Localization of Pituitary Adenylate Cyclase-Activating Polypeptide (PACAP) in the Hypothalamus-Pituitary System in Rats: Light and Electron Microscopic Immunocytochemical Studies

The localization of pituitary adenylate cyclase-activating polypeptide (PACAP) in the hypothalamus-pituitary system in rats was examined in light and electron microscopic immunocytochemistry using a specific antiserum to synthetic PACAP 1–38 (R0831). In light microscopic study, intensely PACAP-immunostained perikarya were observed in the supraoptic and paraventricular magnocellular nucleus in the hypothalamus. In the median eminence, many immunoreactive nerve fibers were observed in the internal layer, but a few immunoreactive terminals were noticed in the external layer. In the pituitary gland, numerous immunoreactive nerve fibers were observed in the posterior lobe. In the intermediate lobe, moderately immunostained cells were observed, but in the anterior lobe no immunostained cells were noticed. In electron microscopic study, PACAP-immunoreactivity was examined by avidin-biotin peroxidase complex method. In the perikarya of the supraoptic and paraventricular magnocellular nucleus, DAB-reaction products were distributed diffusely in the cytoplasmic matrix, frequently attaching to the rough-surfaced endoplasmic reticulum. In the nerve terminals of the posterior lobe, reaction products were observed among the secretory granules, but sometimes upon them. In the cells of the intermediate lobe, reaction products were also distributed in the cytoplasmic matrix.     

Hippocampofugal gamma-aminobutyric acid (GABA)-containing neuron system in the rat: a study using a double-labeling method that combines retrograde tracing and immunocytochemistry

Using a double-labeling method that combines retrograde tracing and immunocytochemistry, we demonstrated the presence of a hippocampofugal gamma-aminobutyric acid (GABA)-like immunoreactive (GABA-LI) neuron system in the rat. As a preliminary experiment, knife-cut of the dorsal fornix was made and it resulted in a remarkable accumulation of GABA-LI fibers caudal to the section. This suggested the presence of a hippocampofugal GABAergic projection. The hypothesis was confirmed by the double-labeling method: injection of the biotinylated wheat germ agglutinin (B-WGA) into the septum resulted in labeling numerous neurons in the hippocampus and some of them also showed GABA-like immunoreactivity by the simultaneous immunostaining with the antiserum against GABA.     

Two cases of Trichosporon beigelii pneumonia in immunocompromised hosts

Two cases of Trichosporon beigelli pneumonia in severely immunocompromised patients are reported. At autopsy, Trichosporon beigelii was detected in all lobes in one patient who had a small cell lung cancer. Polymycotic infection involving Trichosporon beigelii and Aspergillus was proved in the other patient who had a non-Hodgkin's lymphoma. Miconazole therapy was not effective against Trichosporon beigelii infection in both cases. Although diagnosis and management are difficult, Trichosporon beigelii must be considered as a cause of visceral opportunistic fungal infection.     

Percutaneous hepatic venous isolation and extracorporeal charcoal hemoperfusion for high-dose intraarterial chemotherapy in patients with colorectal hepatic metastases

The results of treating 12 consecutive patients with unresectable colorectal hepatic metastases with a hepatic arterial infusion of high-dose Adriamycin, 100–120 mg/m², using hepatic venous isolation (HVI) and charcoal hemoperfusion (CHP) are reported herein. Adriamycin was administered over 5–15 min under extracorporeal drug elimination by HVI-CHP. HVI was percutaneously accomplished by either the double-balloon technique using a Fogarty occlusion catheter (8/22F) or a balloon-tipped catheter (16F). During the infusion, isolated hepatic venous blood was filtered by CHP and pumped into the left axillary vein. There were no lethal complications, and good hemodynamic tolerance to HVI-CHP was confirmed. Tumor liquefaction accompanied by a sharp decrease in serum carcinoembryonic antigen levels by more than 50% of pretreatment levels was observed in 6 of the 12 patients 1 month after treatment. Apart from chemical hepatitis, which developed in 11 (92%) of the patients, the Adriamycin toxicities were well controlled following the development of nausea and vomiting in 2 patients (17%), leukopenia <2,000/mm³ in 3 (25%), and gastric ulcer in 1 (8%). These results indicate that this method is a safe and useful procedure for otherwise hazardous high-dose intraarterial chemotherapy in patients with unresectable hepatic tumors.     

"Necklace olfactory glomeruli" form unique components of the rat primary olfactory system

A distinct subset of rat primary olfactory neurons was identified immunohistochemically by means of a polyclonal antibody against human placental antigen X-P2 (hPAX-P2), an incompletely characterized substance found in all estrogen-biosynthetic organs. The subset of olfactory receptor cells was distributed widely over the olfactory epithelium with some degree of concentration on the dorsocaudal walls of nasal subcavities. The subset formed unique "necklace olfactory glomeruli," which were composed of seven to nine solitary glomeruli located in the caudal end of the olfactory bulb. One of them was located in the "modified glomerular complex" reported to be involved in rat suckling behavior. The projectional patterns of the necklace olfactory system, albeit diffuse, indicated some degree of spatial correspondence between zones of olfactory epithelium and specific glomeruli. Axons emanating from neighboring cells can project to several glomerular loci. From the necklace olfactory system, an average of 150-200 receptor cells were estimated to converge onto a single necklace glomerulus.     

Nuclear magnetic resonance studies on polymer carbanions, 5. Polystyryl carbanion with lithium as counter cation and its dimer model compounds

Oligostyryllithium was synthesized by the reaction of sec-butyllithium with styrene in diethyl ether at ?40°C. Dimer model anions (counter cations: Li and K) of the growing chain ends were synthesized by metallation of 1-methoxy-1,3-diphenylpropane. The structures of the lithium compounds were corroborated by ¹³C NMR spectroscopy and compared with those of the potassium compounds. As for polystyryllithium and its model compounds, the charge density on the α-carbon increased and that on the phenyl ring of the growing end decreased with increasing molecular weight, while, in polystyrylpotassium and its model compounds, the charge density on the α-carbon increased and the one on the phenyl ring did not change with increasing molecular weight. The temperature change of the ¹H NMR spectrum of oligostyryllithium in diethyl ether indicated rotation around the C-1? C_α bond at lower temperatures than in tetrahydrofuran, but the activation energy was the same in both solvents. T₁ values of the phenyl carbons in model compound 2b were one to two orders of magnitude shorter than those of the protonated compound.     

Co-existence of glucagon- and substance P-like immunoreactivity in the chicken retina

This immunohistochemical study of chicken retina using flat-mounts shows that pancreatic glucagon- and substance P-like immunoreactive amacrine cells have more heterogeneous subpopulations than was previously understood to be the case. Using double-staining immunohistochemical procedures we demonstrate that a substantial proportion of all subtypes of glucagon-like immunoreactive cells contain substance P-like immunoreactivity and that the ratio of the amacrine cells containing both peptides to total immunoreactive cells varies according to position in the retinal and cell type. These results suggest that retinal cells may have different functions according to position or cell type.     

Enkephalin fibers synapse on cholinergic neurons in the rat sacral intermediolateral nucleus: a double-immunostaining at the light and electron microscopic levels

Relationships between leucine-enkephalin fibers and cholinergic neurons in the rat sacral intermediolateral nucleus were examined by light and electron microscopy using double-immunostaining method. Cholinergic neurons in the sacral intermediolateral nucleus were labeled by a rat-mouse monoclonal antibody to choline acetyltransferase and stained bluish green with 5-bromo-4-chloro-3-indolyl-beta-D- galactoside reaction products using beta-galactosidase as a marker. On the same sections, leucine-enkephalin fibers were labeled by a rabbit polyclonal antiserum to leucine-enkephalin and stained brown by diaminobenzidine reaction products using peroxidase as a marker. After embedding in Epon, the sections were examined in light and electron microscopes. In the light microscope, choline acetyltransferase-like immunoreactive cells were seen in the sacral intermediolateral nucleus. In the same region, leucine-enkephalin-like immunoreactive cells. In the electron microscope, 5-bromo-4-chloro-3-indolyl-beta-D-galactoside reaction products were in the form of coarse electron dense deposits in the choline acetyltransferase-like immunoreactive structures and could be distinguished from the much finer grained diaminobenzidine reaction products. Choline acetyltransferase-like immunoreactive neurons received synaptic inputs from leucine-enkephalin fibers-like immunoreactive terminals. These findings suggest that leucine-enkephalin fibers may affect the activity of cholinergic parasympathetic preganglionic neurons.     

Retinoblastoma protein prevents staurosporine-induced cell death in a retinoblastoma-defective human glioma cell line.

OBJECTIVE: To investigate the mechanism of staurosporine-induced glioma cell death and cell cycle arrest using adenovirus-mediated gene transfection, as well as the function of retinoblastoma (Rb) and genetic instability induced by staurosporine. METHODS: Cell cycle regulation, cell death and nuclear abnormalities induced by staurosporine were examined using an adenovirus vector expressing Rb, p16 or p21 genes in human glioma cell lines. RESULTS: The Rb-defective SF-539 cell line was resistant to staurosporine compared with cell lines expressing intact Rb. SF-539 glioma cells exposed to staurosporine became multinucleated and then died. Multinucleation was prevented in SF-539 cells transfected with the Rb gene, thus decreasing the death rate of these cells. CONCLUSIONS: These results imply that enforced Rb expression protects cells from genomic instability induced by staurosporine regardless of its upstream molecular effects.