Serial femtosecond and serial synchrotron crystallography can yield data of equivalent quality: A systematic comparison.

For the two proteins myoglobin and fluoroacetate dehalogenase, we present a systematic comparison of crystallographic diffraction data collected by serial femtosecond (SFX) and serial synchrotron crystallography (SSX). To maximize comparability, we used the same batch of micron-sized crystals, the same sample delivery device, and the same data analysis software. Overall figures of merit indicate that the data of both radiation sources are of equivalent quality. For both proteins, reasonable data statistics can be obtained with approximately 5000 room-temperature diffraction images irrespective of the radiation source. The direct comparability of SSX and SFX data indicates that the quality of diffraction data obtained from these samples is linked to the properties of the crystals rather than to the radiation source. Therefore, for other systems with similar properties, time-resolved experiments can be conducted at the radiation source that best matches the desired time resolution.

Visual word form processing deficits driven by severity of reading impairments in children with developmental dyslexia.

The visual word form area (VWFA) in the left ventral occipito-temporal (vOT) cortex is key to fluent reading in children and adults. Diminished VWFA activation during print processing tasks is a common finding in subjects with severe reading problems. Here, we report fMRI data from a multicentre study with 140 children in primary school (7.9-12.2 years; 55 children with dyslexia, 73 typical readers, 12 intermediate readers). All performed a semantic task on visually presented words and a matched control task on symbol strings. With this large group of children, including the entire spectrum from severely impaired to highly fluent readers, we aimed to clarify the association of reading fluency and left vOT activation during visual word processing. The results of this study confirm reduced word-sensitive activation within the left vOT in children with dyslexia. Interestingly, the association of reading skills and left vOT activation was especially strong and spatially extended in children with dyslexia. Thus, deficits in basic visual word form processing increase with the severity of reading disability but seem only weakly associated with fluency within the typical reading range suggesting a linear dependence of reading scores with VFWA activation only in the poorest readers.

Ambulatory blood pressure monitoring on admission in survivors of recent stroke entering inpatient rehabilitation.

Arterial hypertension (AHT) is a major risk factor for stroke, yet blood pressure (BP) goals thereafter remain uncertain. Although additional prognostic value of 24-hour ambulant BP monitoring (ABPM) is acknowledged, its clinical impacts remain limited. We suspected that routine ABPM could identify characteristic circadian BP patterns in different brain lesion types, the knowledge of which might, in turn, be helpful in improving overall BP management in patients after stroke. In our study, we compared cardiovascular parameters derived from ABPM and traditional blood pressure measurements (TBPM) among 105 stroke survivors who entered our inpatient neuro-rehabilitation program. The mean age of mostly male (64.8%) patients was 71 ± 12 years. Ischemic strokes were predominant (75.2%). Despite normotensive systolic BP means in TBPM (133.5 ± 18.2 mmHg) and ABPM (24 h: 122.8 ± 14.7 mmHg), AHT persisted in up to 67.6% of all patients, with ABPM uncovering nocturnal systolic non- or reversed dipping in 89.5% and 53.3%, respectively. The latter was predominant (85.7%) in the hemorrhagic subgroup which also displayed lower daytime SBP than the ischemic one (ABPM: 117.1 ± 11.8 vs 124.7 ± 14.7 mmHg, p = 0.033). Further differences were present among distinct brain lesion types. Sufficient dippers were younger (58 ± 12 vs 75 ± 11 years, p < 0.001), but adjusting for age yielded no independent correlations. In spite of normotensive daytime BP measurements, ABPM detects latent AHT and insufficient nocturnal BP dipping after the acute phase of stroke. Further studies are needed to elucidate the role of increased nocturnal BP in patients after stroke.

High prevalence and functional effects of serum antineuronal antibodies in patients with gastrointestinal disorders.

BACKGROUND: Antineuronal antibodies can be associated with both gastrointestinal (GI) and brain disorders. For example, antibodies against the potassium channel subunit dipeptidyl-peptidase-like protein-6 (DPPX) bind to neurons in the central nervous system (CNS) and myenteric plexus and cause encephalitis, commonly preceded by severe unspecific GI symptoms. We therefore investigated the prevalence of antineuronal antibodies indicative of treatable autoimmune CNS etiologies in GI patients. METHODS: Serum samples of 107 patients (Crohn's disease n = 42, ulcerative colitis n = 16, irritable bowel syndrome n = 13, others n = 36) and 44 healthy controls were screened for anti-DPPX and further antineuronal antibodies using immunofluorescence on rat brain and intestine and cell-based assays. Functional effects of high-titer reactive sera were assessed in organ bath and Ussing chamber experiments and compared to non-reactive patient sera. KEY RESULTS: Twenty-one of 107 patients (19.6%) had antibodies against the enteric nervous system, and 22 (20.6%) had anti-CNS antibodies, thus significantly exceeding frequencies in healthy controls (4.5% each). Screening on cell-based assays excluded established antienteric antibodies. Antibody-positive sera were not associated with motility effects in organ bath experiments. However, they induced significant, tetrodotoxin (TTX)-insensitive secretion in Ussing chambers compared to antibody-negative sera. CONCLUSIONS & INFERENCES: Antineuronal antibodies were significantly more frequent in GI patients and associated with functional effects on bowel secretion. Future studies will determine whether such antibodies indicate patients who might benefit from additional antibody-directed therapies. However, well-characterized encephalitis-related autoantibodies such as against DPPX were not detected, underlining their rarity in routine cohorts.

Brain structural alterations associated with dysfunctional cognitive control of pain in patients with low back pain.

BACKGROUND: Recent evidence has shown that chronic back pain (CBP) can lead to functional alterations in the circuitry underlying the cognitive control of pain. Thought suppression (TS) is a common type of cognitive control of pain. Previous research has shown that TS has paradoxical effects that may increase the awareness of pain. Pain-related TS may also increase individuals' attention to pain, which may also increase pain sensation, but thus far, the relationship between pain-related TS and structural brain alterations is unknown. METHODS: In this study, we investigated a group of 30 patients 6 months after lumbar disc surgery by applying voxel-based morphometry (VBM) to identify brain regions correlated with TS scores. RESULTS: Since pain-related TS has been positively correlated with depression, all calculations were controlled for depression and age. VBM revealed a negative correlation between the TS score and regional grey matter volume (GMV) in the left superior temporal gyrus (LSTG) and the left middle temporal gyrus (L MTG), which is part of the left temporoparietal junction (L TPJ). In addition, a mediation analysis revealed a significant mediation effect of the pain-related TS on the association between GMV of the left TPJ and reported pain intensity in the last 7 days. CONCLUSIONS: These findings are consistent with previous research on the dysfunctional cognitive control of pain and may therefore provide potential insights into the neural substrates of obstructive cognitive control in chronic low back pain, with a special emphasis on pain-related TS. SIGNIFICANCE: The link between pain-related thought suppression and brain morphology may provide a new perspective on the understanding of cognitive control of pain in chronic low back pain, which may help improve cognitive behavioural therapy.

Effects of plant-symbiotic relationships on the living soil microbial community and microbial necromass in a long-term agro-ecosystem.

We examined the impact of arbuscular mycorrhizal fungi and rhizobia on the living microbial community and microbial necromass under different long-term fertilization treatments at the long-term Static Fertilization Experiment Bad Lauchstädt (Germany). Phospholipid fatty acids (PLFA) and amino sugars plus muramic acid, were used as biomarkers for soil microbial bio- and necromass, respectively, and analyzed from six treatments imposed on two crop rotations, varying only in the inclusion/non-inclusion of a legume. Treatments included: two levels of only farmyard manure (FYM), only mineral fertilizer (NPK), the combined application of both fertilizer types and a non-fertilized control. PLFA profiles differed clearly between the investigated crop rotations and were significantly related to labile C, mineral N, and soil pH. This emphasizes the role of carbon, and of mycorrhizal and rhizobial symbioses, as driver for changes in the microbial community composition due to effects on the living conditions in soil. We found some evidence that legume associated symbiosis with arbuscular mycorrhizal fungi and rhizobia act as a buffer, reducing the impact of varying inputs of mineral nutrients on the decomposer community. While our results support former findings that living microbial populations vary within short-term periods and are reflective of a given crop grown in a given year, soil necromass composition indicates longer term changes across the two crop rotation types, mainly shaped by fertilizer related effects on the community composition and C turnover. However, there was some evidence that specifically the presence of a legume, affects the soil necromass composition not only over the whole crop rotation but even in the short-term.

Effect of soluble guanylyl cyclase activator and stimulator therapy on nitroglycerin-induced nitrate tolerance in rats.

Chronic nitroglycerin (GTN) anti-ischemic therapy induces side effects such as nitrate tolerance and endothelial dysfunction. Both phenomena could be based on a desensitization/oxidation of the soluble guanylyl cyclase (sGC). Therefore, the present study aims at investigating the effects of the therapy with the sGC activator BAY 60-2770 and the sGC stimulator BAY 41-8543 on side effects induced by chronic nitroglycerin treatment. Male Wistar rats were treated with nitroglycerin (100mg/kg/d for 3.5days, s.c. in ethanol) and BAY 60-2770 (0.5 or 2.5mg/kg/d) or BAY 41-8543 (1 and 5mg/kg/d) for 6days. Therapy with BAY 60-2770 but not with BAY 41-8543 improved nitroglycerin-triggered endothelial dysfunction and nitrate tolerance, corrected the decrease in aortic nitric oxide levels, improved the cGMP dependent activation of protein kinase I in aortic tissue and reduced vascular, cardiac and whole blood oxidative stress (fluorescence and chemiluminescence assays; 3-nitrotyrosine staining). In contrast to BAY 41-8543, the vasodilator potency of BAY 60-2770 was not impaired in isolated aortic ring segments from nitrate tolerant rats. sGC activator therapy improves partially the adverse effects of nitroglycerin therapy whereas sGC stimulation has only minor beneficial effects pointing to a nitroglycerin-dependent sGC oxidation/inactivation mechanism contributing to nitrate tolerance.

A Comparison of Two Methods for Computing IRT Scores From the Number-Correct Score.

Two estimates for item response theory latent trait scores (θ) based on the summed, number-correct score, X, were compared: (a) the so-called test characteristic curve (TCC) estimates, θTCC , in which the TCC is inverted so that a value of θ can be estimated directly from X and (b) the expected a posteriori-or Bayesian posterior mean-estimates, θEAP . Using data from Tenth-Grade English and Math Tests, the conditional, expected values for θTCC and θEAP (using both normal N(0, 1) and N(0, 10) priors), along with their conditional standard errors, were computed and plotted against a grid of actual θs. Under a normal N(0, 1) prior, it was found that the Bayesian θEAP s showed considerably smaller standard errors of measurement compared with the θTCC s-especially in the tails of the θ-distribution. However, the bias of the θEAP s based on the N(0, 1) prior was substantial in the extremes of the distribution of θ. The normal N(0, 10) prior for computing the θEAP s reduced their bias but increased their standard error-These were not unexpected statistical results, given the nearly universal trade-off between bias and standard error. The choice among the three summed-score θ-estimates examined here depends largely on which of the two major sources of distortion-bias versus standard error-is the more harmful.

Tooth length and incisal wear and growth in guinea pigs (Cavia porcellus) fed diets of different abrasiveness.

Dental diseases are among the most important reasons for presenting guinea pigs (Cavia porcellus) and other rodents to veterinary clinics, but the aetiopathology of this disease complex is unclear. Clinicians tend to believe that the ever-growing teeth of rabbits and rodents have a constant growth that needs to be worn down by the mastication of an appropriate diet. In this study, we tested the effect of four different pelleted diets of increasing abrasiveness [due to both internal (phytoliths) and external abrasives (sand)] or whole grass hay fed for 2 weeks each in random order to 16 guinea pigs on incisor growth and wear, and tooth length of incisors and cheek teeth. There was a positive correlation between wear and growth of incisors. Tooth lengths depended both on internal and external abrasives, but only upper incisors were additionally affected by the feeding of whole hay. Diet effects were most prominent in anterior cheek teeth, in particular M1 and m1. Cheek tooth angle did not become shallower with decreasing diet abrasiveness, suggesting that a lack of dietary abrasiveness does not cause the typical 'bridge formation' of anterior cheek teeth frequently observed in guinea pigs. The findings suggest that other factors than diet abrasiveness, such as mineral imbalances and in particular hereditary malocclusion, are more likely causes for dental problems observed in this species.

Performing radical cystectomy and urinary diversion in regional anesthesia: potential risk reduction in the treatment of bladder cancer.

OBJECTIVE: To assess the feasibility and performance of radical cystectomy with urinary diversion using exclusively regional anesthesia (i.e. combined spinal thoracic epidural anesthesia, CSTEA), avoiding the adverse effects of general anesthesia. MATERIALS AND METHODS: In our hospital, radical cystectomy with extended pelvic and iliac lymphadenectomy and urinary diversion was performed on 28 patients using CSTEA without applying general anesthesia, in 2011 and 2012. Under maintained spontaneous breathing, the patients were awake and responsive during the entire procedure. Outcome measurements included operative time, blood loss, start of oral nutrition, start of mobilization, postoperative pain levels using numerical and visual analog scales (NAS/VAS), postoperative complications according to the Clavien-Dindo classification and length of hospital stay. RESULTS: All surgical procedures were performed without any complications and caused no anesthesiologically or surgically untoward effects. We observed no more severe complications than grade 1 according to the Clavien-Dindo classification. CONCLUSIONS: Our data show that CSTEA is an effective and safe technique for radical cystectomy, whereby spontaneous breathing and reduced interference with the cardiopulmonary system potentially lower the perioperative risks, especially for high-risk patients. We recommend practice of CSTEA for radical cystectomy to further evaluate and monitor the safety, efficacy, outcomes and complications of CSTEA.

How to optimize lipoprotein apheresis treatment--a second look.

BACKGROUND: Lipoprotein apheresis (LA) is used in hypercholesterolemic patients suffering from cardiovascular disease (CHD) if a modified diet and lipid-lowering drug regimens had failed. During the first LA treatments LDL-cholesterol (LDL-C) and lipoprotein (a) (Lp(a)) can be decreased very effectively when using generally accepted formulas for calculating plasma (PV) (e.g. Pearson) or blood volumes (BV) as a basis for calculating treatment volume (e.g. Nadler). With respect to LDL-C and Lp(a) levels after LA treatment not all treated patients on steady state with apheresis treatment procedures may achieve the desired target concentrations for LDL-C (<70 mg/dl) and Lp(a) (<30 mg/dl). Are there further ways to increase the effectiveness of LA? METHODS: Over months or years of LA the treated volumes were stepwise increased in patients to achieve target cholesterol concentrations but not sufficiently in all cases. Therefore the patients' actual LA treatment volumes were compared to the calculated PV or BV. To possibly optimize the treatment capacity of LA procedures independent of calculated PV or BV the capacity threshold was determined in addition. During LA procedures every 20 min cholesterol, triglycerides, LDL-C, HDL-C and Lp(a) concentrations were determined and related to the hematocrit to exclude dilution effects. RESULTS: In patients undergoing regular LA treated volumes vs. calculated volumes were different: for PV 28 ± 18% (n = 7); for BV 28 ± 20% (n = 6). The mean treated volumes were 1.3 fold larger than the calculated volumes to achieve cholesterol target levels in most LA treatments. With respect to the capacity threshold we observed in only 1 of 13 patients an ineffective long treatment time. No LA procedure failed due to exhausted capacity. CONCLUSIONS: Lipoprotein apheresis treatment is a very effective treatment procedure in lowering LDL-C and Lp(a). However, not in all procedures the optimal treatment volume for LA patients may be calculated. However calculations of PV and BV are more or less error-prone. An increase of 1.3 fold in the calculated volumes may be the first step in optimizing individual LA treatment options. In addition, to exclude an exhaustion of LA procedures the determination of the individual capacity threshold in every LA patient may be further helpful to adjust treatment volumes. To substantiate our demand on changed treatment volumes further data are necessary.

Neuronal correlates of impaired habituation in response to repeated trigemino-nociceptive but not to olfactory input in migraineurs: an fMRI study.

INTRODUCTION: Using functional magnetic resonance imaging (fMRI), we aimed to explore the habituation behaviour to trigemino-nociceptive as well as olfactory stimuli in migraine patients. We exclusively focussed on intrasessional behavioural rating patterns and the related blood oxygen level dependent (BOLD) signal changes. FINDINGS: We observed that groups significantly differ in the time course of pain intensity ratings during the stimulation session: whereas interictal migraineurs sensitized (increasing pain ratings), control subjects habituated (decreasing pain ratings). Pain ratings of ictal patients remained unchanged. This behaviour is accompanied by a similar time course of neuronal activity in the bilateral anterior insula, in the middle cingulate cortex and in the thalamus. In these areas, the brain activity increased in migraineurs but decreased in the control group during the session. In contrast to these findings, the rating patterns for the olfactory stimuli (rose odour) did not differ between patients and controls and a gradual decrease of perceived stimulus intensity was found in all three groups. This stimulus specific response may occur because the olfactory system is the only sensory system not passing the thalamus. CONCLUSION: Our data suggest that impaired habituation in functional brain systems in migraine is fundamental only to specific modalities including the trigemino-nociceptive, but, at least, excluding the olfactory system. Our findings further suggest that there is no single neuronal modulator responsible for the altered rating pattern in migraineurs.

Cystectomy and urinary diversion in the treatment of bladder cancer without artificial respiration.

PURPOSE: To assess the feasibility and performance of radical cystectomy with urinary diversion using exclusively regional anesthesia (i.e. combined spinal thoracic epidural anesthesia, CSTEA). MATERIALS AND METHODS: In 2011 radical cystectomy with extended pelvic and iliac lymphadenectomy was performed on 14 patients using urinary diversion without applying general anesthesia. Under maintained spontaneous breathing, the patients were awake and responsive during the entire procedure. Postoperatively, pain management took three days with the remaining epidural catheter before oral analgesics were administered. Mobilization and diet restoration were carried out according to the fast-track concept. Outcome measurements included operative time, blood loss, beginning of oral nutrition, beginning of mobilization, postoperative pain levels using numerical and visual analog scales (NAS/VAS), length of hospital stay. RESULTS: All surgical procedures were performed without any complications. The absence of general anesthesia did not result in any relevant disadvantages. The postoperative progress was normal in all patients. Particularly, cardiopulmonary complications and enteroparesis did not occur. The provided palliative care proved sufficient (NAS max. 3-4). Discharge followed 10 to 22 days after surgery. At the time of discharge, the patients described the procedure to be relatively positive. CONCLUSIONS: Our data show that CSTEA is an effective technique for radical cystectomy, whereby spontaneous breathing and reduced interference with the cardiopulmonary system potentially lower the perioperative risks especially for high-risk patients. We recommend practice of CSTEA for radical cystectomy to further evaluate and monitor the safety, efficacy, outcomes, and complications of CSTEA.

Cystectomy and urinary diversion in the treatment of bladder cancer without artificial respiration

PURPOSE: To assess the feasibility and performance of radical cystectomy with urinary diversion using exclusively regional anesthesia (i.e. combined spinal thoracic epidural anesthesia, CSTEA). MATERIALS AND METHODS: In 2011 radical cystectomy with extended pelvic and iliac lymphadenectomy was performed on 14 patients using urinary diversion without applying general anesthesia. Under maintained spontaneous breathing, the patients were awake and responsive during the entire procedure. Postoperatively, pain management took three days with the remaining epidural catheter before oral analgesics were administered. Mobilization and diet restoration were carried out according to the fast-track concept. Outcome measurements included operative time, blood loss, beginning of oral nutrition, beginning of mobilization, postoperative pain levels using numerical and visual analog scales (NAS/VAS), length of hospital stay. RESULTS: All surgical procedures were performed without any complications. The absence of general anesthesia did not result in any relevant disadvantages. The postoperative progress was normal in all patients. Particularly, cardiopulmonary complications and enteroparesis did not occur. The provided palliative care proved sufficient (NAS max. 3-4). Discharge followed 10 to 22 days after surgery. At the time of discharge, the patients described the procedure to be relatively positive. CONCLUSIONS: Our data show that CSTEA is an effective technique for radical cystectomy, whereby spontaneous breathing and reduced interference with the cardiopulmonary system potentially lower the perioperative risks especially for high-risk patients. We recommend practice of CSTEA for radical cystectomy to further evaluate and monitor the safety, efficacy, outcomes, and complications of CSTEA.

Current view: indications for extracorporeal lipid apheresis treatment.

BACKGROUND: One of the first investigations concerning extracorporeal treatment of hypercholesterolemia was performed in 1967 by plasma exchange in patients with homozygous or severe heterozygous familial hypercholesterolemia (FH). In the following decades, several specific lipid apheresis systems were developed to efficiently eliminate low-density lipoprotein (LDL) cholesterol and Lp(a) cholesterol in hypercholesterolemic patients. In the early 1980s, the main clinical indication has been homozygous FH including mainly children and pregnant women. In consideration of the current development of lipid-lowering regimens and scientific knowledge of preventing progression of cardiovascular diseases, the spectrum of indications to initiate lipid apheresis was extended due to still insufficient lipid-lowering therapy in some clinical cases. However, a generally accepted indication for lipid apheresis treatment is still under discussion. In Germany, the target-oriented distribution of increasingly limited healthcare resources demand data which support the benefit of established treatment procedures such as lipid apheresis. In recent years, the Federal Joint Committee (G-BA), a paramount decision-making body of the German Healthcare System, issued to reassess the approval of chronic lipid apheresis therapy for regular reimbursement. Therefore, in 2005, an interdisciplinary German Apheresis Working Group has been established by members of both the German societies of nephrology. One of the first goals of this working group was a revision of the indications for lipid apheresis corresponding to current guidelines and recommendations for the treatment of lipid disorders. In addition, recently new pathophysiological perceptions of the impact of lipoproteins on atherogenesis and thrombosis were also considered. METHODS AND RESULTS: Since 2005, the working group met on a regular basis to substantiate the first defined goals. The indications for lipid apheresis were critically revised with respect to actual results from clinical investigations, cardiovascular guidelines, and scientific knowledge and were accepted by the members of the apheresis working group. CONCLUSIONS: There is consensus between the medical societies and health insurance funds regarding the need for general accepted guidelines for lipid apheresis. Recommendations for the indications of lipid apheresis were developed, but additionally these results should be confirmed by medical societies to transform them to guidelines. However, due to limited data showing that lipid apheresis has effects on the progression of cardiovascular diseases all members of the apheresis working group support a project for creating a lipid apheresis registry. This apheresis registry has been developed and recently started. The primary goal is to substantiate prospective long-term data on clinical outcome of chronic lipid apheresis treatment and to support additional clinical research activities in this field. In addition, this registry should comply with the actual requests of the Federal Joint Committee (G-BA).