Comparison of mesencephalic free-floating tissue culture grafts and cell suspension grafts in the 6-hydroxydopamine-lesioned rat

Ventral mesencephalon (VM) of fetal rat and human origin grown as free-floating roller-tube (FFRT) cultures can survive subsequent grafting to the adult rat striatum. To further explore the functional efficacy of such grafts, embryonic day 13 ventral mesencephalic tissue was grafted either after 7 days in culture or directly as dissociated cell suspensions, and compared with regard to neuronal survival and ability to normalize rotational behavior in adult rats with unilateral 6-hydroxydopamine (6-OHDA) lesions. Other lesioned rats received injections of cell-free medium and served as controls. The amphetamine-induced rotational behavior of all 6-OHDA-lesioned animals was monitored at various time points from 18 days before transplantation and up to 80 days after transplantation. Tyrosine hydroxylase (TH) immunostaining of the histologically processed brains served to assess the long-term survival of grafted dopaminergic neurons and to correlate that with the behavioral effects. Additional cultures and acutely prepared explants were also fixed and stored for histological investigation in order to estimate the loss of dopaminergic neurons in culture and after transplantation. Similar behavioral improvements in terms of significant reductions in amphetamine-induced rotations were observed in rats grafted with FFRT cultures (127%) and rats grafted with cell suspensions (122%), while control animals showed no normalization of rotational behavior. At 84 days after transplantation, there were similar numbers of TH-immunoreactive (TH-ir) neurons in grafts of cultured tissue (775 +/- 98, mean +/- SEM) and grafts of fresh, dissociated cell suspension (806 +/- 105, mean +/- SEM). Cell counts in fresh explants, 7-day-old cultures, and grafted cultures revealed a 68.2% loss of TH-ir cells 7 days after explantation, with an additional 23.1% loss after grafting, leaving 8.7% of the original number of TH-ir cells in the intracerebral grafts. This is to be compared with a survival rate of 9.1% for the TH-ir cells in the cell-suspension grafts. Immunostaining for the calcium-binding proteins calretinin, calbindin, and parvalbumin showed no differences in the neuronal expression of these proteins between the two graft types. In conclusion, we found comparable dopaminergic cell survival and functional effects of tissue-culture grafts and cell-suspension grafts, which currently is the type of graft most commonly used for experimental and clinical grafting. In this sense the result is promising for the development of an effective in vitro storage of fetal nigral tissue, which at the same time would allow neuroprotective and neurotrophic treatment prior to intracerebral transplantation.

Effects of GDNF pretreatment on function and survival of transplanted fetal ventral mesencephalic cells in the 6-OHDA rat model of Parkinson's disease

Transplantation of fetal dopaminergic (DA) neurons offers an experimental therapy for Parkinson's disease (PD). The low availability and the poor survival and integration of transplanted cells in the host brain are major obstacles in this approach. Glial cell line-derived neurotrophic factor (GDNF) is a potent neurotrophic factor with growth- and survival-promoting capabilities for developing DA neurons. In the present study, we examined whether pretreatment of ventral mesencephalic (VM) free-floating roller tube (FFRT) cultures with GDNF would improve graft survival and function. For that purpose organotypic cultures of E14 rat VM were grown for 2, 4 or 8 days in the absence (control) or presence of GDNF [10 ng/ml] and transplanted into the striatum of 6-hydroxydopamine-lesioned rats. While all groups of rats showed a significant reduction in d-amphetamine-induced rotations at 6 weeks posttransplantation a significantly improved graft function was observed only in the days in vitro (DIV) 4 GDNF pretreated group compared to the control group. In addition, no statistical significant differences between groups were found in the number of surviving tyrosine hydroxylase-immunoreactive (TH-ir) neurons assessed at 9 weeks posttransplantation. However, a tendency for higher TH-ir fiber outgrowth from the transplants in the GDNF pretreated groups as compared to corresponding controls was observed. Furthermore, GDNF pretreatment showed a tendency for a higher number of GIRK2 positive neurons in the grafts. In sum, our findings demonstrate that GDNF pretreatment was not disadvantageous for transplants of embryonic rat VM with the FFRT culture technique but only marginally improved graft survival and function.

A comparative assessment of torque generated by lingual and conventional brackets

The aim of this study was to assess the effect of bracket type on the labiopalatal moments generated by lingual and conventional brackets. Incognito™ lingual brackets (3M Unitek), STb™ lingual brackets (Light Lingual System; ORMCO), In-Ovation L lingual brackets (DENTSPLY GAC), and conventional 0.018 inch slot brackets (Gemini; 3M Unitek) were bonded on identical maxillary acrylic resin models with levelled and aligned teeth. Each model was mounted on the orthodontic measurement and simulation system and 10 0.0175 × 0.0175 TMA wires were used for each bracket type. The wire was ligated with elastomerics into the Incognito, STb, and conventional brackets and each measurement was repeated once after religation. A 15 degrees buccal root torque (+15 degrees) and then a 15 degrees palatal root torque (-15 degrees) were gradually applied to the right central incisor bracket. After each activation, the bracket returned to its initial position and the moments in the sagittal plane were recorded during these rotations of the bracket. One-way analysis of variance with post hoc multiple comparisons (Tukey test at 0.05 error rate) was conducted to assess the effect on bracket type on the generated moments. The magnitude of maximum moment at +15 degrees ranged 8.8, 8.2, 7.1, and 5.8 Nmm for the Incognito, STb, conventional Gemini, and the In-Ovation L brackets, respectively; similar values were recorded at -15 degrees: 8.6, 8.1, 7.0, and 5.7 Nmm, respectively. The recorded differences of maximum moments were statistically significant, except between the Incognito and STb brackets. Additionally, the torque angles were evaluated at which the crown torque fell well below the minimum levels of 5.0 Nmm, as well as the moment/torque ratio at the last part of the activation/deactivation curve, between 10 and 15 degrees. The lowest torque expression was observed at the self-ligating lingual brackets, followed by the conventional brackets. The Incognito and STb lingual brackets generated the highest moments.

An Economic Comparison of Rotational Grazing Systems to Eight Crop Alternatives and the CRP Option for Highly Erodible Land in Southwest Iowa

Economic comparisons of income on highly erodible land (HEL) in Adams County were made utilizing five years of grazing data collected from a 13- paddock intensive-rotational grazing system and a four-paddock rotational-grazing system and four years of data collected from an 18-paddock intensive-rotational grazing system, all at the Adams County CRP Research and Demonstration Farm near Corning. Net income from the average grazing weight-gain of Angus-sired calves nursing crossbred cows was compared to the net income from grazing yearling steers, to the net income of eight NRCS-recommended crop rotations, and to the Conservation Reserve Program (CRP) option. Results of these comparisons show the 13-paddock intensive rotational grazing system with cow-calf pairs to be the most profitable alternative, with a net return of $19.86 per acre per year. The second most profitable alternative is the CRP option, with a net return of $13.09 per acre, and the third most profitable option is the fourpaddock rotation with cows and calves with a net return of $12.53 per acre. An 18-paddock system returned a net income of $2.47 per acre per year with cows and calves in 1993, but lost an average of $107.69 per acre each year in 1994 and 1995 with yearling steers. Each year, the steers were purchased high and sold low, contributing to the large loss per acre. The following recommended crop rotations all show net losses on these 9-14 % slope, Adair-Shelby Complex soils (ApD3): continuous corn; corn-soybean rotation; corn-soybean rotation with a farm program deficiency payment; corn-corn-corn-oats-meadow-meadow rotation with grass headlands; continuous corn to “T” with grass headlands and buffer strips; continuous corn to “T” with grass headlands, buffer strips, and a deficiency payment; corn-corn-oats-meadow rotation to “T”; and corn-soybeans-oats-meadow-meadow-meadow-meadow rotation to “T”. Per-acre yield assumptions of 90 bushels for corn, 30 bushels for soybeans, 45 bushels for oats, and four tons for alfalfa were used, with per-bushel prices of $2.40 on corn, $5.50 on soybeans, and $1.50 on oats. Alfalfa hay was priced at $40.00 per ton and grass hay at $33.33 per ton. The calf weight-gain in the cow/ calf systems was valued at $.90 per pound. All crop expenses except land costs were calculated from ISU publication Fm 1712, “Estimated Costs of Crop Production in Iowa - 1995.” Land costs were determined by using an opportunity cost and actual property tax figures for the land at the grazing site. In preparation for the end of the CRP beginning in 1996, further economic comparisons will be made after additional grazing seasons and data collection. This project is an interagency cooperative effort sponsored by the Southern Iowa Forage and Livestock Committee which has special permission from the USDA Farm Service Agency (FSA) to use CRP land for research and demonstration.

Rotation Grazing Demonstrations With Beef Cows on HEL - Adams County Conservation Reserve Program (CRP) Project

Two grazing systems were demonstrated on Conservation Reserve Program (CRP) land in southwestern Iowa near Corning in the summers of 1991, 1992, 1993, 1994, and 1995. This report summarizes the 1995 data and compares them to results from the four previous years. The systems, a 13-paddock intensive-rotational grazing system and a 4-paddock more traditional rotation, both established in 1991, are aimed at showing economically sustainable grass alternatives for steeply sloping (9-14% slope), highly erodible land (HEL) once the 10-year CRP ends. In a 147-day grazing season in 1995, nursing crossbred calves with no creep gained 2.36 pounds and 2.38 pounds per day on the 13- and 4-paddock systems, respectively. The rotations were stocked at 1.65 acres per cow-calf pair on the 13-paddock system and 1.72 acres per pair on the 4-paddock system. This produced 210.2 pounds of calf gain per acre on the 13-paddock system and 203.2 pounds of calf gain per acre on the 4- paddock system.. Similar calves gained 2.37 pounds and 2.50 pounds per day for 155 days, yielding a total gain per acre of 222.7 pounds on the 13-paddock system and 224.9 pounds on the 4-paddock system in 1994. Results for 1992 remain the highest from both systems in the five years of grazing, with calf gain per head per day at 2.45 for 155 days netting 241.9 pounds per acre on the 13- paddock system and calf gain per head per day at 2.38 for 154 days on the 4-paddock system yielding 263.6 pounds per acre. Cows maintained both their weight and condition scores in both systems again in 1995. A third system, the 18-paddock intensive-rotational grazing system, was stocked with stocker steers in 1995, and the results are reported in a second article in the 1996 ISU Beef Research Report entitled “Intensive- Rotational Grazing Steers on Highly Erodible Land at the Adams County CRP Project.” Concerning grazing management, paddocks were grazed four, five, or six times in the 13-paddock intensive- rotational grazing system during the 147-day grazing season of 1995. This number of times grazed per paddock was nearly equal to times grazed per paddock in 1994. However, several paddocks were subdivided temporarily to equalize paddock size and increase grazing uniformity. This increased the total number of cattle moves in the 13-paddock system from 78 in 1994 to 109 in 1995. The average length of stay on each paddock or subdivision of a paddock per grazing time was 1 to 2.2 days. This was less than in any of the other four grazing years in this project. The principle of not grazing more than half the standing forage during any one grazing period was closely followed in 1995. All paddocks in the 13-paddock system were also rested approximately the recommended 30 days between each grazing cycle in 1995.

Balancing on a slackline: 8-year-olds vs. adults

Children are less stable than adults during static upright stance. We investigated whether the same holds true for a task that was novel for both children and adults and highly dynamic: single-legged stance on a slackline. We compared 8-year-olds with young adults and assessed the following outcome measures: time on the slackline, stability on the slack-line (calculated from slackline reaction force), gaze movement, head-in-space rotation and translation, trunk-in-space rotation, and head-on-trunk rotation. Eight-year-olds fell off the slackline quicker and were generally less stable on the slackline than adults. Eight-year-olds also showed more head-in-space rotation and translation, and more gaze variability around a visual anchor point they were instructed to fixate. Trunk-in-space and head-on-trunk rotations did not differ between groups. The results imply that the lower postural stability of 8-year-olds compared to adults – as found in simple upright stance – holds true for dynamic, novel tasks in which adults lack the advantage of more practice. They also suggest that the lack of head and gaze stability constitutes an important limiting factor in children’s ability to master such tasks

The human vestibular cortex revealed by coordinate-based activation likelihood estimation meta-analysis.

The vestibular system contributes to the control of posture and eye movements and is also involved in various cognitive functions including spatial navigation and memory. These functions are subtended by projections to a vestibular cortex, whose exact location in the human brain is still a matter of debate (Lopez and Blanke, 2011). The vestibular cortex can be defined as the network of all cortical areas receiving inputs from the vestibular system, including areas where vestibular signals influence the processing of other sensory (e.g. somatosensory and visual) and motor signals. Previous neuroimaging studies used caloric vestibular stimulation (CVS), galvanic vestibular stimulation (GVS), and auditory stimulation (clicks and short-tone bursts) to activate the vestibular receptors and localize the vestibular cortex. However, these three methods differ regarding the receptors stimulated (otoliths, semicircular canals) and the concurrent activation of the tactile, thermal, nociceptive and auditory systems. To evaluate the convergence between these methods and provide a statistical analysis of the localization of the human vestibular cortex, we performed an activation likelihood estimation (ALE) meta-analysis of neuroimaging studies using CVS, GVS, and auditory stimuli. We analyzed a total of 352 activation foci reported in 16 studies carried out in a total of 192 healthy participants. The results reveal that the main regions activated by CVS, GVS, or auditory stimuli were located in the Sylvian fissure, insula, retroinsular cortex, fronto-parietal operculum, superior temporal gyrus, and cingulate cortex. Conjunction analysis indicated that regions showing convergence between two stimulation methods were located in the median (short gyrus III) and posterior (long gyrus IV) insula, parietal operculum and retroinsular cortex (Ri). The only area of convergence between all three methods of stimulation was located in Ri. The data indicate that Ri, parietal operculum and posterior insula are vestibular regions where afferents converge from otoliths and semicircular canals, and may thus be involved in the processing of signals informing about body rotations, translations and tilts. Results from the meta-analysis are in agreement with electrophysiological recordings in monkeys showing main vestibular projections in the transitional zone between Ri, the insular granular field (Ig), and SII.

Balancing the mind: vestibular induced facilitation of egocentric mental transformations

The body schema is a key component in accomplishing egocentric mental transformations, which rely on bodily reference frames. These reference frames are based on a plurality of different cognitive and sensory cues among which the vestibular system plays a prominent role. We investigated whether a bottom-up influence of vestibular stimulation modulates the ability to perform egocentric mental transformations. Participants were significantly faster to make correct spatial judgments during vestibular stimulation as compared to sham stimulation. Interestingly, no such effects were found for mental transformation of hand stimuli or during mental transformations of letters, thus showing a selective influence of vestibular stimulation on the rotation of whole-body reference frames. Furthermore, we found an interaction with the angle of rotation and vestibular stimulation demonstrating an increase in facilitation during mental body rotations in a direction congruent with rightward vestibular afferents. We propose that facilitation reflects a convergence in shared brain areas that process bottom-up vestibular signals and top-down imagined whole-body rotations, including the precuneus and tempero-parietal junction. Ultimately, our results show that vestibular information can influence higher-order cognitive processes, such as the body schema and mental imagery.

Training in intensive care medicine. A challenge within reach.

The medical training model is currently immersed in a process of change. The new paradigm is intended to be more effective, more integrated within the healthcare system, and strongly oriented towards the direct application of knowledge to clinical practice. Compared with the established training system based on certification of the completion of a series or rotations and stays in certain healthcare units, the new model proposes a more structured training process based on the gradual acquisition of specific competences, in which residents must play an active role in designing their own training program. Training based on competences guarantees more transparent, updated and homogeneous learning of objective quality, and which can be homologated internationally. The tutors play a key role as the main directors of the process, and institutional commitment to their work is crucial. In this context, tutors should receive time and specific formation to allow the evaluation of training as the cornerstone of the new model. New forms of objective summative and training evaluation should be introduced to guarantee that the predefined competences and skills are effectively acquired. The free movement of specialists within Europe is very desirable and implies that training quality must be high and amenable to homologation among the different countries. The Competency Based training in Intensive Care Medicine in Europe program is our main reference for achieving this goal. Scientific societies in turn must impulse and facilitate all those initiatives destined to improve healthcare quality and therefore specialist training. They have the mission of designing strategies and processes that favor training, accreditation and advisory activities with the government authorities.

Assessment of patient setup errors in IGRT in combination with a six degrees of freedom couch

PURPOSE The range of patient setup errors in six dimensions detected in clinical routine for cranial as well as for extracranial treatments, were analyzed while performing linear accelerator based stereotactic treatments with frameless patient setup systems. Additionally, the need for re-verification of the patient setup for situations where couch rotations are involved was analyzed for patients treated in the cranial region. METHODS AND MATERIALS A total of 2185 initial (i.e. after pre-positioning the patient with the infrared system but before image guidance) patient setup errors (1705 in the cranial and 480 in the extracranial region) obtained by using ExacTrac (BrainLAB AG, Feldkirchen, Germany) were analyzed. Additionally, the patient setup errors as a function of the couch rotation angle were obtained by analyzing 242 setup errors in the cranial region. Before the couch was rotated, the patient setup error was corrected at couch rotation angle 0° with the aid of image guidance and the six degrees of freedom (6DoF) couch. For both situations attainment rates for two different tolerances (tolerance A: ± 0.5mm, ± 0.5°; tolerance B: ± 1.0 mm, ± 1.0°) were calculated. RESULTS The mean (± one standard deviation) initial patient setup errors for the cranial cases were -0.24 ± 1.21°, -0.23 ± 0.91° and -0.03 ± 1.07° for the pitch, roll and couch rotation axes and 0.10 ± 1.17 mm, 0.10 ± 1.62 mm and 0.11 ± 1.29 mm for the lateral, longitudinal and vertical axes, respectively. Attainment rate (all six axes simultaneously) for tolerance A was 0.6% and 13.1% for tolerance B, respectively. For the extracranial cases the corresponding values were -0.21 ± 0.95°, -0.05 ± 1.08° and -0.14 ± 1.02° for the pitch, roll and couch rotation axes and 0.15 ± 1.77 mm, 0.62 ± 1.94 mm and -0.40 ± 2.15 mm for the lateral, longitudinal and vertical axes. Attainment rate (all six axes simultaneously) for tolerance A was 0.0% and 3.1% for tolerance B, respectively. After initial setup correction and rotation of the couch to treatment position a re-correction has to be performed in 77.4% of all cases to fulfill tolerance A and in 15.6% of all cases to fulfill tolerance B. CONCLUSION The analysis of the data shows that all six axes of a 6DoF couch are used extensively for patient setup in clinical routine. In order to fulfill high patient setup accuracies (e.g. for stereotactic treatments), a 6DoF couch is recommended. Moreover, re-verification of the patient setup after rotating the couch is required in clinical routine.

Radialer externer Fixateur zur geschlossenen Behandlung problematischer suprakondylärer Humerusfrakturen Typ III und IV bei Kindern und Jugendlichen

Operationsziel Geschlossene, anatomische Reposition und sichere Fixation von problematischen suprakondylären Typ-III- und Typ-IV-Humerusfrakturen, die mit den herkömmlichen Operationsmethoden nur schwierig geschlossen zu behandeln sind. Indikationen Gemäß der AO-Kinderklassifikation der suprakondylären Humerusfrakturen vom Typ III und IV: Frakturen, welche nicht geschlossen mittels üblicher Repositionsmethoden reponierbar sind sowie Frakturen, die nicht mittels der üblichen, gekreuzten perkutanen Kirschner-Draht-Technik zu fixieren sind. Bei schweren Schwellungszuständen, offener Fraktur oder initial neurologischen und/oder vaskulären Problemen („pulseless pink hand“) sowie bei mehrfachverletzten Kindern, welche eine optimale Rehabilitation benötigen und die Extremität gipsfrei sein sollte. Bei Kindern mit Komorbiditäten (z. B. Anfälle, Spastizität), die eine bessere Stabilität benötigen. Kontraindikationen Prinzipiell keine Kontraindikationen Operationstechnik Im nichtreponierten Zustand unter Durchleuchtungskontrolle Einbringen einer einzelnen Schanz-Schraube in den lateralen (radialen) Aspekt des distalen Fragments, welches sich in der streng seitlichen Röntgenprojektion als „Sand-Uhr“- bzw. Kreisform des Capitulum humeri darstellt. Je nach Größe dieses distalen Fragments kann die Schanz-Schraube rein epiphysär oder metaphysär liegen. Danach in absolut streng seitlicher Projektion des distalen Humerus im Bereich des meta-diaphysären Übergangs Einbohren einer 2. Schanz-Schraube unabhängig von der Ersten, die möglichst rechtwinklig zur Längsachse des Humerus in der a.-p.-Ebene zu liegen kommen sollte, um spätere Manipulationen mittels „Joy-Stick“-Technik zu erleichtern. Sind die beiden Schanz-Schrauben mehr oder weniger in beiden Ebenen parallel, so ist die Fraktur praktisch anatomisch reponiert. Nach erreichter Reposition Feinjustierung aller Achskomponenten. Sicherung der Flexion/Extension mittels einem von radial, distal eingebrachten sog. Anti-Rotations-Kirschner-Drahts, der die Stabilität signifikant erhöht und eine Drehung des distalen Fragments um die einzelne Schanz-Schraube verhindert. Postoperative Behandlung Keine zusätzliche Gipsruhigstellung notwendig. Es sollte eine funktionelle Nachbehandlung erfolgen. Ergebnisse Gemäß unserer Langzeitstudien bewegen die meisten Kinder bereits zum Zeitpunkt der ambulanten Pin-Entfernung in der Frakturambulanz ihren Ellbogen weitgehend normal. Bei einer Follow-up-Zeit über 40 Monate hatten 30/31 Kindern eine seitengleiche Achse und Beweglichkeit.

Hand-held 3D light field photography and applications

We propose a method to acquire 3D light fields using a hand-held camera, and describe several computational photography applications facilitated by our approach. As our input we take an image sequence from a camera translating along an approximately linear path with limited camera rotations. Users can acquire such data easily in a few seconds by moving a hand-held camera. We include a novel approach to resample the input into regularly sampled 3D light fields by aligning them in the spatio-temporal domain, and a technique for high-quality disparity estimation from light fields. We show applications including digital refocusing and synthetic aperture blur, foreground removal, selective colorization, and others.

On restricted families of projections in ℝ3

We study projections onto non-degenerate one-dimensional families of lines and planes in R 3 . Using the classical potential theoretic approach of R. Kaufman, one can show that the Hausdorff dimension of at most 12 -dimensional sets [Math Processing Error] is typically preserved under one-dimensional families of projections onto lines. We improve the result by an ε , proving that if [Math Processing Error], then the packing dimension of the projections is almost surely at least [Math Processing Error]. For projections onto planes, we obtain a similar bound, with the threshold 12 replaced by 1 . In the special case of self-similar sets [Math Processing Error] without rotations, we obtain a full Marstrand-type projection theorem for 1-parameter families of projections onto lines. The [Math Processing Error] case of the result follows from recent work of M. Hochman, but the [Math Processing Error] part is new: with this assumption, we prove that the projections have positive length almost surely.

Estimating Particle Shape and Orientation Using Volume Tensors

We develop statistical procedures for estimating shape and orientation of arbitrary three-dimensional particles. We focus on the case where particles cannot be observed directly, but only via sections. Volume tensors are used for describing particle shape and orientation, and we derive stereological estimators of the tensors. These estimators are combined to provide consistent estimators of the moments of the so-called particle cover density. The covariance structure associated with the particle cover density depends on the orientation and shape of the particles. For instance, if the distribution of the typical particle is invariant under rotations, then the covariance matrix is proportional to the identity matrix. We develop a non-parametric test for such isotropy. A flexible Lévy-based particle model is proposed, which may be analysed using a generalized method of moments in which the volume tensors enter. The developed methods are used to study the cell organization in the human brain cortex.

Computational Tools for Stereo and Light Field Photography

This thesis covers a broad part of the field of computational photography, including video stabilization and image warping techniques, introductions to light field photography and the conversion of monocular images and videos into stereoscopic 3D content. We present a user assisted technique for stereoscopic 3D conversion from 2D images. Our approach exploits the geometric structure of perspective images including vanishing points. We allow a user to indicate lines, planes, and vanishing points in the input image, and directly employ these as guides of an image warp that produces a stereo image pair. Our method is most suitable for scenes with large scale structures such as buildings and is able to skip the step of constructing a depth map. Further, we propose a method to acquire 3D light fields using a hand-held camera, and describe several computational photography applications facilitated by our approach. As the input we take an image sequence from a camera translating along an approximately linear path with limited camera rotations. Users can acquire such data easily in a few seconds by moving a hand-held camera. We convert the input into a regularly sampled 3D light field by resampling and aligning them in the spatio-temporal domain. We also present a novel technique for high-quality disparity estimation from light fields. Finally, we show applications including digital refocusing and synthetic aperture blur, foreground removal, selective colorization, and others.