Correlative microscopy and block-face imaging (CoMBI): a 3D imaging method with wide applicability in the field of biological science.

Correlative microscopy and block-face imaging (CoMBI) is an imaging method, which is characterized by the ability to obtain both serial block-face images as a 3-dimentional (3D) dataset and sections for 2-dimentional (2D) light microscopic analysis. These 3D and 2D morphological data can be correlated with each other to facilitate data interpretation. CoMBI is an easy-to-install and low-cost 3D imaging method since its system can be assembled by the researcher using a regular microtome, consumer digital camera, and some self-made devices, and its installation and instruction manuals are open-source.

Recent technological advances in correlative light and electron microscopy for the comprehensive analysis of neural circuits.

Light microscopy (LM) covers a relatively wide area and is suitable for observing the entire neuronal network. However, resolution of LM is insufficient to identify synapses and determine whether neighboring neurons are connected synapses. In contrast, the resolution of electron microscopy (EM) is sufficiently high to detect synapses and is useful for identifying neuronal connectivity; however, serial images cannot easily show the entire morphology of neurons, as EM covers a relatively narrow region.

Correlative microscopy and block-face imaging (CoMBI) method for both paraffin-embedded and frozen specimens.

Correlative microscopy and block-face imaging (CoMBI), a method that we previously developed, is characterized by the ability to correlate between serial block-face images as 3-dimensional (3D) datasets and sections as 2-dimensional (2D) microscopic images. CoMBI has been performed for the morphological analyses of various biological specimens, and its use is expanding. However, the conventional CoMBI system utilizes a cryostat, which limits its compatibility to only frozen blocks and the resolution of the block-face image.

Loss of VAMP5 in mice results in duplication of the ureter and insufficient expansion of the lung.

Background: Vesicle-associated membrane protein 5 (VAMP5) is a member of the SNARE protein family, which regulates the docking and fusion of membrane vesicles within cells. Previously, we reported ubiquitous expression of VAMP5 proteins in various organs except the brain and small intestine. However, the precise roles of VAMP5 in each organ remain unclear.

A novel imaging method for correlating 2D light microscopic data and 3D volume data based on block-face imaging.

We have developed an imaging method designated as correlative light microscopy and block-face imaging (CoMBI), which contributes to improve the reliability of morphological analyses. This method can collect both the frozen sections and serial block-face images in a single specimen. The frozen section can be used for conventional light microscopic analysis to obtain 2-dimensional (2D) anatomical and molecular information, while serial block-face images can be used as 3-dimensional (3D) volume data for anatomical analysis.

3D lung motion assessments on inspiratory/expiratory thin-section CT: Capability for pulmonary functional loss of smoking-related COPD in comparison with lung destruction and air trapping.

Purpose: To evaluate the utility of three-dimensional (3D) lung motion on inspiratory and expiratory CT for pulmonary functional loss in smoking-related COPD in comparison with lung destruction and air trapping assessments.

Method And Materials: Forty-four consecutive smokers and COPD patients prospectively underwent inspiratory and expiratory CT. A 3D motion vector map was generated from these CTs, and regional motion magnitudes were measured at the horizontal axis (X-axis), the ventrodorsal axis (Y-axis), and the craniocaudal axis (Z-axis).

The correlation between femoral sulcus morphology and osteoarthritic changes in the patello-femoral joint.

Purpose: The purpose of this study was to reveal the correlation between the type of lesion and the depth of osteoarthritic (OA) changes in the patello-femoral (PF) joint and its bony morphological characteristics using computed tomography (CT) data.

Methods: Eighty-seven cadaveric knees were included in this study with median age of 83 years (62-97). OA depth evaluation was performed following Outerbridge's classification.

Organization of organelles and VAMP-associated vesicular transport systems in differentiating skeletal muscle cells.

Vesicular transport plays an important role in the regulation of cellular function and differentiation of the cell, and intracellular vesicles play a role in the delivery of membrane components and in sorting membrane proteins to appropriate domains in organelles and the plasma membrane. Research on vesicular transport in differentiating cells has mostly focused on neurons and epithelial cells, and few such studies have been carried out on skeletal muscle cells. Skeletal muscle cells have specialized organelles and plasma membrane domains, including T-tubules, sarcoplasmic reticulum, neuromuscular junctions, and myotendinous junctions.

Identification of a naturally occurring retinoid X receptor agonist from Brazilian green propolis.

Background: Brazilian green propolis (BGP), a resinous substance produced from Baccharis dracunculifolia by Africanized honey bees (Apis mellifera), is used as a folk medicine. Our present study explores the retinoid X receptor (RXR) agonistic activity of BGP and the identification of an RXR agonist in its extract.

Methods: RXRα agonistic activity was evaluated using a luciferase reporter gene assay.

Filamentous structures in skeletal muscle: anchors for the subsarcolemmal space.

In skeletal muscle fibers, intermediate filaments and actin filaments provide structural support to the myofibrils and the sarcolemma. For many years, it was poorly understood from ultrastructural observations that how these filamentous structures were kept anchored. The present study was conducted to determine the architecture of filamentous anchoring structures in the subsarcolemmal space and the intermyofibrils.

An integrated teaching method of gross anatomy and computed tomography radiology.

It is essential for medical students to learn and comprehend human anatomy in three dimensions (3D). With this in mind, a new system was designed in order to integrate anatomical dissections with diagnostic computed tomography (CT) radiology. Cadavers were scanned by CT scanners, and students then consulted the postmortem CT images during cadaver dissection to gain a better understanding of 3D human anatomy and diagnostic radiology.

Vesicular transport system in myotubes: ultrastructural study and signposting with vesicle-associated membrane proteins.

Myofibers have characteristic membrane compartments in their cytoplasm and sarcolemma, such as the sarcoplasmic reticulum, T-tubules, neuromuscular junction, and myotendinous junction. Little is known about the vesicular transport that is believed to mediate the development of these membrane compartments. We determined the locations of organelles in differentiating myotubes.

Evaluation of ACL mid-substance cross-sectional area for reconstructed autograft selection.

Purpose: The purpose of this study was to compare the size of the native ACL mid-substance cross-sectional area and the size of commonly used autografts. Hypothesis of this study was that the reconstructed graft size with autografts would be smaller than the native ACL size.

Methods: Twelve non-paired human cadaver knees were used.

The localization of VAMP5 in skeletal and cardiac muscle.

Vesicle-associated membrane protein 5 (VAMP5) is a member of the SNARE protein family, which is generally thought to regulate the docking and fusion of vesicles with their target membranes. This study investigated the expression and localization of the VAMP5 protein. Immunoblotting analyses detected the VAMP5 protein in skeletal muscle, heart, spleen, lung, liver, and kidney tissue, but not in brain or small intestine tissue.

Alterations of biochemical marker levels and myonuclear numbers in rat skeletal muscle after ischemia-reperfusion.

Prolonged ischemia-reperfusion results in various damages in skeletal muscle. Following reperfusion, although the damaged muscles undergo regeneration, the precise process and mechanism of regeneration have not yet been fully understood. Here, we show the altered levels of plasma biochemical markers of muscle damage, and the change in myonuclear numbers in adult rat skeletal muscle by ischemia-reperfusion.

Hepatic CT perfusion measurements: a feasibility study for radiation dose reduction using new image reconstruction method.

Objectives: To assess the effects of image reconstruction method on hepatic CT perfusion (CTP) values using two CT protocols with different radiation doses.

Materials And Methods: Sixty patients underwent hepatic CTP and were randomly divided into two groups. Tube currents of 210 or 250 mA were used for the standard dose group and 120 or 140 mA for the low dose group.

ACL footprint size is correlated with the height and area of the lateral wall of femoral intercondylar notch.

Purpose: The purpose of this study was to reveal the correlation between the size of the native anterior cruciate ligament (ACL) footprint and the size of the lateral wall of femoral intercondylar notch.

Methods: Eighteen non-paired human cadaver knees were used. All soft tissues around the knee were resected except the ACL.

VAMP2 marks quiescent satellite cells and myotubes, but not activated myoblasts.

We examined the expression and intracellular localization of vesicle-associated membrane protein 2 (VAMP2) during the differentiation of skeletal muscle cells by immunofluorescence microscopy. In isolated single myofibers, VAMP2 was expressed in quiescent satellite cells, downregulated in proliferating myoblastic cells, and re-expressed with differentiation. In the myoblastic cell line C2C12, VAMP2 was expressed at a low level in the proliferating stage, and then increased after differentiation into myotubes.

Molecular dissection of Otx1 functional domains in the zebrafish embryo.

Otx proteins are involved in the induction of neurectoderm patterning and morphogenetic movements, leading to the formation of the vertebrate central nervous system. Despite lack of homology of sequence outside the homeodomain, a large body of evidence has shown that the Otx/Otd class of proteins has similar functions in many animal phyla. Thus, characterization of functional domains in proteins of this family would help in understanding how this functional equivalence operates.

Expression of protocadherin-19 in the nervous system of the embryonic zebrafish.

We have analyzed the expression pattern of protocadherin-19, a member of the delta2-protocadherins, in the nervous system of developing zebrafish using in situ hybridization methods. mRNA encoding protocadherin-19 (Pcdh19) began to be expressed at about 12 hours post fertilization (hpf) showing a segmental expression pattern in the anterior 1/3 of the neural keel, with strong expression in the presumptive forebrain, cerebellum/rhombomere 1 and rhombomere 4. Pcdh19 expression in the posterior neural keel was continuous and confined to the midline region.

Expression of protocadherin-9 and protocadherin-17 in the nervous system of the embryonic zebrafish.

In this study we analyzed expression patterns of two delta-protocadherins, protocadherin-9 and protocadherin-17, in the developing zebrafish using in situ hybridization and RT-PCR methods. Both protocadherins were mainly detected in the embryonic central nervous system, but each showed a distinct expression pattern. Protocadherin-9 message (Pcdh9) was expressed after 10h post fertilization (hpf).

VAMP2 is expressed in myogenic cells during rat development.

Vesicle-associated membrane protein 2 (VAMP2) is a member of the SNARE family of proteins that regulate the intracellular vesicle fusion process. This study investigated the developmental expression of VAMP2 in the rat embryo. In the trunk, VAMP2 was primarily found in the heart on embryonic day (E) 10.

Staging of disuse atrophy of skeletal muscles on immunofluorescence microscopy.

The Japanese population is rapidly aging, thereby causing excess demand for facilities for elderly invalids. It is imperative that social measures and scientific studies be carried out to enable better care of bedridden elderly people. The purpose of the present study was to review the histological changes that occur in disuse atrophy of skeletal muscles, the primary pathophysiology of bedridden invalids, with the object of developing a staging standard to be used by researchers and clinicians.

Cadherin-6 function in zebrafish retinal development.

Cadherin cell-adhesion molecules play crucial roles in vertebrate development including the development of the visual system. Most studies have focused on examining functions of classical type I cadherins (e.g.

Expression of protocadherin 18 in the CNS and pharyngeal arches of zebrafish embryos.

Here, we report the results of molecular cloning and expression analyses of a non-clustered protocadherin (pcdh), pcdh18 in zebrafish embryos. The predicted zebrafish pcdh18 protein shows 6566% identity and 7879% homology with its mammalian and Xenopus counterparts. It has a Disabled-1 binding motif in its cytoplasmic domain, which is characteristic of pcdh18.