Pristine Early Eocene Wood Buried Deeply in Kimberlite from Northern Canada

We report exceptional preservation of fossil wood buried deeply in a kimberlite pipe that intruded northwestern Canada's Slave Province 53.3±0.6 million years ago (Ma), revealed during excavation of diamond source rock. The wood originated from forest surrounding the eruption zone and collapsed into the diatreme before resettling in volcaniclastic kimberlite to depths >300 m, where it was mummified in a sterile environment. Anatomy of the unpermineralized wood permits conclusive identification to the genus Metasequoia (Cupressaceae). The wood yields genuine cellulose and occluded amber, both of which have been characterized spectroscopically and isotopically. From cellulose d O and d H measurements, we infer that Early Eocene paleoclimates in the western Canadian subarctic were 12-17°C warmer and four times wetter than present. Canadian kimberlites offer Lagerstätte-quality preservation of wood from a region with limited alternate sources of paleobotanical information. © 2012 Wolfe et al.

Temperature and water potential relations of tropical Trametes and other wood-decay fungi from the indigenous forests of Zimbabwe

Three isolates each, of nine different Trametes and five other wood inhabiting basidiomycetes, were collected from the indigenous forests of Zimbabwe, and the impact of temperature (20-60 degrees C), osmotic and matric potential (-0.5 to - 8.0 MPa), and their interactions on in vitro growth compared. Generally, there was no significant difference between growth of isolates of the same species in relation to temperature. Temperature relationships of the species studied correlated well with their geographic distributions. Species occurring in hot, dry regions tolerated a wide temperature range, with some showing unusually high thermotolerance (55 degrees, T. socotrana, T. cingulata and T. cervina). There were significant intra-strain differences for individual species in relation to solute potential on glycerol-modified media. Generally, growth of ail species was better on glycerol- and KCl-modified osmotic media than on a metrically-modified medium (PEG 8000) at 25, 30 and 37 degrees. The limits for growth on the osmotic media were significantly wider than matric medium, being - 4.5 to - 5.0 and - 2.5 to - 4.5 MPa, respectively. An Irpex sp. grew at lower water potentials than all other species, with good growth at - 7.0 MPa. This study suggests that the capacity of these fungi for effective growth over a range of temperatures, osmotic and matric potentials contributes to their rapid wood decay capacities in tropical climates.

Wood degradation, and cellulase and ligninase production, by Trametes and other wood-inhabiting basidiomycetes from indigenous forests of Zimbabwe

Nine species of Trametes and five other wood inhabiting basidiomycetes, were collected from the indigenous forests of Zimbabwe and analysed for cellulases, ligninases, extracellular phenolases and wood degrading ability for the first time. Cellulase enzyme activities varied widely among the species. After 15 d growth exo-glucanase activity had increased in the majority of species whilst Biter paper activity showed the opposite trend, being greatly reduced in all species on day 15 compared to day IO. Endo-glucanase activity was relatively uniform at both sampling times. The fungi were more active against water soluble cellulose derivatives than filter paper cellulase. In all the fungi tested, cellulose activity on filter paper was significantly less than endo- and exo-glucanase activities. The highest cellulase activity was expressed by Cerrena meyenii (683 U mg(-1)) Phaeotrametes decipiens, Trametes modesta, and T. pocas also expressed relatively high cellulase activity on all types of cellulose tested. All Trametes species tested positive for extracellular phenol oxidases whilst Fomotopsis spragueii and Irpex stereoides tested negative. Ail but one of the Trametes species in the study were able to degrade two different lignin preparations in tests for lignin degradation. T. menziesii was unable to degrade both lignin preparations although it had tested positive for production of extracellular oxidase. The species in this study degraded hardwood to a greater extent than softwood. Eight of them caused more than 80% dry weight loss of wood blocks during 70 d incubation. Those fungi that expressed high cellulase activity also caused high weight loss on wood.

Dilute phosphoric acid-catalysed hydrolysis of municipal bio-waste wood shavings using autoclave parr reactor system

The visibility of using municipal bio-waste, wood shavings, as a potential feedstock for ethanol production was investigated. Dilute acid hydrolysis of wood shavings with H3PO4 was undertaken in autoclave parr reactor. A combined severity factor (CSF) was used to integrate the effects of hydrolysis times, temperature and acid concentration into a single variable. Xylose concentration reached a maximum value of 17 g/100 g dry mass corresponding to a yield of 100% at the best identified conditions of 2.5 wt.% H3PO4, 175 degrees C and 10 min reaction time corresponding to a CSF of 1.9. However, for glucose, an average yield of 30% was obtained at 5 wt.% H3PO4, 200 degrees C and 10 min. Xylose production increased with increasing temperature and acid concentration, but its transformation to the degradation product furfural was also catalysed by those factors. The maximum furfural formed was 3 g/100 g dry mass, corresponding to the 24% yield. (C) 2011 Elsevier Ltd. All rights reserved.

Human dental pulp fibroblasts express the "cold-sensing" transient receptor potential channels TRPA1 and TRPM8.

Abstract
INTRODUCTION:
Transient receptor potential (TRP) channels comprise a group of nonselective calcium-permeable cationic channels, which are polymodal sensors of environmental stimuli such as thermal changes and chemicals. TRPM8 and TRPA1 are cold-sensing TRP channels activated by moderate cooling and noxious cold temperatures, respectively. Both receptors have been identified in trigeminal ganglion neurones, and their expression in nonneuronal cells is now the focus of much interest. The aim of this study was to investigate the molecular and functional expression of TRPA1 and TRPM8 in dental pulp fibroblasts.
METHODS:
Human dental pulp fibroblasts were derived from healthy molar teeth. Gene and protein expression was determined by polymerase chain reaction and Western blotting. Cellular localization was investigated by immunohistochemistry, and TRP functionality was determined by Ca(2+) microfluorimetry.
RESULTS:
Polymerase chain reaction and Western blotting showed gene and protein expression of both TRPA1 and TRPM8 in fibroblast cells in culture. Immunohistochemistry studies showed that TRPA1 and TRPM8 immunoreactivity co-localized with the human fibroblast surface protein. In Ca(2+) microfluorimetry studies designed to determine the functionality of TRPA1 and TRPM8 in pulp fibroblasts, we showed increased intracellular calcium ([Ca(2+)](i)) in response to the TRPM8 agonist menthol, the TRPA1 agonist cinnamaldehyde, and to cool and noxious cold stimuli, respectively. The responses to agonists and thermal stimuli were blocked in the presence of specific TRPA1 and TRPM8 antagonists.
CONCLUSIONS:
Human dental pulp fibroblasts express TRPA1 and TRPM8 at the molecular, protein, and functional levels, indicating a possible role for fibroblasts in mediating cold responses in human teeth.

A comparative study of biopolymers and alum in the separation and recovery of pulp fibres from paper mill effluent by flocculation

Recovery of cellulose fibres from paper mill effluent has been studied using common polysaccharides or biopolymers such as Guar gum, Xanthan gum and Locust bean gum as flocculent. Guar gum is commonly used in sizing paper and routinely used in paper making. The results have been compared with the performance of alum, which is a common coagulant and a key ingredient of the paper industry. Guar gum recovered about 3.86 mg/L of fibre and was most effective among the biopolymers. Settling velocity distribution curves demonstrated that Guar gum was able to settle the fibres faster than the other biopolymers; however, alum displayed the highest particle removal rate than all the biopolymers at any of the settling velocities. Alum, Guar gum, Xanthan gum and Locust bean gum removed 97.46%, 94.68%, 92.39% and 92.46% turbidity of raw effluent at a settling velocity of 0.5 cm/min, respectively. The conditions for obtaining the lowest sludge volume index such as pH, dose and mixing speed were optimised for guar gum which was the most effective among the biopolymers. Response surface methodology was used to design all experiments, and an optimum operational setting was proposed. The test results indicate similar performance of alum and Guar gum in terms of floc settling velocities and sludge volume index. Since Guar gum is a plant derived natural substance, it is environmentally benign and offers a green treatment option to the paper mills for pulp recycling.

Long-term mass balance of perennial firn and ice in an Alpine cave (Austria):Constraints from radiocarbon-dated wood fragments

Hundsalm ice cave located at 1520 m altitude in a karst region of western Austria contains up to 7-m-thick deposits of snow, firn and congelation ice. Wood fragments exposed in the lower parts of an ice and firn wall were radiocarbon accelerator mass spectrometry (AMS) dated. Although the local stratigraphy is complex, the 19 individual dates - the largest currently available radiocarbon dataset for an Alpine ice cave - allow to place constraints on the accumulation and ablation history of the cave ice. Most of the cave was either ice free or contained only a small firn and ice body during the 'Roman Warm Period'; dates of three wood fragments mark the onset of firn and ice build-up in the 6th and 7th century ad. In the central part of the cave, the oldest samples date back to the 13th century and record ice growth coeval with the onset of the 'Little Ice Age'. The majority of the ice and firn deposit, albeit compromised by a disturbed stratigraphy, appears to have been formed during the subsequent centuries, supported by wood samples from the 15th to the 17th century. The oldest wood remains found so far inside the ice is from the end of the Bronze Age and implies that local relics of prehistoric ice may be preserved in this cave. The wood record from Hundsalm ice cave shows parallels to the Alpine glacier history of the last three millennia, for example, the lack of preserved wood remains during periods of known glacier minima, and underscores the potential of firn and ice in karst cavities as a long-term palaeoclimate archive, which has been degrading at an alarming rate in recent years. © The Author(s) 2013.

The dynamics of murid gammaherpesvirus 4 within wild, sympatric populations of bank voles and wood mice

Murid gammaherpesvirus 4 (MuHV-4) is widely used as a small animal model for understanding gammaherpesvirus infections in man. However, there have been no epidemiological studies of the virus in wild populations of small mammals. As MuHV-4 both infects cells associated with the respiratory and immune systems and attempts to evade immune control via various molecular mechanisms, infection may reduce immunocompetence with potentially serious fitness consequences for individuals. Here we report a longitudinal study of antibody to MuHV-4 in a mixed assemblage of bank voles (Clethrionomys glareolus) and wood mice (Apodemus sylvaticus) in the UK. The study was conducted between April 2001 and March 2004. Seroprevalence was higher in wood mice than bank voles, supporting earlier work that suggested wood mice were the major host even though the virus was originally isolated from a bank vole. Analyses of both the probability of having antibodies and the probability of initial seroconversion indicated no clear seasonal pattern or relationship with host density. Instead, infection risk was most closely associated with individual characteristics, with heavier males having the highest risk. This may reflect individual variation in susceptibility, potentially related to variability in the ability to mount an effective immune response.

TRP Channels and calcium signalling in dental pulp stem cells

Introduction: Ca2+ ion is an important intracellular messenger essential for the regulation of various cellular functions including proliferation, differentiation and apoptosis. Transient Receptor Potential (TRP) channels are calcium permeable cationic channels that play important role in regulation of free intracellular calcium ([Ca2+]i) in response to thermal, physical and chemical stimuli. Ca2+ signalling in human dental pulp stem cells (hDPSCs) and the ion channels regulating Ca2+ are largely not known. Objectives: Investigate changes in [Ca2+]i and determine the ion channels that regulate calcium signalling in hDPSCs. Methods: DPSCs were derived from immature third molars and cells less than passage 6 were used in all the experiments. Changes in [Ca2+]i were studied with Fura2 calcium imaging. RNA was extracted from DPSCs and a panel of TRP channel gene expression was determined by qPCR employing custom designed FAM TRP specific primers and probes (Roche, UK) and the Light Cycler 480 Probes Master (Roche). Results: hDPSCs express gene transcripts for all TRP families including TRPV1, V2, V4, TRPA1, TRPC3, TRPC5, TRPC6, TRPM3, TRPM7 and TRPP2. Stimulation of cells with appropriate TRP channel agonist induced increase in [Ca2+]i and similar responses were obtained when cell were mechanically stimulated by membrane stretch with application of hypotonic solution. Conclusion: TRP channels mediate calcium signalling in hDPSCs that merit further investigation.

Osteoprotegerin Expression in the Dental Pulp

Objectives: The inflammatory response to pulpal injury or infection has major clinical significance. Osteoprotegerin (OPG) is a soluble decoy receptor for Receptor Activator of NF kappa B Ligand (RANKL), preventing ligand binding to its receptor (RANK), thus inhibiting clastic cell formation. The aim of the study is to investigate the expression of OPG in human dental pulp and the effects of inflammatory mediators. This study will specifically investigate the effects of Transforming Growth Factor Beta-1 (TGF-β1) and Interleukin 1-Beta (IL-1β) on the expression of OPG on pulp fibroblasts in vitro. Method: Five primary pulp fibroblast populations were obtained by explant culture of healthy pulp tissue. Triplicate cultures were grown to confluence in 12-well plates and stimulated for 48 hours with IL-1β (10ng/ml) or TGF-β1 (10ng/ml). The conditioned media was collected and OPG levels detected by ELISA (R+D Systems, UK). Results: All fibroblast populations produced quantifiable levels of OPG in a time-dependant fashion. IL-1β significantly increased the expression of OPG (p<0.05) in all cultures. In contrast, TGF-β1 had no significant effect on OPG expression levels. In addition, previous work in our laboratory demonstrated both TGF-β1 and IL-1β stimulated OPG expression by periodontal ligament fibroblasts. Conclusion: These data indicate that IL-1β-regulated expression of OPG by pulpal fibroblasts may mediate hard tissue turnover in the inflamed dental pulp.

Substance P production by human dental pulp fibroblasts

The inflammatory response to pulpal injury or infection has major clinical significance. Neurogenic inflammation describes the local release of neuropeptides, notably substance P (SP), from afferent neurones, and may play a role in the pathogenesis of pulpal disease. The fibroblast is the most numerous cell type in the dental pulp and recent work has suggested that it is involved in the inflammatory response. Objectives: The aims of the study were to determine whether pulp fibroblasts could produce SP, and to investigate the expression of the SP receptor, NK-1, by these cells. Methods: Primary pulp fibroblast cell populations were isolated by enzymatic digestion from non-carious teeth extracted for orthodontic reasons. Whole pulp tissue was obtained from freshly extracted sound (n=35) and carious (n=39) teeth. Expression of SP and NK-1 mRNA was determined by RT-PCR. The effects of interleukin-1β (IL-1β) and transforming growth factor-β1 (TGF-β1) on SP and NK-1 expression were also determined. The presence of NK-1 on fibroblast cell membranes was established by western blotting. The effects of the cytokines on each parameter were analysed by ANOVA. Radioimmunoassay (RIA) was carried out to quantify SP expression by pulp fibroblasts and in whole pulp tissue. Results: SP was expressed by pulpal fibroblasts both at the mRNA level and the protein level. In addition, NK-1 was detected in fibroblast cultures at the mRNA level and appeared as a double band on western blots of membrane extracts. IL-1β and TGF-β1 significantly stimulated the expression of SP and NK-1. SP levels were significantly greater (p<0.05) in carious compared to sound teeth. Conclusion: Pulp fibroblasts are capable of synthesising and secreting SP, as well as expressing the SP receptor, NK-1. These findings suggest that pulp fibroblasts play a role in neurogenic inflammation in pulpal disease. (Supported by the European Society of Endodontology.)

Co-expression of NPY and VIP in human dental pulp

Introduction: The regulation of pulpal haemodynamics in health and disease involves sympathetic and parasympathetic mechanisms in which both neuropeptide Y (NPY; a sympathetic vasoconstrictor) and vasoactive intestinal polypeptide (VIP; a parasympathetic vasodilator) may play potential pathophysiological roles. We have previously investigated the levels of NPY or VIP present in human dental pulp tissue and shown that their expression is up-regulated in caries induced pulpal inflammation. Objectives: The aim of this study was to investigate the potential correlation between NPY and VIP levels measured in the same dental pulp samples using radioimmunoassay (RIA). Methods: Pulp tissue was obtained from extracted teeth, classified as follows; healthy (n=22), moderately carious (n=20) and grossly carious (n=26). Samples were processed for RIA by boiling in acetic acid as previously described. The levels of NPY and VIP, measured by RIA, were expressed as ng/gram of pulp tissue. The nature of the relationship between NPY and VIP levels in human pulp tissue was tested by calculating Pearson's product moment correlation coefficient using the linear regression test. Results: Calculation of Pearson product moment correlation coefficient showed a significant negative correlation between NPY and VIP levels in pulp tissue samples from non-carious teeth (p = 0.02, r = -.48). This negative correlation in non-carious teeth changed to a significant positive correlation in carious teeth when the levels of NPY and VIP were compared (p = 0.03, r= 0.311). Conclusions: In non-carious teeth, the negative correlation between NPY and VIP levels is in keeping with the previously described modulatory influence of cholinergic nerves on sympathetic function which may be perturbed as caries develops.