Relative density of United States forests has shifted to higher levels over last two decades with important implications for future dynamics.

Tree size-density dynamics can inform key trends in forest productivity along with opportunities to increase ecosystem resiliency. Here, we employ a novel approach to estimate the relative density (RD, range 0-1) of any given forest based on its current size-density relationship compared to a hypothetical maximum using the coterminous US national forest inventory between 1999 and 2020. The analysis suggests a static forest land area in the US with less tree abundance but greatly increased timber volume and tree biomass. Coupled with these resource trends, an increase in RD was identified with 90% of US forest land now reaching a biologically-relevant threshold of canopy closure and/or self-thinning induced mortality (RD > 0.3), particularly in areas prone to future drought conditions (e.g., West Coast). Notably, the area of high RD stands (RD > 0.6) has quintupled over the past 20 years while the least stocked stands (RD < 0.3) have decreased 3%. The evidence from the coterminous US forest RD distribution suggest opportunities to increase live tree stocking in understocked stands, while using density management to address tree mortality and resilience to disturbances in increasingly dense forests.

The use of intraoral local anaesthetic to aid reduction of acute temporomandibular joint dislocation.

Acute temporomandibular joint (TMJ) dislocation is distressing for patients and treatment can often be a challenge for clinicians. This report describes the use of intra oral local anaesthetic (LA) to aid reduction of the joint, reducing the need for conscious sedation, muscle relaxants or general anaesthetic.

A rapidly-reversible absorptive and emissive vapochromic Pt(II) pincer-based chemical sensor.

Selective, robust and cost-effective chemical sensors for detecting small volatile-organic compounds (VOCs) have widespread applications in industry, healthcare and environmental monitoring. Here we design a Pt(II) pincer-type material with selective absorptive and emissive responses to methanol and water. The yellow anhydrous form converts reversibly on a subsecond timescale to a red hydrate in the presence of parts-per-thousand levels of atmospheric water vapour. Exposure to methanol induces a similarly-rapid and reversible colour change to a blue methanol solvate. Stable smart coatings on glass demonstrate robust switching over 104 cycles, and flexible microporous polymer membranes incorporating microcrystals of the complex show identical vapochromic behaviour. The rapid vapochromic response can be rationalised from the crystal structure, and in combination with quantum-chemical modelling, we provide a complete microscopic picture of the switching mechanism. We discuss how this multiscale design approach can be used to obtain new compounds with tailored VOC selectivity and spectral responses.

A novel diamond anvil cell for x-ray diffraction at cryogenic temperatures manufactured by 3D printing.

A new miniature high-pressure diamond anvil cell was designed and constructed using 3D micro laser sintering technology. This is the first application of the use of rapid prototyping technology to construct high-pressure apparatus. The cell is specifically designed for use as an X-ray diffraction cell that can be used with commercially available diffractometers and open-flow cryogenic equipment to collect data at low temperature and high pressure. The cell is constructed from stainless steel 316L and is about 9 mm in diameter and 7 mm in height, giving it both small dimensions and low thermal mass, and it will fit into the cooling envelope of a standard CryostreamTM cooling system. The cell is clamped using a customized miniature buttress thread of diameter 7 mm and pitch of 0.5 mm enabled by 3D micro laser sintering technology; such dimensions are not attainable using conventional machining. The buttress thread was used as it has favourable uniaxial load properties allowing for higher pressure and better anvil alignment. The clamp can support the load of at least 1.5 kN according to finite element analysis (FEA) simulations. FEA simulations were also used to compare the performance of the standard thread and the buttress thread, and demonstrate that stress is distributed more uniformly in the latter. Rapid prototyping of the pressure cell by the laser sintering resulted in a substantially higher tensile yield strength of the 316L stainless steel (675 MPa compared to 220 MPa for the wrought type of the same material), which increased the upper pressure limit of the cell. The cell is capable of reaching pressures of up to 15 GPa with 600 µm diameter culets of diamond anvils. Sample temperature and pressure changes on cooling were assessed using X-ray diffraction on samples of NaCl and HMT-d12.

Toward inventory-based estimates of soil organic carbon in forests of the United States.

Soil organic carbon (SOC) is the largest terrestrial carbon (C) sink on Earth; this pool plays a critical role in ecosystem processes and climate change. Given the cost and time required to measure SOC, and particularly changes in SOC, many signatory nations to the United Nations Framework Convention on Climate Change report estimates of SOC stocks and stock changes using default values from the Intergovernmental Panel on Climate Change or country-specific models. In the United States, SOC in forests is monitored by the national forest inventory (NFI) conducted by the Forest Inventory and Analysis (FIA) program within the U.S. Department of Agriculture, Forest Service. The FIA program has been consistently measuring soil attributes as part of the NFI since 2001 and has amassed an extensive inventory of SOC in forest land in the conterminous United States and southeast and southcentral coastal Alaska. That said, the FIA program has been using country-specific predictions of SOC based, in part, upon a model using SOC estimates from the State Soil Geographic (STATSGO) database compiled by the Natural Resources Conservation Service. Estimates obtained from the STATSGO database are averages over large map units and are not expected to provide accurate estimates for specific locations, e.g., NFI plots. To improve the accuracy of SOC estimates in U.S. forests, NFI SOC observations were used for the first time to predict SOC density to a depth of 100 cm for all forested NFI plots. Incorporating soil-forming factors along with observations of SOC into a new estimation framework resulted in a 75% (48 ± 0.78 Mg/ha) increase in SOC densities nationally. This substantially increases the contribution of the SOC pool, from approximately 44% (17 Pg) of the total forest ecosystem C stocks to 56% (28 Pg), in the forest C budget of the United States.

Attribution of net carbon change by disturbance type across forest lands of the conterminous United States.

BACKGROUND: Locating terrestrial sources and sinks of carbon (C) will be critical to developing strategies that contribute to the climate change mitigation goals of the Paris Agreement. Here we present spatially resolved estimates of net C change across United States (US) forest lands between 2006 and 2010 and attribute them to natural and anthropogenic processes. RESULTS: Forests in the conterminous US sequestered -460 ± 48 Tg C year-1, while C losses from disturbance averaged 191 ± 10 Tg C year-1. Combining estimates of net C losses and gains results in net carbon change of -269 ± 49 Tg C year-1. New forests gained -8 ± 1 Tg C year-1, while deforestation resulted in losses of 6 ± 1 Tg C year-1. Forest land remaining forest land lost 185 ± 10 Tg C year-1 to various disturbances; these losses were compensated by net carbon gains of -452 ± 48 Tg C year-1. C loss in the southern US was highest (105 ± 6 Tg C year-1) with the highest fractional contributions from harvest (92%) and wind (5%). C loss in the western US (44 ± 3 Tg C year-1) was due predominantly to harvest (66%), fire (15%), and insect damage (13%). The northern US had the lowest C loss (41 ± 2 Tg C year-1) with the most significant proportional contributions from harvest (86%), insect damage (9%), and conversion (3%). Taken together, these disturbances reduced the estimated potential C sink of US forests by 42%. CONCLUSION: The framework presented here allows for the integration of ground and space observations to more fully inform US forest C policy and monitoring efforts.

Monitoring Network Confirms Land Use Change is a Substantial Component of the Forest Carbon Sink in the eastern United States.

Quantifying forest carbon (C) stocks and stock change within a matrix of land use (LU) and LU change is a central component of large-scale forest C monitoring and reporting practices prescribed by the Intergovernmental Panel on Climate Change (IPCC). Using a region-wide, repeated forest inventory, forest C stocks and stock change by pool were examined by LU categories. In eastern US forests, LU change is a substantial component of C sink strength (~37% of forest sink strength) only secondary to that of C accumulation in forests remaining forest where their comingling with other LUs does not substantially reduce sink strength. The strongest sinks of forest C were study areas not completely dominated by forests, even when there was some loss of forest to agriculture/settlement/other LUs. Long-term LU planning exercises and policy development that seeks to maintain and/or enhance regional C sinks should explicitly recognize the importance of maximizing non-forest to forest LU changes and not overlook management and conservation of forests located in landscapes not currently dominated by forests.

(C4H12N2)[CoCl4]: tetrahedrally coordinated Co2+ without the orbital degeneracy.

We report on the synthesis, crystal structure and magnetic properties of a previously unreported Co(2+) S = 3/2 compound, (C4H12N2)[CoCl4], based upon a tetrahedral crystalline environment. The S = 3/2 magnetic ground state of Co(2+), measured with magnetization, implies an absence of spin-orbit coupling and orbital degeneracy. This contrasts with compounds based upon an octahedral and even known tetrahedral Co(2+) [Cotton et al. (1961). J. Am. Chem. Soc. 83, 4690] systems where a sizable spin-orbit coupling is measured. The compound is characterized with single-crystal X-ray diffraction, magnetic susceptibility, IR and UV-vis spectroscopy. Magnetic susceptibility measurements find no magnetic ordering above 2 K. The results are also compared with the previously known monoclinic hydrated analogue.

Net carbon flux of dead wood in forests of the Eastern US.

Downed dead wood (DDW) in forest ecosystems is a C pool whose net flux is governed by a complex of natural and anthropogenic processes and is critical to the management of the entire forest C pool. As empirical examination of DDW C net flux has rarely been conducted across large scales, the goal of this study was to use a remeasured inventory of DDW C and ancillary forest attributes to assess C net flux across forests of the Eastern US. Stocks associated with large fine woody debris (diameter 2.6-7.6 cm) decreased over time (-0.11 Mg ha(-1) year(-1)), while stocks of larger-sized coarse DDW increased (0.02 Mg ha(-1) year(-1)). Stocks of total DDW C decreased (-0.14 Mg ha(-1) year(-1)), while standing dead and live tree stocks both increased, 0.01 and 0.44 Mg ha(-1) year(-1), respectively. The spatial distribution of DDW C stock change was highly heterogeneous with random forests model results indicating that management history, live tree stocking, natural disturbance, and growing degree days only partially explain stock change. Natural disturbances drove substantial C transfers from the live tree pool (≈-4 Mg ha(-1) year(-1)) to the standing dead tree pool (≈3 Mg ha(-1) year(-1)) with only a minimal increase in DDW C stocks (≈1 Mg ha(-1) year(-1)) in lower decay classes, suggesting a delayed transfer of C to the DDW pool. The assessment and management of DDW C flux is complicated by the diversity of natural and anthropogenic forces that drive their dynamics with the scale and timing of flux among forest C pools remaining a large knowledge gap.

Development of a sporicidal test method for Clostridium difficile.

BACKGROUND: Disinfectants with claimed activity against Clostridium difficile must be evaluated to ensure efficacy against the spores that comprise an environmental source of patient infection. Unfortunately there is, at present, no generally accepted method for evaluating these disinfectants. In the absence of such a method, laboratories have to adapt protocols that were not designed for products used in medical environments and consequently may use inappropriate test organisms, exposure times, and pass criteria. AIM: To develop and evaluate a method for testing the activity of disinfectants against C. difficile spores using exposure times and pass criteria which are relevant to clinical application. METHODS: A Joint Working Party of the Healthcare Infection Society (HIS) and the Advisory Committee on Antimicrobial Resistance and Healthcare Associated Infections (ARHAI) of the Department of Health in England was assembled. The Working Party adapted a previously described enzyme-based method for spore purification (the Clospore method) using an exposure time of 5 min and a 5 log10 kill as a pass criterion. FINDINGS: Evaluation of the method by three laboratories demonstrated that the method is simple to follow and that the results are repeatable and reproducible. CONCLUSION: The method described by the Working Party produces a clean suspension with a high titre of spores. It is recommended that, for a disinfectant used in the environment, the product should demonstrate a 5 log10 reduction in 5 min under clean or dirty conditions to fulfil the requirements of the test.

Finding a benchmark for monitoring hospital cleanliness.

This study evaluated three methods for monitoring hospital cleanliness. The aim was to find a benchmark that could indicate risk to patients from a contaminated environment. We performed visual monitoring, ATP bioluminescence and microbiological screening of five clinical surfaces before and after detergent-based cleaning on two wards over a four-week period. Five additional sites that were not featured in the routine domestic specification were also sampled. Measurements from all three methods were integrated and compared in order to choose appropriate levels for routine monitoring. We found that visual assessment did not reflect ATP values nor environmental contamination with microbial flora including Staphylococcus aureus and meticillin-resistant S. aureus (MRSA). There was a relationship between microbial growth categories and the proportion of ATP values exceeding a chosen benchmark but neither reliably predicted the presence of S. aureus or MRSA. ATP values were occasionally diverse. Detergent-based cleaning reduced levels of organic soil by 32% (95% confidence interval: 16-44%; P<0.001) but did not necessarily eliminate indicator staphylococci, some of which survived the cleaning process. An ATP benchmark value of 100 relative light units offered the closest correlation with microbial growth levels <2.5 cfu/cm(2) (receiver operating characteristic ROC curve sensitivity: 57%; specificity: 57%). In conclusion, microbiological and ATP monitoring confirmed environmental contamination, persistence of hospital pathogens and measured the effect on the environment from current cleaning practices. This study has provided provisional benchmarks to assist with future assessment of hospital cleanliness. Further work is required to refine practical sampling strategy and choice of benchmarks.

Survival analysis for a large-scale forest health issue: Missouri oak decline.

Survival analysis methodologies provide novel approaches for forest mortality analysis that may aid in detecting, monitoring, and mitigating of large-scale forest health issues. This study examined survivor analysis for evaluating a regional forest health issue - Missouri oak decline. With a statewide Missouri forest inventory, log-rank tests of the effects of covariates on the survivor function and equality of the survivor function among classes were conducted for selected oak species. Additionally, hazard functions were determined for diameter classes for damaged and undamaged oaks. Results indicate that mortality appears to vary significantly among some inventory classes such as oak species, but not among other classes such as ownership class. Indicators of individual tree vigor (i.e., crown class and ratio) were more significant predictors of oak tree mortality than site/stand attributes (i.e., density and aspect). Finally, results indicate that even fast-growing oak trees are at high risk of mortality if damaged by disease. Survival analyses, such as those applied in this study, may enable testing of forest health hypotheses using large-scale inventories. In the context of Missouri's oak forest decline, study results suggest management efforts should focus on limiting the spread of disease damage, increasing the vigor of residual trees, and emphasizing small trees when developing stand prescriptions.

Campylobacter jejuni gene expression in the chick cecum: evidence for adaptation to a low-oxygen environment.

Transcriptional profiling of Campylobacter jejuni during colonization of the chick cecum identified 59 genes that were differentially expressed in vivo compared with the genes in vitro. The data suggest that C. jejuni regulates electron transport and central metabolic pathways to alter its physiological state during establishment in the chick cecum.

Evidence for neuronal localisation of enteroviral sequences in motor neurone disease/amyotrophic lateral sclerosis by in situ hybridization.

Sequences resembling those of human enterovirus type B sequences have been associated with motor neurone disease/amyotrophic lateral sclerosis. In a previous study we detected enteroviral sequences in spinal cord/brain stem from cases of motor neurone disease/amyotrophic lateral sclerosis, but not controls. Adjacent tissue sections to two of those strongly positive for these sequences by reverse-transcriptase polymerase chain reaction were analyzed by in situ hybridization with digoxigenin-labelled virus-specific antisense riboprobes. In one case, a female aged 83 showing 12 month rapid progressive disease, signal was specifically localized to cells identifiable as motor neurones of the anterior horn. In another case, a male aged 63 with a 60-month history of progressive muscle weakness, dysarthia, dyspnoea and increased tendon reflexes, signal was located to neurones in the gracile/cuneate nuclei of the brain stem tissue block that had been analyzed. This case showed loss of neurones in the anterior horn of the spinal cord by histopathologic examination which would account for clinical signs of motor neurone disease/amyotrophic lateral sclerosis. Dysfunction of the gracile/cuneate nuclei might have been masked by the paralytic disease. These structures are adjacent to the hypoglossal nuclei, and suggest either localised dissemination from hypoglossal nuclei or a possible route of dissemination of infection through the brainstem to the hypoglossal nuclei. These findings provide further evidence for the possible involvement of enteroviruses in motor neurone disease/amyotrophic lateral sclerosis.

Granulocyte macrophage colony stimulating factor is elevated in alveolar macrophages from sheep naturally infected with maedi-visna virus and stimulates maedi-visna virus replication in macrophages in vitro.

Infection by maedi-visna virus, a lentivirus of sheep, leads to chronic inflammatory reactions of various tissues. In this report we have analysed the role of specific cytokines in the disease process. A significant increase in expression of interleukin-6, interleukin-10, granulocyte macrophage-colony stimulating factor (GM-CSF) and transforming growth factor-beta1 mRNA was observed in alveolar macrophages isolated from the lungs of naturally infected animals when compared with lungs of seronegative controls. Levels of GM-CSF mRNA expression in alveolar macrophages correlated with the presence of lung lesions, but there was no correlation of interleukin-10, interleukin-6, tumour necrosis factor-alpha and transforming growth factor-beta1 mRNA levels in alveolar macrophages from animals with pulmonary lesions. In vitro investigation showed that GM-CSF in the range 0.1-10 ng/ml induced a significant increase in viral p25 production after 7 days in acutely infected blood monocyte-derived macrophages. The production of p25 peaked between 7 and 14 days exposure to 10 ng/ml of GM-CSF. Quantitative polymerase chain reaction showed that the level of viral DNA in monocyte-derived macrophages was dose-dependent following GM-CSF treatment in the range 0.1-100 ng/ml after 7 days. Viral mRNA expression was also enhanced. These findings indicate a role for GM-CSF in the pathogenesis of lymphoid interstitial pneumonia in infected animals.

A review of bacterial methyl halide degradation: biochemistry, genetics and molecular ecology.

Methyl halide-degrading bacteria are a diverse group of organisms that are found in both terrestrial and marine environments. They potentially play an important role in mitigating ozone depletion resulting from methyl chloride and methyl bromide emissions. The first step in the pathway(s) of methyl halide degradation involves a methyltransferase and, recently, the presence of this pathway has been studied in a number of bacteria. This paper reviews the biochemistry and genetics of methyl halide utilization in the aerobic bacteria Methylobacterium chloromethanicum CM4T, Hyphomicrobium chloromethanicum CM2T, Aminobacter strain IMB-1 and Aminobacter strain CC495. These bacteria are able to use methyl halides as a sole source of carbon and energy, are all members of the alpha-Proteobacteria and were isolated from a variety of polluted and pristine terrestrial environments. An understanding of the genetics of these bacteria identified a unique gene (cmuA) involved in the degradation of methyl halides, which codes for a protein (CmuA) with unique methyltransferase and corrinoid functions. This unique functional gene, cmuA, is being used to develop molecular ecology techniques to examine the diversity and distribution of methyl halide-utilizing bacteria in the environment and hopefully to understand their role in methyl halide degradation in different environments. These techniques will also enable the detection of potentially novel methyl halide-degrading bacteria.

Local lung responses following local lung challenge with recombinant lungworm antigen in systemically sensitized sheep.

BACKGROUND: Chronic mast cell-mediated inflammation may contribute significantly towards the extensive tissue remodelling that is a feature of lungworm infection in ruminants. Understanding the factors that control tissue remodelling is a necessary step toward effective management and treatment of conditions that feature such pathology. OBJECTIVE: We sought to define in a novel ovine model system, the cellular, immune and mast cell phenotypic events that occur following local lung challenge with a recombinant protein antigen, DvA-1, derived from the ruminant lungworm nematode, Dictyocaulus viviparus. METHODS: Two spatially disparate lung segments in systemically sensitized sheep were challenged on three occasions with DvA-1 (3xDVA) and two further segments were challenged with saline (3xSAL). Two months after the third challenge, one of the two segments previously repeatedly challenged with DvA-1 was challenged again with DvA-1 (3xDVA:DVA) whilst the other was challenged with saline (3xDVA:SAL). A similar protocol was followed with the saline challenged segments (3xSAL:SAL and 3xSAL:DVA). Bronchoalveolar lavage fluid (BALF) (n = 16) and tissue (n = 3) were collected after the last challenge. RESULTS: Cellular changes 24 h after the fourth challenge were characterized by an increase in the absolute numbers of neutrophils and eosinophils in BALF from 3xDVA:DVA and 3xSAL:DVA segments. Local antibody production was implied through increased levels of antibody in both 3xDVA:DVA and 3xDVA:SAL segments, with the latter being unaffected by inflammation. Levels of active transforming growth factor beta-1 (TGF-beta(1)) were significantly increased in 3xDVA:SAL segments and a trend towards an increase was apparent in 3xDVA:DVA segments. Total TGF-beta1 levels were significantly correlated with eosinophil counts in all except the 3xDVA:SAL segments. Such changes in the bronchoalveolar space were complemented by increased ratios of sheep mast cell proteinase-1 expressing cells and tryptase expressing cells, to toluidine blue positive cells in airways from 3xDVA:DVA segments. CONCLUSION: Mast cell phenotypic events occurring as a consequence of antigen challenge were limited to segments in which changes in BALF were characterized by neutrophil influx and increased local antibody production.

Identification of methyl halide-utilizing genes in the methyl bromide-utilizing bacterial strain IMB-1 suggests a high degree of conservation of methyl halide-specific genes in gram-negative bacteria.

Strain IMB-1, an aerobic methylotrophic member of the alpha subgroup of the Proteobacteria, can grow with methyl bromide as a sole carbon and energy source. A single cmu gene cluster was identified in IMB-1 that contained six open reading frames: cmuC, cmuA, orf146, paaE, hutI, and partial metF. CmuA from IMB-1 has high sequence homology to the methyltransferase CmuA from Methylobacterium chloromethanicum and Hyphomicrobium chloromethanicum and contains a C-terminal corrinoid-binding motif and an N-terminal methyltransferase motif. However, cmuB, identified in M. chloromethanicum and H. chloromethanicum, was not detected in IMB-1.

Chloromethane utilization gene cluster from Hyphomicrobium chloromethanicum strain CM2(T) and development of functional gene probes to detect halomethane-degrading bacteria.

Hyphomicrobium chloromethanicum CM2(T), an aerobic methylotrophic member of the alpha subclass of the class proteobacteria, can grow with chloromethane as the sole carbon and energy source. H. chloromethanicum possesses an inducible enzyme system for utilization of chloromethane, in which two polypeptides (67-kDa CmuA and 35-kDa CmuB) are expressed. Previously, four genes, cmuA, cmuB, cmuC, and purU, were shown to be essential for growth of Methylobacterium chloromethanicum on chloromethane. The cmuA and cmuB genes were used as probes to identify homologs in H. chloromethanicum. A cmu gene cluster (9.5 kb) in H. chloromethanicum contained 10 open reading frames: folD (partial), pduX, orf153, orf207, orf225, cmuB, cmuC, cmuA, fmdB, and paaE (partial). CmuA from H. chloromethanicum (67 kDa) showed high identity to CmuA from M. chloromethanicum and contains an N-terminal methyltransferase domain and a C-terminal corrinoid-binding domain. CmuB from H. chloromethanicum is related to a family of methyl transfer proteins and to the CmuB methyltransferase from M. chloromethanicum. CmuC from H. chloromethanicum shows identity to CmuC from M. chloromethanicum and is a putative methyltransferase. folD codes for a methylene-tetrahydrofolate cyclohydrolase, which may be involved in the C(1) transfer pathway for carbon assimilation and CO(2) production, and paaE codes for a putative redox active protein. Molecular analyses and some preliminary biochemical data indicated that the chloromethane utilization pathway in H. chloromethanicum is similar to the corrinoid-dependent methyl transfer system in M. chloromethanicum. PCR primers were developed for successful amplification of cmuA genes from newly isolated chloromethane utilizers and enrichment cultures.

In situ-PCR for the detection of Mycobacterium paratuberculosis DNA in paraffin-embedded tissues.

Molecular biological techniques have permitted the rapid and sensitive detection of the Mycobacterium paratuberculosis genome in infected tissues, most commonly by polymerase chain reaction amplification of sequences in the IS900 DNA insertion sequence. The aim of this work was the detection of M. paratuberculosis DNA in ovine tissues by in situ-polymerase chain reaction, which is sensitive and localises the signal within the tissue sample. Paraffin embedded tissues from three acid-fast positive ovine guts with classical lesions of paratuberculosis, and from negative control samples were tested. A 413-bp fragment of the IS900 sequence was amplified in-situ and hybridised to an internal PCR-synthesised digoxygenin-labelled probe. The samples from sheep affected by paratuberculosis clearly showed cell-specific cytoplasmic signals in mucosal and submucosal macrophages. This technique could be useful both in the diagnosis and study of the pathogenesis of infections in which involvement of M. paratuberculosis is suspected.