Reduction of antibiotic-induced biofilm accumulation of Pseudomonas aeruginosa by quaternized phytoglycogen.

Biofilms are oft cited as a factor in the unwanted persistence and recalcitrance of microbial life and a strong research initiative exists to identify, understand, and target vulnerabilities. Phytoglycogen is a biodegradable nanoparticulate biomaterial that is purified from crop plants. Importantly, the highly branched glucan structure provides a scaffold on which to construct novel polymers. Functionalized phytoglycogen (FP) was synthesized using green chemistry principles. Screening of several molecules identified a form of quaternized phytoglycogen which reduced biofilm formation and accretion by Pseudomonas aeruginosa. Exposing P. aeruginosa to modified phytoglycogen and antibiotic in combination not only substantively reduced biofilms, but also prevented increased biofilm formation, a biological response to suboptimal antibiotic concentrations. Treatment of pregrown biofilms with sub-minimum inhibitory concentration antibiotic alone also led to increased proliferation, whereas FP-antibiotic combinations prevented or reduced the extent of this. Swimming, swarming and twitching motility, all critical for biofilm development, were negatively affected by FP. This work supports phytoglycogen as a promising foundational molecule for novel polymers, including those with anti-biofilm function. Critically, in addition to published reports on how suboptimal antibiotic concentrations promote biofilm formation, we demonstrated a similar effect upon pre-existing biofilms, indicating a further route for the failure of antibiotic therapies.

Head motion predictability explains activity-dependent suppression of vestibular balance control.

Vestibular balance control is dynamically weighted during locomotion. This might result from a selective suppression of vestibular inputs in favor of a feed-forward balance regulation based on locomotor efference copies. The feasibility of such a feed-forward mechanism should however critically depend on the predictability of head movements (HMP) during locomotion. To test this, we studied in 10 healthy subjects the differential impact of a stochastic vestibular stimulation (SVS) on body sway (center-of-pressure, COP) during standing and walking at different speeds and compared it to activity-dependent changes in HMP. SVS-COP coupling was determined by correlation analysis in frequency and time domains. HMP was quantified as the proportion of head motion variance that can be explained by the average head trajectory across the locomotor cycle. SVS-COP coupling decreased from standing to walking and further dropped with faster locomotion. Correspondingly, HMP increased with faster locomotion. Furthermore, SVS-COP coupling depended on the gait-cycle-phase with peaks corresponding to periods of least HMP. These findings support the assumption that during stereotyped human self-motion, locomotor efference copies selectively replace vestibular cues, similar to what was previously observed in animal models.

Neuronal network-based mathematical modeling of perceived verticality in acute unilateral vestibular lesions: from nerve to thalamus and cortex.

Acute unilateral lesions of vestibular graviceptive pathways from the otolith organs and semicircular canals via vestibular nuclei and the thalamus to the parieto-insular vestibular cortex regularly cause deviations of perceived verticality in the frontal roll plane. These tilts are ipsilateral in peripheral and in ponto-medullary lesions and contralateral in ponto-mesencephalic lesions. Unilateral lesions of the vestibular thalamus or cortex cause smaller tilts of the perceived vertical, which may be either ipsilateral or contralateral. Using a neural network model, we previously explained why unilateral vestibular midbrain lesions rarely manifest with rotational vertigo. We here extend this approach, focussing on the direction-specific deviations of perceived verticality in the roll plane caused by acute unilateral vestibular lesions from the labyrinth to the cortex. Traditionally, the effect of unilateral peripheral lesions on perceived verticality has been attributed to a lesion-based bias of the otolith system. We here suggest, on the basis of a comparison of model simulations with patient data, that perceived visual tilt after peripheral lesions is caused by the effect of a torsional semicircular canal bias on the central gravity estimator. We further argue that the change of gravity coding from a peripheral/brainstem vectorial representation in otolith coordinates to a distributed population coding at thalamic and cortical levels can explain why unilateral thalamic and cortical lesions have a variable effect on perceived verticality. Finally, we propose how the population-coding network for gravity direction might implement the elements required for the well-known perceptual underestimation of the subjective visual vertical in tilted body positions.

Bacterial contamination of tile drainage water and shallow groundwater under different application methods of liquid swine manure.

A 2 year field experiment evaluated liquid manure application methods on the movement of manure-borne pathogens (Salmonella sp.) and indicator bacteria (Escherichia coli and Clostridium perfringens) to subsurface water. A combination of application methods including surface application, pre-application tillage, and post-application incorporation were applied in a randomized complete block design on an instrumented field site in spring 2007 and 2008. Tile and shallow groundwater were sampled immediately after manure application and after rainfall events. Bacterial enumeration from water samples showed that the surface-applied manure resulted in the highest concentration of E. coli in tile drainage water. Pre-tillage significantly (p < 0.05) reduced the movement of manure-based E. coli and C. perfringens to tile water and to shallow groundwater within 3 days after manure application (DAM) in 2008 and within 10 DAM in 2007. Pre-tillage also decreased the occurrence of Salmonella sp. in tile water samples. Indicator bacteria and pathogens reached nondetectable levels within 50 DAM. The results suggest that tillage before application of liquid swine manure can minimize the movement of bacteria to tile and groundwater, but is effective only for the drainage events immediately after manure application or initial rainfall-associated drainage flows. Furthermore, the study highlights the strong association between bacterial concentrations in subsurface waters and rainfall timing and volume after manure application.

Head position during resting modifies spontaneous daytime decrease of downbeat nystagmus.

BACKGROUND: The intensity of downbeat nystagmus (DBN) decreases during the daytime when the head is in upright position. OBJECTIVE: This prospective study investigated whether resting in different head positions (upright, supine, prone) modulates the intensity of DBN after resting. METHODS: Eye movements of 9 patients with DBN due to cerebellar (n = 2) or unknown etiology (n = 7) were recorded with video-oculography. Mean slow-phase velocities (SPV) of DBN were determined in the upright position before resting at 9 am and then after 2 hours (11 am) and after 4 hours (1 pm) of resting. Whole-body positions during resting were upright, supine, or prone. The effects of all 3 resting positions were assessed on 3 separate days in each patient. RESULTS: Before resting (9 am), the average SPV ranged from 3.05 °/s to 3.6 °/s on the separate days of measurement. After resting in an upright position, the average SPV at 11 am and 1 pm was 0.65 °/sec, which was less (p < 0.05) than after resting in supine (2.1 °/sec) or prone (2.22 °/sec) positions. CONCLUSION: DBN measured during the daytime in an upright position becomes minimal after the patient has rested upright. The spontaneous decrease of DBN is less pronounced when patients lie down to rest. This indicates a modulation by otolithic input. We recommend that patients with DBN rest in an upright position during the daytime. CLASSIFICATION OF EVIDENCE: This study provides Class II evidence that for patients with DBN 2 hours of rest in the upright position decreases nystagmus more than 2 hours of rest in the supine or prone positions (relative improvement 79% upright, 33% supine, and 38% prone: p < 0.05).

Evaluation of Petrifilm EC method for enumeration of E. coli from soil.

AIMS: To evaluate the suitability of commercially available Petrifilm EC plates for enumeration of Escherichia coli from soil. METHODS AND RESULTS: A confirmed E. coli strain isolated from liquid swine manure was inoculated into sterilized sandy clay loam and loam soils at the concentrations of 10(2), 10(3), 10(5) CFU g(-1) of soil. The efficiency of recovery on Petrifilm EC plates for soils spiked with E. coli was compared with standard membrane filtration techniques on m-FC basal medium supplemented with 3-bromo-4-chloro-5-indoyl-beta-D-glucopyranoside (BCIG) and most probable numbers (MPN) techniques in E. coli medium with 4-methylumbelliferyl-beta-D-glucuronide (EC-MUG) broth. Petrifilm EC and m-FC (BCIG) methods were then assessed for the ability to recover E. coli from field soils applied with swine manure. No significant differences (P > 0.05) were observed between Petrifilm EC, m-FC (BCIG) and MPN methods for the recovery of E. coli from spiked samples, irrespective of soil type. However, recovery of E. coli from manure-applied field soil samples showed a significant difference (P < 0.05) between the Petrifilm EC method and the m-FC method in enumerating E. coli possibly as a result of false positives on m-FC. CONCLUSION: The Petrifilm EC method is suitable for the enumeration of E. coli from soil with a detection limit of 10 CFU g(-1) soil. SIGNIFICANCE AND IMPACT OF THE STUDY: The commercially available Petrifilm EC method is comparatively low cost, easy to use method for the enumeration of E. coli from soil without the need for further confirmation tests.

Comparing phosphorus mobilization strategies using Aspergillus niger for the mineral dissolution of three phosphate rocks.

Phosphorus deficiencies are limiting crop production in agricultural soils worldwide. Locally available sources of raw phosphate rock (PR) are being recognized for their potential role in soil fertility improvement. Phosphorus bioavailability is essential for the efficiency of PRs and can be increased by acid treatments. The utilization of organic acid producing micro-organisms, notably Aspergillus niger, presents a sustainable alternative to the use of strong inorganic acids, but acid production of A. niger strongly depends on the mineral content of the growth media. This study compared the phosphorus mobilization efficiency of two biological treatments, namely addition of acidic cell-free supernatants from A. niger cultivations to PRs and the direct cultivation of A. niger with PRs. The results show that addition of PR to cultivations leads to significant differences in the profile of organic acids produced by A. niger. Additions of PR, especially igneous rocks containing high amounts of iron and manganese, lead to reduced citric acid concentrations. In spite of these differences, phosphorus mobilization was similar between treatments, suggesting that the simpler direct cultivation method was not inferior. In addition to citric acid, it is suggested that oxalic acid contributes to PR solubilization in direct cultivations with A. niger, which would benefit farmers in developing countries where conventional fertilizers are not adequately accessible.

MSTd neurons during ocular following and smooth pursuit perturbation.

MSTd neurons in the behaving monkey were investigated during step-ramp smooth pursuit eye movements (SPEM), short perturbations of the small visual target during ongoing pursuit, and large-field visual stimulation inducing ocular following responses (OFR). Neurons responded with short latencies to visual motion during OFR. In contrast the non-retinal responses during SPEM and perturbations followed the eye movements by 100-150 ms and were in the opposite direction to the OFR response. Often neurons were not modulated by the perturbation. Although, both the OFR and the perturbation response are involuntary eye movements due to visual motion, it seems very unlikely that these MSTd neurons with non-retinal responses are involved in their direct control. Based on these responses, we suggest that our MSTd neurons may code for gaze direction in space based on visual estimates of self-motion and extraretinal estimates of eye-in-head motion.

Eye and head torsion is affected in patients with midbrain lesions.

The midbrain has been shown to contain crucial nuclei for the control of vertical and torsional eye movements. Recent studies in monkeys demonstrated that midbrain lesions also affect head movements during gaze saccades, but so far clinical reports on the matter have been missing. We measured 3D eye and head movements in two patients with oculomotor deficits due to unilateral midbrain lesions and in healthy control subjects. Subjects had (1) to perform head-free target directed gaze saccades (head-free task), and (2) to point to the target with a head-laser (head-only task). The patients had vertical eye movement deficits, which were not compensated by the head. Three-dimensional analysis revealed torsional deviations from the normal range of movements for both, the eye and the head movements.

Differences in saccade-evoked brain activation patterns with eyes open or eyes closed in complete darkness.

In this study we attempted to differentiate distinct components of the saccade network, namely cortical ocular motor centers and parieto-occipital brain regions, by means of a "minimal design" approach. Using a blocked design fMRI paradigm we evaluated the BOLD changes in a 2 x 2 factorial design experiment which was performed in complete darkness: while looking straight ahead with eyes open (OPEN) or closed (CLOSED) as well as during the execution of self-initiated horizontal to-and-fro saccades with the eyes open (SACCopen) or closed (SACCclosed). Eye movements were monitored outside the scanner via electro-oculography and during scanning using video-oculography. Unintentional eye-drifts did not differ during OPEN and CLOSED and saccade frequencies, and amplitudes did not vary significantly between the two saccade conditions. The main findings of the functional imaging study were as follows: (1) Saccades with eyes open or closed in complete darkness lead to distinct differences in brain activation patterns. (2) A parieto-occipital brain region including the precuneus, superior parietal lobule, posterior part of the intraparietal sulcus (IPS), and cuneus was relatively deactivated during saccades performed with eyes closed but not during saccades with eyes open or when looking straight ahead. This could indicate a preparatory state for updating spatial information, which is active during saccades with eyes open even without actual visual input. The preparatory state is suppressed when the eyes are closed during the saccades. (3) Selected ocular motor areas, not including the parietal eye field (PEF), show a stronger activation during SACCclosed than during SACCopen. The increased effort involved in performing saccades with eyes closed, perhaps due to the unusualness of the task, may be the cause of this increased activation.

The effect of nicotine on perceptual, ocular motor, postural, and vegetative functions at rest and in motion.

Nicotine has wellknown, unpleasant side effects, e.g., transient dizziness, nausea, and nicotine-induced nystagmus (NIN). To investigate factors influencing these effects, we addressed three questions: (1) Is the intensity of dizziness, nausea, NIN, and unsteadiness dependent on nicotine dosage? (2) Does the intensity of perceptual, ocular motor, vegetative effects, and postural imbalance correlate? (3) Do visual or vestibular motion stimuli produce and/or aggravate distressing dizziness and nausea? Sixty healthy non-smokers or occasional smokers participated; 40 were tested once before and six times after application of a nicotine nasal spray in doses of 1 mg or 2 mg with or without motion stimulation; 20 received a placebo nasal spray. Plasma nicotine concentrations were significantly related to nicotine dosage. Dizziness, nausea, NIN, and unsteadiness also depended on the nicotine dosage (p < 0.01).Nicotine blood concentration was a better predictor for the temporal dependence of nystagmus than nicotine dosage. Dizziness correlated highly with nausea (R = 0.63, p < 0.001). The degree of nicotine-induced nausea significantly correlated with postural imbalance. The time course of postural sway differed according to nicotine dosage and gender: for women, there was no clear relationship between sway magnitude and nicotine dosage, while men showed increased sway with higher dosage. Motion stimulation increased nicotine-induced dizziness and nausea, but did not significantly influence NIN or postural imbalance. Our data support the view that all measured adverse effects reflect dose-dependent nicotine-induced vestibular dysfunction. Additional motion stimulation aggravates dizziness and nausea, i.e., nicotine increases sensitivity to motion sickness.

Structural and functional MRIs disclose cerebellar pathologies in idiopathic downbeat nystagmus.

BACKGROUND: Neurologic disorders in which the etiology and pathogenesis are not yet understood are termed idiopathic. Downbeat nystagmus (DBN) is a frequent eye movement disorder that clinically manifests with oscillopsia and postural instability. Forty percent of patients with DBN are classified as having idiopathic DBN, because no underlying pathology can be demonstrated by conventional MRI or laboratory tests. METHODS: We evaluated gray matter brain volumes of 11 patients with idiopathic DBN and compared them to those of healthy controls using voxel-based morphometry. In a second, functional MRI experiment, patients and controls performed downward smooth pursuit eye movements (DOWN), which were then compared with straight-ahead fixation of a stationary target (MID). RESULTS: Small areas of localized gray matter atrophy were detected in the lateral cerebellar hemispheres (lobule VI) and ocular motor vermis of patients with idiopathic DBN, but not in the flocculus and paraflocculus. The functional imaging data, however, revealed reduced activation in the parafloccular lobule and in the ponto-medullary brainstem of the patients when they performed smooth pursuit eye movements downwards. CONCLUSIONS: The applied specialized imaging and data analysis techniques disclosed pathologies in an idiopathic eye movement disorder. The focal atrophy found in the vermal and lateral cerebellar regions in downbeat nystagmus (DBN) may lead to deficits in smooth pursuit eye movement initiation, which in turn causes hypofunction of the parafloccular lobe, associated with DBN. Our data are in line with experiments in primates showing that ablation of the floccular and parafloccular lobes disrupts smooth pursuit and causes DBN.

Mixed-valence cytoplasmic iron granules are linked to anaerobic respiration.

Intracellular granules containing ferric and ferrous iron formed in Shewanella putrefaciens CN32 during dissimilatory reduction of solid-phase ferric iron. It is the first in situ detection at high resolution (150 nm) of a mixed-valence metal particle residing within a prokaryotic cell. The relationship of the internal particles to Fe(III) reduction may indicate a respiratory role.

Subjective somatosensory vertical during dynamic tilt is dependent on task, inertial condition, and multisensory concordance.

To investigate how visual and vestibular cues are integrated for the perception of gravity during passive self-motion, we measured the ability to maintain a handheld object vertical relative to gravity without visual feedback during sinusoidal roll-tilt stimulation. Visual input, either concordant or discordant with actual dynamic roll-tilt, was delivered by a head-mounted display showing the laboratory. The four visual conditions were darkness, visual-vestibular concordance, stationary visual scene, and a visual scene 180 degrees phase-shifted relative to actual tilt. Tilt-indication performance using a solid, cylindrical joystick was better in the presence of concordant visual input relative to the other visual conditions. In addition, we compared performance when indicating the vertical by the joystick or a full glass of water. Subjects indicated the direction of gravity significantly better when holding the full glass of water than the joystick. Matching the inertial characteristics, including fluid properties, of the handheld object to the glass of water did not improve performance. There was no effect of visual input on tilt performance when using the glass of water to indicate gravitational vertical. The gain of object tilt motion did not change with roll-tilt amplitude and frequency (+/-7.5 degrees at 0.25 Hz, +/-10 degrees at 0.16 Hz, and +/-20 degrees at 0.08 Hz), however, the phase of object tilt relative to subject tilt showed significant phase-leads at the highest frequency tested (0.25 Hz). Comparison of the object and visual effects observed suggest that the task-dependent behavior change may be due to an attentional shift and/or shift in strategy.

Detection of floccular hypometabolism in downbeat nystagmus by fMRI.

The authors evaluated floccular activity with fMRI during the performance of vertical smooth pursuit eye movements in four patients with downbeat nystagmus (DBN) due to cerebellar degeneration and in 16 healthy controls. Region of interest analysis revealed a significantly diminished activation of both floccular lobes during downward but not upward pursuit in DBN. These imaging data support the view that a functional deficiency of the flocculi in downward pursuit causes DBN.

How the eyes move the body.

The increased postural sway of patients with disorders of the vestibular system improves with vision. The suppression of pathologic nystagmus also reduces sway. Because the latter effect cannot be attributed to retinal slip as a relevant feedback for postural control, the authors investigated how eye movements rather than retinal slip affect balance. They found that slow eye movements increase sway, possibly by an efference copy, which explains why spontaneous nystagmus causes postural imbalance.

4-aminopyridine restores visual ocular motor function in upbeat nystagmus.

The effect of the potassium channel blocker 4-aminopyridine (4-AP) on spontaneous upbeat nystagmus (UBN) was investigated with the search coil technique during fixation in different gaze positions and smooth pursuit in a patient before and after ingestion of 10 mg 4-AP. UBN was reduced from 8.6 deg/s to 2.0 deg/s by 4-AP causing subjective relief from distressing oscillopsia, and impaired upward smooth pursuit was restored (gains: before medication 0.38; after medication 0.86). In the dark, UBN was slightly stronger and not affected by 4-AP. We propose that 4-AP improved the function of cerebellar pathways that mediate gaze holding and smooth pursuit by intensifying the excitability of cerebellar Purkinje cells.