Compression socks and the effects on coagulation and fibrinolytic activation during marathon running.

PURPOSE: Compression socks are frequently used in the treatment and prevention of lower-limb pathologies; however, when combined with endurance-based exercise, the impact of compression socks on haemostatic activation remains unclear. OBJECTIVES: To investigate the effect of wearing compression socks on coagulation and fibrinolysis following a marathon. METHODS: Sixty-seven participants [43 males (mean ± SD: age: 46.7 ± 10.3 year) and 24 females (age: 40.0 ± 11.0 year)] were allocated into a compression (SOCK, n = 34) or control (CONTROL, n = 33) group. Venous blood samples were obtained 24 h prior to and immediately POST-marathon, and were analyzed for thrombin-anti-thrombin complex (TAT), tissue factor (TF), tissue factor pathway inhibitor (TFPI), and D-Dimer. RESULTS: Compression significantly attenuated the post-exercise increase in D-Dimer compared to the control group [median (range) SOCK: + 9.02 (- 0.34 to 60.7) ng/mL, CONTROL: + 25.48 (0.95-73.24) ng/mL]. TF increased following the marathon run [median (range), SOCK: + 1.19 (- 7.47 to 9.11) pg/mL, CONTROL: + 3.47 (- 5.01 to 38.56) pg/mL] in all runners. No significant post-exercise changes were observed for TAT and TFPI. CONCLUSIONS: While activation of coagulation and fibrinolysis was apparent in all runners POST-marathon, wearing compression socks was shown to reduce fibrinolytic activity, as demonstrated by lower D-Dimer concentrations. Compression may reduce exercise-associated haemostatic activation when completing prolonged exercise.

Indicator microbes correlate with pathogenic bacteria, yeasts and helminthes in sand at a subtropical recreational beach site.

AIMS: Research into the relationship between pathogens, faecal indicator microbes and environmental factors in beach sand has been limited, yet vital to the understanding of the microbial relationship between sand and the water column and to the improvement of criteria for better human health protection at beaches. The objectives of this study were to evaluate the presence and distribution of pathogens in various zones of beach sand (subtidal, intertidal and supratidal) and to assess their relationship with environmental parameters and indicator microbes at a non-point source subtropical marine beach. METHODS AND RESULTS: In this exploratory study in subtropical Miami (Florida, USA), beach sand samples were collected and analysed over the course of 6 days for several pathogens, microbial source tracking markers and indicator microbes. An inverse correlation between moisture content and most indicator microbes was found. Significant associations were identified between some indicator microbes and pathogens (such as nematode larvae and yeasts in the genus Candida), which are from classes of microbes that are rarely evaluated in the context of recreational beach use. CONCLUSIONS: Results indicate that indicator microbes may predict the presence of some of the pathogens, in particular helminthes, yeasts and the bacterial pathogen Staphylococcus aureus including methicillin-resistant forms. Indicator microbes may thus be useful for monitoring beach sand and water quality at non-point source beaches. SIGNIFICANCE AND IMPACT OF THE STUDY: The presence of both indicator microbes and pathogens in beach sand provides one possible explanation for human health effects reported at non-point sources beaches.

Multiple simultaneous detection of Harmful Algal Blooms (HABs) through a high throughput bead array technology, with potential use in phytoplankton community analysis.

As an alternative to traditional, morphology-based methods, molecular techniques can provide detection of multiple species within the HAB community and, more widely, the phytoplankton community in a rapid, accurate and simultaneous qualitative analysis. These methods require detailed knowledge of the molecular diversity within taxa in order to design efficient specific primers and specific probes able to avoid cross-reaction with non-target sequences. Isolates from Florida coastal communities were sequence-analyzed and compared with the GenBank database. Almost 44% of the genotypes obtained did not match any sequence in GenBank, showing the existence of a large and still unexplored biodiversity among taxa. Based on these results and on the GenBank database, we designed 14 species-specific probes and 4 sets of specific primers. Multiple simultaneous detection was achieved with a bead array method based on the use of a flow cytometer and color-coded microspheres, which are conjugated to the developed probes. Following a parallel double PCR amplification, which employed universal primers in a singleplex reaction and a set of species-specific primers in multiplex, detection was performed in a cost effective and highly specific analysis. This multi-format assay, which required less than 4 h to complete from sample collection, can be expanded according to need. Up to 100 different species can be identified simultaneously in a single sample, which allows for additional use of this method in community analyses extended to all phytoplankton species. Our initial field trials, which were based on the 14 species-specific probes, showed the co-existence and dominance of two or more species of Karenia during toxic blooms in Florida waters.

Molecular sequence analyses of the intergenic spacer (IGS) associated with rDNA of the two varieties of the pathogenic yeast, Cryptococcus neoformans.

The pathogen Crytococcus neoformans has been traditionally grouped in two varieties, C. neoforrmans var. neoformans (serotypes A, D and AD) and C. neoformans var. gattii (serotypes B and C). A recent taxonomic evaluation of C. neoformans var. neoformans described C. neoformans var. grubii as a new variety represented by serotype A isolates. Despite immunological, biochemical, ecological and molecular differences the three varieties are classified within one species. We examined the genetic variability of one hundred and five clinical and environmental isolates that included all varieties and serotypes. Sequence analysis of the intergenic spacer (IGS) associated with rDNA revealed significant differences in nucleotide composition between and within the varieties. Parsimony analysis showed five different genotypes representing distinct genetic lineages. Although there was a high degree of relatedness between serotype and genotype this relatedness was not exclusive as serotypes were not restricted to one particular genotypic group. Serotyping and sequence analyses indicate that C. neoformans var. grubii (serotype A) should not be recognized as a separate variety. Based on this study we propose to accept two separate species, C. neoformans (serotypes A, D and AD) and C. bacillisporus (serotypes B and C synonymous with C. neoformans var. gattii).

Kurtzmanomyces insolitus sp. nov., a new anamorphic heterobasidiomycetous yeast species.

A new anamorphic heterobasidiomycetous yeast species, Kurtzmanomyces insolitus, is described using a polyphasic taxonomic approach. The new species has the salient characteristics of the genus Kurtzmanomyces and, additionally, the ability to produce ballistoconidia. Data derived from comparative micromorphological studies, physiological characterisation, ultrastructure and nucleic acid analyses led to assigning the new species to Kurtzmanomyces rather than to the currently accepted genera of ballistoconidia-forming fungi. An emendation of the genus Kurtzmanomyces is proposed to allow the inclusion of the new species.

Physiological profiles of Australian surf boat rowers.

The physiological profiles of 17 open grade (OG) and 13 reserve grade (RG) male surfboat rowers (SBR) aged 19-44 years were determined and compared. Parameters investigated included anthropometry, agility, isometric strength, flexibility, rowing ergometer performance (MT), peak VO2 and arterialised blood pH, lactate and bicarbonate. Means were compared using t-tests. Multiple regression analyses provided a number of models for the prediction of MT performance in SBR. The mean age, height, mass, and sum of eight skinfolds for SBR are: 26.2 (+/-5.9) years, 180.5 (+/-6.0) cm, 84.4 (+/-9.3) kg and 78.2 (+/-26.2) mm respectively. OG rowers were significantly different from RG for the parameters of ergometer performance (OG: 1360.2+/-42.9 m; RG: 1316.4+/-41.8 m), peak ventilation (OG: 174.2+/-17.2 L x min(-1); RG: 154.8+/-22.1 L x min(-1)), and post exercise blood pH levels (OG: 6.98+/-0.07; RG: 7.04+/-0.07). Performance on a rowing ergometer successfully discriminates between OG and RG rowers with the best predictors of ergometer performance in SBR being height, peak ventilation, and post exercise pH.

Identification of genotypically diverse Cryptococcus neoformans and Cryptococcus gattii isolates by Luminex xMAP technology.

A Luminex suspension array, which had been developed for identification of Cryptococcus neoformans and Cryptococcus gattii isolates, was tested by genotyping a set of 58 mostly clinical isolates. All genotypes of C. neoformans and C. gattii were included. In addition, cerebrospinal fluid (CSF) obtained from patients with cryptococcal meningitis was used to investigate the feasibility of the technique for identification of the infecting strain. The suspension array correctly identified haploid isolates in all cases. Furthermore, hybrid isolates possessing two alleles of the Luminex probe region could be identified as hybrids. In CSF specimens, the genotype of the cryptococcal strains responsible for infection could be identified after optimization of the PCR conditions. However, further optimization of the DNA extraction protocol is needed to enhance the usability of the method in clinical practice.

Microcoding: the second step in DNA barcoding.

After the process of DNA barcoding has become well advanced in a group of organisms, as it has in the economically important fungi, the question then arises as to whether shorter and literally more barcode-like DNA segments should be utilized to facilitate rapid identification and, where applicable, detection. Through appropriate software analysis of typical full-length barcodes (generally over 500 base pairs long), uniquely distinctive oligonucleotide 'microcodes' of less than 25 bp can be found that allow rapid identification of circa 100-200 species on various array-like platforms. Microarrays can in principle fulfill the function of microcode-based species identification but, because of their high cost and low level of reusability, they tend to be less cost-effective. Two alternative platforms in current use in fungal identification are reusable nylon-based macroarrays and the Luminex system of specific, colour-coded DNA detection beads analysed by means of a flow cytometer. When the most efficient means of rapid barcode-based species identification is sought, a choice can be made either for one of these methodologies or for basic high-throughput sequencing, depending on the strategic outlook of the investigator and on current costs. Arrays and functionally similar platforms may have a particular advantage when a biologically complex material such as soil or a human respiratory secretion sample is analysed to give a census of relevant species present.

Rapid identification of yeast species using three primers in a polymerase chain reaction.

Classic methods for identification of yeasts rely on a variety of morphological and physiological tests that often take days to weeks to complete. We have been able to reduce the time to less than one day through the use of multiple segment-specific oligonucleotide priming of a region of the large subunit rDNA in a polymerase chain reaction. The "hot start" reaction was used with two universal external delimiting primers and one internal species-specific primer. Five specific primers were tested: a primer for a biologically similar group of Rhodotorula species, a generic (Cystofilobasidium) primer, and 3 species-specific primers (Leucosporidium scottii, Cryptococcus muscorum, and Rhodotorula mucilaginosa). In the absence of specific target DNA, the universal rDNA segment is amplified; in the presence of target DNA, the specific primer region is amplified. The technique is accurate within two base position differences when a 24 nucleotide-specific primer is used. The technique should be applicable to other marine eukaryotes.

rDNA targeted oligonucleotide primers for the identification of pathogenic yeasts in a polymerase chain reaction.

Species-specific oligonucleotide primers were designed for PCR identification of the basidiomycetous yeasts Cryptococcus neoformans, Trichosporon cutaneum and Rhodotorula mucilaginosa. The procedure uses standard PCR components including DNA from the test species and three primers: two universal external (upstream and downstream) limiting primers and a species-specific internal primer. Species identification requires the formation of a species-specific rDNA nucleotide segment that is significantly smaller (approximately 200 bp) than a non-target segment (approximately 600 bp). The procedure can be used to identify yeasts from single and mixed populations.

Systematic placement of the basidiomycetous yeast Cystofilobasidium lari-marini comb. nov. as predicted by rRNA nucleotide sequence analysis.

Based on similarities in basidial morphology and nucleotide sequences of the V3 variable region in the large sub-unit ribosomal RNA, the yeast Leucosporidium lari-marini is considered phylogenetically related to the genus Cystofilobasidium. Therefore the new combination Cystofilobasidium lari-marini is proposed.

Molecular analyses of the IGS & ITS regions of rDNA of the psychrophilic yeasts in the genus Mrakia.

Species of the genus Mrakia are currently classified as synonyms based on molecular sequence analyses of the large sub-unit ribosomal DNA (LrDNA). Physiological and protein electrophoretic studies, however, reveal possible species differences. To clarify this discrepancy, we undertook molecular sequence analyses of the internal transcribed spacer (ITS) and intergenic spacer (IGS) regions of rDNA from the four psychrophilic Mrakia species and the psychrophilic yeast, Cryptococcus curiosus. Identical ITS sequences were found between C. curiosus, M. nivalis and M. frigida. Although, M. stokesii and M. gelida displayed identical ITS and IGS sequences, their sequences differed from the other three species by 2.3% and 38%, respectively. The results suggest that M. stokesii is a synonym of M. gelida, whereas M. nivalis is a synonym of M. frigida. Sequence differences (1.9%) observed in the IGS region indicates that C. curiosus is a distinct strain of M. frigida.

Separation of strains of the yeasts Xanthophyllomyces dendrorhousand Phaffia rhodozyma based on rDNA IGS and ITS sequence analysis.

The type strains of the anamorph Phaffia rhodozyma (CBS 5905) and the teleomorph Xanthophyllomyces dendrorhous (VKM Y-2786) were analyzed by nucleotide sequence analysis and compared to the sequences found in three additional strains (ATCC 24228, ATCC 24230 and CBS 6938). The results of ribosomal DNA Internal transcribed spacer (ITS) and Intergenic spacer (IGS) region analyses indicate that P. rhodozyma, which was isolated from a beech tree, is a distinct species from the other four strains. The latter that were collected from birch trees are considered to be strains of X. dendrorhous. These individual strains of X. dendrorhous, which have geographically distinct isolation sources, can be distinguished by nucleotide substitutions and deletion/insertion gaps in sub-repeat regions of the Intergenic spacer. The conclusions demonstrate that differences in the IGS region provide molecular markers for denoting strains that may differ in their biochemical and physiological capabilities. The hypothesis is presented that strain differences in the IGS region may be useful to demonstrate geographic and host specificity.

Taxonomy of the yeast genus Metschnikowia: a correction and a new variety.

Metschnikowia isolates from ponds on Chatham Island, New Zealand, previously erroneously reported as M. zobellii, are described herein as M. bicuspidata var. chathamia var. n.

Rapid and pervasive occupation of fallen mangrove leaves by a marine zoosporic fungus.

Samples of leaves of red mangrove (Rhizophora mangle) were incubated on an agar medium selective for pythiaceous oomycetes. Leaves on trees above the water did not contain oomycetes. Marine oomycetes, principally Phytophthora vesicula, had colonized leaves within 2 h of leaf submergence, probably finding them by chemotaxis. The frequency of occurrence of P. vesicula in submerged leaves reached 100% within 30 h of submergence. By 43 h most, if not all, parts of leaves were occupied, and surface treatment with a biocide indicated that leaves were occupied internally. Frequencies of P. vesicula remained near 100% through about 2 weeks of submergence and then declined to about 60% in older (>/=4 weeks) leaves. Leaves of white mangrove (Laguncularia racemosa) were also extensively occupied by P. vesicula after falling into the water column, but decaying leaves of turtlegrass (Thalassia testudinum) were not colonized by oomycetes. Ergosterol analysis indicated that the standing crop of living, non-oomycete (ergosterol-containing) fungal mass in submerged red-mangrove leaves did not rise above that which had been present in senescent leaves on the tree; decaying turtlegrass leaves had an ergosterol content that was only about 2% of the maximum concentration detected for red-mangrove leaves. These results suggest that oomycetes are the predominant mycelial eucaryotic saprotrophs of mangrove leaves that fall into the water column and that for turtlegrass leaves which live, die, and decompose under submerged conditions, mycelial eucaryotes make no substantial contribution to decomposition.

A physiological profile of elite surf ironmen, full time lifeguards & patrolling surf life savers.

The physiological profiles and proficiency of 32 volunteer surf life savers (LS), 15 professional lifeguards (LG), and 8 elite surf ironmen (IM) aged from 18 to 44 were compared. Measurements included anthropometry, muscular power, muscular strength-endurance, flexibility, VO2max, maximum heart rate, peak blood lactate response and proficiency in 3 rescue simulations. Both LG and IM were significantly faster than LS in all rescue simulations. IM had significantly greater VO2max (68.6 ml.kg-1.min-1) than LS (56.3) and LG (57.9). IM had significantly lower heart rates than LS after maximal swimming and running, and significantly lower blood lactate 3 minutes post swim (8.4 mmol/L) in comparison with LS (14.0) and LG (12.2). LG obtained better results than LS in 2 of the 3 muscular strength-endurance tasks. It is concluded that: LG and IM are significantly faster in aquatic rescue simulations than LS; IM have greater aerobic capacities than both other groups; the majority of Australian LS have adequate fitness and aquatic skills for surf rescue, although a small subset of LS do not. This group of poorly performing LS are not identified by current surf rescue screening procedures.

Cryptococcus adeliensis sp. nov., a xylanase producing basidiomycetous yeast from Antarctica.

Cryptococcus adeliensis sp. nov. (CBS 8351) is described based on phenotypic characteristics and molecular sequence analysis of the D1/D2 large subunit and internal transcribed spacer regions of the ribosomal DNA. Molecular comparisons include species closely related to Cryptococcus albidus and several species isolated from the Antarctic. C. adeliensis, which has a cold-adapted xylanase, was isolated from Terre Adelie, Antarctica. ATCC 34633, which has a mesophilic xylanase, was identified as Cryptococcus albidosimilis.

Trichosporon veenhuisii sp. nov., an alkane-assimilating anamorphic basidiomycetous yeast.

A morphological and physiological description of an alkane-assimilating anamorphic basidiomycetous yeast species, named Trichosporon veenhuisii, is presented. The ability to assimilate several aliphatic and aromatic compounds as sole source of carbon and energy is reported. The phylogenetic position within the genus, based on nuclear base sequencing of the D1/D2 region of the large subunit of rDNA, is discussed. The type strain is CBS 7136T.

Diversity in the yeast Cryptococcus albidus and related species as revealed by ribosomal DNA sequence analysis.

Evidence accumulated from studies based on physiological, biochemical, and molecular characteristics has pointed to the heterogeneity of the ubiquitous anamorphic basidiomycetous yeast species Cryptococcus albidus (Saito) Skinner, with its current varieties and synonyms. The taxonomic status of this species has not been reappraised because different studies, mostly involving limited numbers of strains, have not been integrated. To assess species diversity within the clade containing Cryptococcus albidus and other phylogenetically related Cryptococcus and Filobasidium species, we determined ribosomal DNA (rDNA) sequences of 69 strains from the 5' end of the 26S gene, D1/D2 region, and in some cases, the non-coding ITS2 region. Analysis of the sequence data together with available physiological, biochemical, and molecular characteristics, showed the segregation of C. albidus into at least 12 species, leading to the elevation of former varieties to the rank of species (C. aerius, C. diffluens), the reinstatement of synonyms (C. liquefaciens, C. terricola), and the proposal of new species (C. arrabidensis, C. chernovii, C. cylindricus, C. oeirensis, C. phenolicus, C. saitoi, C. uzbekistanensis, C. wieringae). The overall analyses of the results argue in favour of the use of rDNA sequence data to improve species delineation when integrated with other available physiological and molecular characteristics.

Trichosporon guehoae sp.nov., an anamorphic basidiomycetous yeast.

A morphological and physiological description of an anamorphic basidiomycetous yeast species, named Trichosporon guehoae (CBS 8521T), is presented. The ability to assimilate several aliphatic and aromatic compounds as sole source of carbon and energy is reported. The phylogenetic position within the genus, based on nuclear base sequencing of the D1/D2 region of the large subunit of rDNA is discussed.