Update from a 12-Year Nationwide Fungemia Surveillance: Increasing Intrinsic and Acquired Resistance Causes Concern.

New data from the years 2012 to 2015 from the Danish National Fungemia Surveillance are reported, and epidemiological trends are investigated in a 12-year perspective (2004 to 2015). During 2012 to 2015, 1,900 of 1,939 (98%) fungal bloodstream isolates were included. The average incidence was 8.4/100,000 inhabitants, and this appears to represent a stabilizing trend after the increase to 10.1/100,000 in 2011. The incidence was higher in males than females (10.0 versus 6.8) and in patients above 50 years, and those changes were mainly driven by an increasing incidence among 80-to-89-year-old males (65.3/100,000 in 2014 to 2015). The proportion of Candida albicans isolates decreased from 2004 to 2015 (64.4% to 42.4%) in parallel with a doubling of the proportion of Candida glabrata isolates (16.5% to 34.6%, P < 0.0001). C. glabrata was more common among females (34.0% versus 30.4% in males). Following an increase in 2004 to 2011, the annual drug use stabilized during the last 2 to 3 years of that time period but remained higher than in other Nordic countries. This was particularly true for the fluconazole and itraconazole use in the primary health care sector, which exceeded the combined national levels of use of these compounds in each of the other Nordic countries. Fluconazole susceptibility decreased (68.5%, 65.2%, and 60.6% in 2004 to 2007, 2008 to 2011, and 2012 to 2015, respectively, P < 0.0001), and echinocandin resistance emerged in Candida (0%, 0.6%, and 1.7%, respectively, P < 0.001). Amphotericin B susceptibility remained high (98.7%). Among 16 (2.7%) echinocandin-resistant C. glabrata isolates (2012 to 2015), 13 harbored FKS mutations and 5 (31%) were multidrug resistant. The epidemiological changes and the increased incidence of intrinsic and acquired resistance emphasize the importance of continued surveillance and of strengthened focus on antifungal stewardship.

Singlet oxygen luminescence kinetics in a heterogeneous environment - identification of the photosensitizer localization in small unilamellar vesicles.

In vivo measurement of singlet oxygen luminescence kinetics is affected by the heterogeneity of biological samples. Even though singlet oxygen luminescence detection is technically getting easier, the analysis of signals from biological samples is still far from quantitative real time surveillance as it is aspired by the community. In this paper small unilamellar vesicles (SUVs) are used for modelling the general behaviour of heterogeneous samples. The geometry of the SUVs can be determined independently using dynamic light scattering. Therefore an accurate theoretical description of the generation, deactivation and diffusion of the singlet oxygen is possible. The theoretical model developed here perfectly fits the experimental results. Thus the location of the singlet oxygen generating a photosensitizer molecule can be exactly determined from the kinetics of the singlet oxygen luminescence. The application of the used theoretical approach thus allows for accurate quantitative measurements in SUVs.

Inhibition of green algae growth by corrole-based photosensitizers.

AIMS: This study was performed to examine the potential of photodynamic inactivation for growth inhibition of green algae through generation of singlet oxygen. METHODS AND RESULTS: Two cationic and two anionic corroles were investigated according to their photoinhibitive effect on two strains of green algae using visible light for photoexcitation. The development of biomass over the experimental period of 18 days was followed using absorptive properties of the algae samples. The anionic photosensitizers showed no significant phototoxicity, whereas the cationic photosensitizers caused a drastic reduction of biomass on a short time scale and also displayed long-term inhibition of algae growth. CONCLUSIONS: In general, it was proven that photodynamic inactivation of green algae is possible. Concluding from the results of this study, cationic photosensitizers are favourable for this task, while anionic photosensitizers are not suited. SIGNIFICANCE AND IMPACT OF THE STUDY: Phototrophic biofilms are an important factor in biofouling and biodeterioration of building materials, causing great damage to historic and contemporary constructions. Growth inhibition of phototrophic organisms using photodynamic inactivation could pose an alternative to the use of biocides. To this end, successful application of this approach on green algae is a vital step in the development of suitable photosensitizers.

[Risk communication of the Federal Institute for Risk Assessment during a food-related outbreak]. / Risikokommunikation des Bundesinstituts für Risikobewertung bei einem lebensmittelbedingten Ausbruch.

Information about and explanation of risks as well as the initiation of behavioral changes and preventive actions are core tasks of risk communication. During the EHEC/HUS outbreak in spring 2011, the governmental agencies responsible for risk communication mainly focused on these tasks. In general, risk communication is understood as a continuous, long-term process that aims at an adequate handling of risks. In contrast, crisis communication is focused rather on an acute event and aims at timely information and behavioral measures. During the EHEC/HUS outbreak, risk communication partly changed over to crisis communication. The risk communication activities of the Federal Institute for Risk Assessment (Bundesinstitüt für Risikobewertung, BfR) during the EHEC/HUS outbreak are presented here. The results of a representative survey that was conducted in Germany shortly after the outbreak show details of the success of these risk communication activities. Finally, the necessity of communication about scientific uncertainty is addressed and new ways in risk communication with regard to new media are highlighted.

Alpha-1-Fetoprotein (AFP): international proficiency study with different test systems.

BACKGROUND: The present proficiency study aimed to elucidate the comparability and reliability of test systems for the determination of AFP concentrations. METHODS: 25 laboratories using 8 different commercial test systems used liquid BIOREF-AFP control serum in their routine internal quality control over a period of one year. For statistical analysis the results were collected centrally. RESULTS: The statistical analysis of the test results revealed considerable variation for the different laboratories. The deviations of the mean values of different laboratories from the overall mean value varied between 0.1 and 26.1%, and for most of the laboratories the deviation was round about 10%. The precision of measured values in the individual laboratories was in most cases acceptable: Nevertheless, the coefficients of variation of the individual laboratories ranged from 13 to 16.1%. CONCLUSIONS: In conclusion, this study indicates that AFP results vary between different laboratories albeit an international standard for AFP is available. Therefore, every laboratory should participate in external ring studies and should use a quality control serum independent of the test kit manufacturer for the internal quality control.

High incidence of candidaemia in a nationwide cohort: Underlying diseases, risk factors and mortality.

BACKGROUND: Denmark has a high incidence rate of candidaemia. A Nordic study suggested a higher Danish prevalence of haematological malignancies as an underlying reason. This nationwide study ascertained clinical characteristics of Danish candidaemia patients and investigated potential factors contributing to the high incidence and mortality. METHODS: Microbiological and clinical data for candidaemia patients in 2010-2011 were retrieved. 30-day mortality was estimated by hazard ratios (HR) with 95% confidence intervals (CI, Cox regression). RESULTS: Data were available for 912/973 candidaemia episodes (93.7%). Intensive care unit (ICU) held the largest share of patients (43.2%). Prevalent host factors were multi-morbidity (≥2 underlying diseases, 74.2%) and gastrointestinal disease (52.5%). Haematological disease was infrequent (7.8%). Risk factors included antibiotic exposure (90.5%), CVC (71.9%) and Candida colonisation (66.7%). 30-day mortality was 43.4%, and 53.6% in ICU. Mortality was lower for patients with recent abdominal surgery (HR 0.70, 95% CI: 0.54-0.92). CONCLUSION: A substantial prevalence of multi-morbidity and a high 30-day mortality was found. We hypothesise, that an increasing population of severely ill patients with prolonged supportive treatment and microbiological testing may in part explain the high candidaemia incidence in Denmark. Nationwide studies are warranted to clarify this issue.

Lipid nanoparticles for skin penetration enhancement-correlation to drug localization within the particle matrix as determined by fluorescence and parelectric spectroscopy.

With topical treatment of skin diseases, the requirement of a high and reproducible drug uptake often still is not met. Moreover, drug targeting to specific skin strata may improve the use of agents which are prone to cause local unwanted effects. Recent investigations have indicated that improved uptake and skin targeting may become feasible by means of nanoparticular systems such as solid lipid nanoparticles (SLN), nanostructured lipid carriers (NLC) and nanoemulsions (NE). Here we describe techniques to characterize drug loading to carrier systems and skin penetration profiles by using the lipophilic dye nile red as a model agent. Since the mode of drug association with the particle matrix may strongly influence the efficiency of skin targeting, parelectric spectroscopy (PS) was used to differentiate between matrix incorporation and attachment to the particle surface and fluorescence spectroscopy (FS) to solve dye distribution within NLC particles. Nile red was incorporated into the lipid matrix or the covering tensed shell, respectively, of SLN and NLC with all the lipids studied (Compritol, Precirol, oleic acid, Miglyol). In NLC, the dye was enriched in the liquid phase. Next, nile red concentrations were followed by image analysis of vertical sections of pigskin treated with dye-loaded nanoparticular dispersions and an oil-in-water cream for 4 and 8 h in vitro. Following the SLN dispersions, dye penetration increased about fourfold over the uptake obtained following the cream. NLC turned out less potent (

Clinical features of Staphylococcus aureus endocarditis: a 10-year experience in Denmark.

BACKGROUND: Both morbidity and mortality resulting from Staphylococcus aureus endocarditis are known to be high, and the incidence of this disease seems to increase. The Statens Serum Institut, Copenhagen, Denmark, made it possible for us to analyze the clinical features of S aureus endocarditis in a nation-wide population of non-drug addicts. METHODS: Almost all Danish cases of bacteremia due to S aureus are reported to the Staphylococcus laboratory, Statens Serum Institut. The medical records were reviewed in cases reported from 1982 to 1991 in which the diagnosis of endocarditis was reported or suspected. RESULTS: A total of 260 patients, 145 males and 115 females, fulfilled the diagnostic criteria. The median age was 67.5 years. In 83 patients, the diagnosis of endocarditis was not suspected clinically. The overall mortality rate among those patients whose disease was diagnosed clinically was 46%. Among the subset of patients who received medical therapy only and appropriate antistaphylococcal treatment, mortality was significantly associated with late congestive heart failure, age, and involvement of the central nervous system. CONCLUSIONS: A raised awareness of the paucity of clinical findings and a more frequent use of echocardiography as a screening method seem essential to improve the prognosis of patients with S aureus endocarditis. Involvement of the central nervous system constitutes a relative indication of early valve replacement.

Event-related potentials during auditory language processing in congenitally blind and sighted people.

While behavioral studies have documented delayed language acquisition in blind children, other studies have revealed better speech discrimination abilities for blind than sighted adults. Several brain imaging studies have provided evidence for cortical reorganization due to visual deprivation but the cerebral organization of language in blind humans is not known yet. We hypothesized that the increasing specialization of language systems normally observed during development may not take place to the same degree in blind individuals since posterior visual areas do not receive their adequate input. On the other hand, we hypothesized that blind people, due to their greater reliance upon the auditory language signal, may process speech faster than sighted people. To test these assumptions, event-related potentials were recorded while 11 congenitally blind and 11 sighted adults matched in age, gender, handedness and education were engaged in a language task. Participants listened to sentences in order to decide after each sentence if it was meaningful or not. Incongruous sentence-final words elicited an N400 effect in both groups. The N400 effect had a left-lateralized fronto-central scalp distribution in the sighted but a symmetric and broad topography in the blind. Furthermore, the N400 effect started earlier in the blind than in the sighted. Closed class compared to open class sentence middle words elicited a more pronounced late negativity in the blind than in the sighted. These results suggest that blind people process auditory language stimuli faster than sighted people and that some language functions may be reorganized in the blind.

Neurologic manifestations in Staphylococcus aureus endocarditis: a review of 260 bacteremic cases in nondrug addicts.

PURPOSE: To investigate the neurologic manifestations of infective endocarditis caused by Staphylococcus aureus in a population of nondrug addicts with special emphasis on the clinical presentation, epidemiology, and mortality. PATIENTS AND METHODS: During the period from 1982 to 1991 a total of 8,514 cases of bacteremia with S aureus were reported to the Staphylococcus Laboratory, Copenhagen, Denmark. The medical records of cases of suspected infective endocarditis were retrospectively reviewed and classified according to the new diagnostic criteria for endocarditis proposed by Durack. RESULTS: A total of 260 cases from 63 hospitals fulfilled the diagnostic criteria. Overall, 91 patients (35%) experienced neurologic manifestations. Sixty-one presented with neurologic symptoms, whereas 30 patients developed neurologic complications at various intervals (median: 10 days) after the debut of the disease. The most frequent neurologic manifestation was unilateral hemiparesis, which occurred in 41 patients (45%). Forty-two percent of the females had neurologic manifestations compared to only 30% of the males (P = 0.06). Cases with native mitral valve infection had a significantly higher frequency of neurologic manifestations compared with all other valvular involvement (44% versus 29%, P = 0.02) but the frequency of neurologic complications was only nonsignificantly higher in those patients with native mitral valve infection than in those patients with native aortic valve infection (44% versus 31%, P = 0.10). Only two of the patients with tricuspid valve infection and none of those with congenital heart disorder experienced neurologic manifestations. A neurologic manifestation occurred in 22 (35%) of the 63 episodes in which vegetations were detected on the echocardiograms, compared with 17 (26%) of the 65 episodes without vegetations (P = 0.38). The mortality was 74% in patients with major neurologic manifestations and 56% in patients without neurologic manifestations (P = 0.008). In patients with neurologic complications the mortality was significantly higher among those treated with antibiotics alone as compared with those treated surgically (65 of 81, 80% versus 2 of 10, 20%; P = 0.0003). CONCLUSIONS: In a population of nondrug addicts with infective endocarditis caused by S aureus the following main conclusions can be drawn: neurologic manifestations occur with a higher frequency in patients with native mitral valve infection. The presence of vegetations on echocardiograms is not a risk factor for developing neurologic complications but this conclusion is based on the results of transthoracic echocardiograms performed in only one half of the patients. The majority of the neurologic manifestations occur on presentation or shortly thereafter and the risk of recurrent embolism is low. Mortality is increased in patients with neurologic manifestations. A neurologic event per se may constitute an indication for surgical treatment.

The effect of antistaphylococcal agents used alone and in combinations on the survival of Staphylococcus aureus ingested by human polymorphonuclear leukocytes.

The intracellular activity of a number of drugs used alone and in combinations against Staphylococcus aureus was investigated using an experimental design which imitates the clinical situation and differs from other published methods. Staphylococci were phagocytosed by human polymorphonuclear leukocytes and, after differential centrifugation and washing, the granulocytes were incubated in 90% pooled human serum with clinically relevant drug concentrations. When exposed to antibiotics, more than 40-50% of the bacteria were located intracellularly. Fusidic acid (100 mg/l), erythromycin (20 mg/l), and clindamycin (20 mg/l) all had a bacteriostatic effect during the first 6 h of incubation, whereas rifampicin (1 and 5 mg/l), vancomycin (5 and 20 mg/l), and ciprofloxacin (2 mg/l) all acted bactericidally with decreases in viable counts between 1.3-1.9 log10. The greatest bactericidal effect was achieved with tobramycin (10 mg/l), which produced more than a 4 log10 decrease in viable counts at 6 h. Combinations of fusidic acid with other antibiotics all resulted in killing kinetics different from those achieved with the drugs used individually. The bactericidal effect of ciprofloxacin and dicloxacillin during the first 6 h was abolished when these drugs were combined with fusidic acid. However, at 24 h no significant difference was found between the effect of dicloxacillin alone versus the combination dicloxacillin and fusidic acid. The combination of fusidic acid and rifampicin resulted in a killing identical to that achieved with rifampicin used alone during the first 6 h, but at 24 h the killing by the combination was significantly greater. The bactericidal effect of the combination dicloxacillin (20 mg/l) and tobramycin (10 mg/l) equalled that obtained with tobramycin (10 mg/l) used alone. Rifampicin (5 mg/l) antagonized the bactericidal effect of ciprofloxacin (2 mg/l) during the first 6 h of incubation but at 24 h the combination acted synergistically. The results obtained are partly in agreement and partly in conflict with previous results.

Event-related potentials during auditory and somatosensory discrimination in sighted and blind human subjects.

The objective of the present study was to test if and to what extent phasic and tonic event-related potentials of the human EEG may reflect phenomena of cortical plasticity. In particular, it was tested if the occipital cortex of blind subjects participates in the processing of non-visual stimuli. To this end, 12 blind and 12 blindfolded sighted subjects were tested in an auditory and a somatosensory discrimination task with 2 levels of discrimination difficulty. Slow and fast event-related potentials were recorded from 18 scalp electrodes. In addition to the negative slow waves found in sighted subjects over frontal and central sites during auditory and somatosensory discrimination, a pronounced negative wave was revealed in the blind also over occipital brain areas. These negative shifts were time-locked to the train of stimuli which had to be monitored with sustained attention, i.e. they rised and resolved with the beginning and the end of a 20-s discrimination time epoch. The P300 complex, on the other hand, which is a slow positive deflection over the posterior part of the scalp and which follows rare and task-relevant events 200-800 ms after stimulus onset was significantly smaller at occipital electrodes in the blind than in the sighted subjects. Combined with neurophysiological and neuronanatomical evidence originating from studies with visually deprived animals, these data suggest that the occipital cortex of blind human subjects is coactivated whenever the system is engaged in a task which requires sustained attention and is less effectively inhibited at the end of a perceptual time epoch. In total, the data cast doubt on the hypothesis that the occipital cortex of blind subjects participates in modality-specific non-visual information processing.

Auditory memory in congenitally blind adults: a behavioral-electrophysiological investigation.

Blind people must rely more than sighted people on auditory input in order to acquire information about the world. The present study was designed to test the hypothesis that blind people have better memory than sighted individuals for auditory verbal material and specifically to determine whether memory encoding and/or retrieval are improved in blind adults. An incidental memory paradigm was employed in which 11 congenitally blind people and 11 matched sighted controls first listened to 80 sentences which ended either with a semantically appropriate or inappropriate word. Immediately following, the recognition phase occurred, in which all sentence terminal words were presented again randomly intermixed with the same number of new words. Participants indicated whether or not they had heard the word in the initial study phase. Event-related brain potentials (ERPs) were recorded from 28 electrode positions during both the encoding and the retrieval phase. Blind participants' memory performance was superior to that of sighted controls. In addition, during the recognition phase, previously presented words elicited ERPs with larger positive amplitudes than new words, particularly over the right hemisphere. During the study phase, words that would subsequently be recognized elicited a more pronounced late positive potential than words that were not subsequently recognized. These effects were reliable in the congenitally blind participants but could only be obtained in the subgroup of sighted participants who had the highest memory performance. These results imply that blind people encode auditory verbal material more efficiently than matched sighted controls and that this in turn allows them to recognize these items with a higher probability.

Topographic differences of slow event-related brain potentials in blind and sighted adult human subjects during haptic mental rotation.

Twelve blindfolded sighted, nine congenitally blind, and seven adventitiously blind subjects were tested in a haptic mental rotation task while slow event-related brain potentials in the EEG were recorded from 17 scalp locations. The overall topography of the slow wave pattern which prevailed during the task differed for sighted and for blind, but not for congenitally and adventitiously blind subjects. While the tactile stimuli were encoded, the blind showed a pronounced occipital and the sighted a pronounced frontal activation. The task-specific amplitude increment of a negative slow wave which can be understood as a manifestation of the process of mental rotation proper, showed a different topography for sighted and for blind subjects too. It had its maximum over central to parietal cortical areas in both groups, but it extended more towards occipital regions in the blind. In both groups, the effects were very similar to those observed in former studies with visual versions of the mental rotation task, i.e. the slow wave amplitude over central to parietal areas increased monotonously with an increasing angular disparity of the two stimuli to be compared. These results are discussed with respect to the question of whether visual deprivation in the blind can cause a reorganization of cortical representational maps.

Spatial attention to central and peripheral auditory stimuli as indexed by event-related potentials.

Young adult subjects attended selectively to brief noise bursts delivered in free-field via central and peripheral arrays of four loudspeakers each that were arranged along a semi-circle extending from the midline to 90 degrees right of center. Frequent "standard" stimuli (90%) and infrequent "target/deviant" stimuli (10%) of increased bandwidth were delivered at a fast rate in random order and equiprobably from all eight speakers. In separate runs, the subject's task was to selectively attend to the center or rightmost speaker, and to press a button to the infrequent "target" stimuli occurring at the designated (spatial) location. Behavioral detection rates and concurrently recorded event-related potentials (ERPs) indicated that auditory attention was deployed as a finely tuned gradient around the attended source. The attentional gradients were steeper for the central than the peripheral array, indicating that attention can be more sharply focused upon sound sources directly in front of the listener. The ERP data suggested that selection for location is accomplished in two distinct stages, with an initial broadly tuned filtering, followed by a more narrowly focused selection of attended-location deviants.

Inhibition of return and oculomotor control in the blind.

Our attentional systems orient reflexively to novel environmental stimuli. Such attentive orienting is typically followed by a prolonged period of inhibition, known as inhibition of return (IOR), thought to be linked to the eye movement system. It is widely believed that IOR may provide a tagging mechanism that prevents perseveration, and thus facilitates attentional search. Using a tactile variant of the peripheral spatial cuing paradigm, we show IOR in congenitally blind adults and in an individual who had no eyes. These results demonstrate for the first time that spatial IOR can occur in the absence of oculomotor control.

No difference in enterotoxin production among Staphylococcus aureus strains isolated from blood compared with strains isolated from healthy carriers.

The production of enterotoxin A, B, C and D by 196 Staphylococcus aureus strains isolated from blood cultures and 95 strains from nasal carriers was investigated. Half of the bacteraemia strains were from patients who died with or because of their infection, the other half from patients who survived. The nasal strains were selected to match the bacteraemia strains regarding phage types. Overall, 30.6% of the bacteraemia strains and 40.0% of the nasal strains produced enterotoxins; enterotoxins B and C were the toxins produced most frequently in both groups. A similar incidence and pattern of enterotoxin production was found among the bacteraemia strains of S. aureus regardless of acquisition of the infection, the portal of entry, presence or absence of endocarditis and outcome of the infection. Thus, the concept that the enterotoxins play an important role in staphylococcal infections, apart from the diseases caused by the toxins per se such as food poisoning and toxic shock syndrome, cannot be substantiated by the results of the present study.

Effects of interstimulus interval on auditory event-related potentials in congenitally blind and normally sighted humans.

To test the hypothesis of auditory compensation after early visual deprivation, congenitally blind and sighted adults performed an auditory discrimination task. They had to detect a rare target tone among frequent standard tones. Stimuli were presented with different interstimulus intervals (ISIs) (200, 1000, 2000 ms) and the auditory-event related potentials to all tones and reaction times to targets were recorded. Increasing ISIs resulted in an increasing amplitude of the vertex response (N1-P2) in both groups, but this amplitude recovery was more pronounced in the blind. Furthermore, targets elicited larger and more posteriorly distributed N2 responses in the blind than in the sighted. Since target detection times were shorter in the blind as well, these findings imply compensatory adaptations within the auditory modality in humans blind from birth.

The determination of phytosphingosine-containing globotriaosylceramide from human kidney in the presence of lactosylceramide.

Globotriaosylceramide, the natural substrate of alpha-galactosidase A (the enzyme deficient in Fabry's disease) was prepared from human kidney by repeated medium pressure chromatography on Lichroprep Si 60 (E. Merck) before and after peracetylation. The apparently homogeneous preparation migrating as a single band on HPTLC was analysed by fast atom bombardment mass spectrometry and 1H-NMR at 500 MHz. It was found that in this fraction two major molecular species were comigrating: Gal alpha 1-4Gal beta 1-4Glc beta 1-1ceramide with nervonic and lignoceric acid linked to phytosphingosine and Gal beta 1-4Glc beta 1-1 ceramide with palmitic acid linked to sphingosine.

Slow negative brain potentials as reflections of specific modular resources of cognition.

The paper summarizes a series of experiments in which the topography and the amplitude of slow event-related brain potentials (slow ERPs) was studied in cognitive tasks which imposed different amounts of load on functionally distinct processing modules. The results suggest that the topography of slow ERPs reflects the relative activation/inactivation of distinct cortical cell assemblies while the absolute amplitude of the negative maximum seems to reflect how much a particular cell assembly is activated at a particular time. Translated into the terminology of cognitive resource theories one could say that tasks which evoke distinct slow wave patterns draw on independent resources. Likewise, the amplitude of a slow wave pattern could be related to the construct of resource allocation, i.e. the larger the amplitude of a slow wave pattern the more resources of a particular type are allocated to a task. These findings are discussed with respect to possible generator mechanism of slow waves and in relation to possible causes of capacity limitations of the human information processing system.