Alginate Microspheres Containing Temperature Sensitive Liposomes (TSL) for MR-Guided Embolization and Triggered Release of Doxorubicin

Objective

The objective of this study was to develop and characterize alginate microspheres suitable for embolization with on-demand triggered doxorubicin (DOX) release and whereby the microspheres as well as the drug releasing process can be visualized in vivo using MRI.

Methods and Findings

For this purpose, barium crosslinked alginate microspheres were loaded with temperature sensitive liposomes (TSL/TSL-Ba-ms), which release their payload upon mild hyperthermia. These TSL contained DOX and [Gd(HPDO3A)(H₂O)], a T₁ MRI contrast agent, for real time visualization of the release. Empty alginate microspheres crosslinked with holmium ions (T₂* MRI contrast agent, Ho-ms) were mixed with TSL-Ba-ms to allow microsphere visualization. TSL-Ba-ms and Ho-ms were prepared with a homemade spray device and sized by sieving. Encapsulation of TSL in barium crosslinked microspheres changed the triggered release properties only slightly: 95% of the loaded DOX was released from free TSL vs. 86% release for TSL-Ba-ms within 30 seconds in 50% FBS at 42°C. TSL-Ba-ms (76 ± 41 μm) and Ho-ms (64 ± 29 μm) had a comparable size, which most likely will result in a similar in vivo tissue distribution after an i.v. co-injection and therefore Ho-ms can be used as tracer for the TSL-Ba-ms. MR imaging of a TSL-Ba-ms and Ho-ms mixture (ratio 95:5) before and after hyperthermia allowed in vitro and in vivo visualization of microsphere deposition (T₂*-weighted images) as well as temperature-triggered release (T₁-weighted images). The [Gd(HPDO3A)(H₂O)] release and clusters of microspheres containing holmium ions were visualized in a VX₂ tumor model in a rabbit using MRI.

Conclusions

In conclusion, these TSL-Ba-ms and Ho-ms are promising systems for real-time, MR-guided embolization and triggered release of drugs in vivo.     

How important is long‐distance seed dispersal for the regional survival of plant species?

Long‐distance seed dispersal is generally assumed to be important for the regional survival of plant species. In this study, we quantified the importance of long‐distance seed dispersal for regional survival of plant species using wind dispersal as an example. We did this using a new approach, by first relating plant species’ dispersal traits to seed dispersal kernels and then relating the kernels to regional survival of the species. We used a recently developed and tested mechanistic seed dispersal model to calculate dispersal kernels from dispersal traits. We used data on 190 plant species and calculated their regional survival in two ways, using species distribution data from 36,800 1 km²‐grid cells and 10,754 small plots covering the Netherlands during the largest part of the 20th century. We carried out correlation and stepwise multiple regression analyses to quantify the importance of long‐distance dispersal, expressed as the 99‐percentile dispersal distance of the dispersal kernels, relative to the importance of median‐distance dispersal and other plant traits that are likely to contribute to the explanation of regional survival: plant longevity (annual, biennial, perennial), seed longevity, and plant nutrient requirement. Results show that long‐distance dispersal plays a role in determining regional survival, and is more important than median‐distance dispersal and plant longevity. However, long‐distance dispersal by wind explains only 1–3% of the variation in regional survival between species and is equally important as seed longevity and much less important than nutrient requirement. In changing landscapes such as in the Netherlands, where large‐scale eutrophication and habitat destruction took place in the 20th century, plant traits indicating ability to grow under the changed, increasingly nutrient‐rich conditions turn out to be much more important for regional survival than seed dispersal.     

Wind and Water Dispersal of Wetland Plants Across Fragmented Landscapes

Biodiversity in wetlands is threatened by habitat loss and fragmentation, of which agricultural activities often are a cause. Dispersal of plant seeds via wind and ditches (water) may contribute to connecting remnant wetland plant populations in modern agricultural landscapes, and help to maintain and restore biodiversity. We developed a spatially explicit model to assess the relative importance of dispersal by wind and dispersal by water through drainage ditches for two wetland plant species in agricultural landscapes: a typical wind disperser and a typical water-disperser. Simulation results show that the typical wind disperser had a much higher capability to disperse by wind (90th percentile <30 m) than the typical water-disperser (90th percentile <2 m). Surprisingly, the capability to disperse via water was similar for the two species: 90th percentile dispersal distances following a combination of wind and water dispersal were between approximately 100 and 1000 m. Dispersal by water transported more seeds over long distances for both species. The main determinants for dispersal distance by water were roughness of the ditch (determined by, for example, bank vegetation) and the presence of obstructions (for example, culverts). Density or direction of the ditch network did not seem to affect water dispersal distances substantially. From a biodiversity conservation perspective, it would be most useful if areas with suitable riparian wetland habitat were intersected with a network of shallow ditches with a high roughness promoting seed deposition. These areas should then be connected to other suitable areas by a few regularly cleaned ditches with no obstructions and low seed trapping probability.     

The Influence of Texting Language on Grammar and Executive Functions in Primary School Children

When sending text messages on their mobile phone to friends, children often use a special type of register, which is called textese. This register allows the omission of words and the use of textisms: instances of non-standard written language such as 4ever (forever). Previous studies have shown that textese has a positive effect on children’s literacy abilities. In addition, it is possible that children’s grammar system is affected by textese as well, as grammar rules are often transgressed in this register. Therefore, the main aim of this study was to investigate whether the use of textese influences children’s grammar performance, and whether this effect is specific to grammar or language in general. Additionally, studies have not yet investigated the influence of textese on children’s cognitive abilities. Consequently, the secondary aim of this study was to find out whether textese affects children’s executive functions. To investigate this, 55 children between 10 and 13 years old were tested on a receptive vocabulary and grammar performance (sentence repetition) task and various tasks measuring executive functioning. In addition, text messages were elicited and the number of omissions and textisms in children’s messages were calculated. Regression analyses showed that omissions were a significant predictor of children’s grammar performance after various other variables were controlled for: the more words children omitted in their text messages, the better their performance on the grammar task. Although textisms correlated (marginally) significantly with vocabulary, grammar and selective attention scores and omissions marginally significantly with vocabulary scores, no other significant effects were obtained for measures of textese in the regression analyses: neither for the language outcomes, nor for the executive function tasks. Hence, our results show that textese is positively related to children’s grammar performance. On the other hand, use of textese does not affect—positively nor negatively—children’s executive functions.     

Computer-assisted mass spectrometric analysis of naturally occurring and artificially introduced cross-links in proteins and protein complexes

A versatile software tool, VIRTUALMSLAB, is presented that can perform advanced complex virtual proteomic experiments with mass spectrometric analyses to assist in the characterization of proteins. The virtual experimental results allow rapid, flexible and convenient exploration of sample preparation strategies and are used to generate MS reference databases that can be matched with the real MS data obtained from the equivalent real experiments. Matches between virtual and acquired data reveal the identity and nature of reaction products that may lead to characterization of post-translational modification patterns, disulfide bond structures, and cross-linking in proteins or protein complexes. The most important unique feature of this program is the ability to perform multistage experiments in any user-defined order, thus allowing the researcher to vary experimental approaches that can be conducted in the laboratory. Several features of VIRTUALMSLAB are demonstrated by mapping both disulfide bonds and artificially introduced protein cross-links. It is shown that chemical cleavage at aspartate residues in the protease resistant RNase A, followed by tryptic digestion can be optimized so that the rigid protein breaks up into MALDI-MS detectable fragments, leaving the disulfide bonds intact. We also show the mapping of a number of chemically introduced cross-links in the NK1 domain of hepatocyte growth factor/scatter factor. The VIRTUALMSLAB program was used to explore the limitation and potential of mass spectrometry for cross-link studies of more complex biological assemblies, showing the value of high performance instruments such as a Fourier transform mass spectrometer. The program is freely available upon request.     

Hot-water treatment before tissue culture reduces initial contamination in Lilium and Acer

Plant Cell, Tissue and Organ Culture (PCTOC) -     

Non-Invasive Targeted Peripheral Nerve Ablation Using 3D MR Neurography and MRI-Guided High-Intensity Focused Ultrasound (MR-HIFU): Pilot Study in a Swine Model

Purpose

Ultrasound (US)-guided high intensity focused ultrasound (HIFU) has been proposed for noninvasive treatment of neuropathic pain and has been investigated in in-vivo studies. However, ultrasound has important limitations regarding treatment guidance and temperature monitoring. Magnetic resonance (MR)-imaging guidance may overcome these limitations and MR-guided HIFU (MR-HIFU) has been used successfully for other clinical indications. The primary purpose of this study was to evaluate the feasibility of utilizing 3D MR neurography to identify and guide ablation of peripheral nerves using a clinical MR-HIFU system.

Methods

Volumetric MR-HIFU was used to induce lesions in the peripheral nerves of the lower limbs in three pigs. Diffusion-prep MR neurography and T1-weighted images were utilized to identify the target, plan treatment and immediate post-treatment evaluation. For each treatment, one 8 or 12 mm diameter treatment cell was used (sonication duration 20 s and 36 s, power 160–300 W). Peripheral nerves were extracted < 3 hours after treatment. Ablation dimensions were calculated from thermal maps, post-contrast MRI and macroscopy. Histological analysis included standard H&E staining, Masson’s trichrome and toluidine blue staining.

Results

All targeted peripheral nerves were identifiable on MR neurography and T1-weighted images and could be accurately ablated with a single exposure of focused ultrasound, with peak temperatures of 60.3 to 85.7°C. The lesion dimensions as measured on MR neurography were similar to the lesion dimensions as measured on CE-T1, thermal dose maps, and macroscopy. Histology indicated major hyperacute peripheral nerve damage, mostly confined to the location targeted for ablation.

Conclusion

Our preliminary results indicate that targeted peripheral nerve ablation is feasible with MR-HIFU. Diffusion-prep 3D MR neurography has potential for guiding therapy procedures where either nerve targeting or avoidance is desired, and may also have potential for post-treatment verification of thermal lesions without contrast injection.