Search

Modern ocean microbial research utilizes advanced molecular analytical techniques, such as polymerase chain reaction (PCR), DNA and protein probe arrays, and nucleic acid sequencing (etc.). Applying or at least initiating these techniques at the point and time of sample collection can enhance their effectiveness. To that end, in-situ sample processing and real-time molecular detection schemes have been implemented using deployable autonomous systems that can be operated in diverse ocean environments from shallow coastal waters to the deep sea. Such devices have been termed “ecogenomic sensors.” The size of these instruments currently requires that they be moored in a fixed location or passively mobile, drifting at fixed depth and observing microbial communities in a moving frame of reference with ocean currents. With the highly dynamic motion of open water and microbial life, the next frontier of ocean microbial research requires the improved capability of an actively mobile asset. A mobile ecogenomic sensor encompasses a fully maneuverable vehicle with weeks of persistence, environmental data analysis, detection of physical and biological features, autonomous sampling and in situ analysis, and near-real-time data reporting. This system is now being developed by integrating three components: a compact molecular analytical instrument (the 3rd generation Environmental Sample Processor), a long-range autonomous underwater vehicle, and software algorithms for AUV-based feature detection and sampling. A summary of the system and its initial application is presented.

Eating patterns of children have been investigated in the U.S. and have been found to be changeable over extended time periods. Trends can be correlated to changes in the same periods for determinants of overall health such as body mass index (BMI). In Puerto Rico, there have been no such studies so similar correlations cannot be done. Herein, we present baseline information on the number and types of eating occasions in a convenience sample of children from the area of San Juan so that future changes in patterns can be monitored over time.Multiple 24 hour recall questionnaires were administered to school children at 3 different grade levels. Number of eating occasions and type of meal (breakfast, lunch, dinner and snacks) were quantified. Factors considered for analysis were age, gender and BMI of the children, participation in the School Lunch Program (SLP) and if meals were eaten on a weekday or weekend day.Approximately 40% of children were categorized as overweight. There was a trend toward fewer eating occasions in older vs. younger children and fewer eating occasions on weekend days vs. weekdays. Lunch and dinner were consumed more frequently than breakfast and participants in the SLP had more eating occasions than non-participants.The number of eating occasions in Puerto Rican youth is maintained at about 5 for weekdays and about 4.5 per day for weekend days with a trend toward fewer meals as a function of increasing age. This data can be used as baseline information in future studies that wish to correlate changes in dietary patterns with health.

Over 100 years after trypanosomatids were first discovered in plant tissues, Phytomonas parasites have now been isolated across the globe from members of 24 different plant families. Most identified species have not been associated with any plant pathology and to date only two species are definitively known to cause plant disease. These diseases (wilt of palm and coffee phloem necrosis) are problematic in areas of South America where they threaten the economies of developing countries. In contrast to their mammalian infective relatives, our knowledge of the biology of Phytomonas parasites and how they interact with their plant hosts is limited. This review draws together a century of research into plant trypanosomatids, from the first isolations and experimental infections to the recent publication of the first Phytomonas genomes. The availability of genomic data for these plant parasites opens a new avenue for comparative investigations into trypanosomatid biology and provides fresh insight into how this important group of parasites have adapted to survive in a spectrum of hosts from crocodiles to coconuts.

Limited studies have been carried out on the level of vitamin D in non-human primates. Herein, we present data using the preferred vitamin D indicator, serum 25-OH vitamin D, sequentially collected...

Crenarchaeota are ubiquitous and abundant microbial constituents of soils, sediments, lakes, and ocean waters. To further describe the cosmopolitan nonthermophilic Crenarchaeota , we analyzed the genome sequence of one representative, the uncultivated sponge symbiont Cenarchaeum symbiosum . C. symbiosum genotypes coinhabiting the same host partitioned into two dominant populations, corresponding to previously described a- and b-type ribosomal RNA variants. Although they were syntenic, overlapping a- and b-type ribotype genomes harbored significant variability. A single tiling path comprising the dominant a-type genotype was assembled and used to explore the genomic properties of C. symbiosum and its planktonic relatives. Of 2,066 ORFs, 55.6% matched genes with predicted function from previously sequenced genomes. The remaining genes partitioned between functional RNAs (2.4%) and hypotheticals (42%) with limited homology to known functional genes. The latter category included some genes likely involved in the archaeal–sponge symbiotic association. Conversely, 525 C. symbiosum ORFs were most highly similar to sequences from marine environmental genomic surveys, and they apparently represent orthologous genes from free-living planktonic Crenarchaeota . In total, the C. symbiosum genome was remarkably distinct from those of other known Archaea and shared many core metabolic features in common with its free-living planktonic relatives.

The carbon (C) cycle in boreal regions is strongly influenced by fire, which converts biomass and detrital C mainly to gaseous forms (CO2 and smaller proportions of CO and CH4), and some 1–3% of mass to pyrogenic C (PyC). PyC is mainly produced as solid charred residues, including visually-defined charcoal, and a black carbon (BC) fraction chemically defined by its resistance to laboratory oxidation, plus much lower proportions of volatile soot and polycyclic aromatic hydrocarbons (PAHs). All PyC is characterized by fused aromatic rings, but varying in cluster sizes, and presence of other elements (N, O) and functional groups. The range of PyC structures is often described as a continuum from partially charred plant materials, to charcoal, soot and ultimately graphite which is formed by the combination of heat and pressure. There are several reasons for current interest in defining more precisely the role of PyC in the C cycle of boreal regions. First, PyC is largely resistant to decomposition, and therefore contributes to very stable C pools in soils and sediments. Second, it influences soil processes, mainly through its sorption properties and cation exchange capacity, and third, soot aerosols absorb solar radiation and may contribute to global warming. However, there are large gaps in the basic information needed to address these topics. While charcoal is commonly defined by visual criteria, analytical methods for BC are mainly based on various measures of oxidation resistance, or on yield of benzenepolycarboxylic acids. These methods are still being developed, and capture different fractions of the PyC structural continuum. There are few quantitative reports of PyC production and stocks in boreal forests (essentially none for boreal peatlands), and results are difficult to compare due to varying experimental goals and methods, as well as inconsistent terminology. There are almost no direct field measurements of BC aerosol production from boreal wildfires, and little direct information on rates and mechanisms for PyC loss. Structural characterization of charred biomass and forest floor from wildfires generally indicates a low level of thermal alteration, with the bulk of the material having H/C ratios still >0.2, and small aromatic cluster sizes. Especially for the more oxidation-resistant BC fraction, a variety of mainly circumstantial evidence suggests very slow decomposition, with turnover on a millennial timescale (in the order of 5–7 ky), also dependent on environmental conditions. However, there is also evidence that some PyC may be lost in only tens to hundreds of years due to a combination of lower thermal alteration and environmental protection. The potential for long-term PyC storage in soil may also be limited by its consumption by subsequent fires. Degraded, functionalized PyC is also incorporated into humified soil organic matter, and is transported eventually to marine sediments in dissolved and particulate form. Boreal production is estimated as 7–17 Tg BC y−1 of solid residues and 2–2.5 Tg BC y−1 as aerosols, compared to global estimates of 40–240 and 10–30 Tg BC y−1, respectively. Primary research needs include basic field data on PyC production and stocks in boreal forests and peatlands, suitable to support C budget modeling, and development of standardized analytical methods and of improved approaches to assess the chemical recalcitrance of typical chars from boreal wildfires. To accomplish these goals effectively will require much greater emphasis on interdisciplinary cooperation.

The role of accounting in enabling action at a distance has received considerable attention in the accounting literature. In this paper we examine the role of accounts in the actions taken by government officials in Washington, DC to reduce the size of the Navajo livestock holdings on their reservation in the southwestern United States in the 1930s. We examine the role of accounts in three acts: first in enabling action at a distance, second in enacting action in the local setting and finally, in shoring up, or maintaining action when things go wrong. We argue that in order to understand the role of accounts in these three acts, it is necessary to examine the conditions of possibility within which they emerge, the network of agents who adopt and give them life and the way in which they become entwined with various and shifting strategies. We also examine how action taken in Washington, DC had profound consequences for the Navajo when attempts were made to enact actions taken at a distance on the reservation.

Human cytomegalovirus (HCMV) immediate-early (IE) transcription is stimulated by virion phosphoprotein pp71, the product of gene UL82. It has previously been shown that pp71 interacts with the cellular protein hDaxx and, in the studies presented here, the significance of this interaction was investigated for HCMV IE gene expression. In co-transfection experiments, the presence of hDaxx increased the transcriptional response of the HCMV major IE promoter (MIEP) to pp71, but it was not possible to determine whether the effect was due to an interaction between the two proteins or to stimulation of hDaxx synthesis by pp71. The use of small interfering RNA (siRNA) in long- and short-term transfection approaches reduced intracellular hDaxx levels to no more than 3 % of normal. Infection of hDaxx-depleted cells with herpes simplex virus recombinants containing the HCMV MIEP revealed significantly greater promoter activity when hDaxx levels were minimal. Similarly, reducing intracellular hDaxx amounts resulted in greater IE gene expression during infection with an HCMV mutant lacking pp71, but had no effect on IE transcription during infection with wild-type HCMV. The results suggest that hDaxx is not important as a positive-acting factor for the stimulation of HCMV IE transcription by pp71. Instead, it appears that hDaxx acts as a repressor of IE gene expression, and it is proposed here that the interaction of pp71 with hDaxx is important to relieve repression and permit efficient initiation of productive replication.

The natural abundance of 13C (δ13C) generally increases with decomposition of organic matter. This is contrary to the expected decrease, as lignin is hypothesized to accumulate relative to isotopically heavier cellulose. Our objective was to test the hypothesis that 13C depletion should be observed for gymnosperm logs that typically develop advanced brown-rot decay with high lignin content. With increasing lignin concentration [previously determined by nuclear magnetic resonance (NMR)], δ13C tended to become more negative for samples of Pseudotsuga menziesii, Tsuga heterophylla, Thuja plicata, and unidentified species from Coastal Forest Chronosequence sites of southern Vancouver Island. For a larger sample set without NMR analysis, δ13C was significantly more depleted for the highest decay classes, and total C was negatively correlated with δ13C, consistent with the higher total C of lignin than of cellulose. Relationships of total C and δ13C with density were much weaker. We discuss causes for the variability of δ13C in coarse woody debrisfrom these sites, and how the apparent paradox in the predicted change of δ13C with decomposition is largely due to the confusion of lignin, the biopolymer produced by higher plants, with the acid-unhydrolyzable residue (AUR) of the proximate analysis procedure commonly used to assess litter quality in decomposition studies. Key words: Coarse woody debris, decomposition, lignin, 13C NMR, δ13C, proximate analysis

Forest soil organic matter (SOM) is affected by inputs from coarse wood (CW) and fine litter (FL, e.g., leaves, twigs, cones, and needles). The influence of these materials on forest SOM was studied in a lodgepole pine (Pinus contorta) forest in southeastern Wyoming. Organic materials in CW sites were significantly (P < 0.05) more acidic, but contained half the total N of FL sites. Forest floor materials and SOM C contents were significantly greater in CW samples. Lignin decomposition products (CuO analysis) accumulated primarily in organic horizons of both sites, with significantly higher (60–70%) contents in CW materials. Vanillyl compounds were the primary lignin products from both sites, with cinnamyl compounds also important in SOM from FL sites. Vanillic acid to vanillin ratios were significantly higher in mineral soils under CW. 13C-NMR spectra indicated CW materials were enriched in lignin, and that humic acids from both site types were very similar and unusually high in alkyl C. Fulvic acids were also high in O-alkyl and carboxyl C, particularly in the CW sites. Results suggest there are differences in forest C constituents and that removal of CW could possibly alter forest soil dynamics that would impact forest productivity and biodiversity. Key words: Forest, coarse wood, litter, carbon, soil organic matter, humic substances

To reduce soil destruction by urban sprawl, land use planning has to promote the use of soils within cities. As soil functions are now protected by law in Germany, urban soil quality has to be evaluated before soil management. We studied contributions from elemental carbon (EC) and soil organic matter (SOM) quality in topsoil horizons at seven sites in Stuttgart, Germany, differing in impurities by technogenic substrates. The most disturbed site was found at a disused railway area while high-density areas, public parks and garden areas showed varying degrees of disturbance by anthropogenic activities. For most soils, compounds derived from plant litter dominated organic matter (OM) quality characterized by nuclear magnetic resonance (NMR) spectroscopy. Although high contents of EC (up to 70% of soil organic carbon) were indicated by thermal oxidation, this was not confirmed by aromatic C intensities in NMR spectra. Only for the highly aromatic railway soil were results for elemental carbon by thermal oxidation and NMR similar. As other technogenic substrates beside EC like plastics may also contribute in the long-term to OM in urban soils, new analytical techniques are therefore required. This knowledge will promote the evaluation of urban soil properties and their sustainable use.

Microbial life predominates in the ocean, yet little is known about its genomic variability, especially along the depth continuum. We report here genomic analyses of planktonic microbial communities in the North Pacific Subtropical Gyre, from the ocean's surface to near–sea floor depths. Sequence variation in microbial community genes reflected vertical zonation of taxonomic groups, functional gene repertoires, and metabolic potential. The distributional patterns of microbial genes suggested depth-variable community trends in carbon and energy metabolism, attachment and motility, gene mobility, and host-viral interactions. Comparative genomic analyses of stratified microbial communities have the potential to provide significant insight into higher-order community organization and dynamics.

Improving our ability to predict the impact of climate change on the carbon (C) balance of boreal forests requires increased understanding of site-specific factors controlling detrital and soil C accumulation. Jack pine (Pinus banksiana) and black spruce (Picea mariana) stands along the Boreal Forest Transect Case Study (BFTCS) in northern Canada have similar C stocks in aboveground vegetation and large woody detritus, but thick forest floors of poorly-drained black spruce stands have much higher C stocks, comparable to living biomass. Their properties indicate hindered decomposition and N cycling, with high C/N ratios, strongly stratified and depleted δ13C and δ15N values, high concentrations of tannins and phenolics, and 13C nuclear magnetic resonance (NMR) spectra typical of poorly decomposed plant material, especially roots and mosses. The thinner jack pine forest floor appears to be dominated by lichen, with char in some samples. Differences in quantity and quality of aboveground foliar and woody litter inputs are small and unlikely to account for the contrasts in forest floor accumulation and properties. These are more likely associated with site conditions, especially soil texture and drainage, exacerbated by increases in sphagnum coverage, forest floor depth, and tannins. Small changes in environmental conditions, especially reduced moisture, could trigger large C losses through rapid decomposition of forest floor in poorly drained black spruce stands in this region.

Condensed tannins are a major component of litter inputs, but little is known about the effects of tannin structural variations on soil biological processes and organic matter development. Four different condensed tannins (CTs) extracted from balsam fir, western red cedar, kalmia and black spruce were added to Corsican pine litter and subsequently incubated for 16 weeks in order to investigate the effect of the B ring hydroxylation pattern on C, N and P transformations. While for C mineralization the chain length and stereochemistry of the CTs seemed to be a more important parameter, net N and P mineralization rates were clearly reduced compared with non-amended litter. With regard to the B ring hydroxylation, the prodelphinidin (PD) CTs having predominantly three hydroxy groups at the B ring (balsam fir and western red cedar) exhibited significantly lower mineralization rates than the procyanidin (PC) CTs having two OH groups (kalmia and black spruce). The same was true for net nitrification, but this process was only slightly affected by the CTs. Although based on only four CTs, this study indicates that B ring hydroxylation is an important variable determining net N and P mineralization rates. Our results support previous suggestions that PD tannins bind to or react more strongly with soil organic matter. Therefore, more than PC tannins, they reduce the availability of organic N for mineralization as well as their own detectability by standard methods for soil CT.

The plant-colonizing Pseudomonas fluorescens strain SBW25 harbors a gene cluster ( rsp ) whose products show similarity to type III protein secretion systems found in plant and animal pathogens. Here we report a detailed analysis of the expression and regulation of the P. fluorescens rsp pathway, both in the phytosphere and in vitro. A combination of chromosomally integrated transcriptional reporter fusions, overexpressed regulatory genes, and specific mutants reveal that promoters controlling expression of rsp are actively transcribed in the plant rhizosphere but not (with the exception of the rspC promoter) in the phyllosphere. In synthetic medium, regulatory ( rspL and rspR ) and structural ( rspU , plus the putative effector ropE ) genes are poorly expressed; the rspC promoter is subject to an additional level of regulatory control. Ectopic expression of regulatory genes in wild-type and mutant backgrounds showed that RspR controls transcription of the alternate sigma factor, rspL , and that RspL controls expression of gene clusters encoding structural genes. Mutation of rspV did not affect RspR-mediated expression of rspU . A search for additional regulators revealed two candidates—one with a role in the conversion of alanine to pyruvate—suggesting that expression of rsp is partly dependent upon the metabolic status of the cell. Mutations in rsp regulators resulted in a significant reduction in competitive colonization of the root tips of sugar beet seedlings but also caused a marked increase in the lag phase of laboratory-grown cultures, indicating that rsp regulatory genes play a more significant general role in the function of P. fluorescens SBW25 than previously appreciated.

ing the sustainable use of rangelands for livestock production. However, many ecosystem components and Growing interest in the potential for soils to provide a sink for processes like plant community structure, soil properatmospheric C has prompted studies of effects of management on the amountandnatureofsoilorganicC(SOC).Inthisstudy,weevaluated ties, and nutrient cycling are also affected by grazing effects of different grazing management regimes (light grazing [LG], management (Schuman et al., 1999). Density and duraheavy grazing [HG], and non-grazed exclosures [EX]) on amount tion of rangeland stocking can affect plant community and composition of SOC at the USDA–ARS High Plains Grasslands compositionthroughdisplacementofcool-seasonmixed ResearchStation (HPGRS),Cheyenne, WY.Soils (0–5cm) fromeach grassesbywarm-seasonshortgrassesinnorthernmixedtreatment were analyzed for total C and N contents and lignin com- grass prairies (Dormaar and Willms, 1990). position. Soil organic C and N contents were significantly greater in Grazing can also influence the amount and composiLG (SOC–13.8 Mg ha 1 ; total N–1.22 Mg ha 1

A collection of tannins extracted from various plant species was subjected to thermally assisted hydrolysis and methylation (THM) using tetramethylammonium hydroxide. The products obtained included 1,3,5-trimethoxybenzenes, derived from A rings of condensed tannins (CTs), and 1,2-dimethoxybenzene, 1,2,3-trimethoxybenzene, and derivatives as major products from the B ring. 1,2,4-Tri- and two tetramethoxybenzenes were also detected in most analyses. Correlation analyses revealed that they were derived from the B ring, with 1,2,4-trimethoxybenzene being derived from a procyanidin (PC) B ring and 1,2,3,5-tetramethoxybenzene from a prodelphinidin (PD) ring. Tannins from species that contained both CT and hydrolyzable tannin (HT) produced mainly permethylated gallic acid moieties upon THM, and the products derived from CTs were less abundant. Most likely, the ester-bound HTs are more easily transmethylated than the cleavage and methylation of the C-C and C-O linked CTs. Statistical analyses of the THM products and 13C NMR data of the tannins showed good correlations between the B ring hydroxylation and the di- and trimethoxybenzenes observed. Using the ratio of the methyl esters of 3,4-dimethoxybenzoic acid and 3,4,5-trimethoxybenzoic acid only provided good correlations of the percent PC as well. Furthermore, the 1,3,5-trimethoxybenzenes may serve as good markers of tannins in plant, soil, sediment or other samples as analyzed by THM.

Cholesteryl ester transfer protein (CETP) is an important modulator of high density lipoprotein cholesterol in humans and thus considered to be a therapeutic target for preventing cardiovascular disease. The gene encoding CETP has been shown to be highly variable, with multiple single nucleotide polymorphisms responsible for altering both its transcription and sequence. Examining nine missense variants of CETP, we found some had significant associations with CETP mass and high density lipoprotein cholesterol levels. Two variants, Pro-373 and Gln-451, appear to be more stable in vivo, an observation mirrored by partial proteolysis studies performed in vitro. Because these naturally occurring variant proteins are potentially present in clinical populations that will be treated with CETP inhibitors, all commonly occurring haplotypes were tested to determine whether the proteins they encode could be inhibited by torcetrapib, a compound currently in clinical trials in combination with atorvastatin. Torcetrapib behaved similarly with all variants, with no significant differences in inhibition.

Disease-resistant livestock could provide a potentially sustainable and environmentally sound method of controlling tick and tick-borne diseases of livestock in the developing world. Advances in the knowledge and science of genomics open up opportunities to identify selectable genes controlling disease resistance but first, breeds and individuals with distinguishable phenotypes need to be identified. The Bos indicus breed, Sahiwal, has been exploited in dairy breeding programmes, because it is resistant to ticks and has relatively good performance characteristics compared to other indigenous cattle breeds of tropical regions. The analyses reported here show that Sahiwal calves were also more resistant than European Bos taurus (Holstein) dairy breed calves to tick-borne tropical theileriosis (Theileria annulata infection). Following experimental infection with T. annulata sporozoites, a group of Sahiwal calves all survived without treatment, with significantly lower maximum temperatures (P0.01) and lower rates of parasite multiplication (P0.05) than a group of Holstein calves, which all had severe responses. Although the Sahiwals became as anaemic as the Holsteins, other measures of pathology, including enlargement of the draining lymph node and the acute phase proteins, alpha1 acid glycoprotein and haptoglobin, were significantly less in the Sahiwals than in the Holsteins (P0.05). Additionally, the Sahiwals had significantly lower resting levels of alpha1 acid glycoprotein than the Holsteins (P0.05). Production of a third acute phase proteins, serum amyloid A, had very similar kinetics in both breeds. Acute phase proteins are produced in response to systemic release of the kinds of pro-inflammatory cytokines that are thought to be responsible for the pyrexic, cachectic and anorexic responses characteristic of tropical theileriosis. The prolonged production of alpha1 acid glycoprotein in the Holsteins is indicative of chronic production of circulating pro-inflammatory cytokines. In contrast, Sahiwals appear able to overcome infection with T. annulata as well as limit pathology by preventing the over-stimulation of pathways involving these cytokines.

The human cytomegalovirus tegument protein pp71 is important for transactivation of immediate-early (IE) gene expression and for the efficient initiation of virus replication. We have analyzed the properties of pp71 by assaying its effects on gene expression from the genome of in 1312, a herpes simplex virus type 1 (HSV-1) mutant devoid of functional VP16, ICP0, and ICP4. Upon infection of human fibroblasts, in 1312-derived viruses are repressed and retained in a quiescent state, but the presence of pp71 prevented the quiescent state from being attained. Reporter gene cassettes cloned into the in 1312 genome, in addition to the endogenous IE promoters, remained active for at least 12 days postinfection, and infected cells were viable and morphologically normal. Cells expressing pp71 remained responsive to the HSV-1 transactivating factors VP16 and ICP4 and to trichostatin A. The C-terminal 61 amino acids, but not the LACSD motif, were required for pp71 activity. In addition to preventing attainment of quiescence, pp71 was able to disrupt the quiescent state of in 1312 derivatives and promote the resumption of viral gene expression after a lag of approximately 3 days. The results extend the functional analysis of pp71 and suggest a degree of similarity with the HSV-1 IE protein ICP0. The ability to provoke slow reactivation of quiescent genomes, in conjunction with cell survival, represents a novel property for a viral structural protein.

Objectives The objective of this study was to test the predictive value of an oxidative stress biomarker in 634 patients from the Prospective Randomized Evaluation of the Vascular Effects of Norvasc Trial (PREVENT). Background Oxidative stress contributes to mechanisms of atherosclerosis and plaque instability. Biomarkers of oxidation, such as malondialdehyde (MDA), may represent independent indicators of risk for patients with stable coronary artery disease (CAD). Methods Serum MDA levels were measured as thiobarbituric acid reactive substances (TBARS) in 634 patients with documented CAD using reverse-phase high-performance liquid chromatography and spectrophotometric approaches. Results During the three-year study, there were 51 major vascular events such as fatal/nonfatal myocardial infarction, 149 hospitalizations for nonfatal vascular events, and 139 patients underwent a major vascular procedure. At baseline, patients with TBARS levels in the highest quartile had a relative risk (RR) of 3.30 (95% confidence interval [CI] 1.47 to 7.42; p = 0.038) for major vascular events, RR of 4.10 (95% CI 2.55 to 6.60; p Conclusions Serum levels of TBARS were strongly predictive of cardiovascular events in patients with stable CAD, independently of traditional risk factors and inflammatory markers.

Surfactant protein A (SP-A), a member of the collectin family, selectively binds toPneumocystis cariniiand mediates interactions between pathogen and host alveolar macrophages in vitro. To test the hypothesis that mice lacking SP-A have delayed clearance ofPneumocystisorganisms and enhanced lung injury, wild-type C57BL/6 (WT) and SP-A-deficient mice (SP-A−/−) with or without selective CD4+-T-cell depletion were intratracheally inoculated withPneumocystisorganisms. Four weeks later, CD4-depleted SP-A-deficient mice had developed a more severePneumocystisinfection than CD4-depleted WT (P. cariniipneumonia [PCP] scores of 3 versus 2, respectively). Whereas all non-CD4-depleted WT mice were free of PCP, intact SP-A−/−mice also had evidence of increased organism burden.Pneumocystisinfection in SP-A-deficient mice was associated histologically with enhanced peribronchial and/or perivascular cellularity (score of 4 versus 2, SP-A−/−versus C57BL/6 mice, respectively) and a corresponding increase in bronchoalveolar lavage (BAL) cell counts. Increases in SP-D content, gamma interferon, interleukin-4, interleukin-5, and tumor necrosis factor alpha in BAL fluid occurred but were attenuated in PCP-infected SP-A−/−mice compared to WT mice. There were increases in total BAL NO levels in both infected groups, but nitrite levels were higher in SP-A−/−mice, indicating a reduction in production of higher oxides of nitrogen that was also reflected in lower levels of 3-nitrotyrosine staining in the SP-A−/−group. We conclude that despite increases in inflammatory cells, SP-A-deficient mice infected withP. cariniiexhibit an enhanced susceptibility to the organism and attenuated production of proinflammatory cytokines and reactive oxygen-nitrogen species. These data support the concept that SP-A is a local effector molecule in the lung host defense againstP. cariniiin vivo.

Fertilization elicits a dramatic, transient rise in Ca2+ within the egg which is an essential component of egg activation and consequent initiation of development. In the sea urchin egg, three distinct Ca2+ stores have been identified which could, either individually or in combination, initiate Ca2+ release at fertilization. Inositol 1,4,5-trisphosphate (IP3) production by phospholipase C (PLC) has been suggested as the singular signal in initiating the Ca2+ transient. Other studies indicate that Ca2+ stores gated by cyclic adenosine diphosphate ribose (cADPR) or nicotinic acid adenine dinucleotide phosphate (NAADP) are also necessary. We have examined the temporal relationship between the Ca2+ rise and IP3 production at fertilization in vivo within individual eggs using a green fluorescent protein (GFP) coupled to a pleckstrin homology (PH) domain that can detect changes in IP3. Translocation of the probe occurred after the Ca2+ rise was initiated. Earlier, and possibly smaller, IP3 changes could not be excluded due to limitations in probe sensitivity. High IP3 levels are maintained during the decline in cytoplasmic Ca2+, suggesting that later IP3 metabolism might not be related to regulation of Ca2+, but may function to modulate other PIP2 regulated events such as actin polymerization or reflect other novel phosphoinositide signaling pathways.

Access to appropriate healthcare services continues to be a major challenge in rural America. Telehealth technologies offer an opportunity to bridge gaps in health services in rural and remote areas and possibly support rural economic development. Lack of access to healthcare services to a growing population may create barriers to recruitment of businesses and economic growth. Several rural-oriented programs have attempted to leverage these emerging distance technologies, but success has varied despite the application of considerable federal, state governmental, and private resources. Barriers to adoption and sustainability of rural telehealth embody several factors that must be considered when planning, developing, implementing, and evaluating a rural telehealth program. New Mexico, the fifth largest state in the United States, represents many of the issues related to the potential benefits and challenges in developing a telehealth system to serve its rural communities. The Center for Telehealth at the University of New Mexico Health Sciences Center has been supported in large part by state and federal funding. Through our experiences, successes, failures, and lessons learned, we have developed approaches to overcoming barriers to adoption and sustainment of telehealth applications, including the establishment of partnerships with economic development projects in the state. This article describes these experiences and identifies and provides strategies for planning, development, implementation, and sustainment of telehealth in a rural program.

We compare rates of decay of foliar litters of British Columbia tree species in two field studies, and assess which initial litter chemistry parameters best predict the decay rates. Nutrient concentrations, tannins, and carbon fractions (based on proximate analysis and nuclear magnetic resonance spectroscopy) were measured in fresh litter of 14 tree species in one experiment and seven species in a second experiment. Each study was replicated in a different site in order to assess the transferability of results. Broadleaf litters decayed faster than needle litters only during the first year; thereafter, they decayed slower. Lignin concentration was a good predictor of mass loss only during the first year and only in one of the two experiments, which may have resulted from all foliar litters having high lignin concentrations (>170 mg·kg–1). Litter chemistry effects on first-year decay were consistent and transferable among sites. None of the initial litter chemistry parameters were good predictors of mass remaining after 4 or 5 years, because mass loss of most litters was similar by this time. The convergence in mass losses of litters after 4–5 years despite initial differences indicates that decomposition estimates extrapolated from early rates or initial chemistry may not accurately predict long-term decay.

The use of poplar tree systems (PTS) as evapotranspiration barriers on decommissioned landfills is gaining attention as an option for leachate management. This study involved field-testing the Simultaneous Heat and Water (SHAW) model for its ability to reliably estimate poplar transpiration, volumetric soil water content, and soil temperature at a landfill located in southern Ontario, Canada. The model was then used to estimate deep drainage and to ascertain the influence of a young PTS on the soil water balance of the landfill cover. The SHAW model tended to underestimate poplar transpiration [mean difference (MD) ranged from 0.33 to 3.55 mm on a daily total basis] and overestimate volumetric soil water content by up to 0.10 m3 m-3. The model estimated soil temperature very well, particularly in the upper 1 m of the landfill cover (MD ranged from -0.1 to 1.6 ×°C in this layer). The SHAW model simulations showed that deep drainage decreased appreciably with the presence of a young PTS largely through increased interception of rainfall, and that PTS have a good potential to act as effective evapotranspiration barriers in northern temperate climate zones.

Litter decomposition was studied for 2 years in a mixed forest serving as a water protection area (Rhine-Neckar conurbation, SW Germany). Two experiments differing in initial dry weight equivalent in litterbags were set up: one to compare decomposition of European beech leaves (Fagus sylvatica) with common oak leaves (Quercus robur), and the other comparing decomposition of Scots pine needles (Pinus sylvestris) with black cherry leaves (Prunus serotina Ehrh.), respectively. Mass losses were greater for oak litter than for beech (75.0 versus 34.6%), and for cherry litter than for pine (94.6 versus 68.3%). In both experiments, a strong initial loss of soluble compounds occurred. The changes in litter N and P concentrations and the decrease in C-to-N ratio coincided with changes in residual mass. However, neither tannin and phenolic concentrations nor NMR could explain the pronounced variation in mass loss after 2 years. Differences in litter palatability and toughness, nutrient contents and other organic compounds may be responsible for the considerable differences in residual mass between litter types. The fast decay of black cherry leaves appears to play a major role in the present humus dynamics at the studied site. Since black cherry has a high N demand, which is mainly met by root uptake from the forest floor, this species is crucial for internal N cycling at this conurbation forest site. These effects together may significantly contribute to prevent nitrate leaching from the forest ecosystem which is subject to a continuous N deposition on an elevated level.

Phasing out beehive burners and rising costs for landfilling have increased the need to widen options for utilization of the smaller size fractions of woody wastes generated during log handling and sawmilling in British Columbia. We characterized several size classes of logyard fines up to 16 mm sampled from coastal and interior operations. Total C, total N, ash, and condensed tannin concentrations were consistent with properties derived largely from wood, with varying proportions of bark and mixing with mineral soil. Especially for < 3-mm fractions, the latter resulted in high ash contents that would make them unsuitable for fuel. Solid-state 13C cross-polarization magic-angle spinning (CPMAS) nuclear magnetic resonance (NMR) spectra were consistent with the chemical data, with high O-alkyl intensity and similarity to naturally occurring woody forest floor; no samples were high in aromatic or phenolic C. Aqueous extracts of two < 16-mm fines, which accounted for only a small proportion of the total C, were enriched in alkyl C and had low or undetectable tannins. Application to forest sites might cause short-term immobilization of N, but also might include possible longer-term benefits from reduction of N loss after harvesting and restoration of soil organic matter in degraded sites.

Tannins are purported to be an important factor controlling nitrogen cycling in forest ecosystems, and the ability of tannins to bind proteins in protein–tannin complexes is thought to be the primary mechanism responsible for these effects. In this study, we examined the influence of well-characterized tannins purified from five different plant species on C and N dynamics of a forest soil A horizon. Tannic acid, a commonly used and commercially available hydrolyzable tannin (HT), and cellulose were also included for comparison. With the exception of tannins from huckleberry (Vaccinium ovatum), the amendments increased respiration 1.4–4.0 fold, indicating that they were acting as a microbial C source. Tannic acid was significantly more labile than the five purified tannins examined in this study. All treatments decreased net N mineralization substantially, through greater N immobilization and decreased mineralization. The six tannins inhibited gross ammonification rates significantly more than cellulose. This suggests that added tannins had effects in addition to serving as an alternative C source. Tannins purified from Bishop pine (Pinus muricata) were the only tannins that significantly inhibited potential gross nitrification rates, however, rates were low even in the control soil making it difficult to detect any inhibition. Differences in tannin structure such as condensed versus HTs and the hydroxylation pattern of the condensed tannin B-ring likely explain differences observed among the tannin treatments. Contrary to other studies, we did not find that condensed tannins were more labile and less inhibitory than HTs, nor that shorter chained tannins were more labile than longer chained tannins. In addition to supporting the hypothesis that reduced N availability in the presence of tannins is caused by complexation reactions, our data suggests tannins act as a labile C source leading to increased N immobilization.

Acute phase proteins (APP) are synthesised in the liver in response to the systemic presence of high levels of pro-inflammatory cytokines. Bacteria are considered to be strong inducers of APP whereas viruses are weak or non-inducers of APP. Very few reports have been published on APP induction by parasites. Here, we report that the tick-borne protozoan parasite of cattle, Theileria annulata, induced an atypical acute phase response in cattle. Following experimental infection, serum amyloid A (SAA) appeared first, followed by a rise in alpha(1) acid glycoprotein (alpha(1)AGP) in all animals, whereas haptoglobin, which is a major APP in cattle, only appeared in some of the animals, and generally at a low level. All three APP only became elevated around or after the appearance of schizonts in draining lymph nodes and after the first observed temperature rise. Increased alpha(1)AGP levels coincided with the appearance of piroplasms. The production of SAA and alpha(1)AGP correlated strongly with each other, and also with some clinical measures of disease severity including the time to fever, development of leucopaenia, parasitaemia and mortality. These results are consistent with the hypothesis that T. annulata causes severe pathology in susceptible cattle by inducing high levels of pro-inflammatory cytokines.

Phylogenetic and stable-isotope analyses implicated two methanogen-like archaeal groups, ANME-1 and ANME-2, as key participants in the process of anaerobic methane oxidation. Although nothing is known about anaerobic methane oxidation at the molecular level, the evolutionary relationship between methane-oxidizing archaea (MOA) and methanogenic archaea raises the possibility that MOA have co-opted key elements of the methanogenic pathway, reversing many of its steps to oxidize methane anaerobically. In order to explore this hypothesis, the existence and genomic conservation of methyl coenzyme M reductase (MCR), the enzyme catalyzing the terminal step in methanogenesis, was studied in ANME-1 and ANME-2 archaea isolated from various marine environments. Clone libraries targeting a conserved region of the alpha subunit of MCR ( mcrA ) were generated and compared from environmental samples, laboratory-incubated microcosms, and fosmid libraries. Four out of five novel mcrA types identified from these sources were associated with ANME-1 or ANME-2 group members. Assignment of mcrA types to specific phylogenetic groups was based on environmental clone recoveries, selective enrichment of specific MOA and mcrA types in a microcosm, phylogenetic congruence between mcrA and small-subunit rRNA tree topologies, and genomic context derived from fosmid sequences. Analysis of the ANME-1 and ANME-2 mcrA sequences suggested the potential for catalytic activity based on conservation of active-site amino acids. These results provide a basis for identifying methanotrophic archaea with mcrA sequences and define a functional genomic link between methanogenic and methanotrophic archaea.

Polyribonucleotide probes targeting planktonic archaeal (Group I and II) and bacterial rRNA revealed that Ar- chaea comprised a significant fraction of total prokaryote cell abundance in the marine waters west of the Antarctic Peninsula. Determinations of Archaea and Bacteria cell abundances were made during two research cruises to the Palmer Long-Term Ecological Research region during the austral winter and summer of 1999. During the austral summer, surface water abundances of Group I (GI) Archaea were generally low, averaging 4.7 3 10 3 cells ml 21 and comprising 9-39% of the total picoplankton abundance in the meso- (150-1,000 m) and bathypelagic (1,000-3,500 m) circumpolar deep water (CDW). Relative to summertime distri- butions, GI cells were more evenly distributed throughout the water column during the winter, averaging 10% of the picoplankton in the surface waters and 13% in the CDW. Surface water GI abundance increased 44% between the summer and winter, coincident with a fivefold decrease in GI abundance in the deeper waters. The abundance of Group II (GII) Archaea was persistently ,2% of the total picoplankton throughout the water column in both summer and winter. Bacterial abundance was greatest in the upper water column (0-100 m) during the summer, averaging 3.9 3 10 5 cells ml 21 and comprised 89% of the total picoplankton assemblage. Generally, GI Archaea varied seasonally in the deeper waters, whereas bacterial abundance varied more in the upper waters. The observed variability in bacterial and archaeal abundance suggests that these two groups of marine picoplankton are dynamic components of Southern Ocean microbial food webs.

Loss of T cell number and function during HIV infection or secondary to pharmacologic immunosuppression renders individuals susceptible to opportunistic infections, including Pneumocystis carinii pneumonia. Because costimulatory receptors are critical for optimal T cell function, we hypothesized that these proteins would regulate susceptibility to opportunistic infections. We found that despite normal T cell numbers, mice deficient in the costimulatory molecules CD2 and CD28 spontaneously developed P. carinii pneumonia. In experiments using intratracheal injection of P. carinii organisms to induce infection, the loss of CD28 alone was sufficient to render mice susceptible to acute infection; however, the organism was eventually cleared. Examination of inflammatory responses to P. carinii revealed that mice deficient in both CD2 and CD28 accumulated CD8+ T cells in their lungs in response to infection and demonstrated markedly reduced specific Ab titers. Analysis of cytokine profiles suggested that regulation of IL-10 and IL-15 may be important elements of the response to this pathogen. Thus, costimulatory molecule function is critical in determining the initial susceptibility to infection with P. carinii. Analysis of immunologic responses in these mice may provide important insights into the defects that render individuals susceptible to opportunistic infection, and provide opportunities for novel immunologically based therapies.

The protozoon Naegleria gruberi is able to carry out amoeboid locomotion at the water-air interface in a manner indistinguishable from that exhibited on solid substrata with the production of focal contacts and associated filopodia. The speed of locomotion at this interface can be modulated by changes in electrolyte concentrations; these speed changes are identical to those observed at a water-glass interface. The nature of the water-air interface is discussed leading to the hypothesis that surface tension alone could provide suitable properties for the adhesion and translocation of amoebae at this interface without necessitating specific, absorbed molecules. The temporary swimming flagellate stage of Naegleria is able to dock at the interface, make stable adhesions to it, and revert to the amoeboid phenotype. Conversely, amoebae resident at the water-air interface can transform to swimming flagellates and escape into the bulk liquid phase. We report the presence of Naegleria amoebae in the surface microlayers of natural ponds; thus, in freshwater bodies there may be active shuttling of Naegleria amoebae from the benthos to the surface microlayers by means of the non-feeding, swimming flagellate phenotype. The public health implication of this behaviour in the case of the pathogenic relative, Naegleria fowleri, is discussed.

In Philosophy as a Way of Life, Pierre Hadot discusses the understanding of philosophy held by the Greco-Roman ancients. Philosophy was not understood only as an exegetical or analytical exercise, ...

Photosynthetic microorganisms play a crucial role in the marine environment. In vast areas of the oceans, marine primary productivity is performed by cells smaller than 2-3 micro m (picoplankton). Here, we report on molecular analyses of the conserved photosynthetic psbA gene (coding for protein D1 of photosystem II reaction centre) as a diversity indicator of naturally occurring marine oxygenic picophytoplankton. The psbA genes proved to be good indicators of the presence of a wide variety of photosynthetic marine microbial groups, including new cyanobacterial groups and eukaryotic algae (prasinophytes). Furthermore, using environmental bacterial artificial chromosome (BAC) libraries, we were able to correlate psbA genes with small subunit rRNAs and, therefore, to confirm their phylogenetic affiliation.

Observations of surface microlayer material collected from freshwaters demonstrate its ability to assemble rapidly at the water-air interface following experimental disruption. This property is exploited to provide a means for studying and manipulating the in situ locomotory behaviour of some surface microlayer amoebae (Acanthamoeba, Naegleria, Vannella) under laboratory conditions. Amoeboid movement is substratum-dependent and so it is important to know the means by which these organisms attach to a water-air interface. Reflection interference microscopy (RIM) demonstrates that during locomotion Vannella adheres to a glass coverslip by means of an unstable platform of associated contact (parallel to, but ∼100-nm off it) within which stable, focal contacts form attaching to the substratum. These two key features of cell-substratum interactions occur also in Acanthamoeba and Naegleria. Direct examination of cell-substratum interactions during amoeboid movement at the water-air interface by RIM proves that the associated contact is also established here.

Tannins influence ecosystem function by affecting decomposition rates, nutrient cycling, and herbivory. To determine the role of tannins in ecological processes, it is important to quantify their abundance and understand how structural properties affect reactivity. In this study, purified tannins from the foliage of nine species growing in the pygmy forest of the northern California coast were examined for chemical reactivity, protein precipitation capacity (PPC), and structural characteristics (13C NMR). Reactivity of purified tannins varied among species 1.5-fold for the Folin total phenol assay, and 7-fold and 3-fold, respectively, for the acid butanol and vanillin condensed tannin assays. There was about a 5-fold difference in PPC. Variation in chemical reactivity and PPC can be largely explained by differences in structural characteristics of the tannins determined by 13C NMR. In particular, the condensed versus hydrolyzable tannin content, as well as the hydroxylation pattern of the B-ring and stereochemistry at the C-2–C-3 position appear to influence reactivity. Due to the large differences in chemical reactivity among species, it is necessary to use a well-characterized purified tannin from the species of interest to convert assay values to concentrations. Our results suggest that structural characteristics of tannins play an important role in regulating their reactivity in ecological processes.

The objective of this work was to determine the effect of androgen deprivation therapy (ADT) on rates of bone mineral density (BMD) loss in men with prostate cancer. It was a prospective study comparing men receiving ADT to age matched controls for 2 y. Subjects received a history, physical exam, bone mineral density measurement, and laboratory evaluation every 6 months. Thirty-nine subjects receiving continuous ADT for prostate cancer (subjects) were compared to 39 age-matched controls not receiving ADT (controls). Twenty-three subjects and 30 controls completed the study through 24 months. Men in the ADT group demonstrated greater rates of bone mineral density loss than men in the control group at every site except the lumbar spine. Twenty-four month per cent of bone mineral density loss is presented as mean+/-standard error (s.e.). At the distal forearm, the ADT group value was -9.4%+/-1.0% and -4.4%+/-0.3% for controls (P

To aid in understanding black carbon (BC) formation during smoldering combustion in forest fires, we characterized charring of a softwood and hardwood. Charring (150, 340, 480°C) caused mass loss (7-84%), enrichment of organic carbon (OC) (0-32%), and 13C depletion (> 150°C). As determined by 13C MAS NMR, the OC composition of the woods was dominated by (di)-O-alkyl structures, and the chars by alkyl and aromatic structures. With increasing temperature, aromatic structures increased and the chars became more similar, although initial differences in OC concentration and δ 13C of woods persisted. The BC cluster sizes apparently remained small, pointing towards a low resistance against oxidation. Crown Copyright © 2002 Published by Elsevier Science Ltd. All rights reserved.

This paper reports on the common experience of vaginal wetness amongst South African users of progestogen-only injectable contraceptives. The observations emerged in the course of a community-based cross-sectional household survey undertaken in a rural district of KwaZulu-Natal in South Africa. The purpose of the survey was to elicit self-reporting on side effects of injectable contraceptive methods. Eight hundred and forty-eight women aged 15-49 were interviewed and 22.1% reported current use of an injectable contraceptive method, either depot medroxyprogesterone acetate (Depo-Provera) or norethisterone oenanthate (Nur-Isterate). Other modern methods used were oral hormonal contraceptives (4.5%), male condoms (1.3%), the intrauterine device (0.1%), and tubal ligation (0.1%). Vaginal wetness was reported by 18.4% of users and was one of the most common side effects, second only to amenorrhoea (62.5%). It was also what 17.5% of the women liked least about using this method. According to almost half the respondents, men regard women who use the injectable contraceptive as "wet", "cold" and/or "tasteless". These survey findings were supported by participants of 14 focus group interviews held in the sub-district. Since some South African men may prefer dry sex the perception that the injectable contraceptive increases vaginal wetness may be problematic for women who use it. Whilst vaginal wetness can only be classified as a subjective side effect at this stage, further investigations are needed as many South African women opt to use this method.