Effect of Escherichia coli endotoxin on ascorbic acid transport in isolated adrenocortical cells

The active uptake of ascorbic acid by isolated rat adrenocortical cells increases with ascorbic acid concentration, depends on time and calcium, and is inhibited by ACTH concentrations required for maximal steroidogenesis. Lipopolysaccharide of Escherichia coli 0111:B4 modifies the ascorbic acid uptake in a calcium-dependent manner. At low calcium concentrations, lipopolysaccharide exerts a stimulatory effect on ascorbic acid transport and at high concentrations lipopolysaccharide produces a dose-dependent inhibitory effect. This inhibition of the ascorbic acid transport by the endotoxin can alter the ascorbic acid accumulation in the adrenal gland during endotoxin shock.     

Progesterone determinations and clinical examinations of reproductive organs in purebred and crossbred female Zebu cattle

Five experiments were undertaken to investigate variation in progesterone concentrations as related to the collection and handling of samples of milk and blood, to determine reference values for progesterone and to evaluate clinical findings in relation to progesterone data from pure- and crossbred Zebu cattle reared in the Peruvian tropics. Whole-milk progesterone concentrations obtained from 12 Holstein x Nellore pregnant cows at hourly intervals over a 24-h period were highest immediately after milking; this peak was followed by a sharp drop over the next 3 h. Milk-fat content from 28 Brown Swiss x Nellore pregnant cows increased from 2.4% before milking to 6.7% afterwards (P < 0.05), whereas progesterone concentrations in whole milk increased from 18.4 to 59.5 nmol/1 (P < 0.05). Progesterone concentrations in fat-free milk were stable, with the exception of the fore-milk sample, which was lower than subsequent samples collected during milking (P < 0.05). Blood samples collected from 23 purebred, pregnant Nellore cows, were divided into four aliquots, and plasma and serum were harvested periodically over the next 120 h of storage at +4 degrees C, or in the shade at ambient air temperatures. The results indicate that blood samples can be stored unseparated at both temperatures studied for up to 3 h without severe loss of progesterone. Milk samples collected at different intervals during the luteal phase of the estrous cycle and during early and mid-pregnancy from crossbred cows showed no significant differences in progesterone concentrations between Days 9 to 13 of the cycle and Days 9 to 13 of gestation. Progesterone levels during advanced gestation were higher (P < 0.05). Out of 2,607 clinical examinations, the level of agreement between palpatory findings and progesterone determinations was 95.6 and 81.9% in diagnosing non-luteal and luteal structures, respectively. Significant differences in the level of agreement between palpators were found (P < 0.01). It is concluded that milk samples, preferably composite milk or strippings, must be consistently collected at the same stage of milking, and that centrifugation of blood samples should be done as soon as possible and not later than 3 h after collection.     

Milk progesterone profiles in crossbred Brown Swiss x Nellore cattle following natural service

Ovarian activity and estrous behaviour were monitored through milk progesterone determinations and twice daily visual observations in 70 crossbred Brown Swiss x Nellore cows following natural service. Whole milk samples were collected on the day of estrus (Day 1), Day 11, and every 5 d thereafter until the next estrus or pregnancy confirmation. Seventy percent of the cows behaved as expected, i.e. they showed a 19-to 25-d interval between estrus and the next ovulation, or they became pregnant. Estrous cycles of regular length (18 to 24 d) were found in 54% of the cases. Prolonged luteal phases (interval from estrus to next ovulation > 28 d) were found in 15.7% of cows. Short estrous cycles (interestrous interval < 18 d) were found in 7.1% of the cases. Periods of acyclicity (basal progesterone levels for periods >/= 15 d) were found in 5.8% of the cases, and one cow exhibited estrus while pregnant and had a high progesterone concentration. Cows with a prolonged luteal phase and those with a short estrous cycle had an interval between ovulations of 35.0 +/- 6.7 d (x +/- SD) and 9.6 +/- 3.1 d, respectively. Signs of estrus were not detected in 33.3% of the ovulations confirmed by progesterone determinations. Low conception rates, failures in estrus detection and a high frequency of abnormal postbreeding luteal phases were found.     

Long-acting estrogenic responses of estradiol fatty acid esters

Estradiol esters at C-17 and C-3 with palmitic, stearic and oleic acids were chemically synthesized and then evaluated for their long-acting estrogenic responses in ovariectomized rats. The duration of the biological effects was measured after a single subcutaneous dose of 0.1 mumol of each ester and compared with those observed with 17 beta-estradiol, estradiol 3-benzoate and estradiol 17-enanthate. Vaginal citology, uterophyc action, serum gonadotropins inhibition and 17 beta-estradiol levels were measured 0, 5, 10, 20, 30 and 60 days after injection. The results disclosed that most of the estradiol derivatives evaluated exhibited a long-acting estrogenic action. However, the monoesters at C-17 showed longer effects that monoesters at C-3, while the estradiol diesters exhibited the shortest effects. In addition as shown by its low serum levels, all estradiol esters with unsaturated fatty acids show a decreased E2 absorption. The overall results indicated that esterification of E2 with long chain fatty acids provided long-acting properties to it, being higher with C-17 esters. Whether some of these compounds could be employed in substitutive endocrine therapy remains to be established.     

TAR independent activation of the human immunodeficiency virus in phorbol ester stimulated T lymphocytes.

Multiple regulatory elements in the human immunodeficiency virus long terminal repeat (HIV LTR) are required for activation of HIV gene expression. Previous transfection studies of HIV LTR constructs linked to the chloramphenicol acetyltransferase gene indicated that multiple regulatory regions including the enhancer, SP1, TATA and TAR regions were important for HIV gene expression. To characterize these regulatory elements further, mutations in these regions were inserted into both the 5' and 3' HIV LTRs and infectious proviral constructs were assembled. These constructs were transfected into either HeLa cells, Jurkat cells or U937 cells in both the presence and absence of phorbol esters which have previously been demonstrated to activate HIV gene expression. Viral gene expression was assayed by the level of p24 gag protein released from cultures transfected with the proviral constructs. Results in all cell lines indicated that mutations of the SP1, TATA and the TAR loop and stem secondary structure resulted in marked decreases in gene expression while mutations of the enhancer motif or TAR primary sequence resulted in only slight decreases. However, viruses containing mutations in either the TAR loop sequences or stem secondary structure which were very defective for gene expression in untreated Jurkat cells, gave nearly wild-type levels of gene expression in phorbol ester-treated Jurkat cells but not in phorbol ester-treated HeLa or U937 cells. High level gene expression of these TAR mutant constructs in phorbol ester-treated Jurkat cells was eliminated by second site mutations in the enhancer region or by disruption of the tat gene.(ABSTRACT TRUNCATED AT 250 WORDS)     

Seasonal changes in canopy structure in two mediterranean dune shrubs

Abstract. The annual cycle of canopy structure in two mediterranean shrubs in a pioneer zone of the mobile dune system in the Donana National Park, Scrophularia frutescens and Halimium halimifolium, has been analyzed. Destructive methods were used as well as a new non-destructive method, based on frequency analysis of organ distribution within the plant canopy. S. frutescens shows strong seasonal changes of photo-synthetic biomass, but little annual increment in dry weight. In H. halimifolium, seasonal changes are not as strongly differentiated as in S. frutescens, but a higher annual increment is shown. The canopy structure of both species and its temporal changes are compared with existingplant strategy models.     

Numerical analysis of electrophoretic protein patterns of 'Achromobacter'group B, E and F strains from human blood

Thirty-two clinical strains representing ' Achromobacter 'groups B, E and F were characterized by one-dimensional SDS-PAGE of cellular proteins. All the strains were isolated from blood samples from hospital patients in the United Kingdom. The protein patterns, which contained 40 to 45 discrete bands, were highly reproducible and were used as the basis for a numerical analysis which included all the protein bands. The 32 ' Achromobacter ' strains formed two clusters at the 77% S level. The first, phenon 1, included the 28 group B and the two group E strains and the second, phenon 2, contained the two strains of group F. The strains in each phenon were characterized by a clearly distinct pattern of protein bands. Phenon 1 could be further divided at the 87% S level into three subphenons which correlated with differences in the principal bands found between 40.0 and 42.5 kD. Strains of group E clustered with group B strains from which they could not be distinguished by protein patterns. We conclude that high resolution PAGE combined with computerized analysis of protein patterns provides a useful method for the classification of this group of bacteria. Reference strains of each of the PAGE types identified are available from NCTC for inclusion in future studies.     

Neuromuscular development following tetrodotoxin-induced inactivity in mouse embryos

Developmental aspects of the neuromuscular system in mouse embryos chronically paralyzed in utero with tetrodotoxin (TTX) between embryonic days 14 and 18 were studied using biochemical and histological methods. The number of lumbar spinal motoneurons (MNs) was higher in inactive embryos than in controls suggesting a decreased motoneuron cell death. In association with the increase in MN number, choline acetyltransferase activity was significantly increased in both spinal cord and peripheral synaptic sites. Paralyzed muscles exhibited a decreased number of mature myofibers and the nuclei were centrally located. Creatine kinase activity was greatly decreased and total acetylcholine receptor and receptor cluster numbers per myofiber were significantly increased in paralyzed muscles. A similar pattern of changes occurs in the neuromuscular system of the mutant mouse muscular dysgenesis (mdg). However, in contrast to the mdg mutant, tetrodotoxin-treated muscles were similar to controls in their innervation pattern, in the ultrastructural aspects of the excitation–contraction coupling system (i.e., dyads and triads) and in the extent of dihydropyridine binding. Thus, neuromuscular inactivity is not sufficient to impair the pattern of muscle innervation or the appearance of either the triadic junctions or dihydropyridine receptors. These results indicate that alterations of dihydropyridine binding sites and triads in muscular dysgenesis cannot be accounted for by inactivity but rather must reflect a more primary defect involving the structural gene(s) regulating the development of one or more aspects of muscle differentiation.