Off-line pyrolysis was used to liberate lignin moieties from dung and soil and, after trimethylsilylation, the delta(13)C values of these derivatives were determined by gas chromatography-combustion-isotope ratio mass spectrometry. Initial delta(13)C values determined for 4-vinylphenol, syringol, 4-vinylguaiacol, 4-acetylsyringol, 4-vinylsyringol, 4-(2-Z-propenyl)syringol, 4-(2-E-propenyl)syringol and 4-(2-propenone)syringol pyrolysis products of the lignin polyphenol structure from C(4) (delta(13)C(bulk) = -12.6%) and C(3) (delta(13)C(bulk) = -30.1 per thousand) dung confirmed the robust and reproducible nature of the off-line preparation technique. C(4) dung was used as a treatment in a randomised field experiment to assess the short-term sequestration of dung carbon in managed grasslands. Since lignin was on average 3.5 per thousand depleted in (13)C compared with bulk dung delta(13)C values, this may have resulted in an under-estimation of dung C incorporation based on bulk delta(13)C values. Therefore, an investigation of the compound-specific delta(13)C values of dung-derived lignin moieties extracted from soils sampled up to 372 days was undertaken. Delta(13)C values between lignin moieties extracted from treated and untreated soils showed that dung-derived lignin was not especially resistant to degradation and suggested that individual moieties of the lignin macromolecule must: (i) move into soil, (ii) be degraded, or (iii) be transformed diagenetically at different rates. This adds to a gathering body of evidence that lignin is not particularly stable in soils, which has considerable significance for the perceived role of different biochemical components in the cycling of C in soils. Copyright (c) 2008 John Wiley & Sons, Ltd.

Dungait JA, Stear NA, van Dongen BE, Bol R, Evershed RP.

Organic Geochemistry Unit, Bristol Biogeochemistry Research Centre, School of Chemistry, Cantocks Close, Bristol BS8 1TS, UK.

The Editors welcome topical correspondence from readers relating to articles published in the Journal. Responses should be sent electronically via the BJS website (www.bjs.co.uk). All letters will be reviewed and,if approved,appear on the website. A selection of these will be edited and published in the Journal. Letters must be no more than 250 words in length. Copyright (c) 2008 British Journal of Surgery Society Ltd. Published by John Wiley & Sons, Ltd.

Robertson AG, Dunn LJ, Shenfine J, Karat D, Griffin SM.

Northern Oeosphago‐Gastric Cancer Unit, Royal Victoria Infirmary, Newcastle Upon Tyne Hospitals NHS Trust, Queen Victoria Road, Newcastle upon Tyne NE1 4LP, UK.

The effect of a recent vegetation change (<100 years) from C(4) grassland to C(3) woodland in central Queensland, Australia, on soil organic matter (SOM) composition and SOM dynamics has been investigated using a novel coupled thermogravimetry-differential scanning calorimetry-quadrupole.mass spectrometry-isotope ratio mass spectrometry (TG-DSC-QMS-IRMS) system. TG-DSC-QMS-IRMS distinguishes the C isotope composition of discrete SOM pools, showing changes in labile, recalcitrant and refractory carbon in the bulk soil and particle size fractions which track the vegetation changes. Analysis of evolved gases (by QMS) from thermal decomposition, rather than observed weight loss, proved essential in determining the temperature at which SOM decomposes, because smectite and kaolinite clays contribute to observed weight losses. The delta(13)C analyses of the CO(2) evolved at different temperatures for bulk soil and particle size-separates showed that most of the labile SOM under the more recent woody vegetation was C(3)-derived carbon whereas the delta(13)C values in the recalcitrant SOM showed greater C(4) contributions. This indicated a shift from grass (C(4))- to tree (C(3))-derived carbon in the woodland, which was also supported by the two-phase (13)C enrichment with depth, i.e. C(3) vegetation dominated the top soil (0-10 cm), but the C(4) contribution increased with depth (more gradual). This is perturbed by the inclusion of charcoal from forest fires ((14)C age incursions) and by the deep incorporation of C(3) carbon due to root penetration. Copyright (c) 2008 John Wiley & Sons, Ltd.

Lopez-Capel E, Krull ES, Bol R, Manning DA.

School of Civil Engineering and Geosciences, Newcastle University, Newcastle upon Tyne NE1 7RU, UK.

When the Department of Pathology of the Radboud University Nijmegen Medical Centre in The Netherlands moved from its old to its new premises, the demolition of the marble post-mortem tables was impending, taking with it the Department\’s (physical) memory. To keep the importance of this memory and of the post-mortem itself on everyone\’s minds in the new building, the artist Piet Hein Eek was invited to incorporate these post-mortem tables into a work of art. This became a triptych: the three post-mortem tables were stood upright against the wall behind a mounted double sheet of glass, containing screen prints of enlarged microscopic images. The two emblems of pathology-post-mortem tables and double glass slides containing specimens-were thus united into a fascinating work of art: a true memento of the post-mortem, re-embedded in the contemporary world by its design. Copyright (c) 2008 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

van Speybroeck D, van Krieken J, van Daal W, Ruiter D.

Curator of the Radboud University Nijmegen Medical Centre Art Collection, Nijmegen, The Netherlands.

Litter-dwelling (epigeic) Lumbricus rubellus and soil-dwelling (endogeic) Allolobophora chlorotica earthworms were observed aggregating under C(3) (delta(13)C = -31.3 per thousand; delta(15)N = 10.7 per thousand) and C(4) (delta(13)C = -12.6 per thousand; delta(15)N = 7.5 per thousand) synthetic dung pats applied to a temperate grassland (delta(13)C = -30.3 per thousand; delta(15)N = 5.7 per thousand) in an experiment carried out for 372 days. Bulk delta(13)C values of earthworms collected from beneath either C(3) or C(4) dung after 28, 56, 112 and 372 days demonstrated that (i) L. rubellus beneath C(4) dung were significantly (13)C-enriched after 56 days (delta(13)C = -23.8 per thousand) and 112 days (delta(13)C = -22.4 per thousand) compared with those from C(3) dung treatments (56 days, delta(13)C = -26.5 per thousand; 112 days, delta(13)C = -27.0 per thousand), and (ii) A. chlorotica were 2.1 per thousand (13)C-enriched (delta(13)C = -24.2 per thousand) relative to those from C(3) dung (delta(13)C = -26.3 per thousand) treatments after 372 days. Bulk delta(15)N values did not suggest significant uptake of dung N by either species beneath C(3) or C(4) dung, but showed that the endogeic species (total mean delta(15)N = 3.3 per thousand) had higher delta(15)N values than the epigeic species (total mean delta(15)N = 5.4 per thousand). Although the two species exhibited similar fatty acid profiles, individual fatty acid delta(13)C values revealed extensive routing of dietary C into body tissue of L. rubellus, but minor incorporation into A. chlorotica. In particular, the direct incorporation of microbial biomarker fatty acids (iC(17:0), aC(17:0)) from (13)C-labelled dung in situ, the routing of dung C into de novo synthesised compounds (${\\rm C}_{20:4_\\omega 6}$, ${\\rm C}_{20:5_\\omega 3}$), and the assimilation of essential fatty acids (${\\rm C}_{18:1_\\omega 9}$, ${\\rm C}_{18:1_\\omega 7}$, ${\\rm C}_{18:2_\\omega 6}$, ${\\rm C}_{18:3_\\omega 3}$) derived from dung, were determined. Copyright (c) 2008 John Wiley & Sons, Ltd.

Dungait JA, Briones MJ, Bol R, Evershed RP.

Organic Geochemistry Unit, Bristol Biogeochemistry Research Centre, University of Bristol, School of Chemistry, Cantocks Close, Bristol BS8 1TS, UK.

Large old trees are the dominant primary producers of native pine forest, but their influence on spatial patterns of soil properties and potential feedback to tree regeneration in their neighbourhood is poorly understood. We measured stable isotopes of carbon (delta(13)C) and nitrogen (delta(15)N) in soil and litter taken from three zones of influence (inner, middle and outer zone) around the trunk of freestanding old Scots pine (Pinus sylvestris L.) trees, to determine the trees\’ influence on below-ground properties. We also measured delta(15)N and delta(13)C in wood cores extracted from the old trees and from regenerating trees growing within their three zones of influence. We found a significant and positive gradient in soil delta(15)N from the inner zone, nearest to the tree centre, to the outer zone beyond the tree crown. This was probably caused by the higher input of (15)N-depleted litter below the tree crown. In contrast, the soil delta(13)C did not change along the gradient of tree influence. Distance-related trends, although weak, were visible in the wood delta(15)N and delta(13)C of regenerating trees. Moreover, the wood delta(15)N of small trees showed a weak negative relationship with soil N content in the relevant zone of influence. Our results indicate that large old trees control below-ground conditions in their immediate surroundings, and that stable isotopes might act as markers for the spatial and temporal extent of these below-ground effects. Copyright (c) 2008 John Wiley & Sons, Ltd.

Weber P, Bol R, Dixon L, Bardgett RD.

Swiss Federal Research Institute WSL, Zuercherstrasse 111, CH‐8903 Birmensdorf, Switzerland.

BACKGROUND.: By using the age-adjusted Charlson comorbidity index (ACCI), the authors characterized the impact of age and comorbidity on disease progression and overall survival after radical cystectomy (RC) for transitional cell carcinoma of the bladder. Also evaluated was whether ACCI was associated with clinicopathologic and treatment characteristics. METHODS.: The authors evaluated 1121 patients treated by RC for transitional cell carcinoma of the bladder at a single institution (1990-2004). Logistic regression was used to determine the relation between ACCI and clinical features. They evaluated the association between ACCI and overall and progression-free survival by using multivariate survival-time models with pathologic stage and nodal status as covariates. RESULTS.: ACCI scores increased during the study period (P = .009). Extravesical disease was present in 43% of patients with ACCI </=2, 49% with ACCI 3-5, and 56% with ACCI >5 (P = .051). Despite their higher prevalence of extravesical disease, patients with higher ACCI were less likely to have lymph-node dissection (odds ratio, 0.55 and 0.35, respectively, for ACCI 3-5 and >5 vs </=2; P = .005), and when it was performed, fewer lymph nodes were evaluated (P < .0005). Patients with higher ACCI were also less likely to have postoperative chemotherapy (odds ratio, 0.70 and 0.66, respectively, for ACCI 3-5 and >5 vs </=2; P = .04). Higher ACCI was significantly associated with lower overall (P < .005) but not recurrence-free (P = .17) survival after RC. CONCLUSIONS.: Age and comorbidity among patients who underwent RC at a cancer referral hospital increased with time. Both age and comorbidity were associated with treatment selection and survival and should, therefore, be considered when comparing outcomes after RC. Cancer 2008. (c) 2008 American Cancer Society.

Koppie TM, Serio AM, Vickers AJ, Vora K, Dalbagni G, Donat SM, Herr HW, Bochner BH.

Department of Urology, Memorial Sloan Kettering Cancer Center, New York, New York.

Improvement of synthetic bone graft substitutes as suitable alternatives to a patient\’s own bone graft remains a challenge in biomaterials research. Our goal was to answer the question of whether improved osteoinductivity of a material would also translate to better bone-healing orthotopically. Three porous biphasic calcium phosphate (BCP) ceramics (BCPA, BCPB, and BCPC), consisting of hydroxyapatite and beta-tricalcium phosphate, a porous biphasic calcium phosphate ceramic reinforced with a bioresorbable polylactic acid to improve its mechanical properties (BCPC+), a pure hydroxyapatite ceramic (HA), and a carbonated apatite ceramic (CA) were implanted intramuscularly and orthotopically by using a transverse process model in 11 goats for 12 weeks. BCPA and BCPB had similar chemical composition but differed in their microstructure. BCPB was not osteoinductive at all, but BCPA induced ectopic bone formation in 9 of 11 animals. Orthotopically, BCPA performed better than BCPB in both the amount and rate of bone formation. BCPC, similar to BCPA structurally and physicochemically, showed comparable results ectopically and orthotopically. Addition of resorbable polymer to BCPC made the material less osteoinductive (4 of 11 animals) and delayed bone formation orthotopically. Neither HA nor CA were osteoinductive, and their orthotopic performance was inferior to the osteoinductive ceramics. The results of the present study showed that material-derived osteoinduction significantly enhanced bone healing orthotopically, and that this material property appeared more sensitive for predicting orthotopic performance than physicochemical and structural characteristics. (c) 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res.

Habibovic P, Kruyt MC, Juhl MV, Clyens S, Martinetti R, Dolcini L, Theilgaard N, van Blitterswijk CA.

University of Twente, Department of Tissue Regeneration, Enschede, The Netherlands.

BACKGROUND.: The objective of the current study was to assess the staging accuracy and prognostic role of preoperative endoscopic ultrasound (EUS) and computed tomography (CT) in patients with locally advanced gastric cancer (LAGC) after neoadjuvant chemotherapy. METHODS.: Presurgical LAGC patients underwent EUS and CT before and after 3 cycles of neoadjuvant chemotherapy. Chemotherapy was comprised of docetaxel (at a dose of 36 mg/m(2)) and cisplatin (at a dose of 40 mg/m(2)) on Days 1 and 8 of a 3-week cycle. RESULTS.: Forty patients were enrolled in the study. After chemotherapy, the accuracy of EUS and CT was found to be 47% and 57%, respectively for T classification (P = .22) and 39% and 37%, respectively for N classification (P > .99). The 3-year overall survival (OS) rate for patients downstaged with EUS for T and/or N classification was greater than that for nondownstaged patients (69% vs 41%; P = .05). The 2-year recurrence-free survival (RFS) rate was also better for the EUS-downstaged patients than for the nondownstaged patients (77% vs 47%; P = .04). On multivariate analysis, EUS downstaging was found to be correlated with OS (hazards ratio [HR] of 0.12; P = .04), and was correlated with RFS with borderline statistical significance (HR of 0.27; P = .07). The differences in OS and RFS between the patients downstaged with CT and those not downstaged were not found to be statistically significant. CONCLUSIONS.: Restaging by EUS and CT after neoadjuvant chemotherapy in patients with LAGC was found to be inaccurate. However, T and/or N downstaging by EUS was found to be correlated with better OS and RFS. Thus, downstaging by EUS may be a useful clinical parameter with which to predict a better outcome for LAGC patients. Cancer 2008. (c) 2008 American Cancer Society.

Park SR, Lee JS, Kim CG, Kim HK, Kook MC, Kim YW, Ryu KW, Lee JH, Bae JM, Choi IJ.

Research Institute and Hospital, National Cancer Center, Goyang, Gyeonggi, Republic of Korea.

BACKGROUND.: Analysis of different tumor markers by reverse transcriptase-polymerase chain reaction (RT-PCR) in fine-needle aspiration samples of thyroid nodules has been studied with the objective of improving the accuracy of the preoperative diagnosis of thyroid lesions. The aim of the current study was to investigate thyroid fine-needle aspiration samples inadequate for RT-PCR analysis and to determine whether there is a correlation between their proportion and the method of sampling used or the greatest dimension of the nodules. METHODS.: A total of 350 fine-needle aspiration samples from patients with thyroid nodules were analyzed. After the aspirate was smeared for conventional cytology, the leftover material in the needle was used for RT-PCR analysis in 1 group of 175 patients. In another group of 175 patients, a separate puncture was performed to obtain material for RT-PCR analysis only. Samples were considered adequate for RT-PCR analysis if the expression of both glyceraldehyde-3-phosphate dehydrogenase and thyroglobulin was found by RT-PCR. RESULTS.: In total, 61 (17.4%) samples inadequate for RT-PCR were detected. All 12 samples that were inadequate for cytologic diagnosis were also found to be inadequate for RT-PCR analysis. The proportion of inadequate samples for RT-PCR was found to be significantly higher in samples taken from leftover material in the needle (21.7%) then in samples from a separate puncture (13.1%) (P = .049). No statistically significant correlation between the adequacy of samples for RT-PCR and the largest dimension of the nodule was found. CONCLUSIONS.: The proportion of samples inadequate for RT-PCR was found to be higher in samples taken from leftover material in the needle than in samples obtained from a separate puncture. Cancer (Cancer Cytopathol) 2008. (c) 2008 American Cancer Society.

Samija I, Matesa N, Lukac J, Kusic Z.

Department of Oncology and Nuclear Medicine, University Hospital “Sestre milosrdnice,” Zagreb, Croatia.