Neurosecretory and trophic action on fetal rat neuroblasts induced by an amino acid mixture

The effects of a synthetically obtained mixture of amino acids (FACE) were investigated on the trophic and neurosecretory activity of in vitro cultures of fetal rat neuronal cells. The addition of 10(-6) M FACE to the culture medium significantly increased cell DNA content. Secretions of IR-SRIF, IR-VIP, and IR-GRF were also augmented in different proportions by the presence of FACE. Time studies demonstrated that IR-SRIF was significantly increased after 48 (P less than 0.05) and 72 (P less than 0.01) hr of exposure to FACE, and IR-VIP secretion was potentiated after only 24 hr of culture. Dose-response experiments with 10(-7) to 10(-4) M FACE indicated that concentrations of 10(-5) and 10(-4) M significantly increased both somatostatin released to the medium and cell content of IR-SRIF. FACE concentrations as low as 10(-10) M augmented the secretion of IR-GRF, and there was a dose-response correlation between 10(-10) and 10(-5) M FACE. The release and cell content of IR-VIP were also increased by FACE, with a dose-response relation at concentrations of 10(-9) to 10(-6) M. It can thus be concluded that FACE has a powerful effect on the multiplication and survival of fetal cerebrocortical cells and is also an important potentiator of IR-SRIF, IR-VIP, and IR-GRF secretion.     

Phytoplankton dynamics in a deep, tropical, hyposaline lake

The annual variation of the phytoplankton assemblage of deep (64.6 m), hyposaline (8.5 g l^–1) Lake Alchichica, central Mexico (19 ° N, 97° W), was analyzed in relation to thermal regime, and nutrients concentrations. Lake Alchichica is warm monomictic with a 3-month circulation period during the dry, cold season. During the stratified period in the warm, wet season, the hypolimnion became anoxic. N–NH₃ ranged between non detectable (n.d.) and 0.98 mg l^–1, N–NO₂ between n.d. and 0.007 mg l^–1, N–NO₃ from 0.1 to 1.0 mg l^–1 and P–PO₄ from n.d. to 0.54 mg l^–1. Highest nutrient concentrations were found in the circulation period. Chlorophyll a varied from <1 to 19.8 g l^–1 but most values were <5 g l^–1. The euphotic zone (>1% PAR) usually comprised the top 15–20 m. Nineteen algae species were identified, most of them are typical inhabitants of salt lakes. Diatoms showed the highest species number (10) but the small chlorophyte Monoraphidium minutum, the single-cell cyanobacteria, Synechocystis aquatilis, and the colonial chlorophyte, Oocystis parva, were the numerical dominant species over the annual cycle. Chlorophytes, small cyanobacteria and diatoms dominated in the circulation period producing a bloom comparable to the spring bloom in temperate lakes. At the end of the circulation and at the beginning of stratification periods, the presence of a bloom of the nitrogen-fixing cyanobacteria, N. spumigena, indicated nitrogen-deficit conditions. The well-stratified season was characterized by low epilimnetic nutrients levels and the dominance of small single-cell cyanobacteria and colonial chlorophytes. Phytoplankton dynamics in tropical Lake Alchichica is similar to the pattern observed in some deep, hyposaline, North American temperate lakes.     

Urea equilibrium unfolding of the major core protein of the retrovirus feline immunodeficiency virus and its tryptophan mutants

Circular dichroism and fluorescence spectroscopy have been employed to study the urea unfolding mechanism of a recombinant form of the major core protein of feline immunodeficiency virus (FIV-rp24) and its native tryptophan mutants. The equilibrium denaturation curves indicate the existence of two transitions. The first unfolding transition most likely reflects the denaturation of the carboxy-terminal region of FIV-rp24. Consequently, the second transition, where the changes in fluorescence are produced, should reflect the denaturation of the amino-terminal region. If the intermediate observed upon urea denaturation is an on-pathway species, the data described herein can reflect the sequential and independent loss of structure of the two domains that this type of proteins possesses.     

Inwardly rectifying K(+) channels in spermatogenic cells: functional expression and implication in sperm capacitation

To fertilize, mammalian sperm must complete a maturational process called capacitation. It is thought that the membrane potential of sperm hyperpolarizes during capacitation, possibly due to the opening of K(+) channels, but electrophysiological evidence is lacking. In this report, using patch-clamp recordings obtained from isolated mouse spermatogenic cells we document the presence of a novel K(+)-selective inwardly rectifying current. Macroscopic current activated at membrane potentials below the equilibrium potential for K(+) and its magnitude was dependent on the external K(+) concentration. The channels selected K(+) over other monovalent cations. Current was virtually absent when external K(+) was replaced with Na(+) or N-methyl-D-glucamine. Addition of Cs(+) or Ba(2+) (IC(50) of approximately 15 microM) to the external solution effectively blocked K(+) current. Dialyzing the cells with a Mg(2+)-free solution did not affect channel activity. Cytosolic acidification reversibly inhibited the current. We verified that the resting membrane potential of mouse sperm changed from -52 +/- 6 to -66 +/- 9 mV during capacitation in vitro. Notably, application of 0.3-1 mM Ba(2+) during capacitation prevented this hyperpolarization and decreased the subsequent exocytotic response to zona pellucida. A mechanism is proposed whereby opening of inwardly rectifying K(+) channels may produce hyperpolarization under physiological conditions and contribute to the cellular changes that give rise to the capacitated state in mature sperm.     

Bioactive oleanolic acid saponins and other constituents from the roots of Viguiera decurrens

The bisdesmoside oleanolic acid saponin, 3-0-(methyl-beta-D-glucuronopyranosiduronoate)-28-0-beta-D-glucopyranosyl-oleanolate along with nine known compounds (two diterpenic acids, one chromene, three triterpenes, one steroidal glycoside, and two monodesmoside oleanolic acid saponins), were obtained from Viguiera decurrens roots. The chemical structure of the bisdesmoside oleanolic saponin was determined by chemical and NMR spectral evidence. A mixture of monodesmoside saponins displayed cytotoxic activity against P388 and COLON cell lines (ED50= 2.3 and 3.6 microg/ml, respectively). Two of the known compounds showed insecticidal activity against the Mexican bean beetle larvae (Epilachna varivestis).     

Targeting of HIV gp120 by oligonucleotide-photosensitizer conjugates. Light-induced damages

The effects of the lysosphingolipid, sphingosylphosphorylcholine (SPC), on the cardiac ryanodine receptor (RyR) were examined. The open probability of cardiac RyR incorporated in lipid bilayers was decreased by cytoplasmic, but not lumenal side application of micromolar concentrations of SPC. Modification of channel function was characterized by the appearance of a long-lived closed state in addition to the brief channel closings observed in the presence and absence of SPC. Open channel kinetics and ion conduction properties, however, were not altered by this compound. These results suggest that SPC, a putative second messenger derived from sphingomyelin, may regulate Ca(2+) release from the sarcoplasmic reticulum by modifying the gating kinetics of the RyR.     

Kinetic study of phenol hydroxylase and catechol 1,2-dioxygenase biosynthesis by <Emphasis Type="Italic">Candida tropicalis</Emphasis> cells grown on different phenolic substrates

When Candida tropicalis was grown on phenol, catechol or resorcinol, the highest levels of specific activity of phenol hydroxylase (EC. 1.14.13.7) and catechol 1,2-dioxygenase (EC. 1.13.11.1) were attained with phenol. With the three aromatic compounds tested, the yeast cells exhibited sharp peaks of specific activity of both enzymes at particular incubation times. Phenol-induced cells containing high levels of both enzymes were capable of degrading rapidly and without delay 4-chlorophenol and 2,6-dichlorophenol, and to a lesser extend pentachlorophenol. However, the yeast could not grow on chlorophenols as major carbon and energy source.     

Characterization of the First VanB Vancomycin-Resistant <Emphasis Type="Italic">Enterococcus faecium</Emphasis> Isolated in a Spanish Hospital

We report the detection and characterization of the first vancomycin-resistant VanB-type Enterococcus faecium to be isolated in a Spanish hospital. Sequence analysis of the vanB gene showed that this isolate belonged to subtype vanB2. Moreover, PCR amplification analysis indicated that the vanB gene cluster was linked to a Tn5382-like transposon.     

Glycine intake decreases plasma free fatty acids, adipose cell size, and blood pressure in sucrose-fed rats

The study investigated the mechanism by which glycine protects against increased circulating nonesterified fatty acids (NEFA), fat cell size, intra-abdominal fat accumulation, and blood pressure (BP) induced in male Wistar rats by sucrose ingestion. The addition of 1% glycine to the drinking water containing 30% sucrose, for 4 wk, markedly reduced high BP in sucrose-fed rats (SFR) (122.3 +/- 5.6 vs. 147.6 +/- 5.4 mmHg in SFR without glycine, P < 0.001). Decreases in plasma triglyceride (TG) levels (0.9 +/- 0.3 vs. 1.4 +/- 0.3 mM, P < 0.001), intra-abdominal fat (6.8 +/- 2.16 vs. 14.8 +/- 4.0 g, P < 0.01), and adipose cell size were observed in SFR treated with glycine compared with SFR without treatment. Total NEFA concentration in the plasma of SFR was significantly decreased by glycine intake (0.64 +/- 0.08 vs. 1.11 +/- 0.09 mM in SFR without glycine, P < 0.001). In control animals, glycine decreased glucose, TGs, and total NEFA but without reaching significance. In SFR treated with glycine, mitochondrial respiration, as an indicator of the rate of fat oxidation, showed an increase in the state IV oxidation rate of the beta-oxidation substrates octanoic acid and palmitoyl carnitine. This suggests an enhancement of hepatic fatty acid metabolism, i.e., in their transport, activation, or beta-oxidation. These findings imply that the protection by glycine against elevated BP might be attributed to its effect in increasing fatty acid oxidation, reducing intra-abdominal fat accumulation and circulating NEFA, which have been proposed as links between obesity and hypertension.