BACKGROUND

Severe sepsis is characterised by intravascular or extravascular infection with microbial agents, systemic inflammation and microcirculatory dysfunction, leading to tissue damage, organ failure and death. The growth factor angiopoietin (Ang-1) has therapeutic potential but recombinant Ang-1 tends to aggregate and has a short half-life in vivo. This study aimed to investigate the acute effects of the more stable Ang-1 variant matrilin-1-angiopoietin-1 (MAT.Ang-1) on the function of the microcirculation in an experimental model of sepsis, and whether any protection by MAT-Ang-1 was associated with modulation of inflammatory cytokines, angiogenic factors or the endothelial nitric oxide synthase (eNOS)-Akt and vascular endothelial (VE)-cadherin pathways.

METHODS

Aluminium window chambers were implanted into the dorsal skinfold of male C3H/HeN mice (7 to 10 weeks old) to expose the striated muscle microcirculation. Endotoxemia was induced by intraperitoneal injection of lipopolysaccharide (LPS, 1 mg/kg at 0 and 19 hours). MAT.Ang-1 was administered intravenously 20 hours after the onset of sepsis. Microcirculatory function was evaluated by intravital microscopy and Doppler fluximetry.

RESULTS

Endotoxemia resulted in macromolecular leak, which was ameliorated by MAT.Ang-1 post-treatment. LPS induced a dramatic reduction in tissue perfusion, which was improved by MAT.Ang-1. Proteome profiler array analysis of skeletal muscle also demonstrated increased inflammatory and reduced angiogenic factors during endotoxemia. MAT.Ang-1 post-treatment reduced the level of IL-1β but did not significantly induce the expression of angiogenic factors. MAT.Ang-1 alone did not induce leak or increase angiogenic factors but did reduce vascular endothelial growth factor expression in controls.

CONCLUSIONS

Administration of MAT.Ang-1 after the onset of sepsis protects the microcirculation from endotoxemia-induced vascular dysfunction through reducing inflammation but without pro-angiogenic actions, thus representing a novel, potential pharmacotherapeutic agent for the treatment of sepsis.

Dickinsonia is one of the most recognizable forms in the Ediacaran fauna, but its phylogenetic position has been contentious, and it has been placed in almost every kingdom of life. Here, it is hypothesized that the affinities of Dickinsonia lie with the Placozoa (Metazoa), an understudied phylum that is widespread in tropical seas worldwide. Modern placozoans show obvious differences in size and axial organization compared with Dickinsonia, but these differences can be accounted for by the stem-group/crown-group distinction. The affinity with placozoans is evidenced primarily by the unique feeding mode of Dickinsonia, which is demonstrated by a series of feeding traces. These traces indicate that Dickinsonia moved over the Ediacaran matgrounds, and digested the mat using its entire lower sole. The ability of Dickinsonia to move negates an algal, fungal, or sponge affinity, while the feeding mode, external digestion with a ventral sole, rules out placement within any sponge or eumetazoan lineage. The only organisms that both move and feed in this manner are placozoans. Recent molecular phylogenetic studies have demonstrated that placozoans lie above sponges but below Eumetazoa. We hypothesize that Dickinsonia and other externally digesting Ediacaran forms are either stem-placozoans, or a series of extinct lineages above sponges and below eumetazoans on the metazoan tree. We discuss the potential evolutionary transitions between the main metazoan feeding modes in the context of the emerging molecular phylogeny, and suggest that aspects of the sponge and placozoan feeding strategies are relicts of nonuniformitarian Proterozoic ocean conditions.

Essential oil of seeds of Trachyspermum ammi (Linn.) Sprauge and its pure constituent thymol showed promising results when evaluated for larvicidal, oviposition-deterrent, vapor toxicity, and repellent activity against malarial vector, Anopheles stephensi. Thymol was 1.6-fold more toxic than the oil toward fourth-instar larvae of A. stephensi with LD(50) values of 48.88 and 80.77 microg/ml, respectively. Egg laying by female adults of A. stephensi was much significantly reduced when exposed to vapors of thymol compared to the oil of T. ammi seeds, and similar effects were recorded for subsequent egg hatching and larval survival. Vapor toxicity assay showed LC(50) value of 79.5 mg/mat for thymol against adults of A. stephensi, whereas the crude oil exhibited the LC(50) value of 185.4 mg/mat. Thymol provided complete repellency toward A. stephensi adults at the dose of 25.0 mg/mat after 1 h duration, whereas same degree of repellency was obtained by the oil at the dose of 55.0 mg/mat, indicating its double-fold activity than the oil.

The Dutch elm disease fungus Ophiostoma novo-ulmi, which has destroyed billions of elm trees worldwide, originally invaded Europe as a series of clonal populations with a single mating type (MAT-2) and a single vegetative incompatibility (vic) type. The populations then rapidly became diverse with the appearance of the MAT-1 type and many vegetative incompatibility types. Here, we have investigated the mechanism using isolates from sites in Portugal at which the rapid evolution of O. novo-ulmi populations from clonality to heterogeneity was well established. We show by genetic mapping of vic and MAT loci with AFLP markers and by sequence analysis of MAT loci that this diversification was due to selective acquisition by O. novo-ulmi of the MAT-1 and vic loci from another species, Ophiostoma ulmi. A global survey showed that interspecies transfer of the MAT-1 locus occurred on many occasions as O. novo-ulmi spread across the world. We discuss the possibility that fixation of the MAT-1 and vic loci occurred in response to spread of deleterious viruses in the originally clonal populations. The process demonstrates the potential of interspecies gene transfer for facilitating rapid adaptation of invasive organisms to a new environment.

Some of the most remarkable fossils preserve cellular details of soft tissues. In many of these, the tissues have been replaced by calcium phosphate. This process has been assumed to require elevated concentrations of phosphate in sediment pore waters. In decay experiments modern shrimps became partially mineralized in amorphous calcium phosphate, preserving cellular details of muscle tissue, particularly in a system closed to oxygen. The source for the formation of calcium phosphate was the shrimp itself. Mineralization, which was accompanied by a drop in pH, commenced within 2 weeks and increased in extent for at least 4 to 8 weeks. This mechanism halts the normal loss of detail of soft-tissue morphology before fossilization. Similar closed conditions would prevail where organisms are rapidly overgrown by microbial mats.

Worldwide development of cyanobacterial blooms has significantly increased in marine and continental waters in the last century due to water eutrophication. This phenomenon is favoured by the ability of planktonic cyanobacteria to synthesize gas vesicles that allow them to float in the water column. Besides, benthic cyanobacteria that proliferate at the bottom of lakes, rivers and costal waters form dense mats near the shore. Cyanobacterial massive proliferation is of public concern regarding the capacity of certain cyanobacterial strains to produce hepatotoxic and neurotoxic compounds that can affect public health, human activities and wild and stock animals. The cholinergic synapses and voltage-gated sodium channels constitute the targets of choice of cyanobacterial neurotoxins. Anatoxin-a and homoanatoxin-a are agonists of nicotinic acetylcholine receptors. Anatoxin-a(s) is an irreversible inhibitor of acetylcholinesterase. Saxitoxin, kalkitoxin and jamaicamide are blockers of voltage-gated sodium channels, whereas antillatoxin is an activator of such channels. Moreover the neurotoxic amino acid l-beta-N-methylamino-l-alanine was shown to be produced by diverse cyanobacterial taxa. Although controversial, increasing in vivo and in vitro evidence suggest a link between the ingestion of l-beta-N-methylamino-l-alanine and the development of amyotrophic lateral sclerosis/Parkinsonism-dementia complex, a neurodegenerative disease. This paper reviews the occurrence of cyanobacterial neurotoxins, their chemical properties, mode of action and biosynthetic pathways.

A system for genotyping Saccharomyces cerevisiae is described based on a multiplex of ten microsatellite loci and the MAT locus. A database of genotypes has been developed for 246 yeast strains, including a large set of commercial wine yeasts, as well as 35 sequenced natural isolates currently being sequenced. The latter allow us, for the first time, to make direct comparisons of the relationship between DNA sequence data and microsatellite-based genotypes. The genotyping system provides a rapid and valuable system for strain identification as well as studying population genetics of S. cerevisiae.

Despite evidence linking trait mindfulness and mindfulness training with a broad range of effects, still little is known about its underlying active mechanisms. Mindfulness is commonly defined as (1) the ongoing monitoring of present-moment experience (2) with an orientation of acceptance. Building on conceptual, clinical, and empirical work, we describe a testable theoretical account to help explain mindfulness effects on cognition, affect, stress, and health outcomes. Specifically, Monitor and Acceptance Theory (MAT) posits that (1), by enhancing awareness of one's experiences, the skill of attention monitoring explains how mindfulness improves cognitive functioning outcomes, yet this same skill can increase affective reactivity. Second (2), by modifying one's relation to monitored experience, acceptance is necessary for reducing affective reactivity, such that attention monitoring and acceptance skills together explain how mindfulness improves negative affectivity, stress, and stress-related health outcomes. We discuss how MAT contributes to mindfulness science, suggest plausible alternatives to the account, and offer specific predictions for future research.

The purposes of this study were to determine the reliability of unilateral vertical, horizontal, and lateral countermovement jump assessments, the interrelationship between these tests, and their usefulness as predictors of sprint (10 m) and change-of-direction (COD) performance for 80 men and women physical education students. Jump performance was assessed on a contact mat and sprint, and COD performances were assessed using timing lights. With regard to the reliability statistics, the largest coefficient of variation (CV) was observed for the vertical jump (CV = 6.7-7.2%) of both genders, whereas the sprint and COD assessments had smallest variability (CV = 0.8 to 2.8%). All intraclass correlation coefficients (ICC) were greater than 0.85, except for the men's COD assessment with the alternate leg. The shared variance between the single-leg vertical, horizontal, and lateral jumps for men and women was less than 50%, indicating that the jumps are relatively independent of one another and represent different leg strength/power qualities. The ability of the jumps to predict sprint and COD performance was limited (R2 < 43%). It would seem that the ability to change direction with 1 leg is relatively independent of a COD with the other leg, especially in the women (R < 30%) of this study. However, if 1 jump assessment were selected to predict sprint and COD performance in a test battery, the single-leg horizontal countermovement jump would seem the logical choice, given the results of this study. Many of the findings in this study have interesting diagnostic and training implications for the strength and conditioning coach.

Taxonomic studies were performed on four strains (D-1/D-an/IIT, C/C-an/B1T, A-1/C-an/C1T and A-1/C-an/IT) of anaerobic, gram-positive, spore-forming bacteria originally isolated from a mat sample retrieved from a shallow, moated area around Lake Fryxell, an Antarctic freshwater lake. Phylogenetic analyses based on 16S rRNA gene sequence data indicated that these strains are affiliated with cluster I clostridia and form a coherent group with Clostridium estertheticum and Clostridium laramiense. Similarity values among 16S rRNA gene sequences within this assemblage ranged between 96.7 and 99.8%. Despite the close phylogenetic relationship, several distinguishing phenotypic traits were found among the novel strains using a polyphasic approach. All strains were psychrophilic, but the temperature optimum for growth differed markedly, ranging from 4 to 16 degrees C. In addition, substrate utilization patterns, fermentation end products, cellular fatty acid profiles and morphological traits enabled a clear differentiation between the strains. DNA-DNA hybridization experiments revealed that each of the four novel strains represents a distinct species, with DNA-DNA similarity values to related strains in the range 16-62%. In contrast, the type strains of C. estertheticum and C. laramiense shared 79% DNA-DNA similarity, indicating a close relationship at the species level. On the basis of genetic and phenotypic properties, it is proposed to designate four novel species of the genus Clostridium to harbour the newly isolated strains: Clostridium frigoris sp. nov. (type strain D-1/D-an/IIT=DSM 14204T=ATCC BMAA-579T), Clostridium lacusfryxellense sp. nov. (type strain C/C-an/B1T=DSM 14205T=ATCC BAA-580T), Clostridium bowmaniisp. nov. (type strain A-1/C-an/C1T=DSM 14206T=ATCC BAA-581T) and Clostridium psychrophilum sp. nov. (type strain A-1/C-an/IT=DSM 14207T=ATCC BAA-582T). It is also proposed to unite C. laramiense and C. estertheticum under C. estertheticum. The subspecies C. estertheticum subsp. laramiense subsp. nov. is established, represented by strain ATCC 51254T (=DSM 14884T). The type strain of C. estertheticum subsp. estertheticum remains NCIMB 12511T (=DSM 8809T).

Roseiflexus sp. strains were cultivated from a microbial mat of an alkaline siliceous hot spring in Yellowstone National Park. These strains are closely related to predominant filamentous anoxygenic phototrophs found in the mat, as judged by the similarity of small-subunit rRNA, lipid distributions, and genomic and metagenomic sequences. Like a Japanese isolate, R. castenholzii, the Yellowstone isolates contain bacteriochlorophyll a, but not bacteriochlorophyll c or chlorosomes, and grow photoheterotrophically or chemoheterotrophically under dark aerobic conditions. The genome of one isolate, Roseiflexus sp. strain RS1, contains genes necessary to support these metabolisms. This genome also contains genes encoding the 3-hydroxypropionate pathway for CO(2) fixation and a hydrogenase, which might enable photoautotrophic metabolism, even though neither isolate could be grown photoautotrophically with H(2) or H(2)S as a possible electron donor. The isolates exhibit temperature, pH, and sulfide preferences typical of their habitat. Lipids produced by these isolates matched much better with mat lipids than do lipids produced by R. castenholzii or Chloroflexus isolates.

Inferences about the evidence of life recorded in organic compounds within the Earth's ancient rocks have depended on 13C contents low enough to be characteristic of biological debris produced by the well-known CO2 fixation pathway, the Calvin cycle. 'Atypically' high values have been attributed to isotopic alteration of sedimentary organic carbon by thermal metamorphism. We examined the possibility that organic carbon characterized by a relatively high 13C content could have arisen biologically from recently discovered autotrophic pathways. We focused on the green non-sulphur bacterium Chloroflexus aurantiacus that uses the 3-hydroxypropionate pathway for inorganic carbon fixation and is geologically significant as it forms modern mat communities analogous to stromatolites. Organic matter in mats constructed by Chloroflexus spp. alone had relatively high 13C contents (-14.9%) and lipids diagnostic of Chloroflexus that were also isotopically heavy (-8.9% to -18.5%). Organic matter in mats constructed by Chloroflexus in conjunction with cyanobacteria had a more typical Calvin cycle signature (-23.5%). However, lipids diagnostic of Chloroflexus were isotopically enriched (-15.1% to -24.1%) relative to lipids typical of cyanobacteria (-33.9% to -36.3%). This suggests that, in mats formed by both cyanobacteria and Chloroflexus, autotrophy must have a greater effect on Chloroflexus carbon metabolism than the photoheterotrophic consumption of cyanobacterial photosynthate. Chloroflexus cell components were also selectively preserved. Hence, Chloroflexus autotrophy and selective preservation of its products constitute one purely biological mechanism by which isotopically heavy organic carbon could have been introduced into important Precambrian geological features.

Exiguobacterium antarcticum is a psychotropic bacterium isolated for the first time from microbial mats of Lake Fryxell in Antarctica. Many organisms of the genus Exiguobacterium are extremophiles and have properties of biotechnological interest, e.g., the capacity to adapt to cold, which make this genus a target for discovering new enzymes, such as lipases and proteases, in addition to improving our understanding of the mechanisms of adaptation and survival at low temperatures. This study presents the genome of E. antarcticum B7, isolated from a biofilm sample of Ginger Lake on King George Island, Antarctic peninsula.

OBJECTIVE

To review 16 years of National Collegiate Athletic Association (NCAA) injury surveillance data for women's gymnastics and identify potential areas for injury prevention initiatives.

BACKGROUND

In the 1988-1989 academic year, 112 schools were sponsoring varsity women's gymnastics teams, with approximately 1550 participants. By 2003-2004, the number of varsity teams had decreased 23% to 86, involving 1380 participants. Significant participation reductions during this time were particularly apparent in Divisions II and III.

RESULTS

A significant annual average decrease was noted in competition (-4.0%, P < .01) but not in practice (-1.0%, P = .35) injury rates during the sample period. Over the 16 years, the rate of injury in competition was more than 2 times higher than in practice (15.19 versus 6.07 injuries per 1000 athlete-exposures; rate ratio = 2.5, 95% confidence interval [CI] = 2.3, 2.8). A total of 53% of all competition and 69% of all practice injuries were to the lower extremity. A participant was almost 6 times more likely to sustain a knee internal derangement injury in competition than in practice (rate ratio = 5.7, 95% CI = 4.5, 7.3) and almost 3 times more likely to sustain an ankle ligament sprain (rate ratio = 2.7, 95% CI = 2.1, 3.4). The majority of competition injuries (approximately 70%) resulted from either landings in floor exercises or dismounts.

CONCLUSIONS

Gymnasts with a previous history of ankle sprain should either wear an ankle brace or use prophylactic tape on their ankles to decrease the risk of recurrent injury. Preventive efforts may incorporate more neuromuscular training and core stability programs in the off-season and preseason conditioning to enhance proper landing and skill mechanics. Equipment manufacturers are encouraged to reevaluate the design of the landing mats to allow for better absorption of forces.

BACKGROUND

Diagnosis of leptospirosis by the microscopic agglutination test (MAT) or by culture is confined to specialized laboratories. Although ELISA techniques are more common, they still require laboratory facilities. Rapid Diagnostic Tests (RDTs) can be used for easy point-of-care diagnosis. This study aims to evaluate the diagnostic performance of the RDTs LeptoTek Dri Dot, LeptoTek Lateral Flow, and Leptocheck-WB, prospectively.

METHODS

During 2001 to 2012, one or two of the RDTs at the same time have been applied prior to routine diagnostics (MAT, ELISA and culture) on serum specimens from participants sent in for leptospirosis diagnosis. The case definition was based on MAT, ELISA and culture results. Participants not fulfilling the case definition were considered not to have leptospirosis. The diagnostic accuracy was determined based on the 1(st) submitted sample and paired samples, either in an overall analysis or stratified according to days post onset of illness.

RESULTS

The overall sensitivity and specificity for the LeptoTek Dri Dot was 75% respectively 96%, for the LeptoTek Lateral Flow 78% respectively 95%, and for the Leptocheck-WB 78% respectively 98%. Based on the 1(st) submitted sample the sensitivity was low (51% for LeptoTek Dri Dot, 69% for LeptoTek Lateral Flow, and 55% for Leptocheck-WB), but substantially increased when the results of paired samples were combined, although accompanied by a lower specificity (82% respectively 91% for LeptoTek Dri Dot, 86% respectively 84% for LeptoTek Lateral Flow, and 80% respectively 93% for Leptocheck-WB).

CONCLUSIONS

All three tests present antibody tests contributing to the diagnosis of leptospirosis, thus supporting clinical suspicion and contributing to awareness. Since the overall sensitivity of the tested RDTs did not exceed 80%, one should be cautious to rely only on an RDT result, and confirmation by reference tests is strongly recommended.

OBJECTIVE

We sought to determine survival and clinical outcomes of high tibial osteotomy (HTO) with or without articular cartilage surgery and/or meniscal allograft transplantation in patients with medial compartment chondral pathology, varus malalignment, and/or meniscal deficiency, whether there is any difference in survival or clinical outcome between these patient cohorts, and whether there is any difference between opening- (OWHTO) and closing-wedge (CWHTO) techniques.

METHODS

A systematic review of multiple medical databases was performed using PRISMA guidelines. Study quality was assessed via modified Coleman Methodology Scores (MCMS).

RESULTS

Sixty-nine studies were included (4557 subjects). MCMS rating was overall poor. Mean follow-up was 7.1 years. Mean subject age was 53 years. Survival of isolated HTO was 92.4%, 84.5%, 77.3%, and 72.3% at 5, 10, 15, and 20 years of follow-up. At 5 years of follow-up, HTO with articular cartilage surgery had significantly greater survival (97.7%) than either isolated HTO (92.4%) or HTO with MAT (90.9%). Isolated HTO, HTO with articular cartilage surgery, and HTO with MAT all significantly improved subjective and objective clinical outcome scores. At two years of follow-up, survival was significantly greater following OWHTO (98.7%) versus CWHTO (96.7%). However, at all other time points with or without combined articular cartilage surgery and/or MAT, there was no significant survival difference between the techniques.

CONCLUSIONS

Survival and clinical outcomes of isolated HTO were excellent at short- and mid-term follow-ups, but deteriorated with time. HTO with concomitant procedures also demonstrated excellent early survival and clinical outcomes that deteriorated with time (up to 10 years).

Resveratrol is a potent anticancer. However, because of its low half-life (<0.25 h) the molecule is difficult to achieve the therapeutic concentration at the site of action. The aim of this work was to check the brain targeting ability of glyceryl behenate-based solid lipid nanoparticles (SLN) for resveratrol. SLN were prepared by solvent evaporation technique employing high speed homogenization followed by ultrasonication. SLN were designed at varying drug-lipid ratios (1:5, 1:9, 1:10, 1:11, 1:12 and 1:15) using Tween 80 or a combination of Tween 80 and polyvinyl alcohol (PVA) as surfactants. The mean particle size and zeta potential of the optimized formulation (drug-lipid ratio of 1:10) were 248.30 ± 3.80nm and -25.49 ± 0.49mV, respectively. The particle size and the encapsulation efficiency (EE) increased when varying the drug-lipid ratio from 1:5 to 1:15. Scanning electron microscopic (SEM) analysis showed that SLN were spherical in shape and had a smooth surface. The X-ray diffraction (XRD) and differential scanning calorimetry (DSC) analyses revealed that the matrix of drug-loaded SLN was in disordered crystalline phase. The in vitro release study in phosphate buffer pH 7.4 followed a sustained release pattern. The drug release data was found to fit best into Higuchi kinetic model suggesting the diffusion controlled mechanism of drug release. The cytotoxicity assay (MAT) showed that SLN were equally effective (P<0.5) as free resveratrol, as an anti-tumor agent. The in vivo biodistribution study using Wistar rats demonstrated that SLN could significantly (P<0.001) increase the brain concentration of resveratrol (17.28 ± 0.6344 μg/g) as compared to free resveratrol (3.45 ± 0.3961 μg/g). The results showed that our resveratrol-loaded SLN serve as promising therapeutic systems to treat neoplastic diseases located in the brain tissue.

Little information is available concerning the presence of viable Toxoplasma gondii in tissues of goats worldwide. In the present study, hearts of 234 goats obtained from a local USA grocery store were examined for T. gondii infection. Blood clot or fluid removed from each heart was tested for antibodies to T. gondii by using the modified agglutination test (MAT). Antibodies to T. gondii were found in 125 (53.4%) of 234 goats, with titers of 1:5 in 20, 1:10 in 44, 1:20 in 16, 1:40 in five, 1:160 in five, 1:320 in five, and 1:640 or higher in 30 goats. Hearts of 112 goats (46 goats <1:5, and 66 goats 1:10 or higher) were used for isolation of viable T. gondii by bioassays in mice. For bioassays, 50 g of the myocardium were digested in an acid pepsin solution and the digest inoculated into mice; the recipient mice were examined for T. gondii infection. Toxoplasma gondii was isolated from 29 goats; from hearts of one of 46 with titers of <1:5, one of nine with titers of 1:10, one of three with titers of 1:40, and 26 of 40 with titers of 1:160 or higher. Two isolates were highly virulent to outbred Swiss Webster mice; all infected mice died of toxoplasmosis, irrespective of the dose. All T. gondii isolates were subsequently grown in cell cultures. Genotyping of the 29 T. gondii isolates using 10 PCR-restriction fragment length polymorphism markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico) from DNA obtained from cell culture grown tachyzoites revealed 12 genotypes. Nine isolates were clonal Type II lineage, four isolates had type II alleles at all loci except a type I allele at the Apico locus, and four isolates were clonal Type III. The remaining 12 strains were divided into nine atypical genotypes, including five new and four previously identified genotypes. DNA sequences of four introns (EF1, HP2, UPRT1 and UPRT7) and two genes (GRA6 and GRA7) were generated for the five new genotypes. Comparing these sequences with previously published data revealed no unique sequences in these goat strains. Taken together, these results indicate high parasite prevalence and moderate genetic diversity of T. gondii in goats, which have important implications in public health. We believe this is the first genetic analysis of T. gondii isolates from goats in the USA.

The recently isolated strain L21-Fru-ABT represents moderately halophilic, obligately anaerobic and saccharolytic bacteria that thrive in the suboxic transition zones of hypersaline microbial mats. Phylogenetic analyses based on 16S rRNA genes, RpoB proteins and gene content indicated that strain L21-Fru-ABT represents a novel species and genus affiliated with a distinct phylum-level lineage originally designated Verrucomicrobia subdivision 5. A survey of environmental 16S rRNA gene sequences revealed that members of this newly recognized phylum are wide-spread and ecologically important in various anoxic environments ranging from hypersaline sediments to wastewater and the intestine of animals. Characteristic phenotypic traits of the novel strain included the formation of extracellular polymeric substances, a Gram-negative cell wall containing peptidoglycan and the absence of odd-numbered cellular fatty acids. Unusual metabolic features deduced from analysis of the genome sequence were the production of sucrose as osmoprotectant, an atypical glycolytic pathway lacking pyruvate kinase and the synthesis of isoprenoids via mevalonate. On the basis of the analyses of phenotypic, genomic and environmental data, it is proposed that strain L21-Fru-ABT and related bacteria are specifically adapted to the utilization of sulfated glycopolymers produced in microbial mats or biofilms.

Several cold-adapted strains isolated from a variety of algal-rich Antarctic and Southern Ocean samples formed three distinct groups within the class Flavobacteria, phylogenetically distant from other cultivated species. The first taxon, designated Algoriphagus ratkowskyi gen. nov., sp. nov., was isolated from sea ice and from saline lake cyanobacterial mats and includes non-motile, strictly aerobic, saccharolytic rod-like or serpentine strains that were most closely related to the genus Cyclobacterium according to 16S rDNA sequence analysis (sequence similarity 0.85). The second taxon, designated Brumimicrobium glaciale gen. nov., sp. nov., isolated from sea ice and from continental shelf sediment, formed gliding, rod-like cells that were facultatively anaerobic with a fermentative metabolism. The third taxon, designated Cryomorpha ignava gen. nov., sp. nov., isolated from Southern Ocean particulates and from quartz stone subliths, included strictly aerobic, pleomorphic rod-like cells. Brumimicrobium glaciale and Cryomorpha ignava were most closely allied with 'Microscilla aggregans var. catalatica', which, on the basis of its distinctive taxonomic traits, is also proposed as a new genus and species, Crocinitomix catalasitica gen. nov., sp. nov. It is proposed that the three genera Brumimicrobium, Cryomorpha and Crocinitomix belong to a new family, Cryomorphaceae fam. nov. (type genus Cryomorpha), as they possess generally similar morphological and ecophysiological characteristics and form a common and distinct clade within class FLAVOBACTERIA:

OBJECTIVE

Combined meniscal allograft transplantation (MAT) and cartilage repair or restoration is a recognized treatment for patients with painful, meniscus-deficient knees and full-thickness cartilage damage. The purpose of this systematic review was to compare outcomes after combined MAT and cartilage repair/restoration with the outcomes of isolated MAT or cartilage repair/restoration.

METHODS

Multiple databases were searched with specific inclusion and exclusion criteria for clinical outcome studies after combined MAT and cartilage repair or restoration.

RESULTS

Six studies were identified for inclusion. In total 110 patients underwent combined MAT/cartilage repair or restoration (medial compartment in 66 and lateral compartment in 44). Patients underwent MAT and either autologous chondrocyte implantation (n = 73), osteochondral allograft (n = 20), osteochondral autograft transfer (n = 17), or microfracture (n = 3). Thirty-six patients underwent additional concurrent surgeries (high tibial or distal femoral osteotomy, cruciate or collateral ligament reconstruction, and hardware removal). All clinical outcomes were improved at final follow-up (mean, 36 months). In 4 of 6 studies, overall outcomes of combined surgery were equivalent to those of either procedure performed in isolation. In 2 studies outcomes of combined surgery were not as good as those of either procedure performed in isolation. Failure occurred in 12% of patients who underwent combined MAT and cartilage restoration, and they required revision surgery. Most failures (85%) of combined surgery were due to failure of the MAT (as opposed to the cartilage technique). One-half of all patients required at least 1 surgery after the index procedure before final follow-up.

CONCLUSIONS

Clinical outcomes after combined MAT and cartilage repair/restoration are similar to those after either procedure in isolation. Despite low rates of complications and failures, there is a high rate of subsequent surgery after combined MAT and cartilage repair or restoration.

METHODS

Level IV, systematic review of Level IV studies.

BACKGROUND

Most of the current knowledge of leptospirosis epidemiology originates from serological results obtained with the reference Microscopic Agglutination Test (MAT). However, inconsistencies and weaknesses of this diagnostic technique are evident. A growing use of PCR has improved the early diagnosis of leptospirosis but a drawback is that it cannot provide information on the infecting Leptospira strain which provides important epidemiologic data. Our work is aimed at evaluating if the sequence polymorphism of diagnostic PCR products could be used to identify the infecting Leptospira strains in the New Caledonian environment.

RESULTS

Both the lfb1 and secY diagnostic PCR products displayed a sequence polymorphism that could prove useful in presumptively identifying the infecting leptospire. Using both this polymorphism and MLST results with New Caledonian isolates and clinical samples, we confirmed the epidemiological relevance of the sequence-based identification of Leptospira strains. Additionally, we identified one cluster of L. interrogans that contained no reference strain and one cluster of L. borgpetersenii found only in the introduced Rusa deer Cervus timorensis russa that is its probable reservoir.

CONCLUSIONS

The sequence polymorphism of diagnostic PCR products proved useful in presumptively identifying the infecting Leptospira strains. This could contribute to a better understanding of leptospirosis epidemiology by providing epidemiological information that cannot be directly attained from the use of PCR as an early diagnostic test for leptospirosis.

Soil respiration constitutes the second largest flux of carbon (C) between terrestrial ecosystems and the atmosphere. This study provides a synthesis of soil respiration (R(s)) in 20 European grasslands across a climatic transect, including ten meadows, eight pastures and two unmanaged grasslands. Maximum rates of R(s) (R(s(max) )), R(s) at a reference soil temperature (10°C; R(s(10) )) and annual R(s) (estimated for 13 sites) ranged from 1.9 to 15.9 μmol CO(2) m(-2) s(-1), 0.3 to 5.5 μmol CO(2) m(-2) s(-1) and 58 to 1988 g C m(-2) y(-1), respectively. Values obtained for Central European mountain meadows are amongst the highest so far reported for any type of ecosystem. Across all sites R(s(max) ) was closely related to R(s(10) ).Assimilate supply affected R(s) at timescales from daily (but not necessarily diurnal) to annual. Reductions of assimilate supply by removal of aboveground biomass through grazing and cutting resulted in a rapid and a significant decrease of R(s). Temperature-independent seasonal fluctuations of R(s) of an intensively managed pasture were closely related to changes in leaf area index (LAI). Across sites R(s(10) ) increased with mean annual soil temperature (MAT), LAI and gross primary productivity (GPP), indicating that assimilate supply overrides potential acclimation to prevailing temperatures. Also annual R(s) was closely related to LAI and GPP. Because the latter two parameters were coupled to MAT, temperature was a suitable surrogate for deriving estimates of annual R(s) across the grasslands studied. These findings contribute to our understanding of regional patterns of soil C fluxes and highlight the importance of assimilate supply for soil CO(2) emissions at various timescales.

The HO endonuclease was used to introduce a site-specific double-strand break (DSB) in an interval designed to monitor mitotic recombination. The interval included the trp1 and his3 genes inserted into chromosome III of S. cerevisiae between the CRY1 and MAT loci. Mitotic recombination was monitored in a diploid carrying heteroalleles of trp1 and his3. The normal recognition sites for the HO endonuclease were mutated at the MAT alleles and a synthetic recognition site for HO endonuclease was placed between trp1 and his3 on one of the chromosomes. HO-induced cleavage resulted in efficient recombination in this interval. Most of the data can be explained by double-strand gap repair in which the cut chromosome acts as the recipient. However, analysis of some of the recombinants indicates that regions of heteroduplex were generated flanking the site of the cut, and that some recombinants were the result of the cut chromosome acting as the genetic donor.