Monoolein Assisted Oil-Based Transdermal Delivery of Powder Vaccine.

An increasing number of protein vaccines have been researched for cancer, inflammation, and allergy therapies. Most of the protein therapeutics are administered through injection because orally-administered proteins are metabolized by the digestive system. Although transdermal administration has received increasing attention, the natural barrier formed by the skin is an obstacle. Monoolein is a common skin penetration enhancer that facilitates topical and transdermal drug delivery. Conventionally, it has been used in an aqueous vehicle, often with polyhydric alcohols. In the current study, monoolein was dissolved in an oil vehicle, isopropyl myristate, to facilitate the skin permeation of powder proteins. The skin permeabilities of the proteins were examined in-vivo and ex-vivo. Monoolein concentration-dependently enhanced the skin permeation of proteins. The protein permeability correlated with the zeta potential of the macromolecules. Dehydration of the stratum corneum (SC), lipid extraction from the SC, and disordering of ceramides caused by monoolein were demonstrated through Fourier transform infrared spectroscopic analysis and small-angle X-ray scattering analysis. An antigen model protein, ovalbumin from egg white, was delivered to immune cells in living mice, and induced antigen-specific IgG antibodies. The patch system showed the potential for transdermal vaccine delivery.

Large-Area Fluorescence and Electron Microscopic Correlative Imaging With Multibeam Scanning Electron Microscopy.

Recent improvements in correlative light and electron microscopy (CLEM) technology have led to dramatic improvements in the ability to observe tissues and cells. Fluorescence labeling has been used to visualize the localization of molecules of interest through immunostaining or genetic modification strategies for the identification of the molecular signatures of biological specimens. Newer technologies such as tissue clearing have expanded the field of observation available for fluorescence labeling; however, the area of correlative observation available for electron microscopy (EM) remains restricted. In this study, we developed a large-area CLEM imaging procedure to show specific molecular localization in large-scale EM sections of mouse and marmoset brain. Target molecules were labeled with antibodies and sequentially visualized in cryostat sections using fluorescence and gold particles. Fluorescence images were obtained by light microscopy immediately after antibody staining. Immunostained sections were postfixed for EM, and silver-enhanced sections were dehydrated in a graded ethanol series and embedded in resin. Ultrathin sections for EM were prepared from fully polymerized resin blocks, collected on silicon wafers, and observed by multibeam scanning electron microscopy (SEM). Multibeam SEM has made rapid, large-area observation at high resolution possible, paving the way for the analysis of detailed structures using the CLEM approach. Here, we describe detailed methods for large-area CLEM in various tissues of both rodents and primates.

Inhibitory Thoracic Interneurons are not Essential to Generate the Rostro-caudal Gradient of the Thoracic Inspiratory Motor Activity in Neonatal Rat.

The inspiratory motor activities are greater in the intercostal muscles positioned at more rostral thoracic segments. This rostro-caudal gradient of the thoracic inspiratory motor activity is thought to be generated by the spinal interneurons. To clarify the involvement of the inhibitory thoracic interneurons in this rostro-caudal gradient, we examined the effects of 10 µM strychnine, an antagonist of glycine and GABAA receptors, applied to the neonatal rat thoracic spinal cord. The respiratory-related interneuron activities were optically recorded from thoracic segments in the isolated neonatal rat brainstem-spinal cord preparations stained with voltage-sensitive dye, and the electrical inspiratory motor activities were obtained from the third and eleventh thoracic ventral roots (T3VR, T11VR). Although strychnine caused seizure-like activities in all of the ventral roots recorded, the inspiratory motor activities continued. The inspiratory optical signals in the rostral thoracic segments (T2-T5) were significantly larger than those in the caudal thoracic segments (T9-T11) regardless of the existence of strychnine. Similarly, the percent ratio of the amplitude of the inspiratory electrical activity in the T3VR under control and strychnine was significantly larger than that in the T11VR regardless of the existence of strychnine. Strychnine significantly increased the inspiratory activity in both the T3VR and T11VR. These results suggest that the glycinergic and GABAergic inhibitory interneurons are not essential to generate the rostro-caudal gradient in the neonatal rat thoracic inspiratory motor outputs, but these interneurons are likely to play a role in the inhibitory control of inspiratory motor output.

Two new species of photosymbiotic ascidians of the genus Diplosoma from the Ryukyu Archipelago, with partial sequences of the COI gene.

Two new species of Diplosoma are described from coral reefs in the Ryukyu Archipelago, Japan: D. watanabei sp. nov. and D. gumavirens sp. nov. Colonies of both species are green due to algal symbionts of Prochloron, which are distributed in the common cloacal cavity. Both species are characterized by the point of emergence of the retractor muscle and a unique combination of stigma numbers among the four rows of the branchial sac. There are three adhesive papillae in the embryos of D. watanabei, as in most ascidian embryos and larvae, whereas the pre-hatching embryos of D. gumavirens sp. nov. posses 12-16 adhesive papillae. Partial sequences of the cytochrome c oxidase subunit I (COI) gene discriminated the new species from each other and from congeners.

A new species of photosymbiotic ascidian from the Ryukyu Archipelago, Japan, with remarks on the stability of stigma number in photosymbiotic Diplosoma species.

Diplosoma variostigmatum sp. nov. is a photosymbiotic ascidian in the family Didemnidae. This species is characterized by the unique pattern of its stigma number, which is often variable within the same zooid, as well as within the same colony. The total number of stigmata per half branchial sac varies from 22 (five, six, six, and five stigmata from the top to bottom rows) to 27 (seven, seven, seven, and six). In contrast, the stigma patterns are constant in D. ooru, D. simile, D. simileguwa, and D. virens. Thus, the stigma number and its stability (or variability) seem to be important as characters for the taxonomy of the photosymbiotic Diplosoma species.

Medial marginal fracture of the patella following patellar dislocation.

Among patients with patellar dislocation, a small fragment is occasionally found along the medial border of the patella. However, the detailed pathology of this fracture has not been clearly described, especially in relation to the medial patellofemoral ligament (MPFL). The purpose of this study was to describe the pathology of this fracture in relation to the MPFL. The subjects were seven patients with an acute primary patellar dislocation showing a medial marginal fracture of the patella. An open exploration was performed to confirm the relationship between the fragment and the MPFL. The MPFL was repaired by re-attachment to the patella with anchors. The patellar fracture site was determined by the location of anchors in the post-operative plain radiograph. The fragments were continuous with the thick and taut MPFL in all cases. Re-attachment of the MPFL stabilized the patella against lateral shift by manual force improving the tracking. All patients returned to their pre-injury activity level without any episodes of recurrent dislocation or subluxation at the final follow up (av. 31 months). Medial marginal fracture of the patella was an avulsion fracture of the MPFL that occurred at the middle one-third of the patella.

Two new species of diplosoma (Ascidiacea: Didemnidae) bearing prokaryotic algae prochloron from Okinawajima (Ryukyu Archipelago, Japan).

Two new species of didemnid ascidians, Diplosoma ooru sp. nov. and Diplosoma simileguwa sp. nov., are described from coral reefs on Okinawajima (Ryukyu Archipelago, Japan). These two species form green colonies, having a symbiotic association with a prokaryotic alga Prochloron sp. The former species was found at the reef edges in the subtidal zone and the latter was found in a shallow reef lagoon. In these species, the colonies are thinner and the zooids are smaller than those of any other Prochloron-bearing Diplosoma species so far described. Moreover, each of the present new species has a unique combination of stigmatic numbers: 5 stigmata in the first and third rows, 6 in the second row, and 4 in the fourth in D. ooru; 4 stigmata in the first and third rows, 5 in the second row, and 3 in the fourth in D. simileguwa. In both of the new species, the retractor muscle emerges from the underside of the thorax. Larval morphology of D. ooru is also described.