Toluene dioxygenase-catalysed oxidation route to angular cis-monohydrodiols and other bioproducts from bacterial metabolism of 1,2-dihydrobenzocyclobutene and derivatives

A mutant strain (UV4) of the soil bacterium Pseudomonas putida, containing toluene dioxygenase, has been used in the metabolic oxidation of 1,2-dihydrobenzocyclobutene 12 dagger and the related substrates 1,2-dihydrobenzocyclobuten-1-ol 13 and biphenylene 33. Stable angular cis-monohydrodiol metabolites (1R,2S)-bicyclo[4.2.0]octa-3,5-diene-1,2 7, (1S,2S,8S)-bicyclo[4.2.0]octa-3,5-diene-1,2,8-triol 8 and biphenylene-cis-1,8b-diol 9, isolated from each of these substrates, have been structurally and stereochemically assigned. The structure, enantiopurity and absolute configuration of the other cis-diol metabolites, (2R,3S)-bicyclo[4.2.0]octa-1(6),4-diene-2,3-diol 14 and cis-1,2-dihydroxy-1,2-dihydrobenzocyclobutene 16, and the benzylic oxidation bioproducts, 1,2-dihydrobenzocyclobuten-1-ol 13, 1,2-dihydrobenzocyclobuten-1-one 15 and 2-hydroxy-1,2-dihydrobenzocyclobuten-1-one 17, obtained from 1,2-dihydrobenzocyclobutene and 1,2-dihydrobenzocyclobuten-1-ol, have been determined with the aid of chiral stationary-phase HPLC, NMR and CD spectroscopy, and stereochemical correlation. X-Ray crystallographic methods have been used in the determination of absolute configuration of the di-camphanates 27 (from diol 7) and 32 (from diol 9), and the di-MTPA ester 29 (from diol 14) of the corresponding cis-diol metabolites. The metabolic sequence involved in the formation of bioproducts derived from 1,2-dihydrobenzocyclobutene 12 has been investigated.

Early Holocene M similar to 6 explosive eruption from Plosky volcanic massif (Kamchatka) and its tephra as a link between terrestrial and marine paleoenvironmental records

We report tephrochronological and geochemical data on early Holocene activity from Plosky volcanic massif in the Kliuchevskoi volcanic group, Kamchatka Peninsula. Explosive activity of this volcano lasted for similar to 1.5 kyr, produced a series of widely dispersed tephra layers, and was followed by profuse low-viscosity lava flows. This eruptive episode started a major reorganization of the volcanic structures in the western part of the Kliuchevskoi volcanic group. An explosive eruption from Plosky (M similar to 6), previously unstudied, produced tephra (coded PL2) of a volume of 10-12 km(3) (11-13 Gt), being one of the largest Holocene explosive eruptions in Kamchatka. Characteristic diagnostic features of the PL2 tephra are predominantly vitric sponge-shaped fragments with rare phenocrysts and microlites of plagioclase, olivine and pyroxenes, medium- to high-K basaltic andesitic bulk composition, high-K, high-Al and high-P trachyandesitic glass composition with SiO2 = 57.5-59.5 wt%, K2O = 2.3-2.7 wt%, Al2O3 = 15.8-16.5 wt%, and P2O5 = 0.5-0.7 wt%. Other diagnostic features include a typical subduction-related pattern of incompatible elements, high concentrations of all REE (> 10x mantle values), moderate enrichment in LREE (La/Yb similar to 5.3), and non-fractionated mantle-like pattern of LILE. Geochemical fingerprinting of the PL2 tephra with the help of EMP and LA-ICP-MS analyses allowed us to map its occurrence in terrestrial sections across Kamchatka and to identify this layer in Bering Sea sediment cores at a distance of > 600 km from the source. New high-precision C-14 dates suggest that the PL2 eruption occurred similar to 10,200 cal BP, which makes it a valuable isochrone for early Holocene climate fluctuations and permits direct links between terrestrial and marine paleoenvironmental records. The terrestrial and marine C-14 dates related to the PL2 tephra have allowed us to estimate an early Holocene reservoir age for the western Bering Sea at 1,410 +/- A 64 C-14 years. Another important tephra from the early Holocene eruptive episode of Plosky volcano, coded PL1, was dated at 11,650 cal BP. This marker is the oldest geochemically characterized and dated tephra marker layer in Kamchatka to date and is an important local marker for the Younger Dryas-early Holocene transition. One more tephra from Plosky, coded PL3, can be used as a marker northeast of the source at a distance of similar to 110 km.

Determination of geographical origin of distillers dried grains and solubles using isotope ratio mass spectrometry

In recent years distillers dried grains and solubles (DDGS), co-products of the bio-ethanol and beverages industries, have become globally traded commodity for the animal feed sector. As such it is becoming increasingly important to be able to trace the geographical origin of commodities in case of a contamination incident or authenticity issue arise. In this study, 137 DDGS samples from a range of different geographical origins (China, USA, Canada and European Union) were collected and analyzed. Isotope ratio mass spectrometry (IRMS) was used to analyze the DDGS for ²H/¹H, ¹³C/¹²C, ¹⁵N/¹⁴N, ¹⁸O/¹⁶O and ³⁴S/³²S isotope ratios which can vary depending on geographical origin and processing. Univariate and multivariate statistical techniques were employed to investigate the feasibility of using the IRMS data to determine botanical and geographical origin of the DDGS. The results indicated that this commodity could be differentiated according to their place of origin by the analysis of stable isotopes of hydrogen, carbon, nitrogen and oxygen but not with sulfur. By adding data to the models produced in this study, potentially an isotope databank could be set up for traceability procedures for DDGS, similar to the one established already for wine which will help in feed and food security issues arising worldwide.

The prevalence of caries in patients referred for orthodontic assessment in Northern Ireland – A retrospective study

Objectives: To determine the prevalence of untreated carious lesions in permanent teeth in patients (under the age of 18) referred for an orthodontic assessment in specialist practice. In addition, the figures shall be compared with national data for Northern Ireland (as outlined in the recent Child Dental Health Survey 2013)
The Gold standard would be that 100% of patients would be caries free upon presentation.

Methods: The clinical records and radiographs (OPT of quality grading 1 or 2) of 337 patients were reviewed. This encompassed patients who had an orthodontic assessment carried out in specialist practice over a 6 month period (following referral from their general dental practitioner)

Results: A total of 337 patient records were examined. Of these, 149 were male (44.2%) and 188 were female (55.8%), with an age range of 7-17 years at the time of new patient assessment. It was found that 36 patients (10.7%) had evidence (clinical and/or radiographic) of active and untreated dental caries. Of those affected, 14 were male and 22 were female.
Breaking the data down in terms of age, we can also get some indication as to how this cohort compares with national data for Northern Ireland :⃰

7-10 years (Mean = 9.3) = 14.3% caries (versus NI average of 6% for 8 year olds)
11-13 years (Mean = 12.1) = 10.1% caries, (versus NI average of 16% for 12 year olds)
14-17 years (Mean = 15.2) = 9.1% caries (versus NI average of 15% for 15 year olds)

⃰using the diagnostic threshold “Decay into dentine (visual dentine caries excluded)”


Conclusion: In this sample group, a total of 10.7% of patients (9.4% of males, 11.7% of females) presented with evidence of undiagnosed caries upon being assessed as a new patient in specialist orthodontic practice. Hence, the gold standard was not met.

Glucocorticoid induced muscle atrophy is associated with upregulation of myostatin gene expression

The mechanisms by which excessive glucocorticoids cause muscular atrophy remain unclear. We previously demonstrated that dexamethasone increases the expression of myostatin, a negative regulator of skeletal muscle mass, in vitro. In the present study, we tested the hypothesis that dexamethasone-induced muscle loss is associated with increased myostatin expression in vivo. Daily administration (60, 600, 1,200 micro g/kg body wt) of dexamethasone for 5 days resulted in rapid, dose-dependent loss of body weight (-4.0, -13.4, -17.2%, respectively, P <0.05 for each comparison), and muscle atrophy (6.3, 15.0, 16.6% below controls, respectively). These changes were associated with dose-dependent, marked induction of intramuscular myostatin mRNA (66.3, 450, 527.6% increase above controls, P <0.05 for each comparison) and protein expression (0.0, 260.5, 318.4% increase above controls, P <0.05). We found that the effect of dexamethasone on body weight and muscle loss and upregulation of intramuscular myostatin expression was time dependent. When dexamethasone treatment (600 micro g. kg-1. day-1) was extended from 5 to 10 days, the rate of body weight loss was markedly reduced to approximately 2% within this extended period. The concentrations of intramuscular myosin heavy chain type II in dexamethasone-treated rats were significantly lower (-43% after 5-day treatment, -14% after 10-day treatment) than their respective corresponding controls. The intramuscular myostatin concentration in rats treated with dexamethasone for 10 days returned to basal level. Concurrent treatment with RU-486 blocked dexamethasone-induced myostatin expression and significantly attenuated body loss and muscle atrophy. We propose that dexamethasone-induced muscle loss is mediated, at least in part, by the upregulation of myostatin expression through a glucocorticoid receptor-mediated pathway.

Comparisons of plankton at 59 degrees N and 37 degrees N during two Lagrangian drift experiments in the North Atlantic in June and July 1996

Data are summarised for two Lagrangian experiments in the North Atlantic in early summer 1996. At 59 degreesN 20 degreesW, plankton dynamics was studied in an SF, tracer release experiment within a mesoscale eddy over a 9-day period. At 37 degreesN 20 degreesW, a second experiment followed a drifting buoy for 7 days. The data obtained in these two experiments have been averaged for 3 depth strata; the euphotic zone, the surface mixed layer (SML), and the seasonal thermocline immediately beneath the surface mixed layer. At 59 degreesN, the euphotic zone was only marginally deeper than the SML, but at 37 degreesN the SML was ca 30 m and the euphotic depth was ca 110 m. At 37 degreesN, nutrient concentrations in the SML were low but significant new production occurred in the thermocline because of light penetration into the nutricline. The particulate organic carbon (POC) concentration of the SML at 59 degreesN was 13-15 mu mol C kg(-1), but at 37 degreesN POC concentrations were 4 mu mol C kg(-1). These POC measurements include biota and detritus. As a way of investigating latitudinal differences in the plankton communities, estimates have been made of the carbon and nitrogen content of phytoplankton, bacterioplankton, microzooplankton and mesozooplankton. At both 59 degreesN and 37 degreesN, phytoplankton was the largest component, accounting for ca 50% of the planktonic biomass in the SML. At 59 degreesN, microzooplankton was 16% of the planktonic carbon, but at 37 degreesN this reduced to 8% of the total. Mesozooplankton was a relatively constant proportion (ca 20%) of the planktonic carbon in the SML at both 59 degreesN and 37 degreesN. Bacterioplankton was 14% of the biomass at 59 degreesN, increasing to 24% in the microbial loop-dominated system at 37 degreesN. Mean carbon fixation rate in the oligotrophic southern station was 24% of that at the north, with more carbon fixation below the SML at 37 degreesN than at 59 degreesN. Respiration rates showed little variation with latitude, and the rates at 37 degreesN were 80% of those at 59 degreesN. Nitrate and ammonium uptake rates were very low in the oligotrophic conditions in the SML at 37 degreesN, but nitrate uptake in the euphotic zone was comparable to that at 59 degreesN. Ammonium uptake by phytoplankton was also significantly greater at 37 degreesN, in both the euphotic zone and thermocline, but uptake in the SML was only 20% of that in the SML at 59 degreesN. (C) 2001 Elsevier Science Ltd. All rights reserved.

Updated parameters for the transiting exoplanet WASP-3b using RISE, a new fast camera for the Liverpool Telescope

Some of the first results are reported from RISE - a new fast camera mounted on the Liverpool Telescope primarily designed to obtain high time-resolution light curves of transiting extrasolar planets for the purpose of transit timing. A full and partial transit of WASP-3 are presented, and a Markov-Chain Monte Carlo analysis is used to update the parameters from the discovery paper. This results in a planetary radius of 1.29(-0.12)(+0.05) R-J and therefore a density of 0.82(-0.09)(+0.14) rho(J), consistent with previous results. The inclination is 85.06(-0.15)(+0.16) deg, in agreement (but with a significant improvement in the precision) with the previously determined value. Central transit times are found to be consistent with the ephemeris given in the discovery paper; however, a new ephemeris calculated using the longer baseline results in T-c(0) = 2 454 605.55915 +/- 0.00023 HJD and P = 1.846835 +/- 0.000002 days.

An in vitro study of the radiobiological effects of flattening filter free radiotherapy treatments

Flattening filter free (FFF) linear accelerators allow for an increase in instantaneous dose-rate of the x-ray pulses by a factor of 2-6 over the conventional flattened output. As a result, radiobiological investigations are being carried out to determine the effect of these higher dose-rates on cell response. The studies reported thus far have presented conflicting results, highlighting the need for further investigation. To determine the radiobiological impact of the increased dose-rates from FFF exposures a Varian Truebeam medical linear accelerator was used to irradiate two human cancer cell lines in vitro, DU-145 prostate and H460 non-small cell lung, with both flattened and FFF 6 MV beams. The fluence profile of the FFF beam was modified using a custom-designed Nylon compensator to produce a similar dose profile to the flattened beam (6X) at the cell surface but at a higher instantaneous dose-rate. For both cell lines there appeared to be no significant change in cell survival. Curve fitting coefficients for DU145 cells irradiated with constant average dose-rates were 6X: alpha = 0.09 +/- 0.03, beta = 0.03 +/- 0.01 and 6FFF: alpha = 0.14 +/- 0.13, beta = 0.03 +/- 0.02 with a significance of p = 0.75. For H460 cells irradiated with the same instantaneous dose-rate but different average dose-rate the fit coefficients were 6FFF (low dose-rate): alpha = 0.21 +/- 0.11, 0.07 +/- 0.02 and 6FFF (high dose-rate): alpha = 0.21 +/- 0.16, 0.07 +/- 0.03, with p = 0.79. The results indicate that collective damage behaviour does not occur at the instantaneous dose-rates investigated here and that the use of either modality should result in the same clinical outcome, however this will require further validation in vivo.

Prevalence of human papillomavirus in women attending cervical screening in the UK and Ireland:New data from northern Ireland and a systematic review and meta-analysis

There is substantial international variation in human papillomavirus (HPV) prevalence; this study details the first report from Northern Ireland and additionally provides a systematic review and meta-analysis pooling the prevalence of high-risk (HR-HPV) subtypes among women with normal cytology in the UK and Ireland. Between February and December 2009, routine liquid based cytology (LBC) samples were collected for HPV detection (Roche Cobas® 4800 [PCR]) among unselected women attending for cervical cytology testing. Four electronic databases, including MEDLINE, were then searched from their inception till April 2011. A random effects meta-analysis was used to calculate a pooled HR-HPV prevalence and associated 95% confidence intervals (CI). 5,712 women, mean age 39 years (±SD 11.9 years; range 20-64 years), were included in the analysis, of which 5,068 (88.7%), 417 (7.3%) and 72 (1.3%) had normal, low, and high-grade cytological findings, respectively. Crude HR-HPV prevalence was 13.2% (95% CI, 12.7-13.7) among women with normal cytology and increased with cytological grade. In meta-analysis the pooled HR-HPV prevalence among those with normal cytology was 0.12 (95% CIs, 0.10-0.14; 21 studies) with the highest prevalence in younger women. HPV 16 and HPV 18 specific estimates were 0.03 (95% CI, 0.02-0.05) and 0.01 (95% CI, 0.01-0.02), respectively. The findings of this Northern Ireland study and meta-analysis verify the prevalent nature of HPV infection among younger women. Reporting of the type-specific prevalence of HPV infection is relevant for evaluating the impact of future HPV immunization initiatives, particularly against HR-HPV types other than HPV 16 and 18. J. Med. Virol. 85:295-308, 2013. © 2012 Wiley Periodicals, Inc. Copyright © 2012 Wiley Periodicals, Inc.

Differences in cancer awareness and beliefs between Australia, Canada, Denmark, Norway, Sweden and the UK (the International Cancer Benchmarking Partnership):Do they contribute to differences in cancer survival?

Background:There are wide international differences in 1-year cancer survival. The UK and Denmark perform poorly compared with other high-income countries with similar health care systems: Australia, Canada and Sweden have good cancer survival rates, Norway intermediate survival rates. The objective of this study was to examine the pattern of differences in cancer awareness and beliefs across these countries to identify where these might contribute to the pattern of survival.Methods:We carried out a population-based telephone interview survey of 19 079 men and women aged =50 years in Australia, Canada, Denmark, Norway, Sweden and the UK using the Awareness and Beliefs about Cancer measure.Results:Awareness that the risk of cancer increased with age was lower in the UK (14%), Canada (13%) and Australia (16%) but was higher in Denmark (25%), Norway (29%) and Sweden (38%). Symptom awareness was no lower in the UK and Denmark than other countries. Perceived barriers to symptomatic presentation were highest in the UK, in particular being worried about wasting the doctor's time (UK 34%; Canada 21%; Australia 14%; Denmark 12%; Norway 11%; Sweden 9%).Conclusion:The UK had low awareness of age-related risk and the highest perceived barriers to symptomatic presentation, but symptom awareness in the UK did not differ from other countries. Denmark had higher awareness of age-related risk and few perceived barriers to symptomatic presentation. This suggests that other factors must be involved in explaining Denmark's poor survival rates. In the UK, interventions that address barriers to prompt presentation in primary care should be developed and evaluated. © 2013 Cancer Research UK. All rights reserved.

Ages of 24 widespread tephras erupted since 30,000 years ago in New Zealand, with re-evaluation of the timing and palaeoclimatic implications of the Lateglacial cool episode recorded at Kaipo bog

Tephras are important for the NZ-INTIMATE project because they link all three records comprising the composite inter-regional stratotype developed for the New Zealand climate event stratigraphy (NZ-CES). Here we firstly report new calendar ages for 24 widespread marker tephras erupted since 30,000 calendar (cal.) years ago in New Zealand to help facilitate their use as chronostratigraphic dating tools for the NZ-CES and for other palaeoenvironmental and geological applications. The selected tephras comprise 12 rhyolitic tephras from Taupo, nine rhyolitic tephras from Okataina, one peralkaline rhyolitic tephra from Tuhua, and one andesitic tephra each from Tongariro and Egmont/Taranaki volcanic centres. Age models for the tephras were obtained using three methods: (i) C-based wiggle-match dating of wood from trees killed by volcanic eruptions (these dates published previously); (ii) flexible depositional modelling of a high-resolution C-dated age-depth sequence at Kaipo bog using two Bayesian-based modelling programs, Bacon and OxCal's P_Sequence function, and the IntCal09 data set (with SH offset correction-44±17yr); and (iii) calibration of C ages using OxCal's Tau_Boundary function and the SHCal04 and IntCal09 data sets. Our preferred dates or calibrated ages for the 24 tephras are as follows (youngest to oldest, all mid-point or mean ages of 95% probability ranges): Kaharoa AD 1314±12; Taupo (Unit Y) AD 232±10; Mapara (Unit X) 2059±118cal.yrBP; Whakaipo (Unit V) 2800±60cal.yrBP; Waimihia (Unit S) 3401±108cal.yrBP; Stent (Unit Q) 4322±112cal.yrBP; Unit K 5111±210cal.yrBP; Whakatane 5526±145cal.yrBP; Tuhua 6577±547cal.yrBP; Mamaku 7940±257cal.yrBP; Rotoma 9423±120cal.yrBP; Opepe (Unit E) 9991±160cal.yrBP; Poronui (Unit C) 11,170±115cal.yrBP; Karapiti (Unit B) 11,460±172cal.yrBP; Okupata 11,767±192cal.yrBP; Konini (bed b) 11,880±183cal.yrBP; Waiohau 14,009±155cal.yrBP; Rotorua 15,635±412cal.yrBP; Rerewhakaaitu 17,496±462cal.yrBP; Okareka 21,858±290cal.yrBP; Te Rere 25,171±964cal.yrBP; Kawakawa/Oruanui 25,358±162cal.yrBP; Poihipi 28,446±670cal.yrBP; and Okaia 28,621±1428cal.yrBP.Secondly, we have re-dated the start and end of the Lateglacial cool episode (climate event NZce-3 in theNZ-CES), previously referred to as the Lateglacial climate reversal, as defined at Kaipo bog in eastern North Island, New Zealand, using both Bacon and OxCal P_Sequence modelling with the IntCal09 data set. The ca1200-yr-long cool episode, indicated by a lithostratigraphic change in the Kaipo peat sequence to grey mudwith lowered carbon content, and a high-resolution pollen-derived cooling signal, began 13,739±125cal.yrBP and ended 12,550±140cal.yrBP (mid-point ages of the 95% highest posterior density regions, Bacon modelling). The OxCal modelling, generating almost identical ages, confirmed these ages. The Lateglacial cool episode (ca 13.8-12.6cal.kaBP) thus overlaps a large part of the entire Antarctic Cold Reversal chronozone (ca 14.1-12.4cal.kaBP or ca 14.6-12.8cal.kaBP), and an early part of the Greenland Stadial-1 (Younger Dryas) chronozone (ca 12.9-11.7cal.kaBP). The timing of the Lateglacial cool episode at Kaipo is broadly consistent with the latitudinal patterns in the Antarctic Cold Reversal signal suggested for the New Zealand archipelago from marine and terrestrial records, and with records from southern South America. © 2012 Elsevier Ltd.

A narrow-band ultraviolet B course improves vitamin D balance and alters cutaneous CYP27A1 and CYP27B1 mRNA expression levels in haemodialysis patients supplemented with oral vitamin D

Background: Chronic kidney disease (CKD) patients on dialysis are prone to vitamin D insufficiency despite oral vitamin D supplementation. Here, we studied whether narrow-band ultraviolet B (NB-UVB) exposures improve vitamin D balance.

Methods: 14 haemodialysis patients and 15 healthy subjects receiving oral cholecalciferol 20 µg daily got nine NB-UVB exposures on the entire body. Serum 25-hydroxyvitamin D (25(OH)D) was measured by radioimmunoassay. Cutaneous mRNA expression levels of CYP27A1 and CYP27B1, two enzymes required for hydroxylation of vitamin D into its active metabolite, were also measured.

Results: The baseline serum 25(OH)D concentration was 57.6 ± 18.2 nmol/l in the CKD patients and 74.3 ± 14.8 nmol/l in the healthy subjects. The NB-UVB course increased serum 25(OH)D by 14.0 nmol/l (95% CI 8.7-19.5) and 17.0 nmol/l (CI 13.7-20.2), respectively. At baseline the CKD patients showed significantly increased CYP27B1 levels compared to the healthy subjects.

Conclusions: A short NB-UVB course is an efficient way to improve vitamin D balance in CKD patients on dialysis who are receiving oral vitamin D supplementation. The increased cutaneous CYP27B1 levels in the CKD patients suggest that the loss of renal activity of this enzyme is at least partially compensated for by the skin.