Electronic relaxation pathways of the biologically relevant pterin chromophore.

Pterin derivatives are endogenous photosensitizers common to every biological kingdom and are involved in numerous photobiological processes for which they have received widespread attention. Despite the photobiological relevance of the pterin compounds, their excited-state dynamics have not been investigated rigorously-an undertaking that must begin with a thorough understanding of the photophysics of the common core chromophore, 2-amino-1H-pteridin-4-one (a.k.a. pterin). In this contribution, direct observation of the ultrafast excited-state dynamics of pterin in aqueous borate buffered solution at pH 10.5 are revealed using broadband transient absorption spectroscopy with femtosecond time resolution. The time-resolved experiments are complemented with ground- and excited-state calculations that are performed at the time-dependent density functional level of theory. Excitation of pterin at 350 nm initially populates the electronic 1ππ* state with excess vibrational energy. A sizable fraction of the vibrationally-excited 1ππ* state population intersystem crosses to populate a receiver 3nπ* state, while another fraction relaxes to populate the 1ππ* minimum that decays by fluorescence emission to the ground state. The population that reaches the 3nπ* state either internally converts to the 3ππ* state or abstracts a hydrogen atom from the solvent to form a semi-reduced neutral radical species, both of which decay with rates accelerated by molecular oxygen. The neutral radical can also decay through back hydrogen atom transfer to the solvent, as evidenced by the normal kinetic isotope effect observed for this process. The transient species characterized in this study are expected to participate in a wide-range of photochemical reactions that have been documented in the literature between pterin and other biological molecules.

Correction: Photochemical etiology of promising ancestors of the RNA nucleobases.

Correction for 'Photochemical etiology of promising ancestors of the RNA nucleobases' by M. M. Brister et al., Phys. Chem. Chem. Phys., 2016, DOI: .

Photochemical etiology of promising ancestors of the RNA nucleobases.

RNA is a product of chemical and biological evolution and the identification of its heterocyclic ancestors is essential for understanding the molecular origins of life. Among a diverse array of selection pressures thought to have shaped the composition of the nucleobases on prebiotic Earth, protection against intense ultraviolet radiation must have been essential. In this contribution, a detailed spectroscopic and photophysical investigation of barbituric acid and 2,4,6-triaminopyrimidine, two promising candidates for the prebiotic ancestors of RNA nucleobases, is presented in aqueous solution. It is shown that although these pyrimidine derivatives absorb ultraviolet radiation strongly, both compounds possess efficient electronic relaxation mechanisms for dissipating most of the absorbed ultraviolet energy to their aqueous environment as heat within hundreds of femtoseconds, thus safeguarding their chemical integrity. In fact, these two heterocyclic compounds rival the photostability observed in the canonical nucleobases in aqueous solution, thus supporting the recent proposal that both barbituric acid and 2,4,6-triaminopyrimidine are promising ancestors of the RNA nucleobases.

Increase in the photoreactivity of uracil derivatives by doubling thionation.

The ability of 4-thiouracil to strongly absorb UVA radiation and to populate a reactive triplet state in high yield has enabled its use as a versatile photocrosslinker for nearly 50 years. In this contribution, we present a detailed spectroscopic and photochemical investigation of the 2-thiouracil, 4-thiouracil, and 2,4-dithiouracil series in an effort to further advance this chemistry and to scrutinize the photoreactivity of 2,4-dithiouracil. Our results reveal that excitation of 2,4-dithiouracil leads to intersystem crossing to the triplet manifold in 220 ± 40 fs, which enables the population of the reactive triplet state with near unity yield (ΦT = 0.90 ± 0.15) and ultimately leads to a ca. 50% singlet oxygen generation (ΦΔ = 0.49 ± 0.02)-one of the highest singlet oxygen yields reported to date for a photoexcited thiobase. In addition, the long-lived triplet state of 2,4-dithiouracil reacts efficiently with the nucleic acid base adenine 5'-monophosphate through a direct, oxygen-independent photocycloaddition mechanism and at a rate that is at least 3-fold faster than that of 4-thiouracil under equal conditions. The new physico-chemical insights reported for these RNA-thiobase derivatives are compared to those of the DNA and RNA bases and the DNA-thiobase derivatives. Furthermore, the strong near-visible absorption and increased photoreactivity measured for 2,4-dithiouracil lays a solid foundation for developing RNA-targeted photocrosslinking and phototherapeutic agents that are more effective than those currently available.

Periconceptional undernutrition in sheep leads to decreased locomotor activity in a natural environment.

Maternal undernutrition during pregnancy increases offspring obesity and metabolic disease risk. We hypothesized that periconceptional undernutrition in sheep from 60 days before conception through to day 30 of gestation (UN) would decrease voluntary locomotor activity in adult offspring. Distance travelled was measured at 18 months of age for ∼48 h in the paddock. Data were analysed using multiple regression analysis, with explanatory variables including sex, nutrition group, birth weight, average time between GPS measurements and percentage of time during the measurement period spent in daylight. Mean (±s.e.) distance walked (m/h) was greater for control (CON) than UN animals, and greater for females than males [110.2 (6.5), CON females; 110.7 (6.3), CON males; 105.1 (5.3), UN females and 95.5 (5.8), UN males; P = 0.02 for nutrition group effect and for sex effect]. Periconceptional undernutrition may lead to a significant decrease in voluntary physical activity in adult offspring.

Evidence of conserved epitopes in variable region of VP8* subunit of VP4 protein of rotaviruses of P[8]-1 and P[8]-3 lineages.

Although antibody responses to the human rotavirus VP4 protein have been reported, few studies have analyzed the specificity of these responses to the VP8* subunit. This study investigated antibody responses generated against the variable region of the VP4 protein (VP8* subunit) in children infected with rotavirus genotype P[8]. Recombinant VP8* subunit (rVP8*) and truncations corresponding aa 1-102
(peptide A) and 84-180 (peptide B) of rotavirus strains P[8]-1 and P[8]-3 lineages were expressed in Escherichia coli and examined for antibody reactivity using ELISA and Western blot assays. Sera from infected children had IgG antibodies that reacted with full-length rVP8*, peptide A and B of both lineages, with stronger reactivity observed against peptide B. In addition, anti-strain Wa (P[8]-1) and anti-rVP8* (P[8]-3) rabbit polyclonal antiserum reacted against peptide B sequences of both lineages. These data indicate that the VP8* variable region of rotavirus belonging to P[8]-1 and P[8]-3 lineages have conserved epitopes recognized by antibodies elicited during natural infections.

Effects of sex, litter size and periconceptional ewe nutrition on offspring behavioural and physiological response to isolation.

Maternal periconceptional undernutrition alters fetal hypothalamic-pituitary-adrenal (HPA) axis development. However, the effects of this early nutritional insult on postnatal HPA axis function and stress-related behaviours are unknown. We investigated in sheep the effects of different periods of undernutrition, and of sex and litter size, on offspring behavioural and cortisol responses to isolation stress. We studied four nutritional groups: controls well nourished throughout pregnancy (n=39), or ewes undernourished (UN, 10-15% body weight reduction) before mating (-60 to 0d, n=26), after mating (-2 to +30d, n=20) or both (-60 to +30d, n=36). At 4 and 18months of age, offspring were isolated for 5min, their behaviour video recorded, and plasma cortisol concentrations measured. Offspring of all undernourished groups demonstrated 50% fewer escape attempts than controls at 4 months of age, and offspring of UN-60+30 ewes had 20% lower plasma cortisol area under the curve in response to isolation at 18months. Females had higher cortisol concentrations and vocalised more than males at 4 and 18months, and were more active at 18months. After isolation, UN-2+30 males had higher cortisol concentrations than UN-2+30 females whereas in all other groups males had lower concentrations than females. Singleton males made more escape attempts than females, whereas in twins females made more escape attempts than males. These findings suggest that maternal periconceptional undernutrition in sheep can suppress behavioural reactions and cortisol secretion in response to isolation stress in the offspring into adulthood, and that these effects differ between males and females.

Reaction of foster cows to prevention of suckling from and separation from four calves simultaneously or in two steps.

The aim of this study was to investigate if a 2-step method of preventing suckling and cow-calf separation reduces the stress reaction in foster cows compared with a simultaneous separation method. Seven Swedish Holstein and 5 Swedish Red dairy cows were used as foster cows, each having a group of 4 calves. The foster cow-calf group was formed when calves were 1 wk old, and the calves were prevented from suckling at 10 wk of age. In 6 of the cow-calf groups, calves were prevented from suckling by simultaneous separation from the cow (control). In the other 6 groups, calves were fitted with a nose-flap, which prevented them from suckling while they were kept together with the cow for another 2 wk before they were separated (2-step). The behavior of the foster cows was observed at 4 observation periods, 0 to 2, 8.5 to 9.5, 24 to 26, and 72 to 74 h after the calves were prevented from suckling (2-step), after separation (2-step), and after calves were prevented from suckling by simultaneous separation (control). For both treatments, saliva cortisol was sampled once daily for 5 d at wk 10. This was repeated at wk 12 for the 2-step treatment. Heart rate was measured with the behavioral observations. Control foster cows vocalized more (P < 0.001) and walked more (P = 0.005) than the 2-step foster cows after prevention of suckling and after separation from the calves. When control cows were separated from their calves, they more frequently (P < 0.001) held their head out of the pen than was the case with 2-step cows when separated 2 wk after prevention of suckling. The variation in heart rate was larger in the control group compared with 2-step cows at 0 to 2 h after separation/prevention of suckling (P = 0.002). No effect of treatment was found on cortisol concentration. Our conclusion is that separating the 2 events "prevention of suckling" and "separation" reduces the stress experienced by the foster cow at weaning.

Mechanism of formation of guanine as one of the major products in the 254 nm photolysis of guanine derivatives: concentration and pH effects.

The quantum yield of formation of guanine (Gua), one of the major products formed in the 254 nm steady-state photolysis of the following Gua derivatives (9-ethylguanine [9Et-Gua], deoxyguanosine [dGuo], guanosine [Guo], guanosine 5'-monophosphate [GMP]) was determined under different conditions. The formation yield increases in the following order: 9Et-Gua < dGuo approximately Guo < GMP. Electron scavengers or triplet quenchers were incorporated into the irradiated solutions with the purpose of reacting specifically with postulated or previously identified intermediates in the photolysis of Gua derivatives. A decrease in the yield of formation of Gua is observed with increasing electron scavenger concentration or with pH. These results suggest a major contribution from Gua-derivative electron adducts on the process of N(9)-R bond breakage. At an acidic pH a tautomer of the radical cation (E) of Guo is proposed as the precursor for Gua formation. The relative efficiency of the radical cation for initiating the release of free Gua depends on the pH of the solution, being less than 39% in neutral pH. Reactions from OH radicals may also result in base release as shown using N2O as additive. Finally, the formation of aggregates by the bases at concentrations used, plays an important role in the deactivation of the excited states and also in the probability of formation of the free base.

Photochemical and photophysical studies of guanine derivatives: intermediates contributing to its photodestruction mechanism in aqueous solution and the participation of the electron adduct.

The low-intensity steady-state (254 nm), microsecond flash and nanosecond (266 nm) laser photolysis of some guanine (Gua) derivatives in aqueous solution were studied. A photodestruction yield between 10(-3) and 10(-2) at a base concentration of 75 microM was determined for 254 nm irradiation at room temperature using high-performance liquid chromatography. This yield decreases with increasing purine concentration. For a similar concentration of the purine bases (2 +/- 1) x 10(-5) M, the yield increases as follows: Gua approximately 9-ethylguanine < deoxyguanosine approximately guanosine (Guo) < guanosine 5'-monophosphate. At concentrations higher than 2 x 10(-4) M the Gua derivatives' photodestruction yield seems to converge to a limiting value of the order of 10(-4). This behavior is explained in terms of self-quenching and aggregation effects which deactivate the excited states of the bases. The yields of electron photoejection have been determined in the nanosecond laser photolysis (0.083) and in the low-intensity steady-state (5.8 x 10(-3)) for Guo. Competition experiments using electron scavengers suggest that the electron adducts of the bases are one of the principal species participating in the photodestruction mechanism of these monomeric Gua. Close to 75% of the total destruction yield has contributions from initial reactions of the photojected electron at neutral pH. The quantum yield of photodestruction of Guo increases when the pH is increased as follows: 4.7 x 10(-3) (pH 1.1), 6.5 x 10(-3) (pH 2.9), 7.7 x 10(-3) (pH 7.5) and 8.1 x 10(-3) (pH 11.9). This dependence on pH and the electron scavenger experiments provide further evidence for the radical anion or its protonated form as one of the principal species involved in the photodestruction of the bases at the different pH. Under oxygen saturated conditions a 22% increase in the destruction yield is observed for Guo. However, for the dinucleotides adenylyl (3'-->5')-guanosine and thymidylyl (3'-->5')2'-deoxyguanosine, the participation of the electron is 41 and 36%, respectively, suggesting that going into a more DNA or RNA-like structure, the participation of the electron adducts species in the photodamage of DNA and RNA decreases. A mechanism of photodestruction for the Gua derivatives is proposed which takes into account these findings.

Designing and evaluating strategies to recruit African Americans for AIDS/HIV interventions: targeting the African-American family.

The successful recruitment of African-Americans for AIDS/HIV interventions is essential to reducing HIV transmission and the spread of AIDS among this population. However, many high risk groups in the African-American community are reluctant to participate in AIDS/HIV interventions due to factors such as poverty, and cultural and religious values. This paper discusses the basis of these factors and presents a theoretical framework that holds some promise for designing and evaluating AIDS/HIV intervention recruitment strategies and a pilot study implementing this strategy is presented. The results emphasize the importance of targeting and empowering the African-American family in recruitment strategies, as opposed to specific subgroups of high risk individuals. Recruitment messages, which were channeled through a credible source and which stress the role of participants in contributing to the well-being of their families and communities, were most successful.

Infant death rates among the poor and nonpoor in Kentucky, 1982 to 1983.

The mortality rates of infants born in Kentucky during 1982 and 1983 were analyzed to determine whether there were differences between poor and nonpoor infants. We used computer matching with the Social Insurance Food Stamp files and with the Aid for Families With Dependent Children files to define poor and nonpoor Kentucky resident families. The crude death rate for poor infants was 13.7/1,000 births, and the nonpoor death rate was 10.8/1,000 births. After adjusting for several important variables, we found that the poor infant was at a significantly greater risk for death than the nonpoor infant but only during the postneonatal period (relative risk 2.04, P = .0003). Some differences by sex and race were noted. Sudden infant death syndrome and infections were largely responsible for the poor infants' higher postneonatal mortality risk.