Chemical characteristics of wildfire ash across the globe and their environmental and socio-economic implications.

The mobilisation of potentially harmful chemical constituents in wildfire ash can be a major consequence of wildfires, posing widespread societal risks. Knowledge of wildfire ash chemical composition is crucial to anticipate and mitigate these risks. Here we present a comprehensive dataset on the chemical characteristics of a wide range of wildfire ashes (42 types and a total of 148 samples) from wildfires across the globe and examine their potential societal and environmental implications. An extensive review of studies analysing chemical composition in ash was also performed to complement and compare our ash dataset. Most ashes in our dataset had an alkaline reaction (mean pH 8.8, ranging between 6 and 11.2). Important constituents of wildfire ash were organic carbon (mean: 204 g kg-1), calcium, aluminium, and iron (mean: 47.9, 17.9 and 17.1 g kg-1). Mean nitrogen and phosphorus ranged between 1 and 25 g kg-1, and between 0.2 and 9.9 g kg-1, respectively. The largest concentrations of metals of concern for human and ecosystem health were observed for manganese (mean: 1488 mg kg-1; three ecosystems > 1000 mg kg-1), zinc (mean: 181 mg kg-1; two ecosystems > 500 mg kg-1) and lead (mean: 66.9 mg kg-1; two ecosystems > 200 mg kg-1). Burn severity and sampling timing were key factors influencing ash chemical characteristics like pH, carbon and nitrogen concentrations. The highest readily dissolvable fractions (as a % of ash dry weight) in water were observed for sodium (18 %) and magnesium (11.4 %). Although concentrations of elements of concern were very close to, or exceeded international contamination standards in some ashes, the actual effect of ash will depend on factors like ash loads and the dilution into environmental matrices such as water, soil and sediment. Our approach can serve as an initial methodological standardisation of wildfire ash sampling and chemical analysis protocols.

Searching for Chameleon Dark Energy with Mechanical Systems.

A light scalar field framework of dark energy, sometimes referred to as quintessence, introduces a fifth force between normal matter objects. Screening mechanisms, such as the chameleon model, allow the scalar field to be almost massless on cosmological scales while simultaneously evading laboratory constraints. We explore the ability of existing mechanical systems to directly detect the fifth force associated with chameleons in an astrophysically viable regime where it could be dark energy. We provide analytical expressions for the weakest accessible chameleon model parameters in terms of experimentally tunable variables and apply our analysis to two mechanical systems: levitated microspheres and torsion balances, showing that the current generation of these experiments have the sensitivity to rule out a significant portion of the proposed chameleon parameter space. We also indicate regions of theoretically well-motivated chameleon parameter space to guide future experimental work.

Autologous skin cells: a new technique for skin regeneration in diabetic and vascular ulcers.

Diabetic foot ulcers are often difficult to treat due to concurrent infection, neuropathy and vascular compromise. Healing adjuncts, such as negative pressure wound therapy and skin grafts, have been used with good results but eventual healing is often frustrated by slow epithelialisation. Here, we describe a novel therapeutic method to aid epithelial regeneration using autologous skin cells (ReCell; Avita Medical) to aid skin regeneration. We suggest this may provide an alternative to more established therapies used in this patient group.

Intra-operative acute isovolemic hemodilution is safe and effective in eliminating allogeneic blood transfusions during right hepatic lobectomy: comparison of living donor versus non-donors.

BACKGROUND: Multiple studies have shown acute isovolemic hemodilution (AIH) to be safe and effective during liver resection to limit the use of banked blood. However, no studies to date have studied AIH in living donor right hepatectomy. Conventional right hepatectomies for living donors is not identical to non-donor right hepatectomies. Since division of the parenchyma is often performed without devascularization of the right lobe, blood loss may be significantly higher. METHODS: Ten consecutive patients undergoing living donor right hepatectomies (LDRH) and ten consecutive patients undergoing non-donor right hepatectomies (NDRH) were compared using AIH. RESULTS: There was no mortality or morbidity related to the use of AIH. No allogeneic blood transfusions were required in either group, intra-operatively or post-operatively. There was no significant difference in post-operative hematocrit, average estimated blood loss, and average fluid replacement. Average hospital length of stay and operating room time were longer for the LDRH. CONCLUSION: AIH can be performed safely and effectively in both LDRH and NDRH without subjecting patients to unnecessary risks of allogeneic blood transfusions.

Splicing-related catalysis by protein-free snRNAs.

Removal of intervening sequences from eukaryotic messenger RNA precursors is carried out by the spliceosome, a complex assembly of five small nuclear RNAs (snRNAs) and a large number of proteins. Although it has been suggested that the spliceosome might be an RNA enzyme, direct evidence for this has been lacking, and the identity of the catalytic domain of the spliceosome is unknown. Here we show that a protein-free complex of two snRNAs, U2 and U6, can bind and position a small RNA containing the sequence of the intron branch site, and activate the branch adenosine to attack a catalytically critical domain of U6 in a reaction that is related to the first step of splicing. Our data provide direct evidence for the catalytic potential of spliceosomal snRNAs.

Identification and functional characterization of neo-poly(A) polymerase, an RNA processing enzyme overexpressed in human tumors.

Poly(A) polymerase (PAP) plays an essential role in polyadenylation of mRNA precursors, and it has long been thought that mammalian cells contain only a single PAP gene. We describe here the unexpected existence of a human PAP, which we call neo-PAP, encoded by a previously uncharacterized gene. cDNA was isolated from a tumor-derived cDNA library encoding an 82.8-kDa protein bearing 71% overall similarity to human PAP. Strikingly, the organization of the two PAP genes is nearly identical, indicating that they arose from a common ancestor. Neo-PAP and PAP were indistinguishable in in vitro assays of both specific and nonspecific polyadenylation and also endonucleolytic cleavage. Neo-PAP produced by transfection was exclusively nuclear, as demonstrated by immunofluorescence microscopy. However, notable sequence divergence between the C-terminal domains of neo-PAP and PAP suggested that the two enzymes might be differentially regulated. While PAP is phosphorylated throughout the cell cycle and hyperphosphorylated during M phase, neo-PAP did not show evidence of phosphorylation on Western blot analysis, which was unexpected in the context of a conserved cyclin recognition motif and multiple potential cyclin-dependent kinase (cdk) phosphorylation sites. Intriguingly, Northern blot analysis demonstrated that each PAP displayed distinct mRNA splice variants, and both PAP mRNAs were significantly overexpressed in human cancer cells compared to expression in normal or virally transformed cells. Neo-PAP may therefore be an important RNA processing enzyme that is regulated by a mechanism distinct from that utilized by PAP.

Physical and functional interactions between Drosophila TRAF2 and Pelle kinase contribute to Dorsal activation.

Signaling through the Toll receptor is required for dorsal/ventral polarity in Drosophila embryos, and also plays an evolutionarily conserved role in the immune response. Upon ligand binding, Toll appears to multimerize and activate the associated kinase, Pelle. However, the immediate downstream targets of Pelle have not been identified. Here we show that Drosophila tumor necrosis factor receptor-associated factor 2 (dTRAF2), a homologue of human TRAF6, physically and functionally interacts with Pelle, and is phosphorylated by Pelle in vitro. Importantly, dTRAF2 and Pelle cooperate to activate Dorsal synergistically in cotransfected Schneider cells. Deletion of the C-terminal TRAF domain of dTRAF2 enhances Dorsal activation, perhaps reflecting the much stronger interaction of the mutant protein with phosphorylated, active Pelle. Taken together, our results indicate that Pelle and dTRAF2 physically and functionally interact, and that the TRAF domain acts as a regulator of this interaction. dTRAF2 thus appears to be a downstream target of Pelle. We discuss these results in the context of Toll signaling in flies and mammals.

Evolutionarily conserved interaction between CstF-64 and PC4 links transcription, polyadenylation, and termination.

Tight connections exist between transcription and subsequent processing of mRNA precursors, and interactions between the transcription and polyadenylation machineries seem especially extensive. Using a yeast two-hybrid screen to identify factors that interact with the polyadenylation factor CstF-64, we uncovered an interaction with the transcriptional coactivator PC4. Both human proteins have yeast homologs, Rna15p and Sub1p, respectively, and we show that these two proteins also interact. Given evidence that certain polyadenylation factors, including Rna15p, are necessary for termination in yeast, we show that deletion or overexpression of SUB1 suppresses or enhances, respectively, both growth and termination defects detected in an rna15 mutant strain. Our findings provide an additional, unexpected connection between transcription and polyadenylation and suggest that PC4/Sub1p, via its interaction with CstF-64/Rna15p, possesses an evolutionarily conserved antitermination activity.

Early transvaginal ultrasonography versus early cerclage in women with an unclear history of incompetent cervix.

OBJECTIVE: Our aim was to compare outcomes in women with a questionable history of incompetent cervix, followed up with early transvaginal ultrasonography, with outcomes in women who had early cerclage. STUDY DESIGN: Charts were reviewed and patients identified for incompetent cervix from our obstetric database from 1995 through 1997. We included women who had an unclear history of incompetent cervix as follows: second-trimester loss or termination, > or =3 first-trimester terminations, cone biopsy or loop electrosurgical excision, or exposure to diethylstilbestrol. The primary outcome variable was gestational age at delivery. RESULTS: A total of 106 women were included, 45 in the early cerclage group and 61 in the early transvaginal ultrasonography group. The mean gestational age at delivery was 35.1 weeks for the early cerclage group versus 36.1 weeks for the early transvaginal ultrasonography group. CONCLUSION: In women with an unclear history of incompetent cervix, early cerclage does not appear to offer significant benefit over early transvaginal ultrasonography.

Porphyrins with exocyclic rings. 16. Synthesis and spectroscopic characterization of fluoranthoporphyrins, a new class of highly conjugated porphyrin chromophores.

Porphyrins with fused aromatic rings are under detailed investigation due to their unique spectroscopic properties. To gain more insights into the effects due to ring annealation on the porphyrin chromophore, a series of fluoranthoporphyrins have been synthesized. Reaction of 3-nitrofluoranthene with isocyanoacetate esters in the presence of a phosphazene base afforded good yields of the fluorantho[2,3-c]pyrrole esters 8. Cleavage of the ester moiety with KOH in ethylene glycol afforded the parent heterocycle 9, and this condensed with 2 equiv of acetoxymethylpyrroles 10 in refluxing acetic acid-2-propanol to afford tripyrranes 11. Following cleavage of the tert-butyl ester protective groups with TFA, "3 + 1" condensation with pyrrole dialdehyde 12 gave the fluoranthoporphyrins 13 in good overall yields. In addition, reaction of tripyrrane 11 with acenaphthopyrrole dialdehyde 16 gave the mixed acenaphthofluoranthoporphyrin 17 in excellent yields. A difluoranthoporphyrin 18 was also prepared via a "2 + 2" MacDonald condensation. Reaction of fluoranthopyrrole 8a with dimethoxymethane in the presence of p-toluenesulfonic acid gave the symmetrical dipyrrylmethane 19, and following ester saponification, this was condensed with a dipyrrylmethane dialdehyde to afford the adj-difluoranthoporphyrin 18. The UV--vis spectra for these fluoranthoporphyrins gave a series of three broadened absorptions in the Soret band region, although the Q-bands were little effected by ring fusion. The nickel(II), copper(II), and zinc chelates were more unusual, showing strong absorptions near 600 nm. Difluoranthoporphyrin 18 showed many of the same spectroscopic features, although the presence of two ring fusions gave rise to an increase in the spectroscopic shifts. The mixed system 17 gave spectra that showed larger red shifts due to the acenaphthylene unit combined with the features due to the fluoranthene rings. This work further demonstrates the utility of aromatic ring fusion in altering the properties of porphyrinoid systems.

Intracervical fibrin sealants: a potential treatment for early preterm premature rupture of the membranes.

OBJECTIVE: We report our experience with a transvaginally applied intracervical fibrin sealant at <24 weeks' gestation. STUDY DESIGN: This is an observational study of a referred patient population, with preterm premature rupture of the membranes at <24 weeks' gestation. RESULTS: Twelve women consented to our protocol. The mean gestational age at preterm premature rupture of membranes was 19 weeks 4 days (range, 13-23 weeks); the mean gestational age at treatment was 20 weeks 5 days (range, 17-23 weeks). All women had a diminution in the amount of amniotic fluid leakage with an increase in amniotic fluid index. Among the 12 pregnancies (13 fetuses), there were 7 surviving neonates. Two women had apparent "resealing" of the membranes. CONCLUSION: Fibrin sealants in midtrimester rupture of the membranes may lead to improved outcomes and now warrant formal evaluation.

Heterozygous disruption of the TATA-binding protein gene in DT40 cells causes reduced cdc25B phosphatase expression and delayed mitosis.

TATA-binding protein (TBP) is a key general transcription factor required for transcription by all three nuclear RNA polymerases. Although it has been intensively analyzed in vitro and in Saccharomyces cerevisiae, in vivo studies of vertebrate TBP have been limited. We applied gene-targeting techniques using chicken DT40 cells to generate heterozygous cells with one copy of the TBP gene disrupted. Such TBP-heterozygous (TBP-Het) cells showed unexpected phenotypic abnormalities, resembling those of cells with delayed mitosis: a significantly lower growth rate, larger size, more G2/-M- than G1-phase cells, and a high proportion of sub-G1, presumably apoptotic, cells. Further evidence for delayed mitosis in TBP-Het cells was provided by the differential effects of several cell cycle-arresting drugs. To determine the cause of these defects, we first examined the status of cdc2 kinase, which regulates the G2/M transition, and unexpectedly observed more hyperphosphorylated, inactive cdc2 in TBP-Het cells. Providing an explanation for this, mRNA and protein levels of cdc25B, the trigger cdc2 phosphatase, were significantly and specifically reduced. These properties were all due to decreased TBP levels, as they could be rescued by expression of exogeneous TBP, including, in most but not all cases, a mutant form lacking the species-specific N-terminal domain. Our results indicate that small changes in TBP concentration can have profound effects on cell growth in vertebrate cells.

The BARD1-CstF-50 interaction links mRNA 3' end formation to DNA damage and tumor suppression.

The mRNA polyadenylation factor CstF interacts with the BRCA1-associated protein BARD1, and this interaction represses the nuclear mRNA polyadenylation machinery in vitro. Given the suspected role of BRCA1/BARD1 in DNA repair, we tested whether inhibition of mRNA processing is linked to DNA damage. Strikingly, we found that 3' cleavage in extracts from cells treated with hydroxyurea or ultraviolet light was strongly, but transiently, inhibited. Although no changes were detected in CstF, BARD1, and BRCA1 protein levels, increased amounts of a CstF/BARD1/BRCA1 complex were detected. Supporting the physiological significance of these results, a previously identified tumor-associated germline mutation in BARD1 (Gln564His) reduced binding to CstF and abrogated inhibition of polyadenylation. Together these results indicate a link between mRNA 3' processing and DNA repair and tumor suppression.

A randomized comparison of transcervical Foley catheter to intravaginal misoprostol for preinduction cervical ripening.

OBJECTIVE: To compare the efficacy of intravaginal misoprostol tablets with transcervical Foley catheter for preinduction cervical ripening. METHODS: Pregnant women who presented for induction of labor with unfavorable cervices (Bishop score less than 6) were assigned randomly to intravaginal misoprostol (50 microg tablet every 4 hours for a maximum of six doses) or 30-mL Foley catheter placed transcervically with maintenance of traction. RESULTS: Among 111 women, 53 were allocated to misoprostol and 58 to Foley bulb. Contractile abnormalities were more frequent in the misoprostol group (20.4%) than the Foley group (0%) (P <.001). No statistically significant differences were noted between groups in change in Bishop score, preinduction cervical ripening times, and total induction times. There were no statistically significant differences in mode of delivery or adverse neonatal outcomes. Uterine rupture occurred in one woman with two previous cesarean deliveries in the misoprostol group. CONCLUSION: Intravaginal misoprostol and transcervical Foley catheter are equivalent for cervical ripening. Uterine contractile abnormalities and meconium passage are more common with misoprostol.

How echogenic is echogenic? Quantitative acoustics of the renal cortex.

The echogenicity of the cortex is an important parameter in interpreting renal sonograms that suggest changes in cortical structure. Echogenicity is currently measured qualitatively, and no attempts have been made at quantification. We developed a method to quantify renal cortical echogenicity in reference to the liver and evaluated its reproducibility, dependence on scanning variables, and potential utility. Sonograms of the right kidney were digitized, and the mean pixel density of regions of the renal cortex and liver was measured and normalized to the gray scale. Echogenicity was expressed as the ratio of the brightness (inverse of mean pixel density) of the cortex to that of the liver. The mean coefficient of variation among measurements performed on multiple sonograms from the same study was 2.8%, and the coefficient of variation among multiple measurements performed on the same kidney over 1 year was 1.8%. The correlation between measurements obtained by two different individuals on identical images was 0.92, with a mean variation of 3.0%. Echogenicity was not significantly affected by type of scanner or probe frequency, but varied inversely with gain. However, the effect of gain was very small within the useful range. Water loading after an overnight fast increased echogenicity in all cases, with a mean increase of 6.4%. Echogenicity of normal kidneys was significantly less than that of the liver (range, 0.810 to 0.987), and in clinical sonograms analyzed retrospectively but blindly, echogenicity correlated with the qualitative gradations of echogenicity originally assigned. The most echogenic kidneys were 62% brighter than normal kidneys, many times greater than the variability of the measurement. We conclude that quantification of renal cortical echogenicity is feasible and reproducible and may be useful in detecting and following renal disease. Echogenicity of the renal cortex is less than that of the liver in healthy subjects and is influenced by the state of diuresis.

Why is p53 acetylated?

Recent studies suggest that acetylation of the p53 tumor suppressor protein is not important for its DNA binding activity, as was previously thought. We discuss here a number of theories as to how this modification may serve to regulate the protein's functions.

Tehao functions in the Toll pathway in Drosophila melanogaster: possible roles in development and innate immunity.

Toll and related proteins play important roles in innate immunity in both invertebrates and vertebrates. In Drosophila melanogaster, Tehao shares a striking similarity in its intracellular domain with Toll. In this paper, we show that Tehao is expressed throughout development and appears to be glycosylated. In transiently transfected cells, Tehao activated both Dorsal and the transcription of endogenous drosomycin and metchnikowin genes. Purified recombinant Tehao interacted specifically in vitro not only with the Pelle protein kinase, but also with the Toll intracytoplasmic domain. Remarkably, Tehao was found to activate Dorsal-dependent transcription in a synergistic manner with Toll, as well as Pelle in co-transfected cells. Thus, Tehao, alone or with Toll as a multimeric complex, has the potential to participate in both the development and innate immune responses of Drosophila.

Phosphorylation of CPEB by Eg2 mediates the recruitment of CPSF into an active cytoplasmic polyadenylation complex.

The release of Xenopus oocytes from prophase I arrest is largely driven by the cytoplasmic polyadenylation-induced translation of dormant maternal mRNAs. Two cis elements, the CPE and the hexanucleotide AAUAAA, and their respective binding factors, CPEB and a cytoplasmic form of CPSF, control polyadenylation. The most proximal stimulus for polyadenylation is Eg2-catalyzed phosphorylation of CPEB serine 174. Here, we show that this phosphorylation event stimulates an interaction between CPEB and CPSF. This interaction is direct, does not require RNA tethering, and occurs through the 160 kDa subunit of CPSF. Eg2-stimulated and CPE-dependent polyadenylation is reconstituted in vitro using purified components. These results demonstrate that the molecular function of Eg2-phosphorylated CPEB is to recruit CPSF into an active cytoplasmic polyadenylation complex.

Structural basis for signal transduction by the Toll/interleukin-1 receptor domains.

Toll-like receptors (TLRs) and the interleukin-1 receptor superfamily (IL-1Rs) are integral to both innate and adaptive immunity for host defence. These receptors share a conserved cytoplasmic domain, known as the TIR domain. A single-point mutation in the TIR domain of murine TLR4 (Pro712His, the Lps(d) mutation) abolishes the host immune response to lipopolysaccharide (LPS), and mutation of the equivalent residue in TLR2, Pro681His, disrupts signal transduction in response to stimulation by yeast and gram-positive bacteria. Here we report the crystal structures of the TIR domains of human TLR1 and TLR2 and of the Pro681His mutant of TLR2. The structures have a large conserved surface patch that also contains the site of the Lps(d) mutation. Mutagenesis and functional studies confirm that residues in this surface patch are crucial for receptor signalling. The Lps(d) mutation does not disturb the structure of the TIR domain itself. Instead, structural and functional studies indicate that the conserved surface patch may mediate interactions with the down-stream MyD88 adapter molecule, and that the Lps(d) mutation may abolish receptor signalling by disrupting this recruitment.