Histochemical changes of substance P, FRAP, serotonin and succinic dehydrogenase in the spinal cord of rats with adjuvant arthritis

Various histochemical changes were found in spinal segments L4-L5 of rats with adjuvant arthritis, predominantly 30 days after inoculation. A slight to marked increase of substance P immunoreactivity occurred in laminae I, II and X. FRAP activity was enhanced in lamina II. Serotonin immunoreactivity was heavier in laminae I, VIII and IX in a few animals. The intensity of the histoenzymological reaction for succinic dehydrogenase increased in certain laminae VIII and X neurons. At day 15 of the disease the increase of substance P and FRAP activities was chiefly restricted to the medial portion of the superficial dorsal horn. There was a significant positive correlation between the scratching behaviour of arthritic rats and the substance P immunoreactivity in laminae X and I. If one accepts that scratching is pain-related, the data are consistent with a possible role of substance P in the chronic pain associated with adjuvant arthritis. They leave undetermined the significance of the other histochemical changes.     

Action of DCCD on the H+/O stoichiometry of mitoplast cytochrome c oxidase

The mechanistic H+/O ejection stoichiometry of the cytochrome c oxidase reaction in rat liver mitoplasts is close to 4 at level flow when the reduced oxidase is pulsed with O2. Dicyclohexylcarbodiimide (DCCD) up to 30 nmol/mg protein fails to influence the rate of electron flow through the mitoplast oxidase, but inhibits H+ ejection. The inhibition of H+ ejection appears to be biphasic; ejection of 2-3 H+ per O is completely inhibited by very low DCCD, whereas inhibition of the remaining H+ ejection requires very much higher concentrations of DCCD. This effect suggests the occurrence of two types of H+ pumps in the native cytochrome oxidase of mitoplasts.     

Further characterization of [3H]fulunitrazepam binding sites on cultured mouse astroglia

Astroglial cells in primary cultures bind [³H]flunitrazepam with a high affinity on a single type of site and on a number of binding sites which increased during astroglial growth and differentiation. These binding sites show a particular pharmacological spectrum characterized by an inhibition of high affinity by RO-5-4864 (4-chlorodiazepam), an anticonvulsant of the benzodiazepine family and by an inhibition of binding of lower affinities by diazepam clonazepam and clobazam. RO-5-4864 and clonazepam compete for the same binding site in astroglia. The heat stability and the hormonal modulation by thyroxine are similar for astroglia and neuronal-cells. Benxodiazepines modulate the astroglial 5-HT receptor. Such an effect could be a possible physiological response to benzodiazepines for astroglial cells in primary cultures.     

Floristic groups and plant communities of southeastern Santa Fe,Argentina

This paper is a survey of the vegetation of the southeastern departments in the Province of Santa Fe (Argentina). The vegetation was analyzed following Braun-Blanquet's approach modified by Mueller-Dombois & Ellenberg (1974). The most relevant species of the region were placed in 25 groups according to their requirements or general behaviour. Most of the communities are herbaceous, and apart from the woody and some other miscellaneous ones they were grouped into three ecologically and floristically defined sets. The first set, the Stipa grasslands and related communities, which are characterized by the more or less abundant presence of Stipa hyalina, Stipa neesiana and Stipa papposa, comprises five different communities. The second, the halophilous communities, comprises five communities, the two Spartina ssp. grasslands, the halophilous prairies of Distichlis spicata, the short sedge Scirpus americanus communities and the ‘pela-dales’. The third, the hygrophilous communities, comprises nine communities which are not so well defined as the ones in the other sets. Besides, two further communities have been included, the Paspalum quadrifarium and the Melica macra tall grasslands.     

New enzymatic changes of L-cystathionine catalyzed by bovine tissue extracts

L-Cystathionine was used as substrate for enzyme systems prepared by heating bovine tissue extracts in the presence of pyruvate at 60 degrees C for 10 min. Analysis of the products indicated that the systems converted L-cystathionine into the cyclic ketimine form which was detected by its spectral properties and by chromatography on the amino acid analyzer. Alanine, alpha-aminobutyrate and cystine were also produced. Pyruvate and alpha-ketobutyrate enhance the production of the ketimine by liver, kidney and heart extracts, and are necessary for the brain extracts: alpha-Ketoglutarate is much less effective and its presence favors the production of homocystine by all the extracts. Homocystine was found in the brain incubates when any of the ketoacids assayed were added. The overall reaction is explained by the action of heat stable cystathionine gamma-lyase and beta-synthase which produce alpha-ketobutyrate and pyruvate used for the transamination of the remaining cystathionine to the monoketoacid. This last compound cyclizes spontaneously into the ketimine form thus avoiding the removal of the second amino group. This represents a new nontransulfurative path leading to the production of a seven membered etherocyclic product whose biochemical implications are yet unexplored.     

Chronic administration of an organophosphorus insecticide to rats alters cholinergic muscarinic receptors in the pancreas

Male rats were treated for 10 days with the organophosphorus insecticide, acetylcholinesterase inhibitor, O,O-diethyl S-[2-(ethylthio)ethyl]phosphorodithioate (disulfoton, 2 mg/kg/day by gavage). At the end of the treatment, binding of [³H]quinuclidinyl benzilate ([³H]QNB) to cholinergic muscarinic receptors and cholinesterase (ChE) activity were assayed in the pancreas. Functional activity of pancreatic muscarinic receptor was investigated by determining carbachol-stimulated secretion of α-amylase in vitro. ChE activity and [³H]QNB binding were significantly decreased in the pancreas from disulfoton-treated rats. The alteration of [³H]QNB binding was due to a decrease in muscarinic receptor density with no change in the affinity. Basal secretion of amylase from pancreas in vitro was not altered, but carbachol-stimulated secretion was decreased. The effect appeared to be specific since pancreozymin was able to induce the same amylase release from pancreases of control and treated rats. The results suggest that repeated exposures to sublethal doses of an organophosphorus insecticide lead to a biochemical and functional alteration of cholinergic muscarinic receptors in the pancreas.     

Age-related and diurnal changes in Met5-Enk-Arg6-Phe7 and Met5-enkephalin contents of pituitary and rat brain structures

The beta-endorphin, met5-enkephalin-arg6-phe7 (MEAP) and met5-enkephalin (ME) changes related to age and diurnal rhythms were studied in various regions of rat brain and in the pituitary by specific radioimmunoassays. The contents of MEAP, met5-enkephalin and beta-endorphin were higher in the pituitary of old rats (18 months old) than that of young rats (23 days old) while the content of these opioid peptides was higher in the hypothalamus of young rats than in that of old rats. Beta-endorphin was also higher in the striatum of 23 days old rats, but no age-associated changes were observed in the hippocampus, brain stem or cortex. In the diurnal rhythm study, it was found that in the hypothalamus and striatum of the adult rat (2-3 months old), both MEAP and ME contents were higher at mid-dark than at mid-light and that in the intermediate posterior lobe of the pituitary, the ME content was also higher at mid-dark.     

Toxicity oft-butylhydroperoxide in hepatocyte monolayers exposed to hypoxia and reoxygenation

Summary Inasmuch as it is known that the toxicity of anesthetic agents is potentiated by hypoxia and that the reductive metabolism of these agents results in the formation of lipid hydroperoxides, we investigated the toxicity of hydroperoxides under low-oxygen concentrations. We found that hypoxia exacerbates the toxicity oft-butyl hydroperoxide, shifting the dose-response curve oft-butyl hydroperoxide vs. lysis of hepatocytes approximately an order of magnitude to the left. Furthermore, although at the end of a 4-h exposure to 0.5% O₂ hepatocyte monolayers seemed normal by three indices (release of⁵¹Cr and serum glutamate transaminase or exclusion of trypan blue), they were completely lysed after an additional 20 h reoxygenation at 20%. O₂. In contrast, monolayers exposed to 2% O₂ for 4 h seemed normal after 20 h reoxygenation. However, cells exposed to both a subtoxic dose of hydroperoxide and 4 h of 2% O₂, although seeming healthy at the end of the hypoxic period, were completely lysed within 20 h after reoxygenation. The study was supported by grant OH 00978 from the National Institutes for Occupational Safety and Health, Atlanta, Georgia.     

Permanent preservation of whole alizarin red S skeletons by clearing and embedding in polyester resins

A one-step clearing and embedding procedure for alizarin red S stained skeletons is described. Embryos are fixed in formalin, skinned and eviscerated. After staining in a 10 mg/liter solution of alizarin red S in 5% aqueous KOH, specimens are dehydrated in a graded series of acetone-polyester monomer solutions. Finally, the specimens are embedded at room temperature in the polyester resin. A special reusable metallic mold is described for embedment of large fetuses. Specimens previously cleared in glycerol can be processed with this method.