Ultrastructural investigation of substance P-, leucine-enkephalin- and 5-hydroxytryptamine-like immunoreactive terminals in the area of cremaster motoneurons of the male rat

Studies were made on whether substance P-, leucine-enkephalin- and 5-hydroxytryptamine (serotonin)-like immunoreactive fibers exert a direct influence on the cremaster motoneurons of the male rat by immunocytochemistry combined with retrograde tracing at the light- and electron-microscopic levels. Horseradish peroxidase was used as a retrograde tracer. Two days after injection of horseradish peroxidase into the genitofemoral nerves, its accumulation in the cremaster motoneurons was demonstrated by the diaminobenzidine-nickel method. On immunocytochemical examination of the same sections immunoreactive end-products were detected by the peroxidase-antiperoxidase method. Two different kinds of reaction products were distinguishable at both the light- and electron-microscopical levels. The horseradish peroxidase-labeled cremaster motoneurons at the L1 and L2 levels of the spinal cord were surrounded by abundant immunoreactive terminals. Examination at the ultrastructural level showed that substance P-like immunoreactive terminals formed synaptic contacts almost exclusively with the proximal dendrites of these horseradish peroxidase-labeled cremaster motoneurons. On the other hand, in the leucine-enkephalin- and serotonin-like boutons, significant numbers of axosomatic contacts with horseradish peroxidase-labeled cells were seen although axodendritic contacts with horseradish peroxidase-labeled cells were predominant. Occasionally, leucine-enkephalin-like immunoreactive fibers with synapses on horseradish peroxidase-labeled neurons formed axoaxonic contacts with other leucine-enkephalin-like axonal terminals. Thus, substance P-, leucine-enkephalin- and serotonin-like fibers clearly regulate the function of cremaster motoneurons monosynaptically. The varied synaptic contacts of these fibers according to the neuroactive substances involved suggest different actions of these substances on the cremaster motoneurons.     

Osteoclast-type giant cell tumor of the endometrium. An immunohistochemical study

We present a rare case of a 55 year old female with an osteoclast-type giant cell tumor of the endometrium associated with leiomyoma and adenomyosis. Multinucleated giant cells and mononuclear stromal cells reacted with vimentin and alpha-1-antichymotrypsin (AACT) using the immunoperoxidase method. Epithelial membrane antigen (EMA), carcinoembryonic antigen (CEA), and keratin exhibited negative immunoreaction in these tumor cells. Our immunohistochemical results do not support the epithelial origin of an osteoclast-type giant cell tumor and mesenchymal derivation appeared more likely, suggesting histiocytic origin.     

Chitosan hydrogel as a drug delivery carrier to control angiogenesis

An aqueous solution of photocrosslinkable chitosan containing azide groups and lactose moieties (Az-CH-LA) incorporating paclitaxel formed an insoluble hydrogel within 30 s of ultraviolet light (UV) irradiation. The chitosan hydrogel showed strong potential for use as a new tissue adhesive in surgical applications and wound dressing. The fibroblast growth factor (FGF)-2 molecules retained in the chitosan hydrogel and in an injectable chitosan/IO₄-heparin hydrogel remain biologically active, and were gradually released from the hydrogels as they biodegraded in vivo. The controlled release of biologically active FGF-2 molecules from the hydrogels caused induction of angiogenesis and collateral circulation occurred in healing-impaired diabetic (db/db) mice and in the ischemic limbs of rats. Paclitaxel, which is an antitumor reagent, was also retained in the chitosan hydrogel and remained biologically active as it was released on degradation of the hydrogel in vivo. The chitosan hydrogels incorporating paclitaxel effectively inhibited tumor growth and angiogenesis in mice. The purpose of this review is to describe the effectiveness of chitosan hydrogel as a local drug delivery carrier for agents (e.g., FGF-2 and paclitaxel) to control angiogenesis. It is thus proposed that chitosan hydrogel may be a promising new local carrier for drugs such as FGF-2 and paclitaxel to control vascularization.     

Expression of interleukin-6 receptor, leukemia inhibitory factor receptor and glycoprotein 130 in the murine cerebellum and neuropathological effect of leukemia inhibitory factor on cerebellar Purkinje cells

Expression of glycoprotein 130 and the related receptors, including interleukin-6 receptor and leukemia inhibitory factor receptor, was examined in the murine cerebellum at the protein level. Western blot analysis revealed that interleukin-6 receptor, leukemia inhibitory factor receptor and glycoprotein 130 were expressed in the murine cerebellum. Immunoreactivities for interleukin-6 receptor, leukemia inhibitory factor receptor and glycoprotein 130 were strongly localized on the cell body of Purkinje cells, indicating that both interleukin-6 and leukemia inhibitory factor could act directly on Purkinje cells in murine adult mice. The expressions of interleukin-6 receptor, leukemia inhibitory factor receptor and glycoprotein 130 were observed on the cell membranes of Purkinje cells by immunoelectron microscopy. Immunoreactivity for the interleukin-6 receptor was also detected in the cytoplasm of Purkinje cells. Injection of a murine hemopoietic cell line, FDC-P1 cells, transfected with the complementary DNA encoding the leukemia inhibitory factor led to a reduction in calbindin-positive dendrites of the Purkinje cells.

The present results suggest that the leukemia inhibitory factor affects cerebellar functions through Purkinje cells.     

Gene expression of histamine H1 receptor in guinea pig primary sensory neurons: a relationship between H1 receptor mRNA-expressing neurons and peptidergic neurons

Pharmacological studies have suggested that a subgroup of primary sensory neurons is responsive to histamine via the histamine H1 receptor. We addressed this issue using in situ hybridization histochemistry with a cRNA probe for the guinea pig H1 receptor gene. About 15% of the trigeminal and lumber dorsal root ganglion (DRG) neurons, but none of nodose ganglion neurons, were intensely labeled with this probe. The H1 receptor mRNA-positive neurons were exclusively small in size, and were demonstrated to give rise to unmyelinated fibers by ultrastructural analysis of isolectin B4-labeling. However, the H1 receptor mRNA-expressing DRG neurons were not immunoreactive to substance P (SP) and calcitonin gene-related peptide (CGRP). A marked increase in the number of mRNA-positive DRG neurons were observed 1-5 days after a crush injury of the sciatic nerve (3-4-fold of the control value). These neurons turned mRNA-positive after the nerve crush were also mainly small-sized. The mRNA signals were detected in many peptidergic (SP/CGRP) neurons, in contrast to the normal state. On the other hand, in the neurons which showed intense labeling in the normal condition, the mRNA signals were down-regulated. These results suggest that primary sensory neurons include two kinds of H1 receptor-expressing sensory neurons, one expressing H1 receptor mRNAs in the normal state and the other up-regulating the mRNAs following the peripheral nerve damage.     

Up- and down-regulation of BDNF mRNA in distinct subgroups of rat sensory neurons after axotomy

Anterograde transport of BDNF is enhanced by axotomy in the rat sciatic nerve. However, the changes in BDNF gene expression in dorsal root ganglion (DRG) neurons after axotomy are not known. We examined this issue using in situ hybridization histochemistry. BDNF mRNA was detected in 35-40% of DRG neurons (L5) of control rats. Most of these neurons are small. BDNF gene expression in these neurons was down-regulated after application of capsaicin to the sciatic nerve. Transection of the sciatic nerve induced the up-regulation of BDNF mRNA. The intensely labeled neurons were mainly large and immunoreactive for neuropeptide Y. These results suggest that up- and down-regulation of BDNF gene expression in distinct subgroups of rat DRG neurons are caused by damage to the peripheral nerve.     

Endocarditis caused by Candida parapsilosis.

The authors report a case of endocarditis caused by Candida parapsilosis. To the best of our knowledge, a case has not been described previously in Japan in the English literature. A battery of 8 peroxidase-labeled lectins was tested on sections of paraffin-embedded tissue to determine which lectin could be used in the microscopic diagnosis of C. parapsilosis. One lectin, from Archis hypoaea (PNA) was found to react with C. parapsilosis. On the other hand, C. albicans, Aspergillus, Mucor, and Cryptococcus did not react with A. hypoaea (PNA). On fluorescence microscopic study, C. parapsilosis was not fluorescent, but other fungi were fluorescent when exposed to ultraviolet illumination. Therefore, we propose new procedures for identification of C. parapsilosis in tissue sections using lectin histochemistry and fluorescence microscopy.     

Possible involvement of angiotensin-converting enzyme 2 and Mas activation in inhibitory effects of angiotensin II Type 1 receptor blockade on vascular remodeling

We explored the roles of angiotensin-converting enzyme 2 (ACE2), angiotensin-(1-7), and Mas activation in angiotensin II type 1 receptor blockade-mediated attenuation of vascular remodeling. Vascular injury was induced by polyethylene-cuff placement around the mouse femoral artery. After cuff placement, the mRNA level of both ACE2 and Mas was markedly decreased in wild-type mice, whereas ACE mRNA was not changed. Immunostaining of ACE2 and Mas was observed mainly in the media and was reduced in the injured artery. Administration of angiotensin-(1-7) decreased neointimal formation after cuff placement, whereas administration of [D-Ala(7)] angiotensin-(1-7), a Mas antagonist, increased it. Consistent with these results, we also demonstrated that neointimal formation induced by cuff placement was further increased in ACE2 knockout mice. In angiotensin II type 1a receptor knockout mice, mRNA expression and immunostaining of ACE2 and Mas in the injured artery were greater, with less neointimal formation than in wild-type mice. Increased ACE2 expression in the injured artery was also observed by treatment of wild-type mice with an angiotensin II type 1 receptor blocker, olmesartan. These results suggested that activation of the ACE2-angiotensin-(1-7)-Mas axis is at least partly involved in the beneficial effects of angiotensin II type 1 receptor blockade on vascular remodeling.     

Light chain cardiac amyloidosis in a nonagenarian

Cardiac amyloidosis is an infiltrative and restrictive cardiomyopathy caused by the extracellular deposition of amyloid fib-rils within the heart as systemic amyloidosis,lead-ing to heart failure,reduced quality of life,and death.[1]There are two major amyloid fibril proteins that af-fect the heart:amyloid immunoglobulin light chain(AL)and amyloid transthyretin(ATTR).The latter is further subdivided into wild-type ATTR and variant types based on the presence of a mutation in the transthyretin gene.