Granulocyte-macrophage colony stimulating factor induces both HLA-DR expression and cytokine production by human monocytes

The effect of recombinant human granulocyte-macrophage colony stimulating factor (GM-CSF) on the expression of HLA-DR, and the production of the cytokines interleukin-1 (IL-1) and tumor necrosis factor alpha (TNF alpha) by human peripheral blood monocyte-enriched populations was investigated. GM-CSF was shown to induce both the expression of HLA-DR and the cytokines IL-1 and TNF alpha in a dose-dependent manner. In contrast, interferon-gamma (IFN-gamma), which induced major histocompatibility complex (MHC) class II expression, did not induce IL-1 or TNF alpha production. However, IFN-gamma enhanced the cell surface expression of HLA-DR and the production of IL-1 and TNF alpha on monocyte-enriched cells stimulated by GM-CSF. By itself, GM-CSF did not induce surface class II expression on the human monocytic tumour cell line THP-1, whereas it synergized with IFN-gamma to induce surface expression. These cells responded to GM-CSF by producing IL-1 and TNF alpha; Northern blotting showed that mRNA levels of IL-1 and TNF alpha were transiently induced, similar to other cytokines. Our results indicate that GM-CSF is a major macrophage activating factor that is capable of inducing both the expression of HLA-DR and the cytokines involved in T-cell activation by macrophages; therefore, GM-CSF may be of importance in potentiating antigen presenting function.     

Serum-free culture of enriched mouse anterior and ventral prostatic epithelial cells in collagen gel

Summary Sustained growth of mouse ventral and anterior prostatic epithelial cells embedded within collagen gel matrix was achieved in a serum-free medium composed of Dulbecco's modified Eagle's medium and Ham's F12 medium, 1∶1 (vol/vol), supplemented with bovine serum albumin fraction V, epidermal growth factor, transferrin, cholera toxin, prolactin, 5α-dihydrotestosterone, cortisol, putrescine, fibroblast growth factor, and a trace element mixture. Three-dimensional growth of prostatic epithelial cells occurred inside the collagen gel matrix. This serum-free medium allowed cell growth greater than sevenfold over 10 d in culture. Tissue recombination and cell culture techniques were integrated to demonstrate that cultured cells retained prostatic characteristics. Following 10 d of culture, epithelial colonies from mouse ventral and anterior prostatic epithelial cell cultures were isolated and combined with rat fetal urogenital sinus mesenchyme and grown for 4 wk under the renal capsule of intact athymic male mice. These tissue recombinants showed distinctive prostatic histologic characteristic (alveoli and ducts lined with cuboidal or columnar epithelium surrounded by stroma). When histologic sections of recombinants were stained with the Hoechst 33258, epithelial cells of mouse origin were distinguishable from stromal cells of rat origin. Aided by grants CA-05388 and CA-09041 from the National Institutes of Health, Bethesda, MD, and by M. A. R. C. fellowship GM08730 to T. T.     

Age and hypohydration independently influence the peripheral vascular response to heat stress

Seven young (Y, 22-28 yr) and seven middle-aged (MA, 49-60 yr) normotensive men of similar body size, fatness, and maximal oxygen uptake (VO2max) were exposed to a heat challenge in an environmental chamber (48 degrees C, 15% relative humidity). Tests were performed in two hydration states: hydrated (H, 25 ml water/kg body wt 1 h before the test, 2.5 h before exercise) and hypohydrated (Hypo, after 18-20 h of water deprivation). Each test began with a 90-min rest period during which the transiently increased plasma volume and decreased osmolality after drinking in the H condition returned to base line. This period was followed by 30 min of cycle exercise at a mean intensity of 43% VO2max and a 60-min resting recovery period with water ad libitum. Although prior drinking caused no sustained changes in plasma osmolality, Hypo increased plasma osmolality by 7-10 mosmol/kg in both groups. There were no significant age differences in water intake, urine output or osmolality, overall change in body weight, or sweating rate. In the H state, the percent change in plasma volume was less (P less than 0.01) during exercise for the Y group (-5.9 +/- 0.7%) than for the MA group (-9.4 +/- 0.6%). Esophageal temperature (Tes) was higher in the Hypo condition for both groups with no age-related differences. Throughout the 3-h period, mean skin temperature was higher in the Y group and significantly so (P less than 0.05) in the Hypo condition.(ABSTRACT TRUNCATED AT 250 WORDS)     

Preliminary data on rotifers in the interstitial of the Ninnescah river, Kansas, USA

One-hundred five rotifer species were identified from a sandy, plains river in Kansas, USA (the Ninnescah river). Of those rotifers identified, eighty occurred only in the interstitial sand, fifty-six in only the flow as plankton, and thirty-one from both habitats. Most species can be characterized as being more commonly found in littoral habitats, and not the interstitial or plankton. Lecane inermis (Bryce, 1892) had the highest density in the hygropsammon interstitial with an average 401 ind. 10 cm⁻³ at 1–3 cm depth in September 1993. New records for the US include Encentrum wiszniewski Wulfert (1939), E. longidens Donner (1943) ant E. (Parencentrum) longipes Wulfert (1960). A small form of Lecane levistyla Olofsson (1917) is further examined and discussed.     

Characterization of In Vivo Reporter Systems for Gene Expression and Biosensor Applications Based on luxAB Luciferase Genes

Advances in genetic engineering methods have allowed the development of an increasing number of practical and scientific applications for bioluminescence with lux genes cloned from a variety of organisms. Bioluminescence derived from the shortened lux operon (luxAB genes) is a complex process, and applications seem to be proliferating in advance of an understanding of the underlying biochemical processes. In this report, we describe a two-phase kinetic behavior of the light emission which must be properly taken into account in any quantitative measurements of the bioluminescence signal. By using strains of Escherichia coli and Caulobacter crescentus, this behavior was characterized and interpreted in terms of the biochemistry underlying the bacterial luciferase mechanism. We show that the intensity profile of each of the two phases of the luminescence signal is responsive (and exhibits different sensitivities) to the concentration of added decanal and other components of the assay mix, as well as to the order of mixing and incubation times. This study illustrates the importance of appropriate protocol design, and specific recommendations for using the luxAB system as a molecular reporter are presented, along with versatile assay protocols that yield meaningful and reproducible signals.     

Evaluation of Wetland Gain and Loss of Abandoned Agricultural Impoundments in South Louisiana, 1978–1988

At least 43 agriculture fields were built in south Louisiana wetlands between 1900 and 1920 using pumps and levees. All but one project is now abandoned as a result of soil subsidence, financial difficulties, or multiple levee failures. Some are now in flood-protected urban zones, and the remainder are mostly open water. The total area of abandoned agricultural fields in 1990 is large (> 80,000 ha), and their levees continue to deteriorate naturally. Ten failed or abandoned coastal agricultural impoundments (22,680 ha) were examined to determine recent wetland restoration or regression rates from 1978 to 1988. Wetland area and levee length were determined from aerial photography for 1978,1983,1985, and 1988. Average wetland change rates for all areas ranged from ?4.28 to +2.54% per year from 1978 to 1988. One site gained wetland area between 1978 and 1988 (77 ha/yr), and four sites gained wetland area between 1985 and 1988 (range 14–439 ha/yr). Wetland area in the other sites either remained stable or declined during the study period. The results from a multiple regression model indicate that restoration is inversely related to impoundment size and directly related to levee reduction. Results from a multiple regression model indicate that active levee removal will probably enhance wetland restoration rates at a very favorable cost (< $1/ha) and will be sustainable with little additional management.     

Water relations, gas exchange and abscisic acid content of Lupinus cosentinii leaves in response to drying different proportions of the root system

Soil columns in which the root system was divided into threeequal layers, each 24 cm in diameter and 33 cm high were usedto examine the influence of drying different proportions ofthe root system on the water relations, gas exchange and abscisicacid (ABA) concentration of lupin (Lupinus cosentinii Guss.cv. Eregulla) leaves. The treatments imposed were (i) all threelayers adequately watered (control), (ii) the upper layer unwateredwith the remaining layers kept adequately watered, (iii) thetwo upper layers unwatered with the basal layer kept adequatelywatered, (iv) all three layers unwatered. The treatments wereapplied at 56 d after sowing (DAS), and continued for 21 d inthe treatment in which the three layers were dried and for 36d in the other three treatments. After 21 d, the soil matricpotential in the layers that were unwatered had decreased toemdash 1.3MPa, compared to - 0.03 MPa in the adequately-wateredlayers. Within 8 d of cessation of watering, plants with the entireroot system in drying soil had significantly lower stomatalconductances, lower rates of net photosynthesis, and higherleaf ABA contents than did adequately-watered plants. Whilethe leaf osmotic potential decreased within 8 d of cessationof watering, the leaf water potential did not change for thefirst 15 d after water was withheld. After withholding waterfrom all layers, the shoot dry matter was 63% lower than thatin the adequately-watered plants. In the two partially-droughtedtreatments, 17% and 48% of the root length was subjected todrying. Compared to the adequately-watered plants, drying upto 50% of the root system for 36 d, in the two partially-droughtedtreatments, did not reduce stomatal conductance, net photosynthesis,or plant growth. Similarly, there was no significant effecton leaf water potential or osmotic potential. When either theupper or upper and middle layers of soil were dried, the ABAcontent of the leaves for most of the drying period was slightly,but not significantly, higher than in leaves of the adequately-wateredplants. The results suggest that lupins with a well-established rootsystem can utilize localized supplies of available soil waterto maintain leaf gas exchange despite appreciable portions ofthe root system being in dry soil. In contrast to other studies,the results also suggest that when only a portion of the soilvolume is dry and adequate water is available in the wet zone,root signals do not influence stomatal conductance and leafgas exchange of lupin. Key words: Abscisic acid, gas exchange, lupins, split-roots, water deficit     

Analysis of the regulation of penicillin biosynthesis in Aspergillus nidulans by targeted disruption of the acvA gene

To analyse the regulation of the biosynthesis of the secondary metabolite penicillin in Aspergillus nidulans, a strain with an inactivated acvA gene produced by targeted disruption was used. acvA encodes -(l--aminoadipyl)-l-cysteinyl-d-valine synthetase (ACVS), which catalyses the first step in the penicillin biosynthetic pathway. To study the effect of the inactivated acvA gene on the expression of acvA and the second gene, ipnA, which encodes isopenicillin N synthase (IPNS), A. nidulans strain XEPD, with the acvA disruption, was crossed with strain AXB4A carrying acvA-uidA and ipnA-lacZ fusion genes. Ascospores with the predicted non-penicillin producing phenotype and a hybridization pattern indicating the presence of the disrupted acvA gene, and the fusion genes integrated in single copy at the chromosomal argB locus were identified. Both fusion genes were expressed at the same level as in the non-disrupted strain. Western blot analysis (immunoblotting) revealed that similar amounts of IPNS enzyme were present in both strains from 24 to 68 h of a fermentation run. In the acvA disrupted strain, IPNS and acyl-CoA: 6-aminopenicillanic acid acyltransferase (ACT) specific activities were detected, excluding a sequential induction mechanism of regulation of the penicillin biosynthesis gene ipnA and the third gene aat.     

Dry Matter Assimilation and Partitioning in Pepper Cultivars Differing in Susceptibility to Stress-induced Bud and Flower Abscission

Stress-induced abscission of buds and flowers is a constraintto the production of peppers (Capsicum annuum L.). Susceptibilityof pepper cultivars to stress-induced abscission may be relatedto assimilate levels and partitioning. Growth analysis experimentswere conducted in the field to determine whether cultivars differingin susceptibility to stress-induced abscission showed correspondingdifferences in growth rates and dry matter partitioning whensubjected to low light stress. With the stress susceptible cultivar'Shamrock', reduction in net assimilation rate (NAR) and relativegrowth rate (RGR) was greater under low light stress than withthe more tolerant cultivar 'Ace'. Compared to 'Ace', 'Shamrock'partitioned a smaller proportion of dry matter into reproductiveorgans and a larger proportion into expanded leaves. Cultivarsdid not differ in the amount of dry matter partitioned to youngleaves or in the extent to which accumulation of dry matterby young leaves was reduced by low light stress. Other experimentsconducted in growth chambers and greenhouse examined the effectof removal of leaves less than 3 cm long on high temperatureand low light stress-induced bud and flower abscission. Forboth 'Ace' and 'Shamrock', abscission was not altered by removalof these leaves. Preferential partitioning of assimilate toyoung leaves does not appear to be involved in stress-inducedabscission in these pepper cultivars.Copyright 1994, 1999 AcademicPress Pepper (Capsicum annuum L.), abscission, low light stress, high temperature stress, assimilate partitioning, assimilation/growth rate, growth analysis, defoliation