Synthesis and structural investigation of vanadocene(IV) complexes of non-linear pseudohalides

Two series of vanadocene complexes of the type (Cp′ = η⁵-C₅H₅, η⁵-C₅H₄Me; X = dicyanamide, tricyanomethanide, dicyanonitrosomethanide) were prepared by the reaction of appropriate vanadocene dichloride complex with alkali salt of non-linear pseudohalide. The bonding mode of pseudohalide ligands was determined by spectroscopic measurements and X-ray diffraction analyses.     

Synthesis, characterization and cytotoxic effect of ring-substituted and ansa-bridged vanadocene complexes

The cytotoxic effect of vanadocene dichloride (Cp₂VCl₂, 1) and its ring-substituted, (η⁵-C₅H₄Me)₂VCl₂ (2), (η⁵-C₅Me₅)₂VCl₂ (3), (η⁵-C₅H₄R)₂VCl₂ (4: R = MeOCH₂CH₂-, 5: R = 2-MeOC₆H₄CH₂-, 6: R = 4-MeOC₆H₄CH₂-) and ansa-bridged analogs Me₂C(η⁵-C₅H₄)₂VCl₂ (7) and Me₄C₂(η⁵-C₅H₄)₂VCl₂ (8) was investigated. Synthesis of two new methoxy-functionalized compounds (4 and 5) is described. They were characterized by spectroscopic methods and X-ray diffraction analysis. The cytotoxicity studies were performed with leukemic cells MOLT-4.     

Oxidative and phosphorylative activities of the gastric mucosa of animals and humans in relation to the mechanism of stress ulcer

In order to elucidate the mechanism of stress ulceration, which is mainly involved with the corpus mucosa of the stomach, the aerobic energy metabolism of the corpus mucosa of rats, rabbits and humans was investigated and compared with those of the antral mucosa as well as other organs such as heart, kidney and liver. The results showed that the activities of respiration and oxidative phosphorylation and the concentration of the respiratory chain components were far greater in the corpus mucosa than in the antral mucosa, the activities in the former being almost equal to that of the highly aerobic tissue, kidney cortex. Furthermore, the endogenous respiratory rate (in the absence of added substrate) was also much higher in the corpus mucosa than in the antral mucosa, indicating that oxygen demand at resting state (during the period of H+ secretion) is also higher in the corpus mucosa. We conclude that in the corpus mucosa of the stomach the aerobic energy metabolism is predominant. It is suggested that circulatory insufficiency may easily result in cell damage mainly of the corpus mucosa due to energy deficit.     

Studies on the mechanism of action of hexamethylene bisacetamide, a potent inducer of erythroleukemic differentiation.

Hexamethylene bisacetamide (diacetyldiamino hexane) is a potent inducer of erythroid differentiation in murine erythroleukemia cells. Hexamethylene bisacetamide and the closely related pentamethylene bisacetamide were synthesized with radioactive labels in various portions of the molecule and the uptake, metabolism, and intracellular distribution determined. Bisacetamides are taken up by the cell; an intracellular concentration equal to the extracellular concentration is achieved by 6-8 h. Commitment to differentiation is not detected until at least 10 h after equilibration. Both uptake and commitment to differentiate are concentration and temperature dependent. The majority of the compound is deacetylated upon cell entry and the acetate portion incorporated nonspecifically into lipid and protein. Acetate competes with the incorporation of hexamethylene bisacetamide into protein and lipid, but does not affect inducing activity. The diamine portion of the molecule is detected only in the cytoplasm, in a trichloroacetic acid-soluble and acetylated form, whereas the acetate moiety is detected in both cytoplasm and nucleus and in both a trichloroacetic acid-soluble and insoluble form. The cellular uptake of diamines and bisacetamides (acetylated diamines) are similar, but acetylation of the diamine greatly increases inducing activity.     

Oxovanadium complexes with quinoline and pyridinone ligands: syntheses of the complexes and effect of alkyl chains on their apoptosis-inducing activity in leukemia cells

Vanadium complexes with quinoline ligands (1b-g) and pyridinone ligands (2b-d) were synthesized, and the effect of the length and shape of alkyl chains on the antiproliferative activity toward U937 cells was studied. For the synthesis of the vanadium complexes, quinoline and pyridinone ligands were prepared and then treated with VOSO(4) or VO(acac)(2). The vanadyl(IV) complexes were characterized by IR, ESR, and UV-vis spectroscopy and elemental analyses. The antiproliferative activity of 1a-g toward U937 cells showed little dependence on the length and shape of the alkyl chain. In contrast, a good correlation was found between the IC(50) values and partition coefficients (logP) values of 2a-c. Among them, 2c showed the highest inhibitory activity, and its IC(50) value was smaller than that of cisplatin. The apoptosis-inducing ability of 2b and 2c was supported by annexin V-propidium iodide staining experiments and agarose gel electrophoresis analysis. Inhibitors of caspase-3, -8, and -9 did not affect the antiproliferative activity of 2c, indicating that the apoptosis induced by 2c was via a caspase-independent pathway.     

Cacospongionolide and scalaradial, two marine sesterterpenoids as potent apoptosis-inducing factors in human carcinoma cell lines

Apoptosis, a form of programmed cell death, is a critical defence mechanism against the formation and progression of cancer and acts by eliminating potentially deleterious cells without causing such adverse effects, as inflammatory response and ensuing scar formation. Therefore, targeting apoptotic pathways becomes an intriguing strategy for the development of chemotherapeutic agents. In last decades, marine natural products, such as sesterterpenoids, have played an important role in the discovery and development of new drugs. Interestingly, many of these compounds have a strong potential as anticancer drugs by inhibiting cell proliferation and/or inducing cell death. In the present study, we investigated the effects of scalaradial and cacospongionolide, two sesterterpenoids from Cacospongia scalaris and Fasciospongia cavernosa marine sponges, on the apoptotic signalling pathway in three different human tumoral cells. Results were obtained by using DNA fragmentation, comet and viability assays, quantification of the mitochondrial transmembrane potential and Western blot. The T47D (human breast carcinoma), A431 (human epidermoid carcinoma), HeLa (human cervix carcinoma) and HCT116 (human colon carcinoma) cells were incubated for 24 h with scalaradial or cacospongionolide. Treatment of T47D cells with scalaradial or cacospongionolide for 24 h brought about a significant increase in DNA migration as well as fragmentation. Moreover, incubation of HCT116 and HeLa cells with scalaradial or cacospongionolide for 24 h caused an increased expression of pro-apoptotic proteins. Furthermore, scalaradial or cacospongionolide, added to HCT116 and HeLa cells overnight, induced a significant and concentration-dependent loss of mitochondrial transmembrane potential, an early apoptosis signalling event. These effects paralleled with those achieved with p50 and p65, NF-κB subunits, nuclear level. In conclusion, scalaradial and cacospongionolide, by determining human cancer cell apoptosis, may represent new promising compounds to inhibit cancer cell proliferation.     

Studies on the kinetics and mechanism of anation reactions of some six-coordinate complexes of chromium(III) in aqueous solution

Rates of stepwise anation of cis-Cr(ox)₂(H₂O₂)⁻ with SCN⁻/N₃⁻, Cr(acac)₂(H₂O)₂⁺ with SCN⁻ and Cr(atda)(H₂O)₂ with SCN⁻ have been investigated in weakly acidic aqueous solutions. Rate constants, k_I and k_II for the two steps in each system, are composite as k_x = k_x⁰+k_x^X[X⁻] (x = I, II; X⁻ = SCN⁻, N₃⁻). These rate constants have been evaluated also as the corresponding ΔH^≠ and ΔS^≠ values. The results obtained and the plausible I_d mechanism seem to suggest Cr---OOC bond dissociation (hence a strongly negative ΔS^≠) generating the transition state in each system with outer-sphere association forming the precursor complex in the X⁻ dependent paths.     

Studies on the mechanism for entry of vesicular stomatitis virus glycoprotein g mRNA into membrane-bound polyribosome complexes

Glycoprotein mRNA (G mRNA) of vesicular stomatitis virus is synthesized in the cytosol fraction of infected HeLa cells. Shortly after synthesis, this mRNA associates with 40S ribosomal subunits and subsequently forms 80S monosomes in the cytosol fraction. The bulk of labeled G mRNA is then found in polysomes associated with the membrane, without first appearing in the subunit or monomer pool of the membrane-bound fraction. Inhibition of the initiation of protein synthesis by pactamycin or muconomycin A blocks entry of newly synthesized G m RNA into membrane-bound polysomes. Under these circumstances, labeled G mRNA accumulates into the cytosol. Inhibition of the elongation of protein synthesis by cucloheximide, however, allows entry of 60 percent of newly synthesized G mRNA into membrane-bound polysomes. Furthermore, prelabeled G mRNA associated with membrane-bound polysomes is released from the membrane fraction in vivo by pactamycin or mucomycon A and in vitro by 1mM puromycin - 0.5 M KCI. This release is not due to nonspecific effects of the drugs. These results demonstrate that association of G mRNA with membrane-bound polysomes is dependent upon polysome formation and initiation of protein synthesis. Therefore, direct association of the 3' end of G mRNA with the membrane does not appear to be the initial event in the formation of membrane-bound polysomes.     

Structures and mechanism for the design of highly potent glucocorticoids

The evolution of glucocorticoid drugs was driven by the demand of lowering the unwanted side effects, while keeping the beneficial anti-inflammatory effects. Potency is an important aspect of this evolution as many undesirable side effects are associated with use of high-dose glucocorticoids. The side effects can be minimized by highly potent glucocorticoids that achieve the same treatment effects at lower doses. This demand propelled the continuous development of synthetic glucocorticoids with increased potencies, but the structural basis of their potencies is poorly understood. To determine the mechanisms underlying potency, we solved the X-ray structures of the glucocorticoid receptor (GR) ligand-binding domain (LBD) bound to its endogenous ligand, cortisol, which has relatively low potency, and a highly potent synthetic glucocorticoid, mometasone furoate (MF). The cortisol-bound GR LBD revealed that the flexibility of the C1-C2 single bond in the steroid A ring is primarily responsible for the low affinity of cortisol to GR. In contrast, we demonstrate that the very high potency of MF is achieved by its C-17α furoate group completely filling the ligand-binding pocket, thus providing additional anchor contacts for high-affinity binding. A single amino acid in the ligand-binding pocket, Q642, plays a discriminating role in ligand potency between MF and cortisol. Structure-based design led to synthesis of several novel glucocorticoids with much improved potency and efficacy. Together, these results reveal key structural mechanisms of glucocorticoid potency and provide a rational basis for developing novel highly potent glucocorticoids.     

Studies on the growth and phosphatase activities inSuaeda fructicosa

Summary Plants ofSuaeda fructicosa were grown in sand culture with various combinations of excess copper, silicon and sodium. Effects of different concentrations of NaCl on the activities of acid phosphatase and PEP-phosphatase were investigated in the above mentioned plants. Plants which were given extra copper without NaCl showed very poor growth compared with control plants given a normal nutrient whereas plants given extra NaCl alone or with copper displayed luxuriant growth. Enzymes extracted from the plants given extra copper with normal nutrient levels showed stimulation in the activities of both the above mentioned enzymes. In plants supplied with extra sodium chloride the activity of acid phosphatase was inhibited and in that of PEP-phosphatase was stimulated. Sodium silicate in the growth medium eliminated both effects of extra sodium chloride.     

Studies of strontium biokinetics in humans

The radioactive isotopes of strontium, mainly (90)Sr, which are common fission products, may significantly contribute to the internal exposure of the population in case of their accidental release into the environment and transfer to the food chain. For (90)Sr, the internal radiation dose is significantly dependent on the fractional absorption of the ingested activity (f(1)-value). Human data on the absorption of dietary strontium and of soluble forms of the element give values ranging from about 0.15 to 0.45 (up to 1.0) for adults. The International Commission on Radiological Protection (ICRP) has adopted f(1)-values of 0.6 for children of less than 1 year of age, 0.4 for children between 1 and 15 years and 0.3 for adolescents above 15 years of age. This study was aimed at investigating how far these values correspond to the actual uptake of strontium from contaminated foodstuffs. A methodology is presented that has been developed for preparing foodstuffs intrinsically labelled with stable isotopes and that will be used in tracer kinetic investigations. The results show that cress and salad can be adequately labelled, i.e. a strontium concentration of 1.36+/-0.47 g per kg of cress (wet weight) and of 0.29+/-0.04 g per kg of salad (wet weight) may be obtained within 15 days and 24 days, respectively. For the biokinetic investigations on humans, applying stable isotopes of Sr as tracers, about 0.1-1 mg strontium is required per volunteer, i.e. a few grams of the edible parts of the labelled material are sufficient.     

Studies on after-taste of various taste stimuli in humans

This series of experiments was performed in order to evaluatethe physiological characteristics and patterns of after-tasteof various taste substances. The durations of after-taste followingmonosodium glutamate (MSG), inosine-5'-monophosphate (IMP) andguanosine-5'-monophosphate (GMP) (umami substances) were longerthan those for sucrose, NaCl, tartaric acid and quinine-HCIat concentrations corresponding to the recognition threshold.The periods of after-taste of solutions of MSG and IMP, andMSG and GMP, were longer than those for the single componentsolutions. Most subjects recognized sucrose as sweet, NaCl assalty, tartaric acid as sour and quinine-HCI as bitter, bothin terms of immediate taste and after-taste. According to thepatterns of after-taste for umami substances, the subjects weredivided into three groups. In group A, umami (appealing, savorytaste in Japanese cuisine) was the main quality of the after-tastesensation; in group B, an indefinite, equivocal taste was thecharacteristic quality of the aftertaste; and no differencewas reported in group C between the immediate taste and after-taste.These results suggest that the characteristics of after-tastefor MSG, IMP and GMP are different from those of the four basictastes.