Novel cyanoguanidines with potent oral antitumour activity

4-Pyridyl cyanoguanidines with hydrophobic aromatic side chains showed potent antiproliferative activity in the human breast and lung cancer cell lines MCF-7, NYH and H460. In vivo, treatment with N-(6-chlorophenoxyhexyl)-N′-cyano-N″-4-pyridylguanidine (18, 20 mg/kg/day po.), gave a complete remission of tumours in a model of NYH inoculated nude mice.     

Characterization of humoral and CD4+ cellular responses after genetic immunization with retroviral vectors expressing different forms of the hepatitis B virus core and e antigens.

The humoral and CD4+ cellular immune responses in mice following genetic immunization with three retroviral vectors encoding different forms of hepatitis B virus core antigen (HBcAg) and e antigen (HBeAg) were analyzed. The retroviral vectors induced expression of intracellular HBcAg (HBc[3A4]), secreted HBeAg (HBe[5A2]), or an intracellular HBcAg-neomycin phosphoryltransferase fusion protein (HBc-NEO[6A3]). Specific antibody levels and immunoglobulin G isotype restriction were highly dependent on both the host major histocompatibility complex and the transferred gene. Humoral and CD4+ cellular HBcAg and/or HBeAg (HBc/eAg)-specific immune responses following retroviral vector immunization were of a lower magnitude but followed the same characteristics compared with those after immunization with HBc/eAg in adjuvant. Two factors influenced the humoral responses. First, in vivo depletion of CD8+ cells in HBc-NEO[6A3]-immunized H-2k mice abrogated both HBcAg-specific antibodies and in vitro-detectable cytotoxic T lymphocytes. Second, priming of H-2b mice with an HBc/eAg-derived T-helper (Th) peptide in adjuvant prior to retroviral vector immunization greatly enhanced the HBc/eAg-specific humoral responses to all three vectors, suggesting that insufficient HBc/eAg-specific CD4+ Th-cell priming limits the humoral responses. In conclusion, direct injection of retroviral vectors seems to be effective in priming HBc/eAg-specific CD8+ but comparatively inefficient in priming CD4+ Th cells and subsequently specific antibodies. However, the limited HBc/eAg-specific CD4+ cell priming can effectively be circumvented by prior administration of a recombinant or synthetic form of HBc/eAg in adjuvant.     

Formation of domoic acid and fatty acids inPseudonitzschia pungens f.multiseries with scale of culture

Pseudonitzschia pungens f.multiseries was cultured in 20-L polycarbonate carboys, 350-L fibreglass columns and 500-L plastic bags to determine the effects of medium enrichment and scale of culture on cell yield, production of cellular domoic acid and formation of fatty acids, particularly the potential tracer acid 16:4n-1. Cell concentrations were highest in seawater enriched with stock levels of nitrate and phosphate, but with double the stock level of silicate, at all scales of culture. Cellular toxin in 20, 350 and 500-L cultures averaged 0.32, 0.04 and 2.56 pg cell^-1 and was independent of medium used. The order of magnitude difference in levels of cellular toxin was considered to reflect the varying levels of irradiance within the culture vessels. Support was given to this by the significant difference in content of total cellular fatty acids, due principally to the algal storage acid 16: 1n-7, which is known to be influenced by irradiance. Levels of cellular domoic acid correlated significantly with total fatty acids in 350 and 500-L cultures. Bag cultures producing significantly higher levels of cellular domoic acid provided lower relative proportions of 16:4n-1, which limited its use as a tracer for food-web studies.     

The effect of renal vasodilation and hypotonic volume expansion on water excretion in dogs after anesthesia and surgery

The possible effects of renal vasoconstriction from anesthesia and surgery on water excretion after hypotonic volume expansion (HVE) were studied in 18 well conditioned anesthetized dogs, with and without the infusion of phenoxybenzamine and acetylcholine into the renal artery of the cannulated kidney. In 6 dogs (Group 1 - Control) whose renal artery was infused with isosmotic saline, HVE resulted in a bilateral increase in GFR and UV (p < .05). ERPF, Cosm, C_H2O, U_NaV, U_KV, RBF, RVR and MAP did not change significantly. In 6 other dogs (Group 2), whose cannulated kidney was infused with phenoxybenzamine 50 μg/min before and during HVE, GFR increased on the infused side while CH₂O and UV increased bilaterally (p < .05). ERPF, Cosm, U_NaV, U_KV, RBF, RVR and MAP were not affected significantly. The addition of ADH, 2 mu/min into the phenoxybenzamine infusate, decreased ERPF, RBF and RVR bilaterally and CH₂O on the infused side (p < .05). It had no effect upon GFR, Cosm, U_NaV, U_KV and MAP. In another 6 dogs, (Group 3), whose cannulated renal artery was infused with acetylcholine (20 μg/min) before and during HVE, C_H2O, UV and RVR increased bilaterally (p < .05). ERPF and RBF decreased bilaterally (p < .05), whereas GFR, Cosm, U_NaV and MAP were unaffected. U_KV decreased on the infused side (p < .05). The addition of ADH (2 mu/min)_into the acetylcholine infusate, decreased C_H2O bilaterally and increased Cosm and U_KV on the control side (p < .05). It had no effect on ERPF, GFR, UV, U_NaV, RBF, RVR and MAP. These observations suggest that anesthesia and surgery produce renal vasoconstriction and this together with increased ADH release, interfere with water excretion by the kidney. Previous renal vasodilation prevents these influences of anesthesia and surgery.     

Biochemical evidence for the non-inactivation of the steroid sulfatase locus in human placenta and fibroblasts

Summary Steroid sulfatase activities are significantly higher in placentas obtained after the birth of girls than after the birth of boys, and also in female fibroblasts compared to male strains. This constitutes biochemical evidence for the non-inactivation of the X-linked sulfatase locus. No hydrolytic activity is found in the fibroblasts of ichthyotic boys. Heterozygosity is demonstrated in the fibroblasts of the four mothers studied, as they have steroid sulfatase activity of less or equivalent to the normal male value.     

Myrosin cells and dilated cisternae of the endoplasmic reticulum in the order Capparales

Ultrastructural results for different types of protein–rich cells in five families generally accepted as Capparalean (Brassicaceae, Capparaceae, Resedaceae, Tovariaceae and Moringaceae) and two others (Gyrostemonaceae and Bataceae) considered by some workers to be Capparalean, support their alignment in the order Capparales. The term myrosin cell is used for those protein–rich cells which are typically idio–blastic and characterized by a homogenous, granular proteinaceous material in the vacuole and a cytoplasm which is filled with an extensive rough endoplasmic reticulum. This idioblastic myrosin cell type is characteristic for the Brassicaceae, Capparaceae, Tovariaceae, Moringaceae and Gyrostemonaceae. The guard cells of stomata may appear as myrosin cells, in which case they are termed guard–cell myrosin cells; they are found in the Resedaceae, Tovariaceae and Bataceae. Other proteinaceous cells are those with protein–rich dilated cisternae (DC) of the endoplasmic reticulum (ER). One type is the organelle–like DC, utricular or irregular dilations of the ER, filled with protein and ribosome–studded. Utricular DC are characteristic for the Brassicaceae and Capparaceae. Another type of DC is represented by protein–containing vacuoles derived from the ER, protein–rich ER–dependent vacuoles; these are found in the Brassicaceae, Capparaceae, Resedaceae, Tovariaceae and Gyrostemonaceae. The myrosin cells and cells with protein–rich dilated cisternae are here regarded as taxonomic criteria for the order Capparales.     

Dissecting the molecular mechanism of ion-solute cotransport: Substrate specificity mutations in theputP gene affect the kinetics of proline transport

Summary Rare mutations that alter the substrate specificity of proline permease cluster in discrete regions of theputP gene, suggesting that they may replace amino acids at the active site of the enzyme. IfputP substrate specificity mutations directly alter the active site of proline permease, the mutants should show specific defects in the kinetics of proline transport. In order to test this prediction, we examined the kinetics of threeputP substrate specificity mutants. One class of mutation increases theK _m over 120-fold but only decreases theV _max fourfold. SuchK _m mutants may be specifically defective in substrate recognition, thus identifying an amino acid critical for substrate binding. Another class of mutation decreases theV _max 80-fold without changing theK _m .V _max mutants appear to alter the rate of substrate translocation without affecting the substrate binding site. The last class of mutation alters both theK _m andV _max of proline transport. These results indicate that substrate specificity mutations alter amino acids critical for Na⁺/proline symport.     

Factors controlling nitrification and denitrification: A laboratory study with gel-stabilized liquid cattle manure

Nitrification and denitrification were studied in a millimeterscale microenvironment using a two-phase system with a liquid manure-saturated layer. Samples consisted of liquid cattle manure and air-dried soil stabilized with silica gel, placed between two aerobic soil phases with a water content near field capacity. A high potential for NH₄ ⁺ oxidation developed within 0–2 mm distance from the interface, and NH₄ ⁺ diffused only 10–20 mm into the soil. Some NH₄ ⁺ was probably immobilized by microorganisms in the soil between 0 and 4 days, after which nitrification was the only sink for NH₄ ⁺. A potential for denitrification developed within the manure-saturated zone. Maximum rates of both potential and actual denitrification were recorded by Day 4, but denitrification continued for at least 2–3 weeks. The potential for nitrification peaked after 14 days. When the pH of the manure was adjusted to 5.5, nitrification was reduced close to the interface, and NH₄ ⁺ penetrated further into the soil before it was oxidized. The pH adjustment had an inhibitory effect on denitrification: Both potential and actual rates of denitrification were almost eliminated for several days. The size of the manure-saturated layer strongly affected denitrification losses. With layers of 8 and 16 mm thickness, losses equivalent to 33 and 40% of the original NH₄ ⁺ pool, respectively, were estimated. When manure corresponding to a 12 mm layer was homogeneously mixed with the soil, only 0.3% was lost.Offprint requests to: S. O. Petersen.