Uridine enhances the cytotoxic effect of D-glucosamine in rat C6 glioma cells

This paper studies the influence of uridine on the effects exerted by D-glucosamine in rat C6 glioma cells. 2 mM uridine increased markedly both the cytotoxic effect of the aminosugar and the inhibition of thymidine incorporation into acid-insoluble fraction. Furthermore the complete resumption of the capacity to incorporate either 3H-thymidine or 3H-mannose which was observed after the removal of the aminosugar, was impeded when the cells were treated contemporaneously with D-glucosamine and uridine. An exposure for 4 hr to 20 mM glucosamine alone enhanced about 15-fold the cellular pool of UDP-N-acetylhexosamines; the addition of 2 mM uridine intensified the expansion of this pool, which became about 35-fold the control value. The findings suggest a connection between the accumulation of UDP-N-acetylhexosamines in the cells and the appearance of D-glucosamine cytotoxicity.     

“Mixed Inhibitors” of HIV-Reverse Transcriptase: Synthesis and Antiviral Activity

Abstract

Some “AZT-HEPT” and “ddC-HEPT” conjugates were designed, synthesized and evaluated for their anti-HIV activity.     

AFM and electroanalytical studies of synthetic oligonucleotide hybridization

The first and most important step in the development and manufacture of a sensitive DNA-biosensor for hybridization detection is the immobilization procedure of the nucleic acid probe on the transducer surface, maintaining its mobility and conformational flexibility. MAC Mode AFM images were used to demonstrate that oligonucleotide (ODN) molecules adsorb spontaneously at the electrode surface. After adsorption, the ODN layers were formed by molecules with restricted mobility, as well as by superposed molecules, which can lead to reduced hybridization efficiency. The images also showed the existence of pores in the adsorbed ODN film that revealed large parts of the electrode surface, and enabled non-specific adsorption of other ODNs on the uncovered areas. Electrostatic immobilization onto a clean glassy carbon electrode surface was followed by hybridization with complementary sequences and by control experiments with non-complementary sequences, studied using differential pulse voltammetry. The data obtained showed that non-specific adsorption strongly influenced the results, which depended on the sequence of the ODNs. In order to reduce the contribution of non-specific adsorbed ODNs during hybridization experiments, the carbon electrode surface was modified. After modification, the AFM images showed an electrode completely covered by the ODN probe film, which prevented the undesirable binding of target ODN molecules to the electrode surface. The changes of interfacial capacitance that took place after hybridization or control experiments showed the formation of a mixed multilayer that strongly depended on the local environment of the immobilized ODN.     

Synthesis, antimicrobial activity and conformational analysis of novel substituted pyridines: BF(3)-promoted reaction of hydrazine with 2-alkoxy pyridines

Some new 2-alkoxy-3-cyano-4,6-diarylpyridines 3,4 were synthesized by condensation of different alpha,beta-unsaturated ketones 1 with malononitrile 2, followed by cyclization in sodium alkoxide/alcohol system. Lewis acid-catalyzed reaction of 4 with hydrazine afforded the corresponding 1H-pyrazolo[3,4-b]pyridines 5. The potency of the results as antimicrobial agents has been evaluated. The structure of the newly prepared compounds was assessed by microanalysis, IR and NMR spectra. Molecular mechanics (MM2) and semiemperical (AM1) molecular orbital calculations have been performed for the most biologically active compounds 5b and c to get insight into their molecular structures and to learn more about their stable molecular conformations.     

A new specific ageusia: some humans cannot taste L-glutamate

A new specific ageusia was found in human subjects for monosodium L-glutamate (MSG). Four tests were successively applied to discriminate non-tasters and hypotasters from tasters. (i) NaCl and MSG thresholds, and (ii) suprathreshold sensitivity were evaluated using the up-and-down procedure. Only 73% of 109 subjects common to both tests demonstrated a sensitivity for MSG significantly higher than their sensitivity to NaCl, and hence a specific sensitivity to L-glutamate. The remaining 27% who showed no significant difference in sensitivity to MSG and NaCl solutions were considered as putative hypotasters. (iii) Perception profiles (time-intensity) for MSG and NaCl were tested in 58 subjects and appeared significantly different in 47 tasters (81%). This technique helped in identifying among putative hypotasters of tests 1 and 2 a few tasters who perceived equal intensity for isoconcentration of NaCl and MSG but who could discriminate isomolar solutions on other cues. Thus, 19% of subjects, for whom no significant differences were found between MSG and NaCl time-intensity profiles, remained in the hypotaster group. (iv) A discrimination task including 24 triangular presentations per subject of NaCl and MSG 29 mM applied to the eight most severe hypotasters showed that two subjects at least (two of 58; 3.5%) could not discriminate between both stimuli. Moreover, these subjects probably perceived identical sensations for MSG and NaCl solutions. The six other hypotasters (10.3%) could discriminate both stimuli at the limit of significance. None of these eight subjects were able to identify the typical umami taste in 29 mM MSG.     

Corrigendum to "The sow endometrium at different stages of the oestrus cycle: studies on morphological changes and infiltration by cells of the immune system." [Anim. Reprod. Sci. 65 (2001) 95-114

The aim of this study was to investigate the distribution of leukocytes and the morphological changes of the sow endometrium throughout the oestrous cycle. Fifteen crossbred multiparous sows (Swedish Landrace x Swedish Yorkshire), with an average parity number of 3.4+/-0.7 (mean+/-S.D.) were used. Blood samples were collected from the jugular vein 1 h before slaughter for analyses of oestradiol-17beta and progesterone levels. Uterine samples from the mesometrial side of both horns, taken immediately after slaughter at late dioestrus, prooestrus, oestrus, early dioestrus and dioestrus, were fixed, embedded in plastic resin and stained with toluidine blue. The surface and glandular epithelium as well as subepithelial and glandular connective tissue layers were examined by light microscopy (LM). The significantly highest surface and the glandular epithelium were observed at oestrus and dioestrus, respectively. The largest number of capillaries (underneath the surface epithelium) was found at oestrus. In the surface epithelium, the largest number of intraepithelial lymphocytes (IELs, round nucleus) was found at early dioestrus. The largest number of lymphocytes and macrophages within the glandular epithelium were found at early dioestrus and oestrus, respectively. In the subepithelial connective tissue layer, the most common type of leukocytes during all stages was the lymphocyte. The largest numbers of lymphocytes and neutrophils were found at oestrus while the largest number of eosinophils was found at dioestrus. The dominating cells of the immune system in the connective tissue of the glandular layer were lymphocytes and macrophages. The significantly largest numbers of lymphocytes and plasma cells were found at early dioestrus and dioestrus, respectively. The number of lymphocytes in the connective tissue of the glandular layer and the number of plasma cells in the subepithelial layer were positively correlated with the plasma level of progesterone (P < or = 0.05). The numbers of capillaries and neutrophils in the subepithelial layer underneath the surface epithelium as well as the number of macrophages in both surface and glandular epithelium were positively correlated with the plasma level of oestradiol-17beta (P < or = 0.05). In conclusion, the present study showed a variation om the infiltration and distrobution of lymphocytes, neutrophils, eosinophils, macrophages, mast cells, and plasma cells in the sow endometrium during different stages of the oestrous cycle. Also morphological parameters (e.g. height of surface and glandular epithelium, capillaries density and degree of oedema) varied throughout the oestrous cycle.     

Influence of pre-ovulatory insemination and early pregnancy on the infiltration by cells of the immune system in the sow endometrium

The aim of this study was to investigate the distribution of leukocytes in the sow endometrium following insemination and during early pregnancy. Cross-bred multiparous sows (Swedish Landrace x Swedish Yorkshire) were artificially inseminated (AI) once at 20-15 h before ovulation. Blood samples were collected from the jugular vein 1 h before slaughter for analyses of oestradiol-17beta and progesterone levels. The sows were slaughtered at 5-6 h (group I, n = 4) after AI or at different times after ovulation: 20-25 h (group II, n = 4), 70 h (group III, n = 4), day 11 (group IV, n = 3; first day of standing oestrus = day 1) and day 19 (group V, n = 3). Uterine horns were flushed to control for the presence of spermatozoa and neutrophils (groups I-IV) and/or for recovery of oocytes and/or embryos (groups II-IV, control of pregnancy). Mesometrial uterine samples were fixed, embedded in plastic resin and stained with toluidine blue. The surface and glandular epithelia as well as subepithelial and glandular connective tissue layers were examined by light microscopy. A marked number of neutrophils and spermatozoa were observed in the flushings from the uterine horns of sows slaughtered at 5-6 h after insemination. All animals slaughtered after ovulation were pregnant but no morphological effect of pregnancy was observed until day 11. In the surface epithelium, the largest numbers of intraepithelial lymphocytes were found in groups II and III, the smallest number was found in group V. The largest number of lymphocytes within the glandular epithelium was found in group III. The largest number of macrophages within the surface and glandular epithelia were found in group I. Neutrophils were found within the surface epithelium only in groups I and II. In the subepithelial connective tissue layer, a high infiltration of neutrophils was found in groups I and II while the largest number of eosinophils was found in group IV. The largest number of lymphocytes was observed in group V. In conclusion, this study showed a variation in the infiltration and distribution of neutrophils, lymphocytes, macrophages, eosinophils and plasma cells in the endometrium following insemination and during different stages of early pregnancy. Particularly, the pattern of lymphocyte presence on day 19 of pregnancy, indicate that the lymphocyte function may play a role during embryonic attachment in the pig.     

Effects of dexamethasone on human synovial fibroblast-like cells, from osteoarthritic joints, in culture.

The effect of Dexamethasone (DEX) on cell division and macromolecular synthesis was investigated in a line (McCoy cells, A 9) of synovial fibroblast-like cells derived from human osteoarthritic joints. DEX markedly reduced the proliferation of McCoy cells in a time and dose-dependent manner. The maximal inhibition (45%) was found at 500 nM DEX 24 h after incubation and was accompanied by the appearance of giant macrophage-like cells. After DEX treatment cells showed increased content of DNA, proteins and RNA together with the reduction of [3H]-thymidine incorporation into the TCA-precipitable fraction.     

ABSORPTION AND UTILIZATION OF IRRADIANCE BY CYANOBACTERIAL MATS IN TWO ICE-COVERED ANTARCTIC LAKES WITH CONTRASTING LIGHT CLIMATES

We investigated the under-ice light climate and the efficiency with which light was absorbed and utilized by benthic algal mats in Lakes Hoare and Vanda, two perennially ice-covered lakes in the McMurdo Dry Valleys area of Southern Victoria Land, Antarctica. The ice cover and water column of Lake Vanda were much more transparent than those of Lake Hoare (18% vs. 2% transmission though ice and attenuation coefficients for downwelling irradiance of 0.05 vs. 0.12 m ^{− 1}, respectively). In both lakes the under-ice spectra were dominated by blue-green wavelengths. The benthic flora under perennial ice covers of both lakes comprised thick mucilaginous mats, dominated by cyanobacteria. The mats were well suited to absorb the dominant blue-green wavelengths of the under-ice light, with phycoerythrin being present at high concentrations. The pigment systems of the benthic mats absorbed 30%–50% of the light that reached them, varying with depth and lake. There was a tendency for the percentage of absorption to increase as ambient irradiance decreased. The efficiency of utilization of absorbed irradiance was examined by constructing absorbed irradiance/oxygen evolution curves to estimate community quantum yield. Mats from 13 m in Lake Hoare showed the highest quantum yields, approaching 1 mol of carbon fixed for every 8 mol quanta absorbed under light-limiting conditions. Lake Vanda mats had lower quantum yields, but these increased with depth. Calculated in situ irradiance occasionally exceeded the measured saturating irradiance for oxygen evolution in both lakes, thus efficiency in situ was below the maximum at times. As in other environments, optimization strategies allowed efficient capture and utilization of the lower and middle ranges of experienced irradiance but led to a compromised capacity to use the highest irradiances encountered at each depth.