree and rate of bleaching and weathering on the bone, with chi-square tests all indicating a p value of < 0.001. In addition, results indicated that environmental variables that led to the more rapid decomposition of soft tissue, such as temperature, humidity, and the type of plant coverage, may play a greater role in the level of bleaching and weathering achieved by the bones, rather than simply the degree of exposure to UV radiation and light intensity.

No description available.

Much research is being conducted into the causes of human cancer. A number of human autosomal recessive diseases such as Xeroderma Pigmentosum are characterized at least in part by a defect or aberration in one or more forms of DNA repair and at the same time an elevated incidence of cancer. Also, carcinogens cause mutations in DNA and the greater the carcinogenicity, the greater the mutagenicity. As a result, much attention has been focused on DNA repair and its relationship to cancer incidence. The HCR of V antigen formation by UV-irradiated Adenovirus type 2 (Ad 2) was examined using apparently normal human fibroblasts, tumor cells (HeLa CCL2), and cells transformed by Ad 5 DNA (293, 293 N3S). A decrease in the HCR of V antigen formation was found for HeLa CCL2 cells as compared to apparently normal human fibroblasts, but not for the transformed cells. These results are discussed in terms of the characteristics of the cell types. Herpes simplex virus type I encodes a polymerase and thymidine kinase (tk) activity which are involved in viral DNA synthesis. Paa ᷇ 5, an HSV-1 mutant containing one or more mutations in the polymerase gene is an antimutator. If these are also involved in viral DNA repair, then the HSV-1 polymerase, tk activity, and mutant polymerases conferring altered mutation rates should provide excellent tools with which to probe cellular DNA repair processes and mutagenesis. The study of the HCR of plaque forming ability of HSV-1 KOS wild type (WT), Paa ᷇ 5 and PTK3B (lacking thymidine kinase activity ) using VERO cells revealed a decrease in the HCR of Paa ᷇ 5 and increase of surviving fractions of PTK3B with respect to that of HSV-1 KOS WT. Similar studies using apparently normal human fibroblasts, Xeroderma Pigmentosum and Cockayne Syndrome cells also implicated the HSV-1 polymerase in viral DNA repair. The results are discussed in terms of the function of the HSV-1 polymerase and the DNA repair abilities of XP and CS cells. / Thesis / Master of Science (MSc)

No / Green tea catechins (GTC) reduce UV radiation (UVR)-induced inflammation in experimental models, but human studies are scarce and their cutaneous bioavailability and mechanism of photoprotection are unknown. We aimed to examine oral GTC cutaneous uptake, ability to protect human skin against erythema induced by a UVR dose range and impact on potent cyclo-oxygenase- and lipoxygenase-produced mediators of UVR inflammation, PGE2 and 12-hydroxyeicosatetraenoic acid (12-HETE), respectively. In an open oral intervention study, sixteen healthy human subjects (phototype I/II) were given low-dose GTC (540 mg) with vitamin C (50 mg) daily for 12 weeks. Pre- and post-supplementation, the buttock skin was exposed to UVR and the resultant erythema quantified. Skin blister fluid and biopsies were taken from the unexposed and the UVR-exposed skin 24 h after a pro-inflammatory UVR challenge (three minimal erythema doses). Urine, skin tissue and fluid were analysed for catechin content and skin fluid for PGE2 and 12-HETE by liquid chromatography coupled to tandem MS. A total of fourteen completing subjects were supplement compliant (twelve female, median 42·5 years, range 29–59 years). Benzoic acid levels were increased in skin fluid post-supplementation (P= 0·03), and methylated gallic acid and several intact catechins and hydroxyphenyl-valerolactones were detected in the skin tissue and fluid. AUC analysis for UVR erythema revealed reduced response post-GTC (P= 0·037). Pre-supplementation, PGE2 and 12-HETE were UVR induced (P= 0·003, 0·0001). After GTC, UVR-induced 12-HETE reduced from mean 64 (sd 42) to 41 (sd 32) pg/μl (P= 0·01), while PGE2 was unaltered. Thus, GTC intake results in the incorporation of catechin metabolites into human skin associated with abrogated UVR-induced 12-HETE; this may contribute to protection against sunburn inflammation and potentially longer-term UVR-mediated damage.

This study presents the first attempt to combine the fields of ultraviolet (UV) photobiology, plant cell wall biochemistry, aerobic methane production and reactive oxygen species (ROS) mechanisms to investigate the effect of UV radiation on vegetation foliage. Following reports of a 17% increase in decomposition rates in oak (Quercus robur) due to increased UV, which were later ascribed to changes in cell wall carbohydrate extractability, this study investigated the effects of decreased UV levels on ash (Fraxinus excelsior), a fast-growing deciduous tree species. A field experiment was set up in Surrey, UK, with ash seedlings growing under polytunnels made of plastics chosen for the selective transmission of either all UV wavelengths, UV-A only, or no UV. In a subsequent field decomposition experiment on end-of-season leaves, a significant increase of 10% in decomposition rate was found after one year due to removal of UV-B. However, no significant changes in cell wall composition were found, and a sequential extraction of carbohydrate with different extractants suggested no effects of the UV treatments on cell wall structure. Meanwhile, the first observations of aerobic production of methane from vegetation were reported. Pectin, a key cell wall polysaccharide, was identified as a putative source of methane, but no mechanism was suggested for this production. This study therefore tested the effect of UV irradiation on methane emissions from pectin. A linear response of methane emissions against UV irradiation was found. UV-irradiation of de-esterified pectin produced no methane, demonstrating esters (probably methyl esters) to be the source of the observed methane. Addition of ROS-scavengers significantly decreased emissions from pectin, while addition of ROS without UV produced large quantities of methane. Therefore, this study proposes that UV light is generating ROS which are then attacking methyl esters to create methane. The study also demonstrates that this mechanism has the potential to generate several types of methyl halides. These findings may have implications for the global methane budget. In an attempt to demonstrate ROS generation in vivo by UV irradiation, radio-labelling techniques were developed to detect the presence of oxo groups, a product of carbohydrate attack by ROS. Using NaB3H4, the polysaccharides of ash leaflets from the field experiment were radio-labelled, but did not show any significant decrease in oxo groups due to UV treatments. However, UV-irradiation of lettuce leaves showed a significant increase in radio-labelling, suggesting increased UV irradiation caused an increase in the production of ROS. The study shows that the use of this radio-labelling technique has the potential to detect changes in ROS production due to changes in UV levels and could be used to demonstrate a link between ROS levels and methane emissions.

No description available.

O aumento do conhecimento em relação aos danos provocados pela radiação solar, tanto na faixa do UVB quanto na faixa do UVA, o avanço nas tecnologias relacionadas ao produto e às formas de avaliação bem como a disponibilização de novas moléculas no mercado levaram ao aumento da qualidade da proteção proporcionada pelos fotoprotetores. Entretanto, ainda há preocupações em relação à segurança de alguns filtros solares devido a sua fotoinstabilidade e penetração cutânea. Dessa forma, torna-se necessário o desenvolvimento de novos filtros solares mais efetivos e seguros, que apresentem relevância científica e potencial de inovação. Assim, a presente pesquisa teve como objetivo desenvolver novos análogos ao filtro solar benzofenona-3 e determinar o seu potencial fotoprotetor por meio da avaliação da absorção UV e avaliação da fotoestabilidade, fototoxicidade e atividade antioxidante. Para tal, inicialmente seis análogos ao filtro solar químico benzofenona-3 foram sintetizados visando aumentar o tamanho da molécula e aumentar a sua absorção no UVA longo (340-400nm). Foram avaliados os espectros de absorção no UV e a fotodegradação dessas substâncias. A fototoxicidade das substâncias selecionadas foi avaliada por meio do uso de cultura de fibroblastos 3T3, para a determinação da viabilidade celular na presença e ausência da radiação. A atividade antioxidante das substâncias foi avaliada por meio da quimioluminescência gerada pela reação HRP-H2O2-luminol. Os resultados permitiram demonstrar a importância da relação entre a estrutura molecular dos compostos e sua absorção no UV. As moléculas benzofenona-3 fenil amino (B5) e o carbazol da benzofenona-3 fenil amino (B6) apresentaram maior absorção no UVA, longo e curto, e no UVB, quando comparadas com a molécula de partida (benzofenona-3). Apenas as substâncias que apresentavam ponte de hidrogênio intramolecular (B5 e B6) foram consideradas fotoestáveis. Somente a molécula B5 não apresentou potencial para fototoxicidade aguda. Além disso, essa molécula apresentou atividade antioxidante, o que sugere o seu grande potencial para utilização com filtro solar. / The increase of our knowledge of not only UVB but also UVA-induced damages, the advances on product and testing technologies as well as new sunscreen molecules leaded to and enhancement of quality of UV protection provided by topical sunscreens. However there are some safety concerns involving some UV-filters due to their photoinstability and skin penetration. Therefore it is necessary to develop new safer and more effective UV-filters, which also presents scientific relevance and innovation potential. Thus, the aim of the present research was to develop new analogues based on benzophenone-3, and to evaluate their photoprotective potential through their photostability, phototoxicity and antioxidant activity. For this purpose, firstly six new synthetic analogues based on benzophenone-3 were prepared in order to promote molecular weight enhancement as well as improve long-wave UVA absorption (340-400 nm). The UV absorption spectra and photodegradation of these compounds were also analyzed. Phototoxicity of selected compounds was evaluated by using 3T3 monolayer fibroblast culture to determine cell viability in the presence and absence of UVA radiation. The antioxidant activity was evaluated by HRP-H2O2-luminol induced chemiluminescence. The results showed that relationship between the molecular structure and UV absorption. Me molecules phenylamine benzophenone-3 (B5) and phenylamine benzophenone-3 carbazol (B6) showed higher short and long-wave UVA and UVB absorption, when compared to benzophenone-3. Only B5 and B6, which presented intermolecular hydrogen bond, were considered photostable. B5 did not present any acute phototoxicity potential; in addition it has antioxidant activity, which suggests its high UVfilter potential.

A exposição excessiva às radiações UV é capaz de limitar a capacidade dos sistemas de defesa antioxidante em nosso organismo, provocando o estresse oxidativo. Assim, extratos vegetais ricos em compostos antioxidantes são fortes candidatos a serem veiculados em formulações tópicas para a prevenção ou tratamento dos danos causados pela RUV na pele. Dentre a gama de extratos vegetais com atividade biológica, o extrato de açaí, fruto da espécie Euterpe olerecea Mart., tem se destacado por apresentar grande quantidade de antioxidantes na sua composição. Geralmente, a quantidade de ativos que consegue penetrar na pele é pequena, exigindo métodos analíticos muito sensíveis. Assim, o presente trabalho teve como objetivo avaliar o emprego do método de quimioluminescência para a quantificação da penetração dos componentes do extrato de açaí na pele por medida da atividade antioxidante. Os resultados mostraram que o IC50 do extrato de açaí para o ensaio de inibição da quimioluminescência gerada no sistema xantina/luminol/xantina oxidase foi de 0,63 ?g/mL. Além disso, a quantificação do extrato de açaí utilizando esse método foi possível para porcentagens de inibição obtidas até o valor de 60%. Ainda, esse método mostrou-se preciso e exato na determinação da porcentagem de inibição da quimioluminescência do extrato de açaí na concentração próxima a IC50, sendo essa inibição não influenciada pelos componentes das diferentes formulações estudadas. No entanto, o uso da quimioluminescência como um método de quantificação da penetração de componentes antioxidantes de extratos hidrossolúveis torna o processo de extração desses componentes da pele um fator importante, no qual a escolha do solvente extrator é um ponto crítico. Assim, o solvente extrator escolhido para a realização dos estudos de penetração/retenção cutânea foi metanol:água (80:20), visto que esse solvente foi capaz de extrair os componentes antioxidantes do extrato de açaí sem retirar grande quantidade dos compostos inerentes da pele com atividade antioxidante. Por fim, a determinação da inibição da quimioluminescência gerada no sistema xantina/luminol/xantina oxidase mostrou-se um método importante na medida da atividade antioxidante de extratos na pele. Ainda, esse método foi eficaz na quantificação do extrato de açaí nos estudos de penetração/retenção com célula de difusão vertical, apesar de todos os erros inerentes desse método. / The excessive exposure to UV radiation is able to decrease the antioxidant defense systems in the skin leading to the oxidative stress. Thus, plant extracts, rich in antioxidant compounds, are strong candidates to be added in topical formulations for the prevention or treatment of UV radiation induced damages. Among the range of plant extracts with biological activity, the extract of açai, fruit of the Euterpe olerecea Mart. species has become known due to the considerable amount of antioxidants in its composition. Generally, the amount of active compounds that can penetrate in the skin is low requiring sensitive analytical methods. Thus, the present study aimed to evaluate the use of chemiluminescence assay for the quantification of the penetration of açai extract components in the skin by measuring the antioxidant activity. The results showed an IC50 value of 0.63 ?g/mL for the antioxidant activity determined by chemiluminescence assay using the xanthine/luminol/xanthine oxidase system. Moreover, the quantification of acai extract using this method was possible for percentages of inhibition obtained until the 60% of inhibition value. Additionally, this method proved to be precise and accurate in determining the percentage of chemiluminescence inhibition of acai extract at concentrations values close to the IC50 value. Additionally, it was observed that the inhibition was not influenced by the different components of the formulations studied. However, the use of chemiluminescence assay to quantify the penetration of water-soluble extract antioxidant components makes the extraction process of these components from the skin an important step in which the selection of the extractor solvent is a critical point. In the present study it was selected methanol:water (80%) as an organic extractor solvent which was able to extract the antioxidant components of the acai extract without removing large amounts of inherent compounds of the skin with antioxidant activity. Finally, the determination of the chemiluminescence generated in the xanthine/luminol/xanthine oxidase system inhibition showed to be an important method for the measurement of the antioxidant activity of plant extracts in the skin. Still, this method was effective in quantifying the acai extract in cutaneous penetration/retention studies using vertical diffusion cells despite all the intrinsic errors of this used method.

A exposição excessiva às radiações UV é capaz de limitar a capacidade dos sistemas de defesa antioxidante em nosso organismo, provocando o estresse oxidativo. Assim, extratos vegetais ricos em compostos antioxidantes são fortes candidatos a serem veiculados em formulações tópicas para a prevenção ou tratamento dos danos causados pela RUV na pele. Dentre a gama de extratos vegetais com atividade biológica, o extrato de açaí, fruto da espécie Euterpe olerecea Mart., tem se destacado por apresentar grande quantidade de antioxidantes na sua composição. Geralmente, a quantidade de ativos que consegue penetrar na pele é pequena, exigindo métodos analíticos muito sensíveis. Assim, o presente trabalho teve como objetivo avaliar o emprego do método de quimioluminescência para a quantificação da penetração dos componentes do extrato de açaí na pele por medida da atividade antioxidante. Os resultados mostraram que o IC50 do extrato de açaí para o ensaio de inibição da quimioluminescência gerada no sistema xantina/luminol/xantina oxidase foi de 0,63 ?g/mL. Além disso, a quantificação do extrato de açaí utilizando esse método foi possível para porcentagens de inibição obtidas até o valor de 60%. Ainda, esse método mostrou-se preciso e exato na determinação da porcentagem de inibição da quimioluminescência do extrato de açaí na concentração próxima a IC50, sendo essa inibição não influenciada pelos componentes das diferentes formulações estudadas. No entanto, o uso da quimioluminescência como um método de quantificação da penetração de componentes antioxidantes de extratos hidrossolúveis torna o processo de extração desses componentes da pele um fator importante, no qual a escolha do solvente extrator é um ponto crítico. Assim, o solvente extrator escolhido para a realização dos estudos de penetração/retenção cutânea foi metanol:água (80:20), visto que esse solvente foi capaz de extrair os componentes antioxidantes do extrato de açaí sem retirar grande quantidade dos compostos inerentes da pele com atividade antioxidante. Por fim, a determinação da inibição da quimioluminescência gerada no sistema xantina/luminol/xantina oxidase mostrou-se um método importante na medida da atividade antioxidante de extratos na pele. Ainda, esse método foi eficaz na quantificação do extrato de açaí nos estudos de penetração/retenção com célula de difusão vertical, apesar de todos os erros inerentes desse método. / The excessive exposure to UV radiation is able to decrease the antioxidant defense systems in the skin leading to the oxidative stress. Thus, plant extracts, rich in antioxidant compounds, are strong candidates to be added in topical formulations for the prevention or treatment of UV radiation induced damages. Among the range of plant extracts with biological activity, the extract of açai, fruit of the Euterpe olerecea Mart. species has become known due to the considerable amount of antioxidants in its composition. Generally, the amount of active compounds that can penetrate in the skin is low requiring sensitive analytical methods. Thus, the present study aimed to evaluate the use of chemiluminescence assay for the quantification of the penetration of açai extract components in the skin by measuring the antioxidant activity. The results showed an IC50 value of 0.63 ?g/mL for the antioxidant activity determined by chemiluminescence assay using the xanthine/luminol/xanthine oxidase system. Moreover, the quantification of acai extract using this method was possible for percentages of inhibition obtained until the 60% of inhibition value. Additionally, this method proved to be precise and accurate in determining the percentage of chemiluminescence inhibition of acai extract at concentrations values close to the IC50 value. Additionally, it was observed that the inhibition was not influenced by the different components of the formulations studied. However, the use of chemiluminescence assay to quantify the penetration of water-soluble extract antioxidant components makes the extraction process of these components from the skin an important step in which the selection of the extractor solvent is a critical point. In the present study it was selected methanol:water (80%) as an organic extractor solvent which was able to extract the antioxidant components of the acai extract without removing large amounts of inherent compounds of the skin with antioxidant activity. Finally, the determination of the chemiluminescence generated in the xanthine/luminol/xanthine oxidase system inhibition showed to be an important method for the measurement of the antioxidant activity of plant extracts in the skin. Still, this method was effective in quantifying the acai extract in cutaneous penetration/retention studies using vertical diffusion cells despite all the intrinsic errors of this used method.

No description available.