Two of the synaptic receptors involved in the regulation of micturition, P2X(3) receptor, which is operated by ATP, and vanilloid receptor 1 (VR1), which is operated by capsaicin, are regarded as newcomers. To investigate the possibility that these receptors act as therapeutic targets for treatment of an overactive bladder, we investigated their distribution and reciprocal relationship. Eight-week-old Sprague Dawley rats were injected with retrograde nerve tracer within the bladder wall, and 15 rats were injected with 0.5% acetic acid inside the bladder. After a week, the animals were killed, and their dorsal root ganglia (DRG) at the levels of L6 and S1 were harvested. Immunohistochemistry or Western blot analysis of P2X(3) and VR1 were performed on the DRG. The DRG neurons with afferent fibers from the bladder had increased expression of VR1 and downregulated P2X(3) receptors. The P2X(3) receptor and VR1 seemed to account for the important parts of the hypersensitivity of the inflammatory bladder. We conclude that the simultaneous modulations of both P2X(3) receptor and VR1 may have a synergic effect on the treatment of overactive bladder and may produce greater response rates.
Acetic Acid ; Adenosine Triphosphate ; Animals ; Blotting, Western ; Capsaicin ; Diagnosis-Related Groups ; Ganglia, Spinal ; Hypersensitivity ; Immunohistochemistry ; Neurons ; Rats ; Rats, Sprague-Dawley ; Receptors, Neurotransmitter ; Urinary Bladder ; Urinary Bladder, Overactive ; Urination

The anterior cruciate ligament (ACL) and the posterolateral structures are functionally important structures that assist with the stability of the knee joint. Sport-related injuries to these structures and reconstructive surgery are becoming more frequent. However, the anatomic characteristics and the relationship between these structures are not well understood. We measured the morphological characteristics of the posterolateral structures of the knee and the ACL, and we identified the anatomic relationship between the center of the femoral attachment area of the ACL and other femoral structures. We dissected 34 cadaveric knees with no signs of previous surgery, knee abnormality, or disease. The lateral collateral ligament (LCL) and the popliteus tendon (PLT) were present in all knees, although the popliteofibular ligament (PFL) was not present in 20.6% of knees. The mean length of the LCL was 51.99 mm and differed significantly between men and women (P<0.05). The mean length of the PLT was 21.59 mm, and the mean length of the PFL was 18.49 mm. In 67.7% of knees, the PLT was attached to the inferior aspect of the femoral attachment of the LCL, and the femoral attachment of the PLT was covered by the bundle of the LCL. The ACL was distinguished into anteromedial (AM) and posterolateral (PL) bundles in all knees. The mean lengths of the AM and PM bundles were 22.14 mm and 15.98 mm, respectively. The measured lengths in each bundle differed significantly between men and women (P<0.05). The mean distance between the center of the femoral attachment area of the ACL and the intercondylar line was 8.36 mm in the AM bundle, 15.98 mm in the PL bundle, and 10.96 mm in the entire ACL. The mean distance between the center of the femoral attachment area of the ACL and the medial border of the lateral condyle was 8.14 mm in the AM bundle, 7.36 mm in the PL bundle, and 8.45 mm in the entire ACL.
Anterior Cruciate Ligament ; Cadaver ; Collateral Ligaments ; Female ; Humans ; Knee ; Knee Joint ; Ligaments ; Male ; Tendons

Agmatine is a primary amine formed by decarboxylation of L-arginine synthesized in the mammalian brain. Recent studies have shown that agmatine is neuroprotective in models of trauma and ischemia. The purpose of this study was to evaluate the effect of agmatine on the expression of MMP2 and MMP9, which are expressed in reperfusion injury following cerebral ischemia. Mice were subjected to 2 h middle cerebral artery occlusion and 22 h reperfusion. Agmatine (100 mg/kg) was administered intraperitoneally at the start of reperfusion. Agmatine treatment significantly reduced the immunoreactivity of MMP2 and MMP9 in the cortex, striatum, and penumbra on the ipsilateral side. The immunoreactivity of MMP2 and MMP9 was markedly lower in blood vessels of the agmatine-treated group than in the experimental control group. Immunoblot analysis showed that agmatine treatment decreased the expression of MMP2 and MMP9. After exogenous agmatine administration, the expression of agmatine was higher in the striatum and penumbra of the agmatine-treated group than in the experimental control group. The fluorescence intensity was markedly greater in blood vessels in the agmatine-treated group than in the experimental control group. These data suggest that agmatine might decrease the expression of MMP2 and MMP9 by regulating NOS activity, and thereby modulating NO synthesis.
Agmatine ; Animals ; Arginine ; Blood Vessels ; Brain ; Brain Ischemia ; Decarboxylation ; Fluorescence ; Infarction, Middle Cerebral Artery ; Ischemia ; Mice ; Nitric Oxide ; Reperfusion ; Reperfusion Injury

As a nonhistone DNA-binding protein, high mobility group box 1 (HMGB1) is released in large amounts into the extracellular space immediately after ischemic insult and plays a role in the release of proinflammatory cytokines. Here, we the examined cytokine-like or signaling molecule-like function of extracellular HMGB1 in primary cortical cultures. We found that a large amount of HMGB1 was released following zinc-induced neuronal cell death in primary cortical cultures and that this extracellular HMGB1 might aggravate neuronal damage. The conditioned media collected from zinc-treated primary cortical cultures decreased neuronal cell survival to 69.6+/-1.4% of control values when added to fresh primary cortical cultures. In contrast, treatment with HMGB1-depleted conditioned media produced by cultures treated with an HMGB1 siRNA-expression vector suppressed the induction of neuronal death. A mutant HMGB1 siRNA-expression vector did not suppress the induction of neuronal death, demonstrating a role of HMGB1 in neuronal death. Moreover, HMGB1-depletion in media conditioned by cotreatment with anti-HMGB1 antibody or with anti-RAGE antibody, a potential receptor for HMGB1, recovered neuronal cell survival to 81.0+/-4.0% and 79.0+/-4.0%, respectively, when added to fresh primary cortical cultures. These results indicate that extracellular HMGB1 released after zinc treatment induces neuronal death, which might aggravate zinc toxicity.
Cell Death ; Cell Survival ; Culture Media, Conditioned ; Cytokines ; Extracellular Space ; HMGB1 Protein ; Neurons ; Zinc

Hearing loss in adults can stem from damage to the cochlea and the cochlear nerves inflicted by intense noise, mechanical trauma, or disease. Hearing loss is associated with degenerative changes in central auditory pathways, and hearing deficits are often accompanied by changes in the synaptic organization of the central auditory pathways. In addition to structural rearrangements, hearing loss may induce changes in the strength of synaptic transmissions. These effects may alter both transient and persistent regulation of transmitter release from glutamatergic, glycinergic, and GABAergic pathways in the auditory brain stem. The converging excitatory and inhibitory inputs are exquisitely organized topographically and are aligned perfectly with each other. The LSO and MNTB in the mammalian auditory brain stem provide and receive many glycinergic inputs. Thus, this auditory system is a useful model to study inhibitory synaptic development. However, little is known about the inhibitory synapses in the central nervous system. First, we used immunohistochemistry to compare the glycine receptor (GlyR) distribution in the LSO and MNTB, which project glycinergic inhibitory input into the auditory brainstem, in circling mice (P16), which have a spontaneous mutation in the inner ear, with wild-type mice. The relative immunoreactive density of the LSO was 86.4+/-7.2 in wild-type, 76.7+/-10.7 in heterozygous, and 61.1+/-4.1 in homozygous mice. The relative immunoreactive density of the MNTB was 97.6+/-8.7 in wild-type, 91.7+/-8.9 in heterozygous, and 74.9+/-7.8 in homozygous mice. These results reveal a decreased GlyR immunoreactivity in both the LSO and MNTB, which may be attributable to a postsynaptic decrease in GlyR number. Our model uses congenitally deaf mice, in which both spontaneous and evoked auditory nerve activity are disrupted because of dysfunctional hair cell-spiral ganglion cell transmission. This provides a naturally occurring model that may provide valuable insights into the central aspects of human congenital deafness in addition to the central consequences of a lack of auditory nerve activity. Our results are likely to be relevant to our understanding of the central changes underlying human hereditary deafness.
Adult ; Animals ; Auditory Pathways ; Brain ; Brain Stem ; Central Nervous System ; Cochlea ; Cochlear Nerve ; Deafness ; Ear, Inner ; Ganglion Cysts ; Glycine ; Hair ; Hearing ; Hearing Loss ; Humans ; Immunohistochemistry ; Mice ; Noise ; Receptors, Glycine ; Synapses

Brain edema, the infiltration and accumulation of excess fluid causing an increase in brain tissue volume, often leads to a rise in intracranial pressure (ICP) and is a key contributor to the morbidity and mortality associated with brain injury. We examined the mechanisms for vasogenic brain edema produced by focal cerebral ischemia. To test the mechanisms underlying vasogenic brain edema, we compared the expression of endothelial nitric oxide synthase (eNOS) and aquaporin-4 (AQP4) in the penumbra of brain tissues after focal cerebral ischemia. The expression of eNOS and AQP4 in the ischemic penumbra was determined by quantitative immunoblot analysis. In focal cerebral ischemia, eNOS expression increased significantly to 129%+/-6% of the value for sham-operated controls (n=4, P<0.01). In contrast, AQP4 expression decreased significantly to 52%+/-3% of the value for shamoperated controls (n=4, P<0.01). These findings suggest that the expression of eNOS and AQP4 in the focal cerebral ischemia may play different roles in the formation of vasogenic brain edema. Additionally, a larger decrease in the neuronal nuclei (NeuN) (39%+/-7% of the value in sham-operated controls; n=4, P<0.01) than the decrease in glial fibrillary acidic protein (GFAP) (53%+/-14% of sham-operated controls; n=4, P<0.01) in the penumbra after focal cerebral ischemia suggests that the neurons are more susceptible to ischemic injury than are the astrocytes.
Astrocytes ; Brain ; Brain Edema ; Brain Injuries ; Brain Ischemia ; Edema ; Glial Fibrillary Acidic Protein ; Intracranial Pressure ; Neurons ; Nitric Oxide Synthase Type III

It was well known that atrial myocytes systhesize atrial natriuretic peptide[ANP], and secrete it into the atrial lumen through the atrial endocardium. But the mechanism for regulation of ANP secretion has not been clearly elucidated, because there was little information of the atrial morphology concerning basal lamina. Basal lamina is surmised as one of barriers that control the movement of ANP, a large molecule. This study was attempted to elucidate the morphological characteristics of basal lamina and connective tissue fibers of atrial endocardial layer by scanning electron microscopy. Basal lamina was exposed by removal of the overlying endothelium. This was achieved by using OsO4 maceration, immersion in aqueous boric acid or EDTA treatment. After removal of the endothelial cell, the specimens were exposed to ultrasonic vibration in case of need. The external surface of basal lamina showed a fairly smooth appearance on the whole, although a few irregular folds are often encountered. Fenestrations, 0.1-1 micrometer in diameter, were randomly observed on the basal lamina, and they were circular to oval in shape. Margin of fenestrations was somewhat distinct and some was divided into two parts by linear structures. The structural differences of fenestrations between right and left atria were not found. The fibroreticular lamina under the basal lamina was revealed by removal of the endothelial cells and their basal lamina. This layer was consisted of interwoven fine fibers. These fine fibers were repeatedly divided and fused, forming reticular network. Some fine fibers connected with basal lamina. Some connective tissue fibers below fibroreticular layer were collected into thick bundles running parallel to myocytes. Above results may serve as a basis for the physiological and morphological studies of atrium.
Animals ; Atrial Natriuretic Factor ; Basement Membrane* ; Connective Tissue ; Edetic Acid ; Endocardium* ; Endothelial Cells ; Endothelium ; Immersion ; Microscopy, Electron, Scanning ; Muscle Cells ; Rats* ; Running ; Ultrasonics ; Vibration

No abstract available.
Anaphylaxis* ; Histamine Release* ; Histamine* ; Mast Cells*

Many approaches have been adopted to restore function following spinal cord injury (SCI). These have included transplantation of fetal neurons, neuronal progenitor cells, or glial cells, or transplantation of transfected cells which produce a variety of substances. The use of human umbilical cord blood cells (hUCB cells) has recently been reported to alleviate behavioral consequences of stroke injury. We report here that hUCB cells delivered intravenously in rats with compression injury of the spinal cord increase the rate behavioral recovery. Tweny-five rats were divided into 5 groups (laminectomy only, laminectomy+hUCB cells, SCI+hUCB cells devlivered at one day post-injury, SCI+hUCBcells delivered at 5 days post-injury, and SCI only). SCI was produced by compressing the spinal cord for one minute with an aneurysm clip calibrated to a closing pressure of 50 gms. Rats were assessed behaviorally at one, two and three weeks using the BBB behavioral scale, inclined platform, and extension and toe spread tests. Following behavioral testing, spinal cords from these rats were examined immunohistochemically to identify hUCB cells. Spinal cords from SCI+hUCB cells animals contained hUCB cells in the area of SCI: No hUCBcells cells were found in noninjured areas of spinal cord from these animals or in animals in which only a laminectomy was performed. Rats in the SCI+hUCBcells 1 day group were significantly different in recovery of motor function as compared to the SCI+hUCB cells 5 day group and laminectomy groups. By three weeks, SCI+hUCB cells animals were not significantly different from each other. These results indicate that hUCBcells cells may be beneficial in reversing the behavioral effects of SCI.
Aneurysm ; Animals ; Fetal Blood* ; Humans* ; Immunohistochemistry ; Laminectomy ; Neuroglia ; Neurons ; Rats* ; Spinal Cord Injuries* ; Spinal Cord* ; Stem Cells ; Stroke ; Toes ; Umbilical Cord*

We have previously reported that aqueous extract of gall from Rhus chinensis, known as "Obaeja", inhibited rat intestinal alpha-glucosidase and suppressed postprandial hyperglycemia by delaying digestion and absorption of intestinal carbohydrate (Shim et al., 2003). This led us to speculate that obaeja could be involved in ameliorating beta-cell injury by lowering glucotoxicity. In the present study, we thus examined the protective effect of obaeja on pancreatic beta-cell damage along with its anti-diabetic effect in streptozotocin-induced animal models. Streptozotocin was administered to rat pups (neonate/STZ model), or to adult rats with a lower dose using osmotic pump (osmotic pump/STZ model) for inducing beta cell death and diabetes. Obaeja was given to those rat pups after weaning in neonate/STZ model, or 2 weeks before subcutaneous implantation of osmotic pump to rats of the other latter model. In the diabetic control rats of the neonate/STZ model, which were not fed with obaeja, some pancreatic islets demonstrated a destruction of beta cell mass with insulitis 2 weeks after weaning, while some larger and irregular islets were formed by proliferation of alpha cells. In particular, we found some pancreatic lobules showing a severe inflammation and degeneration of islet and acinar tissues in this model. Islets in these inflammatory lobules were smaller in size with only few cells. In contrast, any inflammatory responses and insulitis were not observed in pancreas of the rats fed obaeja in this model. The islets in those rats maintained their normal profiles and islet cell population. Such anti-cytotoxic effect was also monitored in the diabetic rats of osmotic pump/STZ model. Especially, occurrence of hyperglycemia in the obaeja fed rats was delayed by 25~30 days than that of diabetic control rats in this model. Taken together, these results imply that regulation of postprandial blood glucose level by obaeja feeding may ameliorate a secondary injury caused by glucotoxicity.
Absorption ; Adult ; alpha-Glucosidases ; Animals ; Blood Glucose ; Cell Death ; Digestion ; Humans ; Hyperglycemia ; Inflammation ; Islets of Langerhans ; Models, Animal ; Pancreas ; Rats ; Rhus* ; Streptozocin ; Weaning