Evans Blue fluorescence in the cervical spinal cord of G93A mice at early and late stages of disease.

<p>In the cervical spinal cord, EB was clearly detected within the blood vessels (red, arrowheads) in the control C57BL/6J mice at (A, B, C) 12–13 weeks of age or (D, E) in the lumen of vessels (brilliant green) at 19–20 weeks of age. In G93A mice, vascular leakage of EB (red, arrows) was detected (F, G) at early (13 weeks of age) disease symptoms and (H, I, J) at end-stage of disease (17–18 weeks of age) when more EB extravasation was seen. Arrowheads in F and I indicate vessel permeability. Scale bar in A–J is 25 µm.</p

Immunohistochemical staining for endothelial cells (CD146) and astrocytes (GFAP) in the lumbar spinal cord of G93A mice at early and late stages of disease.

<p>(A, B, C) Similar to cervical spinal cord, endothelial cells (green, arrowheads) and astrocytes (red, asterisk) in C57BL/6J mice at 19–20 weeks of age appeared normal. In G93A mice at (D, E) early or (F, G) end-stage of disease, decreased CD146 antigen expression by endothelial cells (green, arrowheads) was observed. Note: increased astrocyte activation in the lumbar spinal cord (F, G, asterisks) was detected in G93A mice at late stage of disease. The nuclei in A, C, D, and F are shown with DAPI. Scale bar in A, C, D, F is 50 µm; B, E, G is 25 µm.</p

Immunohistochemical staining for endothelial cells (CD146) and astrocytes (GFAP) in the cervical spinal cord of G93A mice at early and late stages of disease.

<p>(A, B) Normal appearance of endothelial cells (green, arrowheads) and delineated astrocytes (red, asterisk) was observed in the control C57BL/6J mice at 19–20 weeks of age. Endothelia (green, arrowheads) surrounding capillaries were partially revealed in G93A mice at (C, D) initial or (E, F) late stages of disease. Note: increased astrocyte activation in the cervical spinal cord (F, asterisks) was detected in G93A mice at late stage of disease. The nuclei in A, C, and E are shown with DAPI. Scale bar in A, C, E is 50 µm; B, D, F is 25 µm.</p

Motor neurons in the lumbar spinal cord of G93A mice at early and late stage of disease (cresyl violet staining).

<p>In the lumbar spinal cord, C57BL/6J mice at (A) 12–13 weeks of age and (B) 19–20 weeks of age showed numerous motor neurons with strong Nissl body staining. Most degenerated or swollen motor neurons (asterisks) were found in G93A mice at (C) early (13 weeks of age) and (D) late (17–18 weeks of age) stages of disease; most surviving motor neurons were small. Scale bar on left side is 200 µm, right side is 50 µm.</p

Immunofluorescence staining for Glut-1 in the cervical and lumbar spinal cords of G93A mice at early and late stages of disease.

<p><i>Cervical spinal cord.</i> High expression of Glut-1 (red) was determined in endothelial lining of many blood vessels of various diameters in the cervical spinal cord of the control C57BL/6J mice at (A), (B) 12–13 weeks of age and (C) 19–20 weeks of age. In G93A mice at (D), (E) initial or (F), (G) late stages of disease, immunoreaction for Glut-1 in the endothelial cells appear to be low, or nonexistent. The nuclei in A–G are shown with DAPI. Scale bar in A and E is 50 µm; B, C, D, F, G is 25 µm. Outline of white dots indicates configuration of blood vessels. <i>Lumbar spinal cord.</i> Similar to the cervical spinal cord, most Glut-1-positive endothelial cells (red) were observed in the control C57BL/6J mice at (H), (I) 12–13 weeks of age and (J) 19–20 weeks of age. Less Glut-1 expression was found in G93A mice at (K), (L) early or (M), (N) end-stage of disease. The nuclei in H–N are shown with DAPI. Scale bar in J, M, N is 50 µm; H, I, K, L is 25 µm.</p

Immunofluorescence staining for laminin in the lumbar spinal cord of G93A mice at early and late stages of disease.

<p>Various laminin-positive vessels (red) were observed in the control C57BL/6J mice at (A) 12–13 weeks of age and (B) 19–20 weeks of age similar to cervical spinal cord results. Fewer blood vessels were labeled in G93A mice at (C) early or (D) end-stage of disease. The nuclei in A–D are shown with DAPI. Scale bar in A, B, C, D is 200 µm; inserts a, b, c, d is 50 µm.</p

Evans Blue fluorescence in the lumbar spinal cord of G93A mice at early and late stages of disease.

<p>In the lumbar spinal cord, EB dye (red, arrowheads) was determined intravascularly in the control C57BL/6J at (A, B) 12–13 weeks of age and (C, D) 19–20 weeks of age similar to the cervical spinal cord. EB extravasation abnormalities were found in G93A mice at (E, F) 13 weeks of age (red, arrows). (G, H) Significant EB diffusion (red, arrows) into the parenchyma of the lumbar spinal cord from many blood vessels was detected in G93A mice at end-stage of disease (17–18 weeks of age). Arrowheads in F and G indicate vessel permeability. Scale bar in A–H is 25 µm.</p

Nanorods Formed from a New Class of Peptidomimetics

Although peptide amphiphiles have been explored as nanomaterials for different applications, nanostructures formed by hierarchical molecular assembly of sequence-specific peptidomimetics are much less developed. Such protein-like nanomaterials could enhance the current application of peptide-based amphiphiles by enriching the diversity of nanostructures, increasing <i>in vivo</i> stability for biomedical applications, and facilitating the understanding of biomacromolecular self-assembly. Herein we present a biomimetic γ-AApeptide amphiphile which forms nanorods. Our results demonstrate the capability of γ-AApeptide amphiphiles as a potential scaffold for the preparation of biomimetic and bioinspired nanostructures. The programmability and biocompatibility of γ-AApeptides could lead to novel nanomaterials for a wide variety of applications

Histological analysis for neurons and myelin.

<p>Luxol Fast Blue-Cresyl Violet staining was performed in serial brain sections from control and MCAO rats for examination of myelin and neuron condition. (<b>A</b>) In controls, neurons were well established with distinct cell bodies and nuclei in examined cerebral structures from both hemispheres. Myelin also appeared normal. (<b>B</b>) In the brains of rats 7 days after MCAO, neuronal pyknosis and cellular debris were observed in the border zone of peri-infarct areas of striatum and somatosensory cortex in ipsilateral hemisphere. Fewer pyknotic neurons were seen in ipsilateral motor cortex, however, cell density was decreased compared to controls. Myelin thickness was also reduced. The lateral ventricle was expanded in the ipsilateral hemisphere. In the hemisphere contralateral to MCAO insult, fewer myelin sheaths were determined in somatosensory and motor cortices and striatum. Contralateral striatosomes also showed size decreases similar to striatosomes within infarct area. Neuronal density in motor and somatosensory cortices was diminished compared to controls. Some neurons appeared pyknotic. Scale bar in full brain images is 200 µm, in striatum is 100 µm, in motor and somatosensory cortices is 50 µm. (<b>C</b>) Semi-quantitative analysis of Cresyl Violet stained brain samples demonstrated reduction of neuronal densities in ipsilateral striatum, motor and somatosensory cortices. Neuronal densities were also diminished in analyzed cerebral cortices. Of note, neuronal cell densities in ipsilateral striatum and somatosensory cortex were analyzed outside of perilesional areas. (<b>D</b>) Semi-quantitative analysis of Luxol Fast Blue stained brain samples showed decrease of myelin intensity, mainly, in ipsilateral striatum and somatosensory cortex vs. control (baseline). Reduced myelin staining was also determined in contralateral striatum.</p

Immunohistochemical analysis of activated microglia.

<p>A few OX-6 positive cells (arrowhead) were identified in motor (<b>a</b>) and somatosensory (<b>b</b>) cortices and striatum (<b>c</b>) in control rats. A large number of activated microglial cells were determined in ipsilateral MCAO hemisphere, in brain structures with EB extravasation (<b>d–f</b>). Morphologically, these cells were characterized by large cell bodies and short processes. In the contralateral hemisphere, activated microglia were observed, primarily in the striatum (<b>i</b>) and somatosensory cortex (<b>h</b>). Some OX-6 positive cells were also seen in the contralateral motor cortex (<b>g</b>). Arrowheads indicate microglial cells. Asterisks indicate EB leakage (red). Images of OX-6 expression were converted to grayscale to better display microglia processes in white on black background. Scale bar in a-i is 25 µm. (<b>B</b>) Semi-quantitative analysis showed highest degree of OX-6 immunoexpression in ipsilateral striatum vs. control (baseline).</p