Diversity of bacteria in the marine sponge Aplysina fulva in Brazilian coastal waters.

Microorganisms can account for up to 60% of the fresh weight of marine sponges. Marine sponges have been hypothesized to serve as accumulation spots of particular microbial communities, but it is unknown to what extent these communities are directed by the organism or the site or occur randomly. To address this question, we assessed the composition of specific bacterial communities associated with Aplysina fulva, one of the prevalent sponge species inhabiting Brazilian waters. Specimens of A. fulva and surrounding seawater were collected in triplicate in shallow water at two sites, Caboclo Island and Tartaruga beach, Búzios, Brazil. Total community DNA was extracted from the samples using "direct" and "indirect" approaches. 16S rRNA-based PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analyses of the total bacterial community and of specific bacterial groups--Pseudomonas and Actinobacteria--revealed that the structure of these assemblages in A. fulva differed drastically from that observed in seawater. The DNA extraction methodology and sampling site were determinative for the composition of actinobacterial communities in A. fulva. However, no such effects could be gleaned from total bacterial and Pseudomonas PCR-DGGE profiles. Bacterial 16S rRNA gene clone libraries constructed from directly and indirectly extracted DNA did not differ significantly with respect to diversity and composition. Altogether, the libraries encompassed 15 bacterial phyla and the candidate division TM7. Clone sequences affiliated with the Cyanobacteria, Chloroflexi, Gamma- and Alphaproteobacteria, Actinobacteria, Bacteroidetes, and Acidobacteria were, in this order, most abundant. The bacterial communities associated with the A. fulva specimens were distinct and differed from those described in studies of sponge-associated microbiota performed with other sponge species.

Hungarian hospital antibiotic consumption at the regional level, 1996-2005.

BACKGROUND: Regional variations in antibiotic consumption in outpatients have been reported previously, but nothing is as yet known about the regional distribution of antibiotic consumption in the hospital sector in Hungary. This study was designed to explore regional variations and investigate determinants of antibiotic consumption in hospital care in Hungary. MATERIALS AND METHODS: Regional distribution-based antibiotic sales data were obtained for a 10-year period (1996-2005) for the 20 Hungarian counties. Systemic antibacterial use (Anatomical Therapeutic Chemical code: J01) was expressed as the number of defined daily doses (DDD) per 100 patient-days. The multiple linear regression model was applied to investigate the determinants of regional differences in hospital antibiotic consumption. Independent variables related to health care access, utilization of hospital resources, doctors' workload, type of hospital care provided, and patient's characteristics and infections were considered as possible determinants, and data on these variables were obtained for 2 years (2004, 2005). We also tested the association between hospital and ambulatory care antibiotic consumption in Hungarian regions using the Pearson correlation test. RESULTS: For each year during the 1996-2005 study period, there were large and stable variations in total hospital antibiotic consumption (e.g., min-max(1996): 16.0-28.2; min-max(2005): 15.2-32.2 DDD per 100 patient-days) depending on the region. In the two developed models (Model 1 and Model 2), the number of reported infections accounted for 53% of the observed regional variations in hospital antibiotic consumption (Model 1), and the number of reported infections together with the case-mix index were responsible for 61% (Model 2) . Total antibiotic consumption in hospitals showed a positive correlation (R = 0.71, p = 0.002) with total antibiotic consumption in ambulatory care. CONCLUSION: The case-mix index and the number of reported infections explained some of the observed regional variations. However, the moderate value of the models in explaining these regional variations suggest that determinants which could not be explored in this preliminary study may also contribute to regional differences. Future studies should aim at collecting data for each individual hospital as well as data on possible determinants for hospital antibiotic consumption.

An 8-year evaluation of antibiotic consumption and antibiotic resistance among Streptococcus pneumoniae from in- and out-patients in Szeged, Hungary.

At the beginning of the 1990s, the prevalence of penicillin resistance of Streptococcus pneumoniae strains in Hungary was found to be extremely high (up to 58% non-susceptible) in some studies, while in other publications the percentage of penicillin highly resistant strains was 0-2%. To see whether this was due to differences in methodology or the composition of the patient population studied, a retrospective evaluation was carried out of the penicillin, amoxicillin, ceftriaxone and macrolide resistance of all S. pneumoniae strains isolated from in- and outpatients in our laboratory between 1998 and 2005. Of the 2670 S. pneumoniae isolates only 5.58% was found to exhibit high-level resistance to penicillin, while resistance to amoxicillin, ceftriaxone and erythromycin was 2.62%, 1.12% and 42.06%, respectively. During this period 6 (3.8%) of 155 S. pneumoniae strains isolated from invasive samples displayed high-level resistance to penicillin. Earlier surveillance data on penicillin resistance of S. pneumoniae may have been biased by the age groups affected by the infection, by whether the strain was isolated from an out-patient or an in-patient, and by whether the isolates were obtained from invasive samples. Our 8-year study using the NCCLS/CLSI methodology consequently revealed a low prevalence of high-level resistance to penicillin in S. pneumoniae strains obtained both from adults and children.

Flavopiridol binds to duplex DNA.

Flavopiridol, the first potent cyclin-dependent kinase inhibitor to enter clinical trials, was recently found to be cytotoxic to noncycling cells. The present studies were performed to examine the hypothesis that flavopiridol, like several other antineoplastic agents that kill noncycling cells, might also interact with DNA. Consistent with this possibility, treatment of A549 human lung cancer cells with clinically achievable concentrations of flavopiridol resulted in rapid elevations of the DNA damage-responsive protein p53. In further studies, the binding of flavopiridol to DNA was examined in vitro by four independent techniques. Absorption spectroscopy revealed that addition of DNA to aqueous flavopiridol solutions resulted in a red shift of the flavopiridol lambda(max) from 311 to 344 nm, demonstrating an isosbestic point typical of changes seen with DNA-binding compounds. Reverse-phase high-performance liquid chromatography demonstrated that flavopiridol binds to genomic DNA to a similar extent as ethidium bromide and Hoechst 33258. Nuclear magnetic resonance spectroscopy revealed that DNA caused extreme broadening of flavopiridol 1H nuclear magnetic resonance signals that could be reversed by addition of ethidium bromide or by DNA melting, suggesting that flavopiridol binds to (and likely intercalates into) duplex DNA. Equilibrium dialysis demonstrated that the equilibrium dissociation constant of the flavopiridol-DNA complex (5.4+/-3.4 x 10(-4) M) was in the same range observed for binding of the intercalators doxorubicin and pyrazoloacridine to DNA. Molecular modeling confirmed the feasibility of flavopiridol intercalation into DNA and analysis of the effects of flavopiridol in the National Cancer Institute tumor cell line panel using the COMPARE algorithm demonstrated that flavopiridol most closely resembles cytotoxic antineoplastic intercalators. Collectively, these data suggest that DNA might be a second target of flavopiridol, providing a potential explanation for the ability of this agent to kill noncycling cancer cells.

The relationship between serotypes and PFGE genotypes in isolates of Streptococcus pneumoniae from Hungary.

The relatedness of 112 penicillin-non-susceptible isolates of Streptococcus pneumoniae from Hungary was determined by pulsed-field gel electrophoresis (PFGE), serotyping and antibiotic susceptibility tests. The differences in PFGE patterns closely mirrored the changes in resistance. Some genotypes comprised multiple serotypes, and the genetic diversity among certain serotypes was considerable. Generally, serotyping alone was insufficient for epidemiological mapping of pneumococcal isolates. There was considerable serotype diversity, but the five most frequent international serotypes (6, 9, 14, 23, 19) were the most prevalent. In addition, the presence of some well-defined resistant international pneumococcal clones in the Hungarian population was identified.

Correlation of cytology and cystoscopy.

A total of 152 patients had cytology and cystoscopy performed for either initial or recurrent bladder tumors and postoperative control examinations. Positive cytology was found in 97 per cent of patients with pathologically proved bladder tumors. However, 23 per cent of the patients with negative cytology had positive cystoscopic and pathologically proved findings. Without cystoscopic examination a significant number of recurrent tumors may be missed because of a negative cytology.

Ultrasonically guided percutaneous aspiration biopsy of the pancreas.

One hundred and twenty-six patients had fine needle aspiration of the pancreas under ultrasonic guidance. The aspirates of 66 patients contained malignant cells. Two of 38 patients who had negative cytology subsequently were diagnosed at laparotomy as having carcinoma of the pancreas. The results of this study show that the sensitivity of the cytologic method was 97%, and the specificity and predictive value were 100%.

Chemical and pharmacological characterization of halitoxin from Amphimedon viridis (Porifera) from the southeastern Brazilian coast.

The halitoxin complex from the marine sponge Amphimedon viridis, collected in the São Sebastião channel (southeastern Brazilian coast), was isolated by gel-filtration chromatography on Sephadex LH-20. Spectroscopic data (1H, 13C and 2D-NMR) of halitoxin from A. viridis indicated that it has the same two alkylpyridine monomers composition of the previously isolated halitoxin from Amphimedon compressa (as Haliclona rubens). Nevertheless, analysis by High Performance Gel Permeation Chromatography indicated that the halitoxin complex of A. viridis has a lower molecular weight (500, 2000 and 5000 Da; the major component corresponding to the fraction of 2000 Da MW) when compared to the previously isolated halitoxin complex from A. compressa. Some pharmacological properties of the halitoxin complex from A. viridis were evaluated in terms of lethality, antimitosis, hemolysis and neurotoxicity. The possible chemotaxonomic value of alkyl pyridine alkaloids is discussed.

Data on the contamination of maize with fumonisin B1 and other fusariotoxins in Hungary.

The fumonisin B1, zearalenone, deoxynivalenol and T-2 toxin content of maize samples collected in the period of storage and harvesting was determined by high-performance liquid chromatography (HPLC) methods. Of the mouldy maize samples collected in the period of storage, 70.8% contained fumonisin B1 (0.05-19.8 mg/kg; average concentration: 2.6 mg/kg), 87.5% contained zearalenone (0.01-11.8 mg/kg, average concentration: 1.26 mg/kg), 70% contained deoxynivalenol (0.07-21.2, average concentration: 4.33 mg/kg), and 41.7% contained T-2 toxin (0.06-0.39, average concentration: 0.2 mg/kg). These mycotoxins often occurred together in the samples. In the non-mouldy samples, both the positivity rate and the mycotoxin concentration were markedly lower. In the harvesting period, the mycotoxin content of maize ears more or less affected by moulds (so-called "affected sample") and of the average sample was determined separately for each maize-field involved in the study. Of the affected samples, 70% contained fumonisin B1 (0.095-52.4 mg/kg; average: 6.64 mg/kg), 17% contained zearalenone (0.006-0.079 mg/kg; average: 0.03 mg/kg), 13% contained deoxynivalenol (0.05-0.118 mg/kg; average: 0.09 mg/kg), and 39% contained T-2 toxin (0.05-0.551 mg/kg; average: 0.165 mg/kg). Fumonisin B1 and T-2 toxin often occurred together in the affected maize samples. Of the "average samples", 30% were contaminated with fumonisin B1 (0.06-5.1 mg/kg; average: 1.52 mg/kg) and 9% with T-2 toxin (min.-max.-average: 0.05 mg/kg). The results call attention to the fact that maize cultivated in Hungary is often contaminated with fumonisin B1. High concentrations of fumonisin B1 were found primarily in the mouldy or affected maize ears; therefore, the development of diseases caused by fumonisin B1 should be reckoned with primarily if such maize is fed. Besides fumonisin B1, mouldy or affected maize usually contains also other fusariotoxins, which raises the possibility of mycotoxin interactions. Because of the high prevalence of fumonisin B1 in maize grown in Hungary, the authors suggest that samples of maize used for feeding horses and pigs should be checked for fumonisin B1 content.

Natural deoxynivalenol (DON) contamination of wheat samples grown in 1998 as determined by high-performance liquid chromatography.

A high-performance liquid chromatography--diode array detection (HPLC-DAD) method was developed for determining the deoxynivalenol (DON) content of wheat and other cereals. The samples were extracted with a mixture of acetonitrile and water (84 + 16). Part of the extract was evaporated and purified on Florisil and activated charcoal columns. HPLC separation was performed on a C18 column, using acetonitrile-water (8 + 92) as eluent. Diode array detection (DAD) was performed at 218 and 236 nm, by determination of the UV spectrum. Quantitative analysis was carried out by the external standard method, using the UV spectrum obtained by DAD for confirmation. The recovery rate of DON was 75 +/- 3.1% and the detection limit was 0.05 mg/kg DON. Using this method, the DON content of 99 feeding wheat samples grown in the northeastern part of Hungary in 1998 was determined. Eighty-eight percent of the samples originating from three counties contained 0.94 mg/kg DON on the average. The highest individual value was 4.3 mg/kg. DON contamination of wheat was of higher prevalence (100%) and severity (0.27-4.3 mg/kg) in the southeastern county of Békés than in Szabolcs county located in the northeastern part of Hungary (ratio of positive samples: 82%; DON concentration: 0.05-1.3 mg/kg). The higher than usual DON contamination of feeding wheat can be explained by the rainy summer weather. DON contamination of feeding wheat poses a major risk to the production and animal health status of pig herds.

Phylogeny and systematics of demospongiae in light of new small-subunit ribosomal DNA (18S) sequences.

The most diverse and species-rich class of the phylum Porifera is Demospongiae. In recent years, the systematics of this clade, which contains more than 7000 species, has developed rapidly in light of new studies combining molecular and morphological observations. We add more than 500 new, nearly complete 18S sequences (an increase of more than 200%) in an attempt to further enhance understanding of the phylogeny of Demospongiae. Our study specifically targets representation of type species and genera that have never been sampled for any molecular data in an effort to accelerate progress in classifying this diverse lineage. Our analyses recover four highly supported subclasses of Demospongiae: Keratosa, Myxospongiae, Haploscleromorpha, and Heteroscleromorpha. Within Keratosa, neither Dendroceratida, nor its two families, Darwinellidae and Dictyodendrillidae, are monophyletic and Dictyoceratida is divided into two lineages, one predominantly composed of Dysideidae and the second containing the remaining families (Irciniidae, Spongiidae, Thorectidae, and Verticillitidae). Within Myxospongiae, we find Chondrosida to be paraphyletic with respect to the Verongida. We amend the latter to include species of the genus Chondrosia and erect a new order Chondrillida to contain remaining taxa from Chondrosida, which we now discard. Even with increased taxon sampling of Haploscleromorpha, our analyses are consistent with previous studies; however, Haliclona species are interspersed in even more clades. Haploscleromorpha contains five highly supported clades, each more diverse than previously recognized, and current families are mostly polyphyletic. In addition, we reassign Janulum spinispiculum to Haploscleromorpha and resurrect Reniera filholi as Janulum filholi comb. nov. Within the large clade Heteroscleromorpha, we confirmed 12 recently identified clades based on alternative data, as well as a sister-group relationship between the freshwater Spongillida and the family Vetulinidae. We transfer Stylissa flabelliformis to the genus Scopalina within the family Scopalinidae, which is of uncertain position. Our analyses uncover a large, strongly supported clade containing all heteroscleromorphs other than Spongillida, Vetulinidae, and Scopalinidae. Within this clade, there is a major division separating Axinellidae, Biemnida, Tetractinellida, Bubaridae, Stelligeridae, Raspailiidae, and some species of Petromica, Topsentia, and Axinyssa from Agelasida, Polymastiidae, Placospongiidae, Clionaidae, Spirastrellidae, Tethyidae, Poecilosclerida, Halichondriidae, Suberitidae, and Trachycladus. Among numerous results: (1) Spirophorina and its family Tetillidae are paraphyletic with respect to a strongly supported Astrophorina within Tetractinellida; (2) Agelasida is the earliest diverging lineage within the second clade listed above; and (3) Merlia and Desmacella appear to be the earliest diverging lineages of Poecilosclerida.

Transformation of a cranial fusiform aneurysm into a pseudotumoral-like mass prior to spontaneous occlusion and regression.

This report documents the evolution of a non-ruptured cranial fusiform aneurysm that underwent both spontaneous occlusion and regression. During this process, unique images of the aneurysm as a pseudotumoral-like mass were obtained. The pseudotumoral-like mass most likely reflected inflammation and secondary neovascularization within the aneurysm, supporting the theory that spontaneous aneurysmal healing involves an inflammatory process.