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Acetogenic bacteria: what are the in situ consequences of their diverse metabolic versatilities?

The four decades of the now classic studies by Harland G. Wood and Lars G. Ljungdahl lead to the resolution of the autotrophic acetyl-CoA 'Wood/Ljungdahl' pathway of acetogenesis. This pathway is the hallmark of acetogens, but is also used by other bacteria, including methanogens and sulfate-reducing bacteria, for both catabolic and anabolic purposes. Thus, the pathway is wide spread in nature and plays an important role in the global turnover of carbon. Because most historical studies with acetogens focused on the biochemistry of the acetyl-CoA pathway, the metabolic diversity and ecology of acetogens remained largely unexplored for many years. Although acetogens were initially conceived to be a somewhat obscure bacteriological group with limited metabolic capabilities, it is now clear that acctogens are arguably the most metabolically diverse group of obligate anaerobes characterized to date. Their anaerobic metabolic arsenal includes the capacity to oxidize diverse substrates, including aromatic, C1, C2, and halogenated compounds, and engage a large number of alternative energy-conserving, terminal electron-accepting processes, including classic fermentations and the dissimilation of inorganic nitrogen. In this regard, one might consider acetogens on a collective basis as the pseudomonads of obligate anaerobes. By virtue of their diverse metabolic talents, acetogens can be found in essentially all habitats. This review evaluates the metabolic versatilities of acetogens relative to both the engagement (regulation) of the acetyl-CoA pathway and the ecological roles likely played by this bacteriogical group.

Acetic Acid↗

Electrophoretic resolution versus fluctuations of the lateral dimensions of a capillary.

Because the local electrical resistance is inversely proportional to the local cross-section of a capillary, the intensity of the electric field varies along the migration path if the inner diameter of the capillary is not constant. Therefore, fluctuations of the lateral dimensions of a capillary can directly affect the net elution time as well as the peak width, and, hence the final resolution. In this article, we develop the theoretical framework for the study of such effects. We then examine the simple case where both the mobility and the diffusion coefficient are field-independent; in particular, we demonstrate that resolution can be severely reduced if the inner walls are not flat, and that optimal resolution is always obtained for perfectly flat walls. Generalized to ultrathin gels, our results clearly indicate that both random and systematic variations of the gel thickness can greatly affect the performance of the separation process. Acceptable degrees of flatness are estimated for both geometries. This study thus provides a quantitative understanding of the type of quality control one requires to obtain optimal results with capillaries and ultrathin gels.

Electrophoresis↗

Anaplastic transformation of thyroid cancer: review of clinical, pathologic, and molecular evidence provides new insights into disease biology and future therapy.

BACKGROUND: Anaplastic thyroid cancer ranks among the most lethal of all human malignancies. Its rarity and rapidly fatal course have made it a difficult cancer to both study and treat. Unfortunately, there has been little progress in the management and control of this malignancy. Anaplastic transformation, or the intratumoral evolution of anaplastic carcinoma from pre-existing differentiated thyroid cancer, has become a well-accepted process, despite a limited understanding of its underlying mechanisms. METHODS: It is through review of the literature that an understanding of the aggressive disease biology can be developed. The aim of this review is to evaluate the relevant clinical, pathologic, and molecular studies to develop an insight into the mechanisms that underlie the intratumoral molecular evolution of anaplastic thyroid carcinoma. RESULTS/CONCLUSION: It is based on an understanding of this process that effective treatments for this aggressive malignancy are currently being developed.

Carcinoma↗

Method for predicting fill weight variation when packing powders using vacuum/purge fillers.

The present report demonstrates that for at least four pharmaceutical powders, the variation in fill weight associated with a vacuum/purge filling port is correlated with the length-diameter ratio of that port. This relationship has been mathematically modeled, and a design curve based on production data is presented, which depicts this relationship over a wide range of length-diameter ratios. For powders with properties similar to those presented, the design curve may be used to determine the dimensions of the port which will yield acceptable process weight control. Fill weight variances also can be predicted given a fixed port diameter. For other powders, the model can be used to create design curves with a few data points.

Particle Size↗

Carbohydrate oxidation coupled to Fe(III) reduction, a novel form of anaerobic metabolism.

An isolate, designated GC-29, that could incompletely oxidize glucose to acetate and carbon dioxide with Fe(III) serving as the electron acceptor was recovered from freshwater sediments of the Potomac River, Maryland. This metabolism yielded energy to support cell growth. Strain GC-29 is a facultatively anaerobic, gram-negative motile rod which, in addition to glucose, also used sucrose, lactate, pyruvate, yeast extract, casamino acids or H2 as alternative electron donors for Fe(III) reduction. Stain GC-29 could reduce NO3(-), Mn(IV), U(VI), fumarate, malate, S2O3(2-), and colloidal S0 as well as the humics analog, 2,6-anthraquinone disulfonate. Analysis of the almost complete 16S rRNA sequence indicated that strain GC-29 belongs in the Shewanella genus in the epsilon subdivision of the Proteobacteria. The name Shewanella saccharophilia is proposed. Shewanella saccharophilia differs from previously described fermentative microorganisms that metabolize glucose with the reduction of Fe(III) because it transfers significantly more electron equivalents to Fe(III); acetate and carbon dioxide are the only products of glucose metabolism; energy is conserved from Fe(III) reduction; and glucose is not metabolized in the absence of Fe(III). The metabolism of organisms like S. saccharophilia may account for the fact that glucose is metabolized primarily to acetate and carbon dioxide in a variety of sediments in which Fe(III) reduction is the terminal electron accepting process.

Journal Article↗

[Changes in the concept of fracture healing and callus formation].

Although the treatment of fractures has repeatedly been documented throughout history, little attention has been paid to the historical development of the understanding of fracture healing and callus formation in itself. In antiquity, the capability of bone to regenerate was doubted, and Galen attributed the consolidation of a bone fracture to a sap (succus ossificus) that came from the food ingested. Later, in the eighteenth century, a haughty debate broke out as to the seat of the organ that controlled the by now accepted process of new bone formation. While Duhamel du Monceau (1700-1781) believed the periosteum was responsible for the callus formation--this idea being based on his observations of trees--Albrecht von Haller (1708-1777) and his scholars and disciples ascribed this function to the succus ossificus and to the bone marrow. The studies on experimental fracture healing that began during the seventeenth century resulted in the eighteenth-century discovery of the complex processes involved in the formation of callus with the aid of madder staining. The conciliation of the two major schools of thought was finally brought about when in the nineteenth century both periosteal and endosteal callus formation were shown to occur. The anatomical reposition of the fractured components and the stable fixation through osteosynthesis led to a novelty of the twentieth century: callus-free healing of bone fractures. Based on key texts and their authors, the zigzag course taken by our understanding of bone fracture healing and callus formation is illustrated.

Bony Callus↗

Geochemical and physiological evidence for mixed aerobic and anaerobic field biodegradation of coal tar waste by subsurface microbial communities.

We used geochemical analyses of groundwater and laboratory-incubated microcosms to investigate the physiological responses of naturally occurring microorganisms to coal-tar-waste constituents in a contaminated aquifer. Waters were sampled from wells along a natural hydrologic gradient extending from uncontaminated (1 well) into contaminated (3 wells) zones. Groundwater analyses determined the concentrations of carbon and energy sources (pollutants or total organic carbon), final electron acceptors (oxygen, nitrate, sulfate), and metabolic byproducts (dissolved inorganic carbon [DIC], alkalinity, methane, ferrous iron, sulfide, Mn2+). In the contaminated zone of the study site, concentrations of methane, hydrogen, alkalinity, and DIC were enhanced, while dissolved oxygen and nitrate were depleted. Field-initiated biodegradation assays using headspace-free serum bottle microcosms filled with groundwater examined metabolism of the ambient organic contaminants (naphthalene, 2-methylnaphthalene, benzothiophene, and indene) by the native microbial communities. Unamended microcosms from the contaminated zone demonstrated the simultaneous degradation of several coal-tar-waste constituents at the in situ temperature (10 degrees C). Lag phases prior to the onset of biodegradation indicated the prevalence of both aerobic and anaerobic conditions in situ. Electron acceptor-amended microcosms from the most contaminated well waters demonstrated only aerobic naphthalene degradation. Collectively, the geochemical and microbial evidence show that biodegradation of coal-tar-waste constituents occurs via both aerobic and anaerobic terminal electron accepting processes at this site.

Bacteria, Aerobic↗

Effect of Supplemental Electron Donors on the Microbial Reduction of Fe(III), Sulfate, and CO(2) in Coal Mining-Impacted Freshwater Lake Sediments.

In acidic mining-impacted lake sediments, the microbial reduction of Fe(III) is the dominant electron-accepting process, whereas the reduction of sulfate seems to be restricted to a narrow sediment zone of elevated pH and lower amounts of total and reactive iron. To evaluate the microbial heterogeneity and the commensal interactions of the microbial community, the flow of supplemental carbon and reductant was evaluated in four different zones of the sediment in anoxic microcosms at the in situ temperature of 12 degrees C. Substrate consumption, product formation, and the potential to reduce Fe(III) and sulfate were similar with both upper and lower sediment zones. In the upper acidic iron-rich sediment zone, the rate of Fe(II) formation 204 nmol ml(-1) d(-1) was enhanced to 833 nmol ml(-1) d(-1) and 462 nmol ml(-1) d(-1) by supplemental glucose and H(2), respectively. Supplemental lactate and acetate were not consumed under acidic conditions and decreased the rate of Fe(II) formation to 130 nmol ml(-1) d(-1) and 52 nmol ml(-1) d(-1), respectively. When the pH of the upper sediment increased above pH 5, acetate-dependent reduction of sulfate was initiated even though the pool of Fe(III) was not depleted. In deeper sediment zones with elevated pH, the rapid consumption of acetate was always coincident to a decrease in the concentration of sulfate and soluble Fe(II), indicating the formation of Fe(II) sulfides. Although the reduction of Fe(III) was still an ongoing process in deeper sediment zones, the formation of Fe(II) was only slightly enhanced by the consumption of glucose or cellobiose, but not by H(2) or acetate. H(2)-utilizing acetogens seemed to be involved in the consumption of H(2). These collective results indicated (i) that the reduction of Fe(III) predominated over the reduction of sulfate as long as the sediment remained acidic and carbon-limited, and (ii) that the sulfate-reducing microbiota in this heterogeneous sediment were better adapted to the geochemical gradients present than were other neutrophilic dissimilatory Fe(III) reducers.

Journal Article↗

Biological preservation of foods with reference to protective cultures, bacteriocins and food-grade enzymes.

A review is presented on the present status of biological preservation of foods. Recent developments are discussed with respect to underlying mechanisms of inhibition by 'protective' cultures, and special reference is made to lactic acid bacteria (LAB) and their 'food-grade' safety. The role of bacteriocins, their limitations and potentiating role in biological systems, is also addressed. The use of enzymes (e.g. lysozyme) for food preservation is mainly restricted by economic factors, their inactivation by endogenous food components and their limited activity spectrum. Practical applications of protective cultures refer to particular food commodities that either constitute novel systems with respect to packaging and/or composition, or represent special hygienic risks. It is concluded that biological preservation cannot substitute GMP; it, however, offers an additional (and acceptable) processing parameter for improving the safety and assuring the quality of a given food.

Antibiosis↗

Drugs or drums: what relieves postoperative pain?

The cybernetic model of analgesia is a simple servo-loop: altered sensory input--complaints--medication--temporary restoration of acceptable processing of the sensory input--return of altered sensations. Under ordinary clinical circumstances the functioning of this mechanism is masked by "noise" generated by the interaction of several persons who provide the medication and time factors prolonging the interval between the onset of complaints and the action of the medication. Demand analgesia (DA) has been designed to provide prompt pain relief under the patient's direct control. The technique turned out to be useful also for the investigation of acute pain's course. The present protocol was designed to study the precision with which patients respond to minute changes in the level of a narcotic (hydromorphone) at the target site: this was achieved by varying in a double-blind fashion the amount of narcotic delivered in response to triggering the apparatus and noting how the patient adapts the dosing interval in response to the imposed changes. Thirty-four patients were studied following extensive surgery. Observation times ranged from 16 to 75 h (median 45 h) during which 89 dose changes were initiated. While the majority of subjects adapted successfully to up to 4-fold variations in drug delivery, a sizeable minority (7/34) triggered the apparatus in response to some clue other than the amount of drug received. These non-drug responders seem to correspond to the "placebo reactors" of other therapeutic settings. Our calculations allowed the estimation of how large was the contribution of drug action and non-drug action to the overall therapeutic effect. In the drug-responsive group, the mean contribution of drug action was 78%, while among the non-drug responders this amounted to only 40%. The difference between these two figures is statistically significant (P less than 0.02), at the same time it also reveals important contributions of non-drug factors in drug responders and vice versa. Whether a subject behaves as a placebo reactor or not might depend on the circumstances rather than on innate mechanisms.

Analgesia↗

Italian version of the International Knee Documentation Committee Subjective Knee Form: cross-cultural adaptation and validation.

PURPOSE: Patient-oriented measures, represented by self-administered questionnaires, have become an important aspect of clinical outcome assessment. To be used with different language groups and in different countries, questionnaires must be translated and adapted to new cultural characteristics, and then validated by a widely accepted process to evaluate reliability and validity, fundamental characteristics for each measure. The aim of the study was to perform the cross-cultural adaptation and to assess the reliability and validity of the Italian version of the International Knee Documentation Committee (IKDC) Subjective Knee Form. TYPE OF STUDY: A cross-cultural adaptation and cross-sectional study of a sample of patients undergoing anterior cruciate ligament (ACL) reconstruction with a subsample followed up prospectively for retest reliability. METHODS: The IKDC Subjective Knee Form was culturally adapted for Italian-speaking people, following the simplified Guillemin criteria. Reliability and validity were assessed in a cross-sectional study of 50 consecutive patients undergoing ACL reconstruction. A subsample of 20 patients was followed up prospectively for retest reliability. RESULTS: The results were compared with other validated patient-oriented measures. The principal IKDC scale showed a high correlation with other patient-oriented measures as hypothesized, and it also showed good values with regard to reproducibility, consistency, and validity, compared with the versions of IKDC published in other languages. CONCLUSIONS: These findings suggest that the evaluation capacities of the IKDC Italian version are equivalent to those of other language versions of the IKDC. LEVEL OF EVIDENCE: Level II.

Adolescent↗

Anaerobic degradation of naphthalene in a fluvial aquifer: a radiotracer study.

A radiotracer study was conducted in a creosote-contaminated aquifer beneath the Fraser River, British Columbia Canada to investigate the in situ degradation of naphthalene. The groundwater is anaerobic, with abundant methane, ferrous iron and carbon dioxide. This study followed earlier work at the site where the contaminant distribution could only be explained by invoking a mass loss through degradation, even though extensive field and laboratory microcosm studies closer to the source zone onshore could not confirm degradation. Accordingly, 14C-naphthalene was injected into the aquifer offshore, further from the source zone where modeling suggested degradation was occurring. During the 230-day monitoring period, 14CO2 was detected, confirming the degradation of the radio-labeled naphthalene tracer. A zero-order degradation rate of naphthalene of 5 microg/L-day was estimated based on the decrease in 14C-naphthalene concentration with time. While the degradation pathway could not be determined from the radiotracer study alone, the geochemistry of the site suggests that either iron reduction or methanogenesis is the terminal electron accepting processes responsible for naphthalene oxidation.

Anaerobiosis↗

Methyl-coenzyme M reductase genes: unique functional markers for methanogenic and anaerobic methane-oxidizing Archaea.

In many anoxic environments, methanogenesis is the predominant terminal electron accepting process involved in the mineralization of organic matter, which is catalyzed by methanogenic Archaea. These organisms represent a unique but phylogenetically diverse guild of prokaryotes, which can be conveniently tracked in the environment by targeting the mcrA gene as a functional marker. This gene encodes the alpha subunit of the methyl-coenzyme M reductase (MCR), which catalyzes the last step in methanogenesis and is present in all methanogens. Cultivation-independent analysis of methanogenic communities involves the polymerase chain reaction (PCR) amplification of the mcrA gene from extracted community DNA, comparative analysis of mcrA clone libraries, or PCR-based fingerprinting analysis by terminal restriction fragment polymorphism analysis (T-RFLP). It has also been suggested that anaerobic methane-oxidizing Archaea possess MCR, which facilitates detection of this novel group of "reverse methanogens" as well using the mcrA gene as a functional marker.

Archaea↗

An assessment of natural biotransformation of petroleum hydrocarbons and chlorinated solvents at an aquifer plume transect.

Field biogeochemical characterization and laboratory microcosm studies were performed to assess the potential for future biotransformation of trichloroethylene (TCE) and toluene in a plume containing petroleum hydrocarbons and chlorinated solvents at the former Wurtsmith Air Force Base in Oscoda, MI. In situ terminal electron accepting processes (TEAPs), contaminant composition and microbial phylogeny were studied at a plume transect 100 m downgradient of the source. The presence of reduced electron acceptors, relevant microbial communities, and elevated dissolved methane and carbon dioxide concentrations at the transect, as well as downgradient accumulation of BTEX metabolites and dechlorination products, indicated that past or current reductive dechlorination at the transect was likely driven by BTEX biodegradation in the methanogenic zone. However, TCE and toluene mineralization in sediment-groundwater microcosms without added electron acceptors did not exceed 5% during 300 days of incubation and was nearly invariant with original sediment TEAP, even following amendments of nitrogen and phosphorus. Mineralization rates were on the order of 0.0015-0.03 mumol/g day. After 8 months, microcosms showed evidence of methanogenesis, but CH4 and CO2 production arose from the degradation of contaminants other than toluene. Cis-dichloroethylene was observed in only one methanogenic microcosm after more than 500 days. It appears likely that spatially and temporally dynamic redox zonation at the plume transect will prevent future sustained reductive dehalogenation of highly chlorinated solvents, for during the course of a year, the predominant TEAP at the highly contaminated water table shifted from methanogenesis to iron- and sulfate-reduction. It is recommended that biotransformation studies combine considerations of long-term, spatially relevant changes in redox zonation with laboratory-scale studies of electron donor utilization and cometabolic substrate transformation to yield a more accurate assessment of natural bioattenuation of specific pollutants in aquifers contaminated by undefined organic waste mixtures.

Biotransformation↗

Quantitative myocardial perfusion SPECT.

In recent years, there has been much interest in the clinical application of attenuation compensation to myocardial perfusion single photon emission computed tomography (SPECT) with the promise that accurate quantitative images can be obtained to improve clinical diagnoses. The different attenuation compensation methods that are available create confusion and some misconceptions. Also, attenuation-compensated images reveal other image-degrading effects including collimator-detector blurring and scatter that are not apparent in uncompensated images. This article presents basic concepts of the major factors that degrade the quality and quantitative accuracy of myocardial perfusion SPECT images, and includes a discussion of the various image reconstruction and compensation methods and misconceptions and pitfalls in implementation. The differences between the various compensation methods and their performance are demonstrated. Particular emphasis is directed to an approach that promises to provide quantitative myocardial perfusion SPECT images by accurately compensating for the 3-dimensional (3-D) attenuation, collimator-detector response, and scatter effects. With advances in the computer hardware and optimized implementation techniques, quantitatively accurate and high-quality myocardial perfusion SPECT images can be obtained in clinically acceptable processing time. Examples from simulation, phantom, and patient studies are used to demonstrate the various aspects of the investigation. We conclude that quantitative myocardial perfusion SPECT, which holds great promise to improve clinical diagnosis, is an achievable goal in the near future.

Algorithms↗

Implementing information systems in health care organizations: myths and challenges.

Successfully implementing patient care information systems (PCIS) in health care organizations appears to be a difficult task. After critically examining the very notions of 'success' and 'failure', and after discussing the problematic nature of lists of 'critical success- or failure factors', this paper discusses three myths that often hamper implementation processes. Alternative insights are presented, and illustrated with concrete examples. First of all, the implementation of a PCIS is a process of mutual transformation; the organization and the technology transform each other during the implementation process. When this is foreseen, PCIS implementations can be intended strategically to help transform the organization. Second, such a process can only get off the ground when properly supported by both central management and future users. A top down framework for the implementation is crucial to turn user-input into a coherent steering force, creating a solid basis for organizational transformation. Finally, the management of IS implementation processes is a careful balancing act between initiating organizational change, and drawing upon IS as a change agent, without attempting to pre-specify and control this process. Accepting, and even drawing upon, this inevitable uncertainty might be the hardest lesson to learn.

Delivery of Health Care↗

'Can I put--I want a slippers to put on': young children's development of request forms in a code-switching environment.

This paper concerns the development of semi-modal want/want to, and modals may, can and could in two Trinidadian children, whose Verb Phrase development was studied in naturalistic settings, between the ages of 2;3 and 4;1, and 2;4 and 4;9. The similarities and differences in development between the two are important for underlining a number of key factors in the acquisition process. Accepting the salience of requests/demands in early child language, the study draws attention to the child's propensity for using the means most readily available to him/her in the input, for expression of this function. Additionally, there is evidence of the child's ability to make stylistic discriminations at a very early age and of the propensity for making semantic distinctions among forms. The study indicates that work on the development of grammatical features is incomplete without consideration of sociolinguistic aspects of usage as integral to grammatical acquisition from the outset.

Child↗

Assessment of natural attenuation of chlorinated aliphatics and BTEX in subarctic groundwater.

We examined biogeochemistry and microbiology associated with natural attenuation of trichloroethene (TCE), trichloroethane (TCA), and benzene in a subarctic aquifer. Identification of a predominant terminal electron-accepting process (TEAP) and characterization of typical natural attenuation footprints was difficult. Hydrogen and ferrous iron concentrations suggested that iron reduction was the predominant TEAP; calculated in situ Gibbs free energies for iron reduction were energetically feasible at all wells although a source of ferric iron has not been conclusively determined. The presence of dissolved sulfide and favorable free energies for sulfate reduction provided support of concurrent iron and sulfate reduction. Methanogenesis from H2/CO2 was generally not energetically favorable. The presence of TCE and TCA degradation intermediates suggested that biological reductive dechlorination occurred, although proportions of intermediates relative to parent compounds remained stable. By September 2000, contaminant concentrations were within regulatory standards at most sampling points. However, low rates of microbial activity and incomplete degradation imply that intrinsic bioremediation did not likely represent an important contribution to contaminant removal atthis site, where dilution appeared to be the primary attenuation mechanism.

Biodegradation, Environmental↗