Modulating effect of cerulein on benzodiazepine receptors

Subcutaneous administration of caerulein (100-500 micrograms/kg) significantly reduced the development of picrotoxin (8 mg/kg) seizures in male mice. The same doses of caerulein inhibited 3H-flunitrazepam binding in in vivo experiments. Proglumide, an antagonist of cholecystokinin receptors, in low dose (5 mg/kg) potentiated the effects of caerulein (100 micrograms/kg), whereas the administration of proglumide in high dose (25 mg/kg) reduced the action of caerulein on 3H-flunitrazepam binding and picrotoxin seizures. Caerulein (5-1000 nM) decreased 3H-flunitrazepam binding in in vitro experiments only after supplementation of the binding medium with 120 mM NaCl and 5mM KCl. The results suggest the possible interaction of caerulein with chloride ionophor. It seems probable that the direct interaction of caerulein with chloride ionophor in involved in the inhibitory effect of caerulein on picrotoxin seizures and 3H-flunitrazepam binding.     

Analogs of oxytocin containing a modified peptide bond

Analogs of deamino-oxytocin and deamino-oxypressin containing a CH2-NH group instead of an amide bond between positions 8 and 9 were synthesized. All tested compounds exhibit significantly lowered biological activities.     

Reinnervation of experimental superficial wounds in rats

Sensory reinnervation of a superficial skin wound in the rat was studied by labeling sensory axons with anterogradely transported wheat germ agglutinin-horseradish peroxidase. Reinnervation starts after 3 days from the edge of the wound as well as from beneath the wound. About 2 weeks after the production of the wound, some hyperinnervation appears to be present, but after a few additional weeks, the innervation pattern is essentially normal. The results indicate that structural recovery of sensory axons is rapid and probably complete when skin wounds heal with no or minimal scar formation.     

Purification and properties of thyrotrophin releasing hormone (TRH)-containing particles from the rat hypothalamus

The subcellular distribution of the TRH-like immunoreactivity in the rat hypothalamus and brain was studied. In differential centrifugation, the 900 g for 10 min supernatant (S1) of the hypothalamus or brain contained 61--79% of the total TRH. At 11,000 g for 20 min, 51--73% of the TRH in S1 was sedimented. When the hypothalamic S1 was fractioned under non-equilibrium conditions at 25 degrees C, two populations of TRH-containing particles were observed in several types of continuous linear density gradients. Metrizamide and sucrose gradients affected TRH-assay. TRH-particles were very light in Percol-gradients. Isotonic dextran 40,000-sucrose gradients gave the most reproducible results. In these gradients, the large TRH-particles (35%) equilibrated at 1.055--1.060 kg/l and the small ones (23%) at 1.041--1.047 kg/l. Working at 4 degrees C decreased the amount of large TRH-particles. The apparently larger particles contained cytoplasmic and mitochondrial enzymes and were sensitive to hypoosmotic shock like synaptosomes. Electron micrographs confirmed that these particles were synaptosomes. The true nature of the small particles remained unclear but morphologically a part of them were also synaptosomes. Treatment of the animals with reserpine (10 mg/kg i.p., 24 h), with 6-hydroxydopamine (100 microgram/rat i.c.v.) or with 5,7-dihydroxytryptamine (200 microgram/rat i.c.v.) did not affect significantly TRH-recovery or distribution in the hypothalamus.     

Syngeneic anti-idiotypic antisera to murine monoclonal antibodies to monomorphic and polymorphic determinants of HLA class I antigens

BALB/c mice were immunized with syngeneic anti-HLA class I monoclonal antibodies. The latter included the anti-HLA-A2, A28 monoclonal antibody (MoAb) CR11-351, the MoAb Q6/64 to a determinant restricted to HLA-B antigens and the MoAb CR10-215 and CR11-115 to the same (or spatially close) monomorphic determinant. Anti-idiotypic antibodies could be detected in bleedings obtained 3 days after the first booster, increased in titer in bleedings obtained after the second booster, and persisted at high levels in subsequent bleedings. The four anti-HLA class I MoAb did not differ in their ability to elicit syngeneic anti-idiotypic antibodies. Cross-blocking studies with a panel of anti-HLA class I, anti-HLA class II, and anti-human melanoma-associated antigen (MAA) MoAb showed that the anti-MoAb CR10-215 and anti-MoAb CR11-115 antisera contain only antibodies to private idiotopes, whereas the anti-HLA MoAb CR11-351 and anti-MoAb Q6/64 antisera also contain antibodies to public idiotopes. The latter are expressed by the anti-HLA class I MoAb CR11-351, Q1/28, Q6/64, and 6/31, and by the anti-HLA class II MoAb Q5/6, Q5/13, 127, and 441. Public idiotopes were not detected on the nine anti-MAA MoAb tested. Public idiotopes do not interfere with the binding of anti-HLA MoAb with the corresponding antigenic determinants. On the other hand private idiotopes are located within the antigen-combining site, because anti-idiotypic antisera specifically inhibit the binding of the corresponding immunizing anti-HLA class I MoAb to cultured human lymphoid cells in a dose-dependent manner. Analysis by isoelectric focusing of the anti-HLA class I MoAb antisera showed that the spectrotype of the anti-MoAb CR11-351 antiserum comprises four components that focus in the pH 6.9 to 6.2 range, the spectrotype of anti-MoAb Q6/64 antiserum comprises three components that focus in the pH 6.5 to 6.1 range, the spectrotype of the anti-MoAb CR10-215 antiserum comprises three components that focus in the pH 6.4 to 6.1 range, and the spectrotype of the anti-MoAb CR11-115 antiserum comprises three components that focus in the pH 6.6 to 6.4 range.     

Membrane-bound and secreted IgA contain structurally different alpha-chains

Three different forms of alpha-chains are synthesized by BF0.3 and 615.2, two cloned cell lines derived from the murine B lymphoma 1.29. The three forms of alpha-chains differ in size, pI, cellular location, and rate of turnover. They were identified by means of lactoperoxidase-catalyzed radioiodination, internal 14C or 35S labeling, and immunofluorescence techniques as membrane-bound(alpha m), secreted (alpha s), and intracellular (alpha ic) proteins. Comparison of immunoglobulin products of the two lymphoma lines with those of a hybridoma cell line, Id 150, which secretes IgA of the 1.29 idiotype but lacks membrane IgA, confirmed the assignments of alpha m, alpha s, and alpha ic. Results of biosynthetic labeling of BF0.3, 615.2, and Id 150 in the presence and absence of tunicamycin suggest that the difference in m.w. and charge observed between alpha m and alpha s can be attributed to differences in primary amino acid structure rather than different degrees of glycosylation.     

Structure of the lysosomal sphingolipid activator protein 1 by homology with influenza virus neuraminidase

The sphingolipid activator protein 1 (SAP-1) increases the rate of hydrolysis of sphingolipids in the lysosome by apparently bringing together the substrate and the corresponding hydrolytic enzyme. This implies specific recognition of both the substrate and enzyme by SAP-1. However, binding domains in SAP-1 and recognition mechanisms involved are unknown. Amino acid sequence comparison of SAP-1 with influenza virus neuraminidase (EC 3.2.1.18, FLU NA) indicates that functional amino acid residues in or near the sialic acid binding site of FLU NA are also found at equivalent positions in the first 48 N-terminal amino acids of SAP-1. This region of homology allows to propose folding of the SAP-1 polypeptide chain by comparison with known crystallographic structure of FLU NA and identify a potential domain for lysosomal enzyme recognition through sialic acid binding. There is also a region of 10 amino acid residues near the C-terminal end of SAP-1 which has a strong propensity to form an alpha-helix with amphiphilic properties of lipid-binding helices. This domain in SAP-1 is probably responsible for the lipid(substrate)-binding function of SAP-1.