Methylene blue administration in patients with refractory distributive shock - a retrospective study.

Hemodynamic effectiveness of methylene blue (MB) was tested in patients with refractory distributive shock. A retrospective analysis of 20 critically-ill patients who developed refractory shock was performed. Patients were divided into two study groups as responders with positive hemodynamic response to MB administration (defined as 10% decrease of norepinephrine dose) and non-responders. Hemodynamic, outcome data and baseline tissue hypoxia-related parameters including ratio of central venous-to-arterial carbon dioxide tension to arterio-venous oxygen content (P(v-a)CO2/C(a-v)O2) were compared between the groups. There were 9 (45%) responders and 11 (55%) non-responders to single bolus of MB administration. Dose of MB did not differ between responders and non-responders (1.3 ± 0.5 vs. 1.3 ± 0.4 mg/kg respectively, P = 0.979). MB responders had lower baseline P(v-a) CO2/C(a-v)O2 (1.79 ± 0.73 vs. 3.24 ± 1.18, P = 0.007), higher pH (7.26 ± 0.11 vs. 7.16 ± 0.10, P = 0.037) and lower lactate levels at 12 hours post MB administration (3.4 ± 2.7 vs. 9.9 ± 2.2 mmol/L, P = 0.002) compared to non-responders. Methylene blue represents a non-adrenergic vasopressor with only limited effectiveness in patients with refractory distributive shock. Profound tissue hypoxia with high degree of anaerobic metabolism was associated with the loss of hemodynamic responsiveness to its administration.

Spontaneous vegetation succession at different central European mining sites: a comparison across seres.

We performed detrended correspondence analysis (DCA) ordination to compare seven successional seres running in stone quarries, coal mining spoil heaps, sand and gravel pits, and extracted peatlands in the Czech Republic in central Europe. In total, we obtained 1,187 vegetation samples containing 705 species. These represent various successional stages aged from 1 to 100 years. The successional seres studied were more similar in their species composition in the initial stages, in which synathropic species prevailed, than in later successional stages. This vegetation differentiation was determined especially by local moisture conditions. In most cases, succession led to a woodland, which usually established after approximately 20 years. In very dry or wet places, by contrast, where woody species were limited, often highly valuable, open vegetation developed. Except in the peatlands, the total number of species and the number of target species increased during succession. Participation of invasive aliens was mostly unimportant. Spontaneous vegetation succession generally appears to be an ecologically suitable and cheap way of ecosystem restoration of heavily disturbed sites. It should, therefore, be preferred over technical reclamation.

Living together: the marine amoeba Thecamoeba hilla Schaeffer, 1926 and its endosymbiont Labyrinthula sp.

Two protists isolated simultaneously from the same sample of gill tissue of Psetta maxima (L.) were identified as Thecamoeba hilla Schaeffer, 1926 and Labyrinthula sp. A Labyrinthula strain (LTH) derived from a mixed culture of both organisms was well established in a short time, while subcultures of T. hilla continued to be associated with Labyrinthula cells despite all efforts to eliminate them. Ultrastructural examination, repeated several times in the course of long-lasting subculturing of amoebae, revealed that trophozoites of T. hilla host in their cytoplasm multiplying labyrinthulid cells. Comparison of SSU rDNA sequences of the Labyrinthula strain LTH and those from labyrinthulid endosymbionts from T. hilla verified the assumption that the extra- and intra-cellularly multiplying Labyrinthula cells are identical organisms. The association of the marine amoeba T. hilla and Labyrinthula sp. displayed signs of mutualistic symbiosis.

Phylogeny of Flabellulidae (Amoebozoa: Leptomyxida) inferred from SSU rDNA sequences of the type strain of Flabellula citata Schaeffer, 1926 and newly isolated strains of marine amoebae.

New strains of non-vannellid flattened amoebae isolated from fish, an invertebrate and the marine environment were studied together with Flabellula citata Schaeffer, 1926 selected by morphology as a reference strain. The study revealed a paucity of features distinguishing individual strains at the generic level, but clearly evidenced mutual phylogenetic relationships within the assemblage of strains as well as their affiliation to the Leptomyxida. In this study, the SSU rDNA dataset of leptomyxids was expanded and a new branching pattern was presented within this lineage of Amoebozoa. Sequences of three newly introduced strains clustered in close relationship with the type strain of F. citata, the type species of the genus. Three strains, including one resembling Flamella sp., were positioned within a sister-group containing Paraflabellula spp. Results of phylogenetic analysis confirmed doubts of previous authors regarding generic assignment of several Rhizanmoeba and Ripidomnyxa strains.

Phylogeny of Neoparamoeba strains isolated from marine fish and invertebrates as inferred from SSU rDNA sequences.

We characterised 9 strains selected from primary isolates referable to Paramoeba/Neoparamoeba spp. Based on ultrastructural study, 5 strains isolated from fish (amoebic gill disease [AGD]-affected Atlantic salmon and dead southern bluefin tuna), 1 strain from netting of a floating sea cage and 3 strains isolated from invertebrates (sea urchins and crab) were assigned to the genus Neoparamoeba Page, 1987. Phylogenetic analyses based on SSU rDNA sequences revealed affiliations of newly introduced and previously analysed Neoparamoeba strains. Three strains from the invertebrates and 2 out of 3 strains from gills of southern bluefin tunas were members of the N. branchiphila clade, while the remaining, fish-isolated strains, as well as the fish cage strain, clustered within the clade of N. pemaquidensis. These findings and previous reports point to the possibility that N. pemaquidensis and N. branchiphila can affect both fish and invertebrates. A new potential fish host, southern bluefin tuna, was included in the list of farmed fish endangered by N. branchiphila. The sequence of P. eilhardi (Culture Collection of Algae and Protozoa [CCAP] strain 1560/2) appeared in all analyses among sequences of strain representatives of Neoparamoeba species, in a position well supported by bootstrap value, Bremer index and Bayesian posterior probability. Our research shows that isolation of additional strains from invertebrates and further analyses of relations between molecular data and morphological characters of the genera Paramoeba and Neoparamoeba are required. This complexity needs to be considered when attempting to define molecular markers for identification of Paramoeba/Neoparamoeba species in tissues of fish and invertebrates.

Didymium-like myxogastrids (class Mycetozoa) as endocommensals of sea urchins (Sphaerechinus granularis).

This paper sums up the results of light microscopical, ultrastructural and molecular studies of five strains of amoeboid organisms isolated as endocommensals from coelomic fluid of sea urchins, Sphaerechinus granularis (Lamarck), collected in the Adriatic Sea. The organisms are reported as Didymium-like myxogastrids. Of the life-cycle stages, the attached amoeboids, flagellated trophozoites, cysts and biflagellated swarmers are described. Formation of fruiting bodies was not observed. Although phylogenetic analyses of SSU rDNA sequences indicated a close relationship with Hyperamoeba dachnaya, our sea-urchin strains have not been assigned to the genus Hyperamoeba Alexeieff, 1923. The presence of either one or two flagella reported in phylogenetically closely related organisms and mutually distant phylogenetic positions of strains declared as representatives of the genus Hyperamoeba justify our approach. Data obtained in this study may be useful in future analyses of relationships of the genera Didymium, Hyperamoeba, Physarum and Pseudodidymium as well as in higher-order phylogeny of Myxogastrea.

Fish-isolated Naegleria strains and their phylogeny inferred from ITS and SSU rDNA sequences.

Effort was made to identify Naegleria strains isolated from organs of fish, using phylogenetic analyses of SSU rDNA and ITS sequences. Eighteen fish-isolated strains studied enlarged substantially the so far available set of Naegleria strains characterized by both molecular markers. The phylogenetic analyses of separate and concatenated SSU rDNA and ITS sequences revealed phylogenetic relationships of strains under study; however, they failed to solve classification of fish-isolated strains into species. The sequence similarity of strain-representatives of Naegleria species as well as data obtained on intragenomic variation of ITS sequences discouraged the authors from the definition of new species. The results of the present study provide evidence of a need to re-evaluate the current practice of setting boundaries between species of the genus Naegleria. Sequences obtained in this study have been deposited in GenBank with accession numbers DQ768714-DQ768743.

The effect of motor cortex stimulation in deafferentated rats.

OBJECTIVES: The aim of the study was to describe the effect of motor cortex stimulation (MCS) on pain thresholds in deafferentated rats. SETTINGS AND DESIGN: The effect of MCS was studied in 18 deafferentated and 14 intact laboratory rats, using a standardised plantar test and tail-flick latencies. Two inoxious stimulation electrodes were implanted subdurally over the cerebral cortex and a C5-Th1 dorsal root rhizotomy was performed on the left side. Pain thresholds were measured before and after cortical stimulation. The data were analysed with ANOVA for repeated measures. RESULTS: MCS in intact animals evoked no changes in pain thresholds except for the contralateral forelimb, in which the pain threshold increased after MCS. Following deafferentation, pain thresholds increased in both plantar test and tail-flick in comparison to baseline values. When MCS was applied to the deafferentated animals, the pain thresholds returned to baseline levels. The effect of MCS disappeared within 24 hours. MAIN FINDINGS: 1. MCS in intact animals evoked hypoesthesia in the corresponding contralateral forelimb; 2. deafferentation itself increased pain thresholds in the unaffected limbs; 3. under MCS, pain thresholds in deafferentated rats were not different from pre-dafferentation values; 4. the effect of MCS disappeared in 24 hours and oscillated. CONCLUSIONS: Our results show a similar effect of the stimulation in man and experimental animals despite the differences in the organisation of the cerebral cortex. The use of laboratory animals is promising for further studies in the field of involved antalgic mechanisms of MCS.

Fish-isolated strains of Hartmannella vermiformis page, 1967: morphology, phylogeny and molecular diagnosis of the species in tissue lesions.

Based on morphological and molecular characterisation, four amoeba strains isolated from organs of freshwater fish were identified as Hartmannella vermiformis Page, 1967. Small subunit rRNA gene sequences of these strains expand the set of corresponding complete and almost complete sequences of this species to twelve. A new species-specific oligonucleotide probe inferred from recently available SSU rRNA gene sequences was designed and successfully tested in tissue lesions produced by one strain of H. vermiformis in experimentally infected fish.